in the event that I'm off on this mall. Oh, I just tripped. Let's try that again. Anyways, what's up guys? It's another beautiful day in Paris. I'm running really, really late right now. So I'm like speed walking to the metro station. I just shoved an entire bowl of French cereal down my throat, rushed out of there, and hopefully I won't be too late to class. I do have a test today and tomorrow. So I really need to be there. Also, do you hear that? No honking, no screaming, no singing. This is honestly like the first quiet day since the World Cup. Even most of the trash has been picked up. Oh. Nobody compares to you. Nobody compares to you. The really big one is one card game. The really big one. Don't get that, that job moment is nasty. Wait, is this six euros? Yeah, so by the one-zero-one, this is something that gets about three times Why should I get this? She's looking at it. It's a bit, it's a bit. Put the bread away. You know it's dry when it's wet. Put the brush. It's dry when it's wet. Feel this dry when you're tearing up. This thing feels like a brick. He could met us in one with this. I think it's good. This is so boring. I wonder if one of those moosewishes is going to have to grab it. I wonder if this is like a, how'd you just go like with one card game. I actually think all of from-pre is going to bunsevon. Every from-pre in France is going to bunsevon. I'm pretty tired. And we had an exam today that I still need a lot for. We have another one tomorrow. I'm a little tired if I haven't said that. But hey, we're going to- But we're going to get some bread and go to the Rodin. I bet. We're going to the Musée Rodin. Gonna see some thinker. Le Penseur. Le Penseur, oui. Oui. I didn't make my own day for the baguette. After taking, who knows, like 12 minutes to decide which type of bread she should get. We finally got out of the store. This one, this particular one is not that good. I think the other bread would have been better. So don't always succumb to peer pressure. You should cut there. You'll be better off. There it is. Where's the towel? Oh, there it is. Right there. Sponsor me Diet Coke. This is what I'm talking about. What? Look at that. Oh yeah. It looks like I give birth to the Eiffel Tower. That's what, yeah. That's exactly what we're looking for over there. Right there. That's where the bag was stolen. That's where it was all stolen. Oh, right on that bench. It wasn't the bench's fault, but still salty. I'm just saying, I've gotten so much more used to just talking to the camera in public and having people look at me weirdly. Yeah, I mean, for someone who posts so many YouTube videos, you should have gotten used to that by now. Especially in Paris. We're like speaking English, distinguishing you as like immediately American and people stare at you when it was. People can tell even if we don't speak Spanish. Yeah. Especially when I'm walking around with like my phone held up to my face. But yeah, just got to have a French magic shop. Pretty cool. Didn't see David Stone there.  Nothing can be Tannins. But it was pretty cool. There's some people doing some stuff. They had a large collection like old magic props. A lot of cool cards. Something I wanted to check out before I headed out. But now we're on our way to... Le Pont Mirabeau. Le Pont Mirabeau. Exactly. Which was a poem we had to memorize during the year. Can you try reciting the poem? Le Pont Mirabeau. Cool as thin. In os amour. Faut-il qu'il m'en souvienne? We're now sur Le Pont Mirabeau. We are sure. Sur Le Pont Mirabeau. Yeah, I don't think we're the one that gets soo... We're not soo Le Pont Mirabeau. We're almost there though. Look at that view. The Eiffel Tower. When we got the OG Statue of Liberty right there. Sur Le Pont Mirabeau. Faut-il qu'il m'en souvienne? Après Le Pont. I was in Guillaume signature. Bien le nuit sonne leur. Les jours sont vants. Shit. French fast food place of all time. And it is right there. You can't see the sign, but it is a place called Otakos. That my French host brother actually introduced me to. Basically, you pick your meat. And then they take people who are looking at me. Because I was speaking English in the middle of a French street corner. But you put whatever meat you want. And then they add fries and cheese and put it in a tortilla. It's absolutely amazing. Notre Dame. I was here in two other videos that I filmed previously. We actually came here the very first day after class. We walked to the Pantheon and then the Notre Dame. We were also here the night of the 4th of July. Which was pretty cool. Except for at night the bushes around here are just absolutely filled with rats. So it's quite strange actually. I think I have video, or my friends might have video. I'll throw it in if I have it. I keep saying that I'm going to go to Mass here. But I haven't made it yet. So mom, if you're watching this, I'm going to try it. Speaking of rats.


All right, so today, Vince Fior from Rockfeller University will tell us about morphogenesis, mechanics and malignancy about development of cancer cells in skin. So, Vince, please take the lead. Fantastic. Yeah, pleasure to speak with you all. Please feel free to interrupt as we go if there are questions and I'll try to go maybe a bit more rapid because of the late start. But yeah, it's a pleasure to share the work and hear your feedback. So, you know, one of the things that really fascinates me is the diversity of forms that biological systems take and in particular, within the human body. If we look at our tissues at the micro scale at the cellular scale, we recognize that all these different tissues throughout our body have these different structures and these structures are intimately linked to their function. How these arise are really still questions that we want to understand. And if we kind of zoom in and think about things that may be a bit of a conceptual level, what we understand is that the tissues have this arrangement of different cell types in different spaces. So in the middle panel here, I'm showing just cartoons of on the top, the skin, and it's a appendage, the hair follicle, or on the bottom, the intestine, and its distinct structure, which includes a crypt and the villus, the villus is where nutrients are absorbed. Crypt is where stem cells reside. And they produce all of the the other cell types of the tissue. So we I just color code cells, you know, for, for example, but we can think that, okay, red cells different than an orange cell, which is different than a yellow cell. And these cells have different functions. And really, the question is how do these cells come to be in the right place at the right time? And how does that influence the the whole function of the tissue? These are big questions. But I think the way kind of I think about it, perhaps, in engineering terms, is there's some sort of a signal, and there's some output. And there's this black box that the cell interprets the signal, and it makes a choice about what what sort of behavior will it will undergo, whether it's motility, growth, or death. And this can change the organization of the tissue. And this probably goes back into an iterative feedback loop sort of manner to ultimately give rise to these complex architectures. And one of the important things to think about is that each cell state, so how in what say flavor of cell you exist in at any given time, probably colors the this input output relationship. So what do we know about this process, we know quite a bit about how tissue tissues change their shape. And a lot of this comes from work in a simple model organisms such as the Drosophila, which is a monolayered epithelium, showing here the the the ectodermal cells here that will give rise to these folds, these dorsal folds, which is one of the one of the say prototypical mechanisms of shape change that have been observed. And so how does this happen? There's I would say two major mechanisms that have been described quite quite abundantly in the literature, which are ideas of differential growth and differential tension. So in differential growth to two tissues or two structures that are growing at different rates will give rise to potentially deformations within a confined area. Or you can just have growth, like extended growth within an unconfined space that will give rise to to shape change. The other is differential tensions. And this has been widely described in the context of the actomycin contractility, where say, if we're looking at a monolayer, the apical side of a cell might contract and change its shape, preferentially to the basal side, and this can give rise to shape changes like these dorsal folds that are seen here. So these are really these are ubiquitous mechanisms throughout the animal kingdom. And they've really been largely studied in simple epithelia. But some questions that have been really instrumental to my work is what other mechanisms could there be? As we know that there is really a diversity of architectures that of tissues that are seen throughout more higher order complex mammalian animals. Lastly, I'll just say that this this architecture is important because one of the hallmarks and maybe maybe the the synphonone characteristic of cancer is a loss of normal tissue architecture. And so we see in the schematic here on on the screen that as as cancers initiate and then progress to a real problematic state, a normal architecture say of this epithelium, where you have cells at the bottom, which are different from cells at the top, that really gets destroyed and disorganized and understanding that process is we think very important to understanding cancer. So some of these overarching questions are what are biophysical drivers of these tumor architectural changes? Do these changes influence tumor progression? And if so, how? And are say similar developmental mechanisms that we're that we're learning about are they co-opted during tumor genesis? Or are there similar mechanisms between tumor genesis and normal tissue development? So my work in the past few years has focused on the skin as a model tissue. This is a very interesting tissue, not only for its relevance in terms of disease and homeostasis of humans, but also its inherent architectural features. What I'm going to tell you about today is how cancers arise in the skin. And one of the things that really interested me when I joined a lab here at Rockefeller that focused on skin is that normally you have this architecture that's what we call a complex epithelium. There's cells at the base of the tissue that give rise to all of the differentiated cell layers above it. When those cells, what we call basal cells, when they get a mutation that can confer cancer, they give rise to two distinct shapes in terms of the pre, what I'll call premalation lesion. So before they totally invade the tissue and become really problematic, they have two different shapes. And those are schematized here. So if a basal cell gets a mutation in the hedgehog pathway, it gives rise to what are called basal cell carcinomas. And what I realized was that they have these distinct features in terms of their shape, which we've termed budding. And I'll describe that more on the next slide. Whereas if a basal cell gets a mutation in the brass or map kinase pathway, they give rise to premalignant lesions that we term folding. They both originate from those basal stem cells, but they drive distinct tumor architectures. And it really wasn't clear why. Importantly, however, these two cancer causing mutations give rise to very different prognoses and outcomes in terms of tumor progression. So while BCCs or basal cell carcinomas are rarely really invasive and metastatic, squamous cell carcinomas are. And so this kind of set up a nice question for us to try to ask the question of how do these distinct architectures arise? And might they be important for tumor progression? So to study this, we set up a system where we could introduce these lesions within the skin of mice and follow the growth and morphology of these lesions over time. And so to do this, we I'll spare some of the details, but we inject lentivirus