OK, thank you. Professor Gao received his specialist degree from Xi'an Jiao Dong University of China in 1982. And his master in PhD degrees in engineering science from Harvard University in 1984 and 1988, respectively. He served on a faculty of Stanford University between 1988 and 2002, when he was promoted to full professor with tenure in 1994 and full professor in 2000. He served as the director of the Max Plan Institute for Metals Research between 2001 and 2006. And before joining the faculty of Brown University in 2006, at present, he is the water and number professor of engineering at Brown. Professor Gao's interest focused on understanding of basic principles that control mechanic properties and behaviors of materials in those engineering and biological systems. He has been elected to National Academy of Engineering, German National Academy of Science, and Chinese Academy of Sciences. And he is the editor-in-chief of the Journal of Mechanisms, Francis Solis, the leading journal of his field. He has received numerous academic awards from John Simon and Guggenheim Fellowship in 1995 and to recent to the Rodney Heel Prize in Solid Mechanics from the International Union of Theoretical and Applied Mechanics in 2012, Prager Medal from Society of Engineering Science in 2015, Nada Medal from American Society of Mechanical Engineers in 2015, and the Theodor von Kamen Medal from American Society of Civil Engineering in 2017. So let's welcome Professor Gao. Thank you. Thank you. OK, can you hear me? This is working. Thank you, Ting, for your kind introduction. And thank you all for having me here, giving this prestigious lecture. It's my great honor to share with some of you some of the work in my group on nanomaterial cell interactions. So let me first acknowledge two graduate students, one postdoc in my group, who did actually the work. And I also benefit with a lot of faculty collaborators. So Bob Hurd, he's a chemical engineer. I've been working for over a decade. Professor Agnes King from Brown Medical School and Professor Manu Nakis from Brown Medical School, Professor Pada Flahovska. She actually used to be a Brown. She moved to a Northwestern University this year, and also two faculty from Harvard Medical School. So I've been working in this field for the last decade. Basic idea was looking at the mechanical aspect of how nanomaterial interact with cell. Of course, there's a huge chemical aspect as well. They redox the reactions. But those aspects, I'm not addressing today. I'm addressing from a mechanical engineering point of view today. So the kind of problem I'm interested in, so-called mechanics of cell nanomaterial interactions. Here's a couple of review papers we wrote in the last couple of years. Mostly we're interested in how a special cell membrane interacts with nanoparticles of various geometries, such as a sphere, or a cube, or a tube, fiber, or a soft or stiff pad. These are called one-dimensional nanomaterial, or two-dimensional nanomaterial, or sometimes this is called zero dimension. Also the surface chemistry sometimes come in, but not always. So also we're interested if this nanomaterial get into cell, what are the destiny? How does the material interact with intracellular vesicles, such as lysosome? What are the biological consequences, their biological effects? So here's some summary. Cell-optic pathways of nanomaterial with different geometrical properties, such as size, shape, orientation. Mechanical properties, such as stiffness. Or chemical properties, such as surface functionalization. So how these different properties influence the cell-optic pathways of the nanomaterials? Second of all, the intracellular packaging of nanomaterials, so after the nanomaterial get into cell, they have a package of issues. So how are they going to go into the cell? What are the consequences? Toxicity damage of nanomaterial to cells and membranes. So these are the issues we're interested in. Let me show you a few backgrounds again. What is the driving force? What is the background motivation for this type of work? First background I mentioned is so-called nanotoxicology. Essentially, this is an emerging field, especially in the last decade or so, since the National Nanotechnology Initiative. President Clinton initiated this almost 20 years ago now. So essentially addressing the biological environmental impact of nanomaterials, huge field. People from different fields can all contribute understanding. But there's a need because the engineer nanomaterials. So these are not a result of natural evolution, but because we made them in the laboratory, or we fabricated them, there have become a significant fraction of material flowing in a global economy. It is estimated about the majority of them. This is like an estimate a few years ago. 100,000 tons of this nanomaterial, most of them will end up in landfill. They will end up in environment. No one knows their long-term