[Music] first of all I would like to thank the organizers and specifically er Andre from a colleague of mine at the College of Agriculture of Rennes who I didn't met for maybe 20 25 years it's been a real pleasure to be here today in Rome okay so now we are coming close to the conclusion of today and you've got a huge amount of information on butyrate so it's quite a challenge to try to to add-on visa outstanding presentation I will try I will reduce my talk from many slides I'm afraid but it's a way it is okay so initially I'd plan to to give you some information on butyrate and get barrier function because this is a key function in keeping Alfea not only the gate but the entire organism the second point and I will really develop this one is a link between beauty right butyrate and a very important intestinal enzyme which is called also intestinal alkaline phosphatase it's a very potent anti-inflammatory enzyme and as also links with the gut microbiota get pro-inflammatory components etc the third aspect was about butyrate and get inducible each shot proteins and I would guess that this could add to the discussions one group at earlier this afternoon on eat eater stress in poultry and even in other species and fourth point on butyrate and epigenetics but just as a flavor because this is already a big story and so the time is too short for doing this so I can skip this a free major function of the guts of physiology microbiology immunology then the kind of messy situation at the gut interface I would say between the luminal contents with food nutrients died components the organism cannot digest the microbiota and a number of metabolites and even constitutive components of the microbiota and then the epithelium layer very very major to protect the organism and we get underneath the lamina propria with all these components of the immune system and then the smooth muscle layer so with a combination also of enteric nerves so we've seen with the previous speakers or links between the gut and and the brain so I can skip this all of you know this but I can skip this one as well this one as well because we are microbiologist in the room that know this better than myself but there are a few key beauty white producers that one should remember okay so given the very many function so I'd plan to focus on value functions I will go very fast and then on proliferation and linked with this enzyme and each shock proteins so I skip this one so further gut barrier function I just made a very quick review of her recent literature on this because say ten years ago there were not very many data I will not command this in detail the previous speaker did a lot on it I just want to draw your attention on that these effects have been shown in different types of get epithelial cells of different species and origins which means that this is probably a kind of universal mechanism okay so I will skip this one and then moving to the intestinal alkaline phosphatase story so first of all so a group in Chicago now or more than fifteen years ago showed that among the short chain fatty-acids butyrate was strongest in inducing this enzyme intestinal alkaline phosphatase so AIP and so why is this enzyme so important so this is a kind of summary of many of its words it's a figure I made in the first review I did on this topic nine years ago nearly ten years ago and since then have been many additional information showing the strong protective effects of this enzyme and have published two other reviews on this topic one in 2014 and another one that was accepted last December that should be released early but you can see I will not comment all the functions of this enzyme that you can see that it can contribute to regulate energy through fatty acid absorption through the small intestine it regulates suffice pH at the epithelial lumen surface but the major points are it's a I would say fantastic role in being able to detoxify pro-inflammatory bacterial components here when I made this picture there was only data on LPS but since then we have been data on also flagellin and of the nuclear components from from bacteria on the other right-hand side so it's a moral more or less corner of this is also its ability to regulate intestinal bacterial translocation which is another story that it has been shown and now this enzyme has profound aunt inflammatory effects not only at the gut level but also at the systemic level and part of it is related to the fact that this enzyme is is produced at the article side of the internal side but also at the basolateral side I will show it in more detail so I'm talking of the intestinal as a form of the visions I'm not very different isoform one is a tissue nonspecific isoform which is found nearly everywhere but this one is produced exclusively by the internal side so mostly at the apical brush border and part of it down also at basolateral level in the large intestine normally the expression and activity of this enzyme are very low and when when we are I it's quite often synonymous with inflammation so when one looks at the small versus large intestine in interpreting the data it's very important to know which isoforms are have been produced not to make mr. interpretation and when at the plasma level or serum level usually I plasma are total alkaline phosphatase activity is synonymous of inflammation even generalized inflammation and might be considered as a marker of inflammation so coming back to the internal side so this enzyme is mostly so produced by the by the cell exported at the apical side and released through hydrolysis of the unsharp part released so in the lumen part of it is produced or release at the basolateral membrane so probably explaining the small contribution of this isoform to the total plasma serum alkaline phosphatase is low is below ten percent of the total activity so interesting point here is that visions i'm when releasing the small intestine will go down to the large intestine and will resist partly to digestion which means that it's still active in the column in terms of mechanisms it's an enzyme that acts with very very awful a very simple chemical reaction it's a difference relation of a number of moieties like LPS and once with LPS molecules are they phosphorylated we are no longer recognized by a receptor like tlr4 receptor for LPS so this breaks inflammatory cascade but at the same time at the Ontario side level this enzyme is able by itself to inhibit various steps of this inflammatory cascade I will not go in further details you can find the details on the slide beside this difference relating and detoxifying activity of LPS I said fragile in CPG DNA motif but also non-bacterial phosphorylated moieties or molecules like for example extracellular ATP ADP am P and visa kernel known as a dangerous signals very strong in use of immune responses so this enzyme is also able to 2d forceful relate with moieties to go down to the adenosine molecule which is bias ins I would say anti-inflammatory this is kind of summary I have no time to detail everything you can go to my reviews but it's a summary I published as an editorial in American Journal of physiology some years ago showing that aap by acting on one side LPS fragile in etc and on the other side extracellular ATP is able to modulate the composition of the gut microbiota and I would say unfortunate so it's strongly in anti inflammatory that's a big message to remember and fortunately when