into the amniotic sacs of mice. This will infect the earliest layer of cells they see, which happens to be those basal cells. We can then, by using this method, introduce any variety of oncogenic or genetic changes to those cells in a mosaic fashion. So in this case, we are introducing these cancer causing mutations, which are in the background of the mouse, either atras or smoothened through a through a cremated excision of their floxed alleles. The power of the system is we can also introduce other types of genetic alterations like knockdown of a specific gene of interest or CDNAs to overexpress genes. And so this gives rise to these these distinct folds, which features, which I'll characterize by these these morphometric features. But let me let me show you first that this works. So if we introduce these mutations, we'll give rise to if we introduce mutation in the hedgehog pathway, which I'll abbreviate as SMO or SMO M2, which is a which the details aren't critical, but that will be activating mutation in the hedgehog pathway. We get these butted like features. And if we introduce a mutation in the RASMAP kinase pathway, we get these folded features. And so I'll characterize them by a few different morphometric parameters. But what we what we decided was a very easy non dimensional way to parameterize the system was this shape factor s, which is essentially the indentation depth of the of the growth from the basal plane of the tissue over the curvature radius of the distal tip of that tissue. And so these look these look in our system, if you let them develop, they look like the lesions that we see in humans. And so he and the the morphometric parameters are here. So so we see that there's differences in indentation depth where these butted smoke smoke lesions made a little bit further have a very have a much smaller curvature radius, and that gives them a higher shape factor compared to the RAS holds. So so that so I'll say that the take home here is that for this shape factor s, these butted lesions that we get from from from hedgehog pathway activation, or if a high shape factor, and these folded lesions that we get from RASMAP kinase pathway activation have this low shape factor. Okay, so this was the system that we we set up, and we wanted to try to start to understand, you know, how how are these different architectures being driven? One of the first things we looked at was proliferation as these are oncogenic mutations. So if we look at how quickly cell cycle once they've been activated with the mutation, we find is that not surprisingly, the smoothen the BCC's and the SEC's, which I'll use for for shorthand, but think BCC is the smoothen activating mutant and the SEC is this RAS activating you. In both cases, cell proliferation is activated. And interestingly, we see that the density of cells within these mutant clones is elevated as are some of their shape parameters like the aspect ratio of a cell is actually increased. So these cells are getting kind of packed in within these clones, their density is increasing and they're becoming more columnar. They also experienced this anisotropy actually around the periphery. But this was all kind of well expected within and has been demonstrated within the literature as a function of growth within a confined space. So basically we have cells, they're proliferating higher than their neighbors. And so they're experiencing these compressive forces that are changing their shape and density. Is this what's giving rise to differences between these architectures as we see it really in both cases? To do this, we derived a functional genetic experiment to try to test this, which is if we diminish the proliferation of cells within these mutant tissues, do we change the architecture of these lesions, i.e. do we make a bud into a full or vice versa? And the answer is no. So what we see is that actually when we decrease the proliferation rate of cells within these mosaic tissues, the main take home that we see, the effect we see is actually, of course, the growth area of the individual clones diminishes. And this is just by titrating the amount of the cell cycle inhibitor within the tissue as does the indentation depth or general deformation of these lesions within the tissue. So in a dose dependent manner, if you decrease proliferation, you decrease the growth of these lesions and you decrease their ability to deform the out of plane of the normal epithelial plane, but we don't see changes in, this is, I should say, this is the same irrespective of the mutation that's being activated. So this kind of told us... Vince, what about the shape factor? So indentation depth sort of clear that it decreases, but what about shape factor? Yeah, so okay, so good question. And the answer is that, well, the shape factor actually, it diminishes slightly for both cases, but it doesn't switch the architecture from a budding to folding. So the shape factor will diminish slightly as a function of decreasing indentation depth, but it's not the drastic, it's not what is changing or dictating budding or folding. Yeah, but that's a good question. So what about, so it seems like differential growth is certainly happening and we think that's what's, you know, the important thing for driving these deformations, right? You need the growth to actually get out of plane deformation, but not the difference in shape. What about differential tension? This would be another hypothesis. So to do this, we did what are pretty classic experiments. We do a laser ablation at the junctions, in this case either within a mutant clone at the periphery of a mutant clone and wild type cells or say just normal wild type, wild type neighbors. And what we find is that, and so this is quite a bit of data, but I'll walk through one of the most important parts. If you look at, excuse me, could you explain a little more detail what is the effect of laser ablation? How does it influence the tension? Yeah, great, great. So the idea here is that we'll bisect a junction, it's kind of schematized here with the red dotted line, and then we'll follow the vertices of two junctions and we'll see how they change in their position over time. The idea is that as there's more tension on a junction, their displacement over time increases at a faster, at a higher rate or velocity than a junction that has lower tension on it. So this is purely a way to measure whether there is differential tensions on junctions within these mutant clones. There are some caveats to this, which is basically, you know, how are the material properties, let's say, of the tissue slightly affected between these cases, which could also influence retraction velocity, but for a good, for first proxy, these very initial reaction velocities, you know, seem to be a pretty well accepted and pre-robust measure of tension on a junction. Essentially, you cut it and you see how fast it moves apart. If you look at mutant junction, so the junction between two mutant cells in both smoothened or wrasse, it's very low, and we think this is because as these cells are compacting together, basically their tensions, their lateral tensions are being diminished. This is compared to two wild-type cells that are distilled to a clone where they actually do have a retraction velocity. That is quite substantial. The interesting thing here was that if you look at the retraction velocity between a mutant wild-type cell and a wild-type wild-type cell within the smoothened mutant tissue, there we do see a significant difference in their retraction velocity between these two cell types. So this really does suggest to us that at the edge of these mutant clones, there is this differential tension that could perhaps be giving rise to that shape, whereas in the case of wrasse, we don't see a difference in the interfacial tension between a wild-type mutant cell and a wild-type wild-type cell. Wins, can we also conclude from this that mutant cells push on the wild-type cells in this picture? So basically there is a tension more in the non-mutant tissue, right? Yes, yes, this is correct. This is correct. So if you look at, and yeah, within the center of the clone, there's compressive forces. Within the periphery of the clone, there's extensile forces, if that makes sense. If you looked at a wild-type cell right next to the mutant clone there, you would also be experiencing these differences in compression and tension, this pushing of mutant non-wild-type cells. In these cases where we're looking at these wild-type cells, they're actually quite distilled to the clones. We wanted to have a condition where it's kind of the base case, if you will. So I should also say that there is this slight trend where in totally wild-type tissue it is a bit elevated, though not statistically from the in the mutant tissue, which again gets to that point. This was corroborated by, if you look, if you're staying for things like actin and myosin, which are these major force-generating components of the tissue. If we look at the border of these smoothened clones, you can see these differences in concentration of these molecules that we don't see in wild-type or in the RAS case, and this is quantified here. So indeed it seems like the smoothened lesions, they exhibit differential interfacial tensions that are driven by actin myosin. At this point and throughout the rest of my talk, I will be going kind of back and forth between simulation and experiment, which hopefully I think for the audience will be some of the really interesting points of the work. Because yeah, maybe I should have prefaced this. Obviously there's a lot going on within these tissues in vivo, so it's quite a complex system, and we're able to make these genetic manipulations, but still if we want to try to maybe gain from some first principles what we think is going on, we thought it would be very beneficial to have a simulation component to see how well we were with our hypotheses and our measurements compared to what we could actually derive from first principles. So at this point I'll introduce you to the vertex model, which hopefully most are familiar with, but this is essentially describing the tissue with cellular resolution as with really two terms at this point that describe the work of the tissue that we minimize, which is this cell incompressibility term and these active line tensions that are at the interface between these cells. So incompressibility, we have this constant that is quite high, cells don't like to be incompressible, so this is essentially saying cells aren't going to disappear into nothing, they're going to maintain their area in this case, and the line tension saying that this gamma term can describe some differential tension or force on these junctions between cells like we had measured actually in the real tissue. The cool thing about this model is that like the skin is these, we built this model in this multi-layered fashion such that there's these blue cells at the bottom which are the proliferative cells in a normal case. If we introduce a mutation then that actually is going to drive this differential proliferation between the mutant and basal cells, and it's only the basal cells that'll proliferate, but they can rearrange their junctions, they can move upwards in the tissue, and eventually within this this bounded box they'll give rise to tissue deformations. So hopefully any, if there's any questions please, please. Yes, I wonder, but this vertex model, the number of vertices is not fixed, is it right? So it's not all hexagonal. So you don't use all hexagonal cells, right? No, no, it's not all hexagonal, and as the simulation runs, cells grow, divide, and once they reach a certain area, we bisect them, and so then they give rise to two cells. Yeah, so not all perfect hexagonal, and also, yeah, the number, these junctions are constantly rearranging as more cells are added throughout the simulation. So if we run the simulation and we, now I'll show you for cases where we're driving different tensions between the mutant and wild-type cells, as we observed in the smooth-in case, what is the effect? What