consequences. So there's a need to know how to influence the biological world, the environmental ecology. Two specific examples, carbon nanotube production exceeded several thousand tons per year in 2013. These are really man-made nanomaterials who are chemically very stable, ultra stable. They're not easily degraded by biological enzymes. So they will be there for a long time. And there have been many applications, or have been used in many applications, such as energy storage, automotive parts, boat hose, sporting goods, water filter, thin film electronics, coatings, actuator, lots of applications of this material. This is a booming nanotechnology. And so a lot of this material will eventually end up in environment. So what are the effects on our life, on our species, not so clear? And graphene is a newer, even newer nanomaterial. Graphene was around 15 times annual production, 2009. And five years later, it's grown by a factor of 10. Today it's even much higher. So all this just shows a rapid growth of nanotechnology, nanofabricated material. Their environmental biological impacts are unclear at this moment. OK. Another motivation for this type of study is called nanomedicine. A lot of people are using nanoparticles, nanotechnology, for diagnosing cancers and treating cancers. So a lot of people. So I'm giving an example of antibiotic. How this nanomaterial can also be used for the benefit of addressing bacteria issues. As we know, since 1940s, since they emerged in the penicillin and the various antibiotics, this has made a huge difference in the quality of life. A lot of today's population, a large fraction, exist because of the antibiotic. This is a big advance in medicine. However, bacteria also been evolving. So there's a war between the human development of various antibiotics and the evolution in the biological world. They keep getting new species evolving, mutations, and things. And currently, the looming crisis means the human pace of innovation has not been able to keep up the pace with the emergence of new bacteria. In fact, the World Health Organization issued the crisis, the warning. The most recent release was a year ago, February 27, 2017. They identified a couple of bacteria in a critical stage, wider critical. This antibiotic called copper panium. Copper panium is the last resort. It's not prescribed generally for antibiotics. It's only for the last resort if everything else failed. So these emerging couple of bacteria become resistant to copper panium. That means we have no cure for them if they break out. So the challenges here in this field is the development of a new antibiotic has not been able to keep up with the pace of antibiotic resistance. Most existing antibiotic targets the protein, or cell wall, DNA synthesis pathways. They do not work on non-growing bacteria. If the bacteria, they're called persisters, some of the bacteria stop synthesizing, they become sleeping, they go in the sleep. So the current antibiotic have no effect on them. So the potential solution to this is two questions. Can we use modeling simulation to speed up the pace of drug innovation discovery? First, we need to speed up the innovation pace. Second, can we seek membrane active antibiotic that kill persisters? Because these persisters, they're not synthesizing so traditional antibiotic on the market, they target some synthesis pathway, they do not work. Can we develop such things? So one of my topic today is I'll give examples of this application. Another background is the rapid advance in bioimaging. Traditionally, optical microscopy is the prime tool in using medical school. Well, material science, we use electron microscopy. But electron microscopy do not work on live cells.   So optical microscopy, the resolution is limited to optical wavelengths. So in the last 20 years, the last two decades, there are various near-field scanning microscopy and other fluorescence techniques emerged. So to bring the lens dimension into nanoscale. In fact, 2014 Nobel Prize in Chemistry was awarded to three scientists, Eric Betzig, Warner and Stephen Hale for developing super-resolved fluorescence microscopy, which brings optical macro into nanodimension. So many of the things we can only speculate, maybe a decade ago, can today be visualized. This is one example. Showing the phyllo-podia is a structure on a cell membrane. Phyllo-podia, it's called pseudo-legs of cells, play an important role in the cell migration. You can see these structures are typically 100 nanometer or even thinner. But this movie actually is not an electron microscopy, it's optical microscopy, it's a nanostructure evolution. This is only possible, this is a paper from a paper published in Science in 2014. So not even possible five years ago, but today, using these highly-resolved optical microscopy, you can begin to study the mechanical aspect of nanostructures in the drug with the cell. In a limited time, I thought today, this