 Okay, thank you. Good afternoon, everyone. First of all, I would like to thank the organization for the opportunity to present my work today. I decided to present the progress of our research work concerning the functional analysis of post biotics that analyze time biotic relationships between the gut microbiome and the host genome. So, as you know, the gut microbiota is a complex system of mutualistic microorganisms. Hosting 100 trillion bacteria representing 1 to 3% of body mass, encoding for maybe four millions of genes. And it sits at the interface between the environment and the host organism. And it contributes to nutritional processes through many bacterial-specific metabolic reactions. So, our objectives were to identify biological relationships between the gut microbiome and the host genome through the identity metabolic profiling and physiological phenotyping. As I have a question, I'm not sure it's important. You said 100 trillion bacteria. I read several places 10 times more bacteria than cells in the body. But according to this, it's less. I agree. So, where is this number coming from? The 100 trillion. Wikipedia. No, not Wikipedia, maybe. Oh, it's the same thing, you know, per meta. No, I found it in one papers. And so, yeah, I will. How it was counted, the different way to count. How it was counted. What is the protocol? There are several counting protocols, it's not obvious. How do you count either of them? I don't know. So, for your answer, it wasn't only recently the number was correctly estimated, right? Well, I don't think this is a major point of the talk. Thank you. I will try. And you have a good, good point of view about how. But I don't know. So, first, I try to continue. And we established correlation analysis in human. We used untargeted gas chromatography, sorry, of this machine, and we obtained peaks corresponding to metabolite signals. We acquired around 15,000 metabolites, and we obtained this kind of figure, which gave us a cluster of co-regulated metabolites. This metabolite from the gun bacteria? Yeah. No, microbiome metabolites from the serum of patients. The serum of patients? Ah, yes, that's the point of view. It's a serum, serum. Yeah, OK, so it's not microbiome, it's serum. Yes, and we, there are more than 15,000 in the human serum, there are about 25,000 in the P-State. And you only found 15,000, which you already know, or the chromatography, you know, they are there, but don't have their names. But 15,000, they are known, which they are, or only, you know, they're different. Let me see. I'm sorry, I'll be back. These are maybe more metabolite signals. Only signals, OK, you don't know what they are. They're signals, OK. That's all right, that's all right, that's all right. We didn't identify them. Yes, OK. And we used the semi-quantitative data from non-metabolites, and we tested them for association with clinical... They're non-metabolites, not all of them. So they were known, OK. So how many knowns? OK, 100. How many? On these methods, we have 100 metabolites, no. 101, I think, it's 101. OK, but in future, 1,000 just signals, OK. This is the Russo-Cardian, which is a non-metabolite. And we demonstrated that one metabolite, the fork-resol, is negatively correlated with obesity and diabetes. So fork-resol is a phenol. Then you can eat this poor kind of diabetes. Type 2 diabetes, type 2 diabetes, type 2 diabetes. Type 2, this is a little divided into five groups again. Yeah, yeah. Genetically. Sorry. OK, fork-resol is the phenol metabolized from tyrosine by Mycobiotta. Here is the structure of the phenol. We can also find chrysalis at low levels in food, such as vegetables, tomatoes, asparagus, cheeses, butter, bacon. And also in some drinks, such as coffee, black tea, wine, brandy and rum. So we are physiologists. That's why we developed a pipeline to investigate the different role of microbial metabolites in vivo in mouse. For this reason, we used C67 Black 6 mice on show diet or on IFAT diet to study the obesity and insulin resistance. In fact, we used these osmotic pumps from RLZ. We filled this reservoir with a metabolite solution, and we put the osmotic pumps here in mice. Under the skin. Under the skin, under the skin, under the skin. It's a little incision here in dorsal lateral, and we put the pumps under the skin. The experimental design was four groups of six male mice, control groups with pump-containing saline on show diet or on IFAT diet, and experimental group with the metabolites of interest today, the Cresol on show diet or on IFAT diet. Here you can see the experiment timeline. But what level of concentration, comparable to what? It's a very good question. We used 0.5 milligrams per kilo of mice per day. I think the concentration is 1,000 times smaller than toxic concentration. We performed a toxicity analysis. There's no concentration in blood compared to what you have observed in blood. In blood, it's very, very, very, very, very complicated because I saw in the literature maybe 0.8 micromolar to 150 millimolar. It's a big range. It's a big range. I don't have the answer, unfortunately, but we're going to check in a larger diet bank. So we tested several molecules, and today we're going to focus on the chronic infusion of four Cresol. So first, we wanted to check six. Six? Six small. Yes. Wow. Good. So we wanted to check the effect of this metabolite, the four Cresol, on glucose homostasis. We performed an IPGT. IPGT is for entrapuretinal glucose tolerance test. In fact, we injected a glucose solution to grams per kilo to animals and to control their response to this glycemic stress over time. In fact, we measured the glycemia during the time. First, we wanted to measure the impact of the diet. Here is the black curves. Dotted line corresponding to control on show diet and the solid line for control on IFAT diet. And as expected, we demonstrated that the IFAT diet deteriorates glucose tolerance. Now, we had red curves corresponding to groups under four Cresol treatments. And we demonstrated that the four Cresol improves the glucose tolerance, as confirmed by the calculation