is the effect? No, here, let me, let me, let me see if, yeah, it runs, it runs. It runs, okay. So, so, and I'll run it a few times, but basically on the top, there's no difference in tension between the mutant and wild-type cells, and as we move downward, we are increasing this parameter, this gamma parameter to a case where at the bottom, it's actually quite super physiologic, but we did want to explore the parameter space, and the middle is what we, what we actually, we measure in the tissue. So if I run it again, essentially what we can see is, yeah, cells are dividing, rearranging, and in, in all cases, we're arriving to this, this really a butted shape that, that looks most like the smooth-in lesions. So this for us was quite interesting, which essentially told us that, well, in this case, where we're just having cells divide, and we're having this multi-layered model, whatever parameters, the system is always giving rise to these butted shapes. So, so while we have this differential tension in the smooth-in lesions, which is associated with budding, it's really, we're still not able to figure out what is different between the budded and folded architecture. So this was kind of one of the outcome. I mean, it didn't, we didn't, we didn't solve our problem of what was distinguishing the two. And the main features that we do see is that you, you tend to see these cells become more excluded from their neighbors as their tensions get higher. So this would end, and yeah. And this is, I will say, very different from what we know about in a simple monolayer epithelium. So in this case, we're running the simulation, increasing the tension, either on the base or the basal side or the apical side of the tissue. And here we do really get a diversity of shapes, like just periodic folds or these kind of more apical or basely-oriented folded structures. And this is known throughout the literature for a long time that really, all you need is these differences in tension at a junction to get a real pretty, pretty large diversity of shapes. So clearly there was something about the multilayer aspect of the tissue that was different in being able to generate a diversity of shapes. And this is just, this is just shown here. This is just a graph of these results where we're showing, sorry, I'm showing the result of shape factor from our in silico simulations for a variety of interfacial tension parameters. And you really, you see that indeed the shape factor is not changing dramatically. And it's not getting us to distinguish between these two mutants. The last thing we did was, we did indeed do a genetic manipulation of this, where we knocked down myosin within these smoothen lesions, which is the main isoform that is generating these actomyosin tensions. And while, again, we see this subtle decrease in shape factor, which actually we predict from in silico as well, it's really still, it's not drastically changing the shape. So we can see the differential tension is involved, but it's not dictating the difference between the buds and folds. So what is, we kind of then took an agnostic approach, we decided to isolate these cells between these smoothened or ras embryos and see what kind of molecules are differentially expressed. Maybe that gives us a clue. And this is where we were quite excited by the fact that we found that the most highly enriched terms, if you compared smoothened to ras mutants, were genes involved in the extracellular matrix. And in particular this specialized extracellular matrix that's the basement membrane. So just a very quick, a very quick background on this basement membrane is this this polymer network that underlies epithelial cells. It's this network of laminin and collagen molecules that really provides this structural separation between epithelia and mesonkine. And these networks can additionally be crosslinked to change their biophysical properties by a certain number of accessory molecules. And in the skin, this membrane connects to the epithelial cells through these specific adhesion components and their termed hemidesmosomes. The interesting thing was that the, you know, really we know a lot about how these proteins behave biophysically by purifying them from tissue, maybe self-assembling them in vitro and measuring their properties. But what is the, over the biophysical properties of these networks really in the in vivo tissue was largely still a mystery when we started this work. So that was what we set out to do next. First I'll describe the model, the modeling aspect of it, and then we'll go into the measurements. So then in the model we implemented, we basically implemented terms for the stiffness, so both the stretching and bending modulus of the basement membrane, as well as a timescale for remodeling or assembly, which I'll talk about more in general. But basically how quickly is this basal membrane adapting to the growth timescale of the cell? So how quickly does that, when a cell is growing, how quickly does that membrane grow with it? I hope that makes sense. But okay, so let me show some of the simulations. And so here's where we started to get excited, as we could see really getting a more diversity of shapes by including the material properties of this basement membrane. And here if we look at the full kind of spectrum, stiffness on the Y and assembly on the X axis, we see that by bearing these parameters we get a quite rich spectrum of shapes, where you really need a high bending modulus to get folded type architectures, a lower modulus is giving rise to buds, and really with high assembly rates, we get these budded features to dominate. Sorry, Wins, can you comment a little about the model? So tau is the characteristic time of, and in your model, and lambda is connected, you're connecting the area, right? Yes, correct. Exactly. Yeah, exactly. So with respect to the previous model that you discussed, the area term, the effect of the area of the cell is still included, but through this lambda times square root of thing, right? Yeah. Okay. And it's not clear, how is the bending included in a way? How do you, what is C? Is it the curvature? C is the, yes, correct. The curvature of the Yeah, the curvature of this local. Yeah, the local, the element, yes. Yeah. So B, what is B? So B is our bending modulus, which we show for thin film, really, bending is dominating over stretching, and so that is our characteristic bending modulus of that network. Okay. And then what of all these quantities, so you can measure bending modulus in tissues experimentally, like a curvature, you already explained that you have been measuring to determine the shape factor, correct? Yes, thank you. Yeah. So now I'll get to the measurement of modules, but yes, this is the good point. So it was, we did have to devise a way to actually measure the local bending modulus, or we use actually small indentation and Young's modulus and converts to, it really is a good proxy for bending modulus in this case, but essentially we use a atomic force microscopy to, once we separate the epidermis from the base of membrane, we can go and locally measure the mechanical property of this tissue, which just is schematized here, but it was a headache, but we were able to do it. We find is that the base of membrane is much stiffer than the underlying dermis or the stroma, so we think by first proxy, that's the most stiff mechanically important element of this tissue, and as the model predicted, if you look at the smoothen lesions, actually those have the lower modulus, whereas the brass folds have a higher modulus. If we look at these proximal regions versus the distal tips, actually they have a high modulus, even higher modulus, which if I have time I'll get into describing why we think that is, but essentially at least to a first approximation, the model correctly predicted that you have a lower bending modulus in the smoothing case versus in the brass case. So is this really important? Does this influence the shape, architecture of these tissues? Again, we use a functional approach to basically vary the modulus by knocking down components of the base of membrane. If we knock down a collagen trimer molecule, we decrease the modulus. If we knock down one of these proteoglycans, we can actually increase the modulus, and if we knock down a crosslinking enzyme, we can also decrease the modulus, although to a bit lesser extent. So we went and then measure the effects of these knockdowns on the tissue, and we I'll just kind of point your attention to the quantification here, but here we do see that indeed if we change the stiffness, we can affect the shape of these tissues in a way that is kind of consistent with the model, where if we decrease the stiffness, we increase the shape factor, and if we increase the stiffness, we can we decrease the shape factor. And this is actually the trend is similar for both types of lesions. And so it seems like this modulus is indeed important in driving the shape, but we're not all the way there in terms of describing what is different or what is really the distinguishing features of these two. But I think we're getting close. So the last thing I mentioned was this, which was that assembly of the basin membrane is important. So we went to we went to investigate this. Essentially, if we look at types of molecules that are incorporated, either in an actively actively assembling basin membrane versus a more mature basin membrane, these are enriched within these distal tips of smoothened buds versus folds. We derived an assay to actually measure this in an X plant. So X vivo, we can take these tissues, we can culture them with just a small amount of fluorescently labeled basement membrane molecules and see how quickly they get incorporated into the existing basin membrane. And what we find is that indeed a high basin membrane assembly is is characteristic of these smoothened buds. Lastly, if we knock down laminin to decrease basin membrane assembly, we actually diminish the shape factor of these of these tissues, which is again, what the model predicted. So so here, I'll kind of describe what what what we really in in measuring and varying both the stiffness and the assembly of the basin membrane. We found that again, we were able to quite describe the morphometry of these lesions in ways that really, I think, started to make a lot of sense. So if we look at these regions here with a very so here shape factor is in cut in a heat map scale here. So you really you get these high shape factor lesions where we have a a high assembly rate and moderate to low stiffness. The shape factor decreases as we decrease assembly rate of the basin membrane. And it also decreases as we increase the stiffness. And so this was and so this is really this is, you know, quite consistent with with the results of our experiments. Yeah, the things I just want to want that in five minutes, we should go to the questions sessions. Yeah. So what I'm going to do is I'm going to skip through a a part here, I'll get to the conclusions. And I think, yeah, we can do questions and discussion. And I think that would be the most most fruitful. And I'll just go to a summary slide, which will kind of, yeah, I think get us there. But essentially, what we did was there's one, there's one more aspect that is important here. So we did uncover that these tumors, what I'll call tumor specific basin membrane mechanics are important in dictating these tumor architectures, as is a tumor specific cell differentiation program, which I'll just get into the summary of that on the final slide. So I'll skip all that. And I'll tell you and I'll just tell you all the summary, which is that the architecture, we think really is dictated by interactions between expansive forces, which are driven by the proliferation of those basal cells. The assembly dynamics and stiffness of that basin membrane that I talked about. And also the material properties of the super basal cells. And this is due to their differential differentiation program and the expression of care and molecules. And just to describe it briefly, it is that as these cells, as they