is a huge field. So I'm just selecting three problems to share with you. Select a problem in mechanics of cell nanomaterial interaction. First problem, first class problem, I say morphology. This is an area where we question the mechanics of cell entry of a 1D nanomaterial. And we can just give you an example here. Second problem I'm sharing with you is called packaging. How a nanoscale material is packaged in an intracellular vesicle. So this has to do with cytotoxicity of a flexible nanofiber. Third problem I'm sharing with you is the membrane active antibiotic. There we do energy mapping, showing how physical science can help. This is really a biological problem, but I'm just giving you an example of how physical science is helping us understanding the mechanism by mapping the energy evolution as this nanomaterial interact with cell membrane. So the first problem, I'm showing a little bit of mathematics, so only two slides. So bear with me. So here, for example, I'm just giving you this example. Suppose we want to know how a nanofiber, such as carbon nanotube intercells. So we can use a physical science approach by writing down the free energy of interaction of the two objects. One is a cell membrane, as I draw here. One is a nanofiber, which is the intercell angle theta. Theta is orientation. So there are the free energy. We have two term. The free energy consists of, first term is the bending energy, deformation energy. Because in order to bring this nanomaterial inside a cell, the cell membrane has to bend, curved around its nano-objects. There's a deformation energy. So here, the curvature, H is the mean curvature of this membrane. Kappa is called bending-rigidative membrane. So this term represents the deformation energy of the membrane. The second term is the membrane tension energy. So the membrane subject to some tension is maintained by the cell. Because the nanomaterial, stick of nanomaterial, change the area, change the membrane surface area so that has a tension energy. Anyway, so you're looking for the minimum energy configuration of these two objects, assembly of these two objects. It turns out this problem is governed by a single parameter. This parameter is a non-dimensional parameter. It's called normalized tension. It's defined as the real membrane tension times the square of the diameter of the fiber divided by the bending rigidity, a single dimensionless parameter. That parameter is given. You know the behavior. You know what is the minimum energy configuration. It turns out the critical number is this 2 pi over 5. If this dimensionless number is bigger than 2 pi, which is 1.26, if the normalized membrane tension is bigger than this number, then the minimum energy configuration is shown here, having this fiber just like a tree, like a tree planted on the ground. This is the minimum energy. The nanomaterial like to stick perpendicular to the cell membrane. Another case is, as soon as the normalized membrane tension is bigger than this number, 1.26, then the favorable energy configuration, we call paralleled hearing, is having this nanomaterial just fall down. This tree just fall down on the ground with a very shallow angle. This is very simple. The conclusion is very simple. So I'm not going to go into the depths of mathematics, but it's just simple. So here's two examples of free energy evolving. So here shows one example is normalized tension for the 1, which is in a subcritical range. So here I'm plotting the red is the bending energy. The first term, the membrane bending energy, how the bending energy change with the entry angle, with the orientation of this nanomaterial, which clearly shows the bending energy is minimized at 90 degree when this is normal to the cell membrane. The tension energy is showing the blue curve. The tension energy prefers a shallow angle, not 90 degrees. Actually, the energy increase. But total energy, the net, if you add these two terms together, the bending energy dominates. So this is total energy, the black curve. So total energy favors this mode, perpendicular entry mode. For on the other hand, if this normalized tension equal to 3, which is a supercritical case, which is this case, you see the bending energy still prefers 90 degree entry. But tension energy become more stronger so it dominates total energy. Total energy now prefers the paralleled hearing mode. And there's a lot of math details I'm going to skip in here. But let me give you a simple explanation how to use in the mathematical geometry. To understand why there's a perpendicular mode, or interaction mode, when the normalized tension is smaller than this 1.26 critical number, consider the bending energy. Bending energy of a membrane deformation is given by the bending rigidity of the membrane times the mean curvature. Mean curvature. If you still remember the calculus, the analytical geometry in your college class, there's a special mathematical surface called