information is going up a number of inflammatory cytokines are inhibitors of the expression of the join of fat enzyme so inflammation is really key in the is a landscape and then it's also an enzyme that connects with the immune system and for example it has been shown to inhibit tissue in infiltration by neutrophils this time in fish which means that again it's a kind of a universal enzyme that as a fundamental role in in in the body so moving now to get inducible it shock proteins so the the shock proteins are highly conserved proteins or like intestinal like a landfill photos with wide variation in molecular weights between 10 and more than 100 kilo Dalton depending on the class of HSPs and many of these recipes are constitutive so we're turning the cells and their concentration for given cells excluding epithelial cells do not do not vary a lot their molecular chaperones involved in a protein protein folding and also mom bran translocation of transannular trans Marana mum brand etc but one unique feature of epithelia and specifically the gut epithelium is whether some glasses of iced recipes that are unusable they can be in usable under different conditions of stress like it's stress because we had the question earlier this afternoon but they can also be induced I would say preventively at the end rampage of the organism for example we've a glutamine and this can help to help animals to go through critical periods whatever pigs birds probably humans etc so in this case with inducible Foreman of HP's contribute to protein stabilization and prevent them from cellular denaturation and degradation ok I showed only two example of this inducible it shock protein on the Left HSP 27 or 25 depending on the austere species which is an enzyme which which is key in the stabilization and reinforcements I would say of the tight junctions I will not go again different eye junctions that VHS piece for the smaller molecular weight one act at the tight Junction to strengthen the gut barrier on the right hand side to show that the other class of inducible HSP hsp70 is involved so in a self protein trafficking and in interferes with misfolded protein to prevent them from proteasome degradation in kind of emergency situations okay and it also as a stronger anti-apoptotic effects so now our visa HSPs are induced by your sacred knowledge and metabolites of bacterial origin I will comment only the frame pardon in red in fact and butyl normal be shared is one of the shots and fatty acid well that are really strong in users of a chess piece which is shown in different types of cells I will not comment into detail that most of these are intestinal epithelial cells against non epithelial cells do not react to beauty right for example by inducing with a chess piece so sometimes depending on the type of cells and the conditions of challenge with Gujarat of over short chain fatty acid I've only one HSP or both classes of SSP will be induced what do we know on the sensors and the cell signaling pathways involved in bacteria in this case short chain fatty acid and butyrate induced HSP induction in fact we don't know a lot of thing I found only one paper back in 2007 but this was not intestinal cells it was human breast cancer cells showing that GPR receptors were involved on visca gpr-43 and map kinase pathways also so I would guess but this world's true for the internal side that wishes to be us to be shown okay so in summary the gut microbiota and probably particular bachelor species in strain are responsible forget epithelial protection mediated by in inducible each shock proteins butyrate is a major bacterial metabolite stimulating inducible a chess piece in the gut so as part of the answer to the question about each stress I think was were involved in this business please look at a chess piece it could contribute to to explain differences between treatments so now regarding your butyrate and epigenetics I will not go very deep in this story just to remind you that it has already been said that so epigenetics involved a different modification like Eastern acetylation with the acetyl group coming from acetyl consume a and the acetylation for Eastern density deacetylase okay and in fact has been told the beauty rat is an inhibitor of this enzyme which increases the access to the genes and and their expression one important point is the second part of the slide on the left lower side is these so-called epigenetic marks can be transferred from generationally and this this type of question early programming and long term effects was raised in our group of discussion earlier this afternoon so this I finger is placed for four experiments and for hypothesis in in this in this area I will not detail this just to say that in many studies we saw with the previous speaker that Eastern h3 is quite often a target for betray action and okay so again making the link between butyrate and epigenetics of the gene for intestinal alkaline phosphatase it's been shown in the early 2000 still by the group in Chicago that Eastern acetylation is involved in in the x over expression of this enzyme and against it it links to butyrate I came to this story of interest in alkaline phosphatase precisely with findings coming from early programming experiment in rats so the idea is to was to see how the gut or some gut function would be affected with this because ten years ago most of the experiments and knowledge were related for example to the first to the pancreas and and second and so glucose regulation etc and second to the brain but at that time there was nothing on the gut and by doing I would say rather a simple experiment in rats we could show that this enzyme was the only one of the gut we tested with tested nearly ten enzymes but was the only one but was imprinted by the early environment of the offspring looked at at the adult age maybe I have no time to go into details if you have question I can explain that cid to to to return is that there was an imprinting in this case and when we crossed this early conditioning and late or other life nutrition we're having two treatments normal fat died versus eyford dies we could show that with an well-balanced diet there was no difference in the exam activity between the two groups of offspring but when they move to a part of them move to I fat died so one having been treated in a special way in utero and during the early infancy freedom mothers died was not able to adapted to increase the level of his enzyme in order to control inflammation is set in short okay and more or less in the peak with another model a model of providing antibiotics to sow in a window around farrowing we went with this enzyme more or less to the same story not exactly the same because one can imagine that there would be interspecies differences of course but the basic idea is that there are a number of fundamental functions that can be programmed very early in life even in utero and with maternal influence but also with paternal influence which are far less studied so coming to the conclusion I will not add too much on top of this so I would say to answer the question yes a bit rate makes part of this complex landscape at the gut level and is involved not only in providing energy to the system but also in stimulating different systems whatever they are I would say our targeting our rain forcing the earth differences or targeting a number of toxic compounds from the gut microbiota okay thank you [Music]