differentiate and move upwards, they actually change their material properties as well, which all, which influences the folding versus budding shapes of the tissue. These mechanisms, at least these last two, thinking about basin membrane and super basal cells, these are distinct from super simple epithelia, where it's really been shown that this differential actinized intention is was driving things. Although, what we find is that actinized intention does kind of further quantitatively or subtly also these architectures, which we could think maybe they're really critical and normal morphogenesis. And what I didn't have time to talk about, but we think is really important, which is that actually the tension, the the extensile strain that's experienced by this basal membrane is elevated in a case where we have lower assembly of basal membrane and stiff super basal cells, which is exactly what we see within these swamous cell carcinomas that are more prone to invasion and basal membrane breakdown. So we think this could be a new mechanism by which this basal membrane is actually disrupted during tumor genesis and then leads to worse prognosis. So just kind of getting back to the general kind of questions, you know, I think that really these networks, them being the the extracellular matrix, the the base membrane and these cytoskeleton, these in particular intermediate filaments are really key, key networks, key drivers of how this this what I'll call mechanotransduction kind of feedback loop arises to give these different tissue architectures. And I think we've kind of at least understood a bit about how different architectural diversity can arise due to the nature of a complex versus a simple epithelium. And that is really this combination of basal membrane and differentiation of those super basal cells that expands this architectural diversity. So with that, I'll take questions. And if we want to go back through any parts of the data itself, I'm happy to. But I'd just like to acknowledge Elaine Fuchs, who's my postdoctoral mentor here at Rockefeller contributors, in particular, Mette Prank, who was at Princeton during the time of this work in the lab of Stops Schwarzman, who were really the drivers of the simulations and the computational work, as well as other collaborators that were involved in the study. So with that, I'll be happy to take questions. Thank you. Thank you, Vince. All right, we, because of the late start, we'll take a few questions over the time that was allocated. And for the talk, so if anyone wants to ask a question, you're free to connect your audio microphone, to connect your microphone and ask. Otherwise, if you have issues with your microphone, you can also type in the chat. All right, just while the audience thinks of the question, I will ask a question of my own. So you mentioned that the end of the talk is that while the cancer tissue proliferates, and it may also some layers of this tissue may change properties. So they will have different, so it doesn't mean that they will have different like bending models and other mechanical properties that you need for your simulation model. Is it correct? And if it is correct, what is the mechanism that drives this change? Yeah, yeah, so I'll just show a bit of the data. But yeah, essentially, it's just that. One of the things that we noticed was if we look, so this is doing this RNA-seq, right, and looking at different genes and how they're expressed. One of the other things, so I showed you what was really highly expressed in the smoothing case, which was these extracellular matrix proteins. What's highly expressed in the RAS case are these, we call them their differentiation genes of the skin, but it's essentially these proteins that make up the barrier. And that barrier, the outside of our skin obviously needs to be robust mechanically to keep water in and keep microbes and such out. And so this is again one of the major differences between these two states. And what I'll, yeah, I guess to more, these are like, you know, these looking at different keratin molecules, which are known to be important for this making the barrier. But if we just think about the material properties, again, now we could go in and actually measure these with AFM, looking at the layers, and we see that it's really quite beautiful. You see this fast change in the material properties of those cells as you move upwards in the layer.  And this also happens in the tumors. So to answer the question, there is this change in material properties. It seems to be due to just the normal process of barrier formation that the stem cells undergo. I see. Thank you. All right. Do we have questions? Yes, I have a question. Yes. Thank you for your presentation. I have some question. Why is it important to study the architecture of skin cancer? If it's to define the stages of all the skin cancer, is it to provide the medical treatment or what? Yeah. Good question. So, yes, I mean, at the end of the day, what we do when we uncover is kind of novel mechanisms of how cancer progresses. And whether it's, you know, skin cancer per se, or whether these are more generalizable principles to other forms of cancer, that's, you know, that's like the next set of questions and where we want to go. But we really, you can think about how these principles can inform mechanisms of progression and how it depends on the architecture of the normal tissue. So there's multiple, multi-layered complex epithelia within the body. We like to know what are these same mechanisms are at play within those tissues. But the bottom line is just that, yes, we still don't have mechanisms to stop even that initial process of barrier breakdown, the base of membrane, and stopping cells from spreading from that initial primary tumor site. So, yeah, the goal is to actually use these findings to inform, to inform treatment strategies, although I mean, we can talk about what they are. They're not, they're, they're not easy, but that's what we're thinking about. Yeah, well, I would add that to Boris' question that, first of all, architecture is very important also in the design of drug delivery systems. So one of the things is that when you treat the cancer, you need to ensure that drugs that need to, that fight the cancer will, will be delivered there and diffused through these tissues. So architecture of those cancer is important for designing cures and medications. That means when, when we find the architecture, we know exactly almost the kind of treatment we are going to apply to the skins. Well, there are several questions to that. Okay, first there are, there is a search for drugs and drug delivery. So for that, you need to know mechanical properties of the tissues and architecture. But there is also a question, or why this, right, why, why this cancer form this architecture, these features in the skin. All right, so, so to answer your question, I mean, this, this is new. So what we do know is that when, when a pathologist would go for a cancer, look at skin cancer, let's say specifically, they would see, okay, it's got this budded histology or this folded histology, or at some maybe later stage of that, but really then they will classify them as two different cancers and they'll, they will treat them differently. You know, again, when we set out to start this, we didn't know why they looked different. But, you know, now that we understand that and what some of the, the kind of constraints are that drive these different shapes, I think, you know, nobody, we don't have the treatment that says, okay, we need more base of membrane assembly in these brass driven squamous cell carcinomas, which again, before we even identified the, the, the cancer causing gene, we, we, we treated them differently. But now that we know that, you know, that that would be the goal is to, yeah, we're working on some screens to try to increase the expression of these important molecules that help to halt the, the next stage. Thank you. You do have another question, Boris? Yes, I have the last one. Yeah, go, even if it's not.  Sorry. Can you go back? Yeah. Could you say again to, to, to, to where is it? In the figure, I think it's this slide video though. No, no, no, no. The beginning of the presentation. Yes. About slide number 10 means one. I'll just, I'll just keep clicking. You can say, no, no, it's, it's before, it's before. Before. Yeah. Go back. That's the second slide. Ah, no. Move, or why is it not here? Could you just formulate the question without the slide? The problem is in the figure, he, he used two, two rules. One is like reversible. And I would like to know if, if I move from the beginning of the, the skin cancer, that means the, the first stage and move from the, the second stage card, is it possible to go back to the, to the first stage? Because in the figure, I have two rules. I think you want this kind of schematic maybe? Uh, it, yes, but yeah. The, the rule, because here you used to, you have, for instance, male stage and moderate stage. Can we move from the moderate stage to the male stage? Yeah. I mean, yeah, I think in principle, those are, those are the ideas that, that, that we're trying to explore, which is, yeah, I mean, clearly you can. So clearly you can push, obviously one thing is to eliminate the tumor cells, but then to push, push those cells to say differentiate, differentiation therapy to promote them to kind of be removed from the tissue through normal means. Is, you know, a potential way to move from, say, a disease state back to a more normal tissue architecture. So I think if that's, if that's the question is, can we and how can we, that would be an example of, of an approach. But I think we're, you know, we're still a long way from actually being able to do that. But, but that would be a goal. Obviously, you know, if it's, if it's a tumor that you're able to see and operate on, then you would move it. But if it's not, and it's, you know, say, small or it's, you know, there are many, many of them, then you do have to find a way to, to get rid of the tumor genetic tissue while still maintaining the normal tissue. Yeah. Thank you. Thank you. Are the questions, do we have? Not at the time? Okay, just


Good morning and welcome to this week's edition of Encompass Live. I am your host, Krista Burns, here at the Nebraska Library Commission. Encompass Live is the Commission's weekly online event. We're a webinar, a webcast, an online show, whatever you want to call us. We are online and we're live here every Wednesday morning at 10 a.m. Central Time. We do live sessions on Wednesdays and then we record our shows every week. So if you're unable to join us on Wednesdays, that's fine. You can always come back to our webpage and go into our archive sessions and see recordings of any of our shows that we've done. We put up the recordings, we put up any slides or presentations or documents that people may have, and also we link to any websites that may be relevant to a session. We do a mixture of things here, presentations, interviews, mini-training sessions, demos, anything that is library related we will have on the show. And we do bring in guest speakers sometimes from outside the Nebraska Library Commission or the Nebraska area, and we sometimes have commission staff. This morning we have a commission-centric session, Nebraska-centric, definitely. Next to me is Susan Nicely, who at our online services librarian here at the Nebraska Library Commission. Hello, Susan. Hi, everybody. And she's going to tell us about discount shopping with the Nebraska Library Commission. Some great deals and things you can get on all sorts of stuff, library related things through the commission. And I had mentioned earlier, and I'll mention it again here while we're on the recording. You'll notice if you are a regular viewer of Encompass Live, we do not have a camera view. We had some technical difficulties with our software for running our camera this morning. So unfortunately, you will not have our faces on the screen for you. It'll just be the slides today. We hope that we'll have things back up to speed and working