of Ariander curve. And here is the color code for the rest of my presentation, saline control group in white and in black for four Cresol treated animals. So we demonstrated that the four Cresol improves glucose tolerance. In parallel, we collected blood samples and we measured insulin secretion using LSA-SA. And we demonstrated that here with the black bars that the four Cresol successfully stimulate the glucose induced insulin secretion. The most striking observation in our experiment was concerning the pancreas weight. We did ratio pancreas weight on body weight. And we observed. This correspond to more than one. The phenomenon you observed would rather correlate to be the one, not the two. I think it's a bit too. For that big mistake, they have high level of insulin and they control the blood glucose. Later on, the pancreas is overexhusted. Insulin level goes down and glucose level go up. But this is what is occurring in patients before they show up high blood, high glucose level in the blood. Normally, with the IFAD diet, we induced type 2 diabetes. So chronic administration of four Cresol strongly increases the pancreas weight when compared to control mice. That's why we analyzed the histology of pancreas to test whether the islet structure was affected by four Cresol. We performed hematoxiline eosin staining, which stain in pink here islets which produce insulin, you can find here too. And we confirmed this with immunochemistry against insulin, with an antibody against insulin, here in Brown. And with statistical analysis, we observed that the four Cresol administration is associated with a strong increase in both islet density and insulin positive area. To investigate the possible cause of this increase of beta cell area and islet density, we performed the collocalization experiment against insulin, like in the previous experiment, and against K67, because K67 is the biomarker of cell proliferation. So we reproduced the same thing here with the statistical analysis that the four Cresol increases the size of islets. And here with the K67 approach, we demonstrated that the four Cresol increases the number of proliferative nuclei in islets. So we have proliferation with a four Cresol. The next and last step was to understand or to document the mechanism that four Cresol can impact the cell proliferation. So one kinase, the Dir-Kwane, sorry for the name, because it's very long, it has been demonstrated that these kinases can be inhibited by the armin, this, in fact. And this inhibition increases the human pancreatic beta cell proliferation. In another paper here, we can also find that the inhibition of Dir-Kwane stimulates the beta cell proliferation. So our hypothesis was that the four Cresol can mimic the effect of armin on metabolism and beta cell proliferation through the inhibition of Dir-Kwane. Preliminary results in biochemical assay show that the four Cresol and Dir-Kwane can interact and here with the half maximal inhibitory curve, we demonstrated that the four Cresol is a Dir-Kwane inhibitor. Finally, we performed a QPS analysis in our animals and we demonstrated that the four Cresol don't regulate the eminrin expression of Dir-Kwane. So in conclusion, the animals chronically treated with four Cresol exhibit a significant don't regulation of pancreatic Dir-Kwane transcription. To conclude, we wanted to illustrate the effect of this gut microbiome activity on human health. In fact, tyrosine is metabolized by microbiota in four Cresol. Four Cresol can interact with Dir-Kwane and the result of this interaction has an impact on pancreas. The four Cresol increases the beta cell proliferation into an secretion and don't regulate the eminrin level of Dir-Kwane. Our data illustrates the health benefits of spostbiotics which involve communication between bacterial metabolites and the host through the regulation of Ocellular proteins. So four Cresol is affecting the transcription of Dir-Kwane? Yes, yes, yes. What's their level of the protein, the transcription? We don't know yet, we have to... You are showing the eminrin expression. The eminrin expression, but we hope that it's correlated with the protein, but we don't know. No, no, no, I'm saying that the mechanism is through eminrin expression, not the protein. Transcription is versus transcription. Right. OK. Yeah? OK, we'd like to thank just my lab, my supervisor, Dominique Gugier, present here and all my collaborators from Kyoto University and McGill. Thank you. Questions? Do you have any idea which bacteria you may make exactly? Many bacteria, principally Clostridium difficile. E. coli can make it? If... E. coli. E. coli? No, no, no, I think it's a special name with lactobacillus, blah, blah, blah. I don't know the real time. I think this is quite amazing because you can use it to cure type 2 diabetes. I hope. For Cresol. Instead of giving them insulin. Yeah, I think the idea is that it's something we discussed this morning. It's how we can change the bacteria ecosystem. Yeah, yeah, yeah. And drive the ecosystem to produce something. The fortress over there itself can replace insulin. This is the major thing because it's very difficult to transplant a pancreatic beta cell in patients with type 2 diabetes. But this can do that because it increases the proliferation of the cells. Yes, exactly. If I can answer. We performed an experiment on a type 2 diabetes model in rat, GK-RAT, and we restored the beta cell proliferation. So it's... Well, yeah, it's a wow, but we have to be... So glucose tolerance test. Yes, yes. And then we... Yes, and we demonstrated that the structure of islets, because normally the pancreas of GK-RAT are totally disrupted. And the... Yes, yes, yes, yes, yes, while we are trying to patent now. Okay, yeah. What's the solubility for Cresol and how do you think it gets from the intestine into the pancreas? The avability? Solubility. Soluble compounds. So I'm curious whether there has to be a carrier or something that gets it... I think that Cresol can interact with Abimin. That's the only thing I know. With no more questions, we'll go to the second part.