catenoid. Catenoid is given by this surface here. It's drawn exactly the shape. As I draw here in this figure, this is the shape of the membrane called catenoid. Catenoid is a very famous curve, soap bubble related to, and things. This mathematical curve has two principal curves. Every point has two principal curvature, kappa 1 and 2. They're equal in magnitude, but opposite in sign, which means if I have a mathematical surface like this, then any point along the surface have zero mean curvature. Because a biological membrane, the deformation energy is proportional to the mean curvature of the surface. So in other words, this curved membrane have zero deformation energy. Even though it's curved, it's highly deformed. It looks like deformed, but it actually have zero deformation energy. This is a tractor existing nature. So provided a perfect mechanism for our cell to take in food and nutrition, it has this special shape. You can curve in so the food and everything can be transported in and out of the cell without causing too much deformation energy. This is an attractive state. But unfortunately, this profile dominates only when tension is not too big, but a little critical normalized tension. If tension is sufficiently big in a supercritical stage, then tension energy dominates. Tension energy is proportional to the increase in membrane area as a result of sticking fiber into the cell membrane, which prefers a shadow, a parallel adherent node. So that's why our food particle is always break down our body, our stomach, right? Our body process a breakdown food particle nanoscale so that we have this mode. So a cell absorb all the nanoparticle nanomaterials through this channel by deform itself into a catanoid. So it can receive this material at a low cost, a zero cost, in deformation energy. But if the material is big, if the nanofiber is big, then it actually can evade this pathway and become a parallel adherent. So here's some experimental evidence. Sorry, let me just say one word here. So this condition, when sigma smaller than sigma c, when they normalize tension smaller than this one, is equivalent to, say, a critical size effect. If the nanofiber, if the fiber is smaller than AC, where AC is given by this, square root of five, membrane tension divided by pi times bending rigidity, this is about a length scale of 40 to 150 nanometer, depends on the cell type, depends on the cell type, in various, so the critical dimension. So this critical state is also equivalent to a critical size for the fiber. If the fiber is thin enough, if the nanofiber, really nanofiber, if the radius of the fiber is smaller than this critical number, then it prefers the normal perpendicular entry. If it's big fiber, then you prefer to lie down. So this is what I'm gonna interpret. So experimentally, we did experiments on my collaborators. We feed the cell into various nanomaterials with different chemistry. So here showed, the micrograph shows a carbon nanotube. Okay, when A is, we consider small enough fibers, small enough radius. So here shows a carbon nanotube, right? And so you can see they're nearly perpendicular. This is a scanning electron microscopy. Okay, showing these nanotubes are standing perpendicular to the cell surface, as we expected from theory, right? And when our paper was submitted, the reviewer was saying you gotta check this is not a chemistry fact, right? So we changed the gold nanowire. Okay, here's actually a bundle of two gold nanowire, but still thin, okay? You can see it's a normal perpendicular to cell surface. Asbestos fiber of completely different chemistry from gold and carbon, also stand in perpendicular to cell membrane, okay? Now, for bigger bundles, okay? We find that when a fiber bundle together, you form a large enough bundles, when they exceed the critical size, then they fall down, they lie down on the cell surface, okay? So 1D nanomaterial, for example, CNT's nanowires, asbestos, fibers, intercellular tip-first perpendicular entry mode, as predicted by our theory, and fiber bundles adopt the horizontal motive interaction. This is kind of consistent with what theory predicts. But ideal experiment, ideally, if we can design experiment, we should have a growing fiber. The fiber growing diameter, our theory predicts initially fiber would stick perpendicular. Now it will lie down at a critical size. But unfortunately, we couldn't do the experiments. So we took a look at this again, this phylo-podia, phylo-podia. Phylo-podia is like a growing fiber. It's not, except this phylo-podia protruding out of the cell membrane instead of going from outside to in, right? Instead of entering cell, actually it's poking out of the cell. But the free energy function is the same, okay? We write down the free energy function for two problems, even though they're physically different problem, but they're mathematically, they're analogous to each other. Okay, the phylo-podia are