correctly for next week's show. So I will just hand over to Susan to take it away. Okay, thanks, Christa. As Christa said, my name is Susan Nicely, and I work in the Technology and Access Services Department of the Nebraska Library Commission. And one area that I'm responsible for is discount opportunities on databases, books and supplies, conferences. And so that is kind of what constitutes our discount shopping program and what I'm going to talk about today. There we go. Christa and I were just talking before the show started, and we've done a version of this program before, back in 2009. In many ways, the program has continued on in the same vein as it has been chugging along in for quite some time. But we did want to revisit this program on Encompass Live because we do know that library directors, library staff members come and go. And so people who've been around for a while know about this program and the opportunities that are available through it. But we want to make sure that we periodically get the word out to catch new people that might not be as aware of what to look for. When I talk about our discount program, I really like to start with the Nebraska Library Commission mission statement. It's two sentences long, and I have it on this slide. And I just want to go ahead and read it to you. The mission of the Nebraska Library Commission is statewide promotion, development and coordination of library and information services. As the state library agency, the commission is an advocate for the library and information service needs of Nebraska. I really like to pair the mission statement with this program because I think this program really clearly fulfills this mission and works towards meeting it. One of the big challenges in Nebraska is that we have a relatively small population. We have lots of small libraries, which means small budgets, small staff, not a lot of staff members sometimes, not necessarily a lot of library hours open every week. And so that does provide some special challenges. And so if we are able to make people's jobs easier and help their budgets go farther, then that directly contributes to helping them meet the needs of their patrons, Nebraska residents in terms of making services available to them and getting that information out to them. So we try to work to get pricing that is affordable for our libraries. We try to get information out to libraries about discounts that they may want to take advantage of. And we also really do strongly advocate for Nebraska libraries with the vendors who don't often have an understanding of what we mean in Nebraska when we say small library. And that really comes into play when we talk about cost and some of their pricing strategies. We've had vendors who consider a small library, any library serving 20,000 pop served or smaller. And you just don't lump in Nebraska, you don't lump a library that serves 20,000 and has a budget that is supported by a pop served of 20,000 into the same category as a library that serves 1,000. They're not going to be able to afford and pay for the same price is not going to work for both of those libraries. And so that's one of the ways that I think we really do strongly advocate and try to educate vendors. And that's an ongoing process because as many of you know, there's a lot of staff turnover with library vendors and sales representatives. And so oftentimes we'll work with one sales representative for a couple of years and they'll start getting the picture. They'll try to sell a product in Nebraska at a price that's not affordable. And then finally they'll start getting the idea that they need lower prices if they want to drum up business. So we'll get them educated and then they move on and we get another vendor in and start the process over again. But we are in a good position to really try to drive that point home with vendors and so sometimes with greater and sometimes with lesser success, we're able to sometimes get them to modify their pricing to be more affordable in Nebraska. I thought I'd give a little bit of history about this program. And so what I did was I have to admit I wasn't energetic enough to actually go down into our paper archives, but I did poke around online in some of our online newsletters archives. And I did find our department technology and access services used to be the network services department. We used to have a newsletter called Nebraska Library Commission Network Services News, abbreviated as N-Cubed. And I found a July, August 1996 newsletter online and in that newsletter there was an article talking about referencing summer 1995, talked about the Nebraska Library Commission Steering Committee discussing the possibility of offering statewide first search database trials. So this was kind of the beginning of offering database trials, trying to promote products to libraries and see what we could do cooperatively to make services available. And in that article when they were discussing the Steering Committee's recommendations, they did directly tie this trial and the first search service to the Library Commission's mission statement. So they were thinking about this program in terms of our mission from the very beginning. The first first search trial ran from February 1st through April 30th, 1996, and they reported that 155 libraries participated. Back then it looks like people actually had to sign up to participate so they really were able to track those numbers. So this was a big deal. This was really the start of that sort of broadcasting database trial opportunities statewide and trying to get libraries interested, getting feedback, etc. I didn't include this information on the slide, but it was interesting in the January 1997 issue of the n-cubed newsletter. I also found references to discount on registration fees for the Computers and Library Conference, which is a conference discount that we still offer today. There was also an article talking about the CD-ROM group purchase that had just taken place. Back in the day we used to work with another library network based in Colorado and they would coordinate actual CD-ROM purchases and we'd get the physical CD-ROMs and we would package them up and send them out. You remember using CD-ROMs in the library? I worked in you. That's the way to do it. So back in the mid-1990s we were really getting started with this program and it has grown since that time. So there are really, I would say, three strands to the discount purchasing program. We offer discounts on databases and electronic resources and I would say this is by far the most part of the program that the most libraries participate in and take advantage of. It's where the most savings derives from and it's also where the most staff time gets devoted. We spend more time on this strand of the program than on the others. We also have discounts that we offer on books and supplies and we also regularly offer discounts on conferences. So I'm going to go ahead and move through these three different types of discounts throughout the program today and give you a little bit more information. So what I want to do now is actually direct you towards the pages on our website where you can get information about the program. So this page is just a screenshot of our discounts on databases and eResources page and I'm going to go ahead and jump out to that page now. I'm going to go ahead and start out on our main Nebraska Library Commission webpage because I want to show you how you can navigate from this point to the discount pages. Over on the left side menu we do have a discount group purchases category and as you'll see we have a flyout menu. And over on the right under discounts we've got our three different strands of the program books and supplies, conferences, databases and eResources. So I want to go ahead and start out with the databases and eResources part of the program. This is an InVex page and you'll see we've also got links that let you jump directly to discounts on books and supplies and discounts on conferences. So we've tried to make it a little bit circular so you can move from one part of the program to the other. This page is an alphabetical list and what we try to do is we try to list both vendor names and database online resource names. So depending on whether you're thinking about a database by its specific name, for instance, maybe you're thinking of Grove Music Online. You can find it alphabetically under G. That's actually a product that's sold by Oxford University Press now and so you'll also find a listing for Oxford University Press. So we've tried to provide cross-referencing so that you can find products that you're interested in. This page is particularly useful I think for new staff members, new directors that move to Nebraska and they want to get an overview of what products we have discounts on, what vendors we have relationships with. It kind of gives a good overview for what opportunities are already in place. Let me go ahead and just jump to one of the specific pages so you can see the type of information that we provide. So I'm going to go ahead and go to Britannica Online. On a vendor specific page, the information that we include varies but it usually includes some version of the following. We'll have a list of products that are available at discounted prices through the program. Descriptions of the product will often link to the vendor's website for more information about the product. We'll have information on pricing. In this particular case, pricing is based on weighted number of users and so we'll talk about how to determine the number of users you're going to base your pricing on. So for instance, four-year colleges count 100% of their full-time equivalent students. Two-year colleges count 75%. Public libraries use 7% of their total population served. It's not at all unusual for vendors to not want us to include pricing on the public website and so you will not always find pricing actually listed on the website. Sometimes you'll just see a statement that says, contact Susan Misley for pricing and there's a link to my email so that you can email me to request pricing. And when you do that, it's usually a good idea to provide me with what your FTE is if you're an academic institution, what your population served is if you're a public library or what your student enrollment is. And then I can get pricing to you either based on a priceless that I already have in my files or in some cases I have to contact the database vendor individually each time an institution is interested in pricing in order to get a quote. In this particular case, Britannica has provided us with pricing and they don't object to us putting it up on the website and so you actually have the pricing up here on the website. And because the pricing is predictable and it's uniform depending on whether you're a new subscriber or a renewed library, we actually have been able to put up an online order form. This is one of the few products that we actually still use online order forms for so in this case you can just fill out an online order form and submit it online. Usually, again, there's information here saying contact Susan Nisley if you want to order and I'll send you as an email attachment paper order form to sign and fax or scan an email back to me. We usually also have information on the subscription term that the vendor is working with and the order deadline. I'll often try to track down information on how to contact tech support. If the information is easily available and I'm able to track it down, I'll provide information on how to access usage statistics. And in this case, I've got access URLs that you use. So whatever information I'm able to find, I will link to it from this page. It does vary from vendor to vendor though. But that's kind of a good overview of the type of information you can find on a particular page for a vendor. Just to give you a little bit of some background information on the types of discounts we are able to arrange. And I've kind of broken these into categories to give you some background information. A lot of times, vendors are just going to offer a standard percentage discount off-list price. This can vary anywhere from 5% to up to 25%. Offer times will work with a vendor and they're hoping that there will be a lot of new subscribers. And if that's the case, they may offer a bigger discount off-list price if we're able to bring a bunch of new subscribers to the table. Sometimes if there's only one or two libraries that are interested in a particular product, which again is not unusual in a smaller state like Nebraska, they may say after we've promoted the product, offer the trial on it, they may say, okay, well, we'll give anyone who wants to subscribe 5% off. I'm usually fairly disappointed in a 5% discount. That's kind of the... Is it even worth the hassle? You kind of think if the subscription cost is low, 5% isn't very much money saved. If subscription cost is high, then 5% can be significant. 5% isn't great, but it's better than nothing and so that's when you sort of think, well, anything we can do, if we can help save a little bit of money, it's still worth the effort. 25% then you're really starting to talk significant savings, so it really varies. Oftentimes vendors do break pricing into tiers based on population served or number of students. This is where we can sometimes really negotiate on behalf of smaller Nebraska libraries. As I said, sometimes libraries, sometimes vendors will have pricing tiers that are way too broad or their lowest tier really is not conducive to our smaller Nebraska libraries. Maybe they'll say, any library that serves 1,000 or fewer students, we're going to charge this price to. A school that has 1,000 students enrolled usually has a much larger budget than a school that has 150 students enrolled. That's a case where sometimes we've been able to get vendors to offer special tiers that they just offer to Nebraska libraries. ProQuest has done this with their SIRS databases. They have for a number of years offered special pricing for school libraries serving fewer than 300 students. This has really been a significant savings opportunity and as a result we have quite a few Nebraska schools that do subscribe to SIRS products through us because they serve fewer than 300 students. And it's one of the few products that's really affordable to them in the state. We often do have vendors who are going to say that the discount is contingent on the number of libraries that participate in a subscription. This is particularly the case if they're trying to introduce a new database into the market. EBSCO is a good example of this. As a rule, I think EBSCO gives everyone a standard 5% discount off this price in the state of Nebraska just because Nebraska does so much business with EBSCO. Oftentimes they'll have a new product that they're introducing or a product that they want to promote and try to drum up business for. And so they'll approach us and say we'd like to offer a trial of this product and we'd like to also offer special discounting depending on a number of participants. And so they'll give us a scale, something like 1 to 5 new subscribers will give you the standard 5% discount, 6 to 10 new subscribers will give you 10%, 11 to 15, 15%, etc. And this is a model used by other vendors too. This model tends to be a little bit difficult to achieve in Nebraska just because finding, especially if it's a specialized database, finding enough libraries to enter into a new subscription at any one time and order it to achieve the higher discount sometimes is a challenge. I was wondering how many libraries do we really still have lots of libraries that haven't already jumped on to the databases that they already know about and need? Well, and that's the thing. Some of that sort of basic all-purpose databases already have quite a few subscribers and so some of these tend to be the more unique narrowly focused products which means again you're going to have a smaller pool of interested libraries. These discounts can also be a little bit challenging because you can't always tell libraries exactly what they're going to pay because you don't know how many libraries are going to subscribe so you don't know what your discounts are going to be ahead of time. And so there's a little bit more back and forth communication between me and the libraries and the vendors just saying, okay, are you willing? If we get a 10% discount, this would be your price. Who's interested? Depending on the numbers, we may have to go back then and say, okay, we only got 5%. Are you still interested? So there's a little bit of back and forth there. But again, that is the staff time we put in to try to make these deals happen. We're also able to sometimes piggyback on discounts that other library networks in other states have organized. If you've been around the library business for a while, you may be familiar with other library networks. For example, Lyrisis is a library network that provides services to libraries. Primarily, it used to be the southeast and the east coast, central eastern coast. Minitex is a library network based out of Minneapolis and they provide services to libraries in Minnesota, North Dakota, South Dakota. There's also MCLS, which is the Midwest Collaborative for Library Services, something like that. And it started out serving just Michigan and now it serves Michigan and Indiana. In any event, these networks are usually nonprofit organizations that have been formed in order to coordinate services for libraries. They arrange discounts, they offer training, et cetera. So in some ways, many of the services that we offer, the difference is they're not affiliated necessarily with a state government. They're freestanding. Sometimes libraries have to pay dues in order to be considered members of these networks. These networks, although they're not nonprofit, they do have to generate income in order to pay for salaries, et cetera. And so sometimes these networks do also keep commissions. So a network may arrange a discount of 10%. This is an example for the Mango Languages discount. I think MCLS receives a 10% discounted off list price and then they pass 5% on to the libraries and they keep 5% for themselves, which is something that we as a state agency don't have to do. Nevertheless, it's still really valuable for us to partner with some of these networks on discounts that they've arranged because they are able to bring so many libraries together into a group that they're often able to achieve bigger discounts. The Oxford University Press discount is coordinated by Lyrisis, and that is one of those discounts where some of the products are available at 25% off list price to libraries. Lyrisis is already going to be keeping some sort of commission as a result of that discount, but libraries are still getting 25% discount off list price. So that's an example of why it sometimes behooves us to partner with other libraries because they've got a library network, because they've got a larger pool of libraries that they can bring together and therefore they can negotiate larger discounts. So when our small group in Nebraska libraries can link up with their giant group of states that they cover, yeah. And our members don't usually have to pay any kind of membership fee to take advantage of that discount. And we do benefit the network by contributing libraries to the pool of libraries that are subscribed. So we added their numbers too. So it can help them as well if it's one of those situations where the more libraries can get the cheaper everyone can get. Absolutely. And they only have to deal with us at the library commission instead of the multiple individual libraries. So we'll organize it. So we serve as that central point so they don't have to email back and forth with, say, 15 libraries. They just email back and forth with us and then we coordinate the communication on our end. So again, that works out well. Finally, sometimes vendors are willing to give us a price quote based on the pool of population served of all the interested libraries. And they then let us allocate the pricing. So for example, and this is not a real example because numbers never come out nicely rounded like this. But for example, say a vendor usually says, okay, if your library serves a thousand, zero to a thousand pop served, we're going to charge you $800. Say you've got four libraries that serve populations of 250 each, normally they each have to pay $800 because they're in that zero to 1,000 pop served category. So they each have to pay $800 in order to subscribe to a particular product. Occasionally a vendor will say, okay, if we just have to deal with you, just let us know the total pop served of all the libraries that want to participate. And so we'll say, okay, we've got four libraries and their combined pop served is a thousand and they'll say, okay, that's $800. And then we're able to proportionately divide it up and each library pays then $200. So when we're able to get a vendor to offer us a deal like that, that's when we really see some really low prices. So there are products out there that we have some libraries subscribing to for under $200 because the vendors offer deals like this. Those are situations where I kind of sometimes hope that the vendor doesn't realize how little some libraries are paying because that sometimes makes them uncomfortable. But occasionally we're able to get a deal like that and that then is really beneficial. That's one thing that I'll say about working on these kinds of discount deals is sometimes you don't know going into it what kind of deal you're going to get, whether it's going to be whether you're going to wind up with a really great deal or sort of a disappointing 5% off. So we go into it not knowing always sometimes we're really pleasantly surprised with what we're able to come away with. Other times we get the 5% discount and we take what we can get and we continue to work on getting better deals in the future. So that's kind of a snapshot of the types of discounting we're able to arrange and how we're able to get discounts. A lot of the information on this slide has probably already been touched on again, but I want to make sure that I point it out. As I said, if we can publish pricing on our pricing pages on the website, we will do so. If not, I will send it out individually by email to interested libraries or sometimes I can send it out over the trial mailing list, which I will talk about in a minute. Part of the discount program is us doing centralized billing if more than one library subscribes. So that's one of the things that we're able to bring to the table when we talk to vendors. We'll say if you'll offer our library's discounts, then we will do the billing and the order processing for you and then you only have to deal with us. So for instance, if we were going to do that for a vendor in order to get a discount for our libraries, we will send out all the subscriber agreements, renewal agreements to the individual libraries. We'll collect them from the libraries. We'll let the vendor know in one email here are all the libraries that are subscribing. When it comes time for the vendor to bill out the subscription cost, they send one bill to us. Sometimes they'll have it broken down by library. Sometimes they'll just send just one lump sum amount bill, not even broken down. We pay the vendors. They only have to worry about dealing with us. And then we turn around and we bill the libraries, the individual broken out cost. Again, it's us doing a lot of paperwork and administrative activity with the intention of it benefiting you. That also sometimes gives you a little bit more flexibility in terms of payment. Sometimes we'll have libraries that actually want to pay early. And so we can bill them early if they want to get some funds encumbered before the end of fiscal year. Sometimes they may need to wait a little bit before they pay. So of course, a vendor would send them a bill and want to be paid in 30 days. Sometimes we can maybe wait a little bit longer and let them get into their next budget period or something. So we are able to offer a little bit more flexibility that way. To try to keep our