considerably protruding microtubular proteins, right? You can see this paper, it was 15 years ago, showing how phylo-podia, the forming process phylo-podia, have this microtubular protruding out, then they merge together, right? So it's like a fiber protruding out of the cell membrane, and that fiber is growing, right? But before, nobody was able to explain why phylo-podia, the size is always limited. They don't grow too big, okay? Phylo-podia grow to be like 100 nanometers or so, then they stop, you know, it's pretty stable, right? So here's a movie I showed already. Okay, if you look at, they do transition, right? They do, they do actually, right? You can see they actually fall down. They have the transition between standing up and lying down. Okay, so again, our theory indicates the 1D protruding nanostruction cells, they should become unstable at a critical condition, which is given by this critical diameter. So this movie combined with this explains why these fibers are unstable, right? These fiber structures, phylo-podia, initially they want to protrude out, right? But then when they grow thick enough, they lose their mechanical stability, okay? Okay, so, of course we can also, we have a theory, we can always check by simulation because these days, computer simulation becomes so powerful, we can conduct simulation to check this prediction. So in one simulation, here's a molecular dynamic simulation, we have a cell membrane with receptor molecules interact with carbon nanotube. In one case, we pick the membrane tension, normalized tension to be subcritical, okay? In this case, we see, we give it the initial condition as the fiber is oriented some angle with respect to the membrane. We see that the interaction, the physics, okay, just the physical force alone. There's no biological motor in this model, right? The fiber is spontaneous oriented to be normal, perpendicular to the membrane. If you take a supercritical case, normalized tension is bigger than critical than they actually lie down, right? So here's a movie I show in a subcritical case. We try to explain, okay? So we try to explain this experimental observation for various type of nanomaterial that tend to stand perpendicular, right? So in this simulation, we randomly orient a fiber with respect to cell membrane. We show, so biophysical energy, just purely physical forces, the membrane's spontaneous orient them. Again, this is because of the catanoid, right? The biological membrane prefer in this catanoid configuration, which is a minimum energy, minimum deformation energy. Okay, so let me show you a little bit a connection to disease, okay? For many years, it's known asbestos, nanofibers, asbestos fiber, they cause lung cancer, right? Because asbestos was used a lot last century. For construction material, they're cheap, they're fireproof, they can use insulation, they have very good insulation. But today we don't use them anymore. Most of the manufacturers asbestos have become bankrupt because of lawsuits, right? So this connection is known, they're connected to disease. But why? The mechanism, the mechanistic origin is not understood. So our model shed some light, right? So we said, there's a critical dimension, okay? If you make the fibers thin enough in a nanoscale, nanofibers lower than this critical size range, then the cell cannot distinguish a fiber from a particle. The cell only identified a tip of the fiber, the thing is a nanoparticle. So the cell tried to take it in, okay? Without worrying how long it is. Now imagine the fiber is very long, right? The fiber is actually very long, right? So you get it in this state, right? This is a famous paper probably in 10 years ago, Nature Nanotechnology. This is actually not asbestos fiber. This is a carbon nanotube. Here's an immune cell, okay? Immune cell, right? So you can think what is happening, right? Because this important size scale exists in nature, cell interacts with a 1D nanomaterial only with its tip if the fiber is thin enough because it's catenoid, right? Try to take it in. Actually immune cell is trying to kill it. It's trying to remove it out of our body, right? But this is a huge fiber, right?  So you're getting into trouble, right? So cells overeating, so cell getting in trouble. When the immune cell get into the stuck like this is called frustrated endocytosis or frustrated phagocytosis. The immune cell what it does is the immune cell will call for help, right? You activate your inflammatory mechanism cause a permanent inflammation in the target body many years later evolving to many health issues, okay? So this is like a priest try to arrest a criminal but the criminal turn out to be too strong, right? So the priest constantly call for help. For enemy that shouldn't, couldn't be killed so you get lots of problems. Your body turn into an agitated state for years. Okay, so no surprisingly, so this paper studied carbon nanotubes to the different chemistry