own sanity, we try to keep subscriptions on some standard terms. July 1st through June 30th has always been our preferred term because that's our fiscal year. But we also have a number of subscriptions that run on January 1st through December 31st term. Oftentimes, if somebody wants to subscribe midterm, we'll try to get them on prorated subscriptions. So they might pick up a subscription in October and then it'll be October through June 30th subscription with a prorated price. And then they'll be on the regular subscription cycle in the future. So we're able to do that. Sometimes our nice, neat subscription terms don't work out. And so we do have some subscriptions that run on odd terms. For example, Britannica Online is always on June 1st through May 30th term. That's just the way it is. So we do try to be flexible. But when we can, we push vendors to let us have everybody on the same term. Again, to reiterate, one nice thing about being a state agency is that we don't have to try to raise money to exist. And so we are able to pass entire discounts on to libraries, whereas other networks that do similar activities, discount programs, have to keep some of it themselves. So that's another benefit that we're able to pass on. Just to give you a little bit of an idea of what we do, I tried to capture the numbers of subscriptions, database subscriptions that we worked on from a January 1st, 2014 to December 30th, 2014 period. So a 12-month cycle. We processed here at the Library Commission 538 individual subscriptions for 161 unique databases. We had 128 different libraries participate in the program and the total cost of all those subscriptions was almost $2.5 million. Now the one number that we can't always easily track is how much money was saved since it's often hard to really pin down vendors on. What would you have charged outside of the consortium? What would you have charged? It would be hugely time consuming to try to pin that information down. So we don't know exactly how much is saved, but you can see $2.5 million is a lot of business going through us. I want to go ahead now and move on to books and supplies. This is a lot more static of a discount program with the discounts on databases. As you can imagine, there's always annual renewals, so there's always activity going on, new pricing coming in. We are constantly communicating about that program. Discounts on books and supplies is kind of more set, so you don't hear as much about it. That's a really good reason for us to be having this Encompass live, so I can promote it and remind people that it exists. Again, we've got a page on our website that has information on this program, so I'm going to go ahead and jump back to our live website. Again, I can come up here and go to the discounts menu on the left and go to books and supplies. The way this program works is that vendors for supplies or books will contact us and say, we want to offer a discount to your members, and they'll often use the word members, because they're used to dealing with networks like Lyrasys or Minitex, where you actually as a library have to pay a membership fee and become a member. We use that term member very loosely. In some instances, a particular vendor might only be offering a discount to public libraries. Sometimes this happens because of the way they break their business down and a certain division only works with public libraries, so they are not able to offer that discount through their channel to a school library. Occasionally, we interpret members as any library in Nebraska whenever we can. Occasionally, a vendor will be a little bit more specific about which market they work with, and that's just because of their internal business model. Again, sometimes we're able to partner with other networks, so here's the Midwest Collaborative for Library Services, and they work with 3M Tatl Tape Security Strips, and so there's information that lets you take advantage of that discount. We've got Baker and Taylor, Nebraska Barnes & Noble stores have a discount program that they wanted us to promote on this page. Broadart offers savings. One thing to be aware of is some of these discounts on books and supplies, they contact us, they send us information about the discount they want to offer, and it's not time limited. They don't have any sort of expiration term on the discount, so they send us the information, we put it up, the information's available to you, we try to publicize it. From that point on, you work directly with the supplier, so we are not involved in that process at all, and so we don't always, it's not in the front of our minds even as much as the discounts are, because we don't have an intermediary role once you start working with that supplier. And sometimes when they don't have a particular end date for their discount, we don't always know if terms change, so it's not bad if something's not working for you, if you contact us and let us know, we can sometimes touch base with the supplier again. Every couple years I try to go through and reach out to the contacts at these companies just to make sure that they're still offering the discount. Again, when you've got staff turnover sometimes, the person that I was dealing with two years ago is no longer there, and so I need to reach out to the company again and get updated contact information, etc. Some of them are really good at reaching out to me every year, including the updates, others not so much, so that's something that I try to do every couple years. In some instances, for example, we're looking right now at BroDart, they always offer discounts for annual term, and you'll see right now this particular one is effective through January 1st, 2015. I actually had, a month or so ago, reached out to the company and have been trying to get updated discounts and have been assured, for example, that they will continue to offer this discount until they get us revised information. They actually said that they've been out of the office so much because of the bad weather on the east coast that they're incredibly backlogged as far as getting new discounts out. I actually did get this information earlier this week, so I have it and I just need to post it up here.  DEMCO, they've got a code for instance that you need to input on orders either on their online order form or if you're doing it by phone, making sure that they're aware of that. This program, like I said, I put the information up there and then it sort of takes care of itself except maybe annually when I need to update information or if I need to reach out to companies to double check to make sure they're still offering the discount. Again, it's not something that's as active as the database discounts, so I think sometimes we don't publicize it as much just because there's nothing new there. That's one of the reasons why this Encompass Live is good to be able to remind people that it's there. Yeah, I've already covered all that information, so this slide is there before review if you view the slides later. We also are able to offer discounts on information today conferences, so you can kind of assume if you're going to be attending computers and libraries, internet, librarian, that they're going to be offering a discount to Nebraska Libraries through the Nebraska Library Commission. This has been in place since obviously at least as far back as 1997 when I saw it. You first found the information, something about it. It was interesting because at that time the discounted rate to attend computers and libraries was $99. So that's interesting to see how cheap it was back then. But as you can see, there's absolutely no reason to pay the standard rate to attend when you can get the discount. It's over $200 off. It used to be over 50% off. The discount is not quite as steep now as it used to be, but still that's pretty significant. So when these discounts become available, I post them to the conference discount page on our website. Jump up here and discounts on conferences. So I will put the discount here. There's usually a promotional code that you have to input into the online registration form in order to get the discount rate. I don't put that on the public website, but what I do do is I will send out mailing list messages, usually to the trial mailing list and systems mailing lists that includes that code. I also post information on our Encompass blog, and I will say in their contact, if you didn't receive the mailing list message that included the promotional code, contact Susan nicely to get that code. I try to get that information out there so you know it's available.  Staying informed, we do have a trial mailing list, and this is the primary vehicle we use to communicate about database trials that are being offered. Asking for libraries that are interested in subscribing after a trial, seeing what kind of discounts we can arrange. I will often post pricing to the trial mailing list. I'll post user names and passwords for trials on the trial mailing list. So this is really a mailing list that we encourage you to make sure at least one staff member signed up for if you're interested in taking advantage of any of the discounts. This is the primary way we communicate detailed information that can't necessarily go out on our public website to libraries. The trial mailing list is really, you really need to be on there if you're going to be participating in some of these programs. We also try to post information to the Encompass blog which is fed to the Library Commission homepage, so if you follow that blog and there's something special going on, I try to post to it, but I don't post every single little message that I post to the trial mailing list to the Encompass blog. That's really not the purpose of the blog, so I try to catch your attention on the blog, but within my blog posts I usually say to make sure you get information like this in the future, sign up for our trial mailing list and I'll point you to the trial mailing list from my blog post. I'm on the list just to keep track of what we do here and there can be quite a few. This trial is happening, this one started, this one's coming up, this one's ending, so let us know if you want it. There's a lot of reminder type things like that. You don't necessarily want to see on the blog as a regular feed, so we always try to post enough to the blog to catch people who might be new to the state, who might not have heard about the trial mailing list, but we really are trying to get people to the trial mailing list because that's where the most information and the most detailed information is going to go out. I've already showed you, we've got the three pages in our website that are sort of our jumping off points for the discounts, database and e-resources, discount page, books and supplies page and conferences page, and I think that you've seen the ways you can get to those through our menu system. I just want to clarify something to ask, this trial mailing list that we have is just for Nebraska Webber. The trial mailing list is just... We do have people that watch the show or do the recordings from elsewhere, this would be something that only Nebraska librarians would get on because it's just trials for our Nebraska people. And we do monitor who signs up for it. Occasionally we've had vendors try to sign up for it because they view it as a wonderful vehicle for communicating with all the librarians in the state about their particular products and we do not allow that. We're going to be the intermediary between them. We are going to be the intermediary. But again, that's one of the reasons why they will sometimes offer us a discount because they know that we have really good ways to communicate with all of you. So we will send out the information but sometimes we won't include all the superlatives that they include such as, oh, this is the best, most exclusive, absolute must-have product. Sometimes they'll send us that kind of language and if it's a brochure or something that's clearly something that is from the vendor and we send it as an attachment, that's one thing. But if we're going to be typing something into a mailing list message that they send us will often leave out some of the superlatives. What we always tell librarians is we really can't necessarily evaluate these products in terms of which is the best, which you should subscribe to. We try to give you opportunities to try them out but we don't know what your patrons need, what your curriculum is, et cetera. So we don't always have any specialized expertise. Yeah, we don't give an opinion or evaluation, that's what the trials are for. If you guys figure that out for yourself, is this going to work for my students, my users, or not? Absolutely, yeah. Finally, I wanted to list contact information. Debra Dabragos is the head of our department and so sometimes she is the primary contact, the first contact that a vendor will make. You don't contact her and say, we really want to work with you to make some deal available to Nebraska libraries. Depending on the situation, someone's Debra will negotiate with them until something's established and then she'll pass it on to me and I'll do all of the communication, putting information up on our website, getting it out to you, doing subscriptions, renewals, et cetera. Sometimes vendors just contact me directly because they've worked with me in the past and so they just go straight to me. The other person that works with the program is Jennifer Rampe and she helps me with sending out renewal agreements and she's also the person that does actual invoices so she's the one that sends out invoices to libraries. If you have questions after you've already ordered something and you want to know about invoicing or billing or have you paid, she's the person that's on top of that. When in doubt, I'd say call me. I'm the person that has my fingers in the most places and can get you the information you need but I wanted to make sure that you aware that Debra Dabragos can answer questions and then also Jennifer Rampe is also involved in the process and Jennifer's pretty good about actually knowing where I keep paperwork in my office too so if I'm out of my office sometimes she's been able to answer questions just because she's been able to poke around and find folders on my desk, et cetera. Just like Jennifer can sometimes answer questions about things that I work with, Jennifer does the invoicing but I can get into QuickBooks and see if she's invoiced to you. I can resend invoices. I can see if we have a record of you being paid or not. Even though we do slightly different things, there's overlap. But again, when in doubt, I'm probably the first person to call unless you specifically know what you're dealing with is something best addressed to somebody else. That's all I have today. If anybody has any more questions, I'd be glad to answer them. Otherwise, it's just great to have another opportunity to make sure that people are aware of this program. Great. Thank you so much, Susan. This is something we've been trying to do for a while, is get you back on to update if there's anything new or just get the information out there to remind people that we do this and we've been doing it for so long and all the great discounts. Even if we only had a small number of people attending, just the fact that we're advertising that we're having this, the recording will go out. Maybe send it to the trial mailing list. That's a good idea. The information's updated and out there and hopefully it crosses people's radar again that this program is available. Does anybody have any questions? Nobody really typed in anything during the show when she was talking, so that's good. If you do have any questions, let us know here. Type in your questions or you can use your microphone. I think it was a pretty comprehensive coverage of what we've been doing, history of it and everything. That was great. It doesn't look like anything urgent is coming in now. Well, if you do have questions, you know who to call. Absolutely. Susan, obviously. Yeah. Or anyone else, if you need to get attached to someone quickly, either Deborah or Jennifer as well. That, I think, since nothing is coming in. Oh, we just have that. Susan took it to 11. I guess that's good. Yes. It's probably one of the library commission staff that was always saying, I don't know if I've got more than 30 minutes, but I don't know if I can make it up. Yeah. And just some thank yous coming through. Yes, great. Thanks for the information. It's very useful. All right. So I'm going to pop back over to our website as well. So thank you very much everyone for attending. Thank you very much Susan for getting all this information out there. The show has been recorded as usual, so that will be available most likely later this afternoon. I'll get it all together and put out there. And thanks to Krista for doing all of the online webinar parts of the presentation. Yeah. Sometimes I do these on my own outside of Encompass, and it's much nicer when Krista has all of the controlling parts. I do it every week, and I just kind of mindlessly go through the motions, but just because I know it, yeah. And then we have the technical issues of the camera. Sorry about that.  Yeah, I think so. So that will wrap it up for today's show, but I'm going to pop us over to our Encompass Live website here, just like Susan did to her sessions. It will be posted onto the website here, down here underneath our upcoming sessions is our archive sessions. I'll post the link there. It will have, let's see if this last one had yet. The recordings available, we put them up on our YouTube channel. The presentation will go up on our slide share, so all these slides will be there. And then all the links to the different places on our website that Susan mentioned will be available as well. So that will wrap it up for today, as I said. I hope you join us next week when it is our monthly tech talk with Michael Sowers. Michael is our technology innovation librarian here at the Nebraska Library Commission. And once a month he comes on, usually the last week of the month to talk about something more techy. And next week he's going to be doing a demo of the Amazon Echo. It's this little device that's a voice controlled device from Amazon that you can use to do all sorts of things in your house. And you can talk about that and other voice control type technologies coming up. So definitely do sign up for that. And any of our other shows that we have here on our schedule through April are all up here, all the different topics we have coming up. And if you are a Facebook user, Encompass Live is also on Facebook. Definitely do go over there and like us on Facebook. We'll post when the recordings are available there. I remind people every week to log in for the live show on the fly if they want to. So definitely give us a like over on Facebook and keep up to date on what we are doing here. Other than that, I don't see no urgent questions last minute have come in. So that's great. We are good for today. Thank you very much and we'll see you next week on Encompass Live. Bye-bye everybody.


I bet you didn't know why Michael Jordan wore number 23 on his jersey. MJ was motivated as a child by his older brother Larry, so they decided to both wear the jersey number 45 and when they became partners on the same team, they couldn't wear the same number, so he decided to split the number 45 in half and rounded it up to 23 and that's how he got the number 23. He then wore that number in college and then went on to the NBA to create one of the most iconic jersey numbers in history. So subscribe so you can tell your friends.


We just made this two-tiered cake stand with only two cuts and no other tools. We even added some image transfer to really make it pop. This thing was a piece of cake. We'll show you how we did it right now. What is up? A welcome back? Would you like to do it, Builder or make it? A great... That's what we do on this channel every week. This week we're making a two-tiered cake stand. That's like double the cakes half the cost. Maybe it's cupcakes. Quadruple the amount of cupcakes half the cost. We got a candle holder that we found at Hobby Lobby like half off. It was over there in the clearance. So I decided we should make some cakes. Yeah, hold cupcakes and cookies and pastries. We found two scrap pieces of wood. We have a 16 by 12 and an 8 by 8. Just happened to be there. And we picked up these little candle holders from Hobby Lobby. We're going to use them as feet. Step one! We're going to glue the feet to the bottom one. Look at these cute little guys. Can you see them? Cute little candlesticks. They got a flat bottom. That's going to go against the board. And we're going to leave the open piece. Is that the other one? Yeah, I think so. It's already sticking out. I don't know. I think we're good. Okay. So that's setting up. We're going to glue this piece to the top piece. But we're going to use Gorilla Glue. Where did the Gorilla Glue go? Oh, it's right in front of me. We're using Gorilla Glue because there's a little lip on here. So I'm hoping that the Gorilla Glue will expand in here. And stick the top to it. We'll see. Put a little on the rim. But yes, that stuff. I don't know if you've ever used this Gorilla Glue. But it definitely expands. You want to put any like... Yeah. And then I'm going to triple glue. Okay. You want to show them what you've got? All right. See? It's my paradise of glue. Is that center? Let's see. It's good. Yeah. I don't like that. We have it centered. It's straight as a center. As a look. We're going to give this a few minutes to dry. Two hours later. Glue seems dry. Now we're going to paint it. Yes. I think I'm going to clean up this glue. We all would have. The Gorilla Glue. I got a little fuzzy. You got to foam it out. So I'm going to trim that up real quick. Step two. Now we paint. Now we paint. Oh my goodness. There's these images down on the boards. I'm going to photo transfer. Although we don't have the photo transfer Mod Podge, which they do make. So we're just going to use our matte finish Mod Podge. Paint the front. Wait a minute. Now we wait. It's going to stay in here. On this project ever. Yeah. Tomorrow. Everything is jewelry. Everything is dry. See now we just got these random pieces of paper on here. We actually gave it like 12 hours to dry. We're actually pretty good. I don't think I'm lasting the whole 24 hours.  It feels really dry. I think it's dry. How much dryer can it get? Dry is dry, right? Dry is dry. I'm going with dry is dry. So now we're going to start to get it wet and rub the paper off. See if it works. Got me a cup of water. Can you see how great those turned out? Look at that. I am impressed. Impressed. I was very doubtful that it would actually work so good. I was... He was. I didn't think it would work so good. We're going to distress it. We're going to scuff it up a bit. Make it look aged. Make it look used, loved. I think it looks good. You can't really see the distressing I don't think. Because we didn't put a coat of brown down first. It's just distressing right through to the pine. But we have a fix for that. Step... We wax it. And we're not talking the Brazilian kind. We're talking the furniture stuff. The decorative wax. The key is the dry brush it. You can go... Oh wait. Oh yeah. There you go.  You brush almost all of it off. Just like poop. Yeah. It does something. Now it's a two year old. Yeah. Now. Now who's it? Say I got it real dry. Definitely Garrett's favorite part. Poke a poke up in here. Dirty bird over here.  That's a dirty bee. Dirty bee. I think this will work. I don't have a cloth. Other than this one. I think it looks great. That's what we got. Tilt it forward so they can see. There you go. There you go. Right side up. Let me see all the little things. See little feets. Alright. Alright. Looking good. Looking good. If you like this week's project, hit that like button. If you're not yet subscribed, hit that subscribe button. And hit that bell because we do this thing every week. And if you need another video to hold you over till next week, check out this one. Or that one. Maybe that one. Definitely this one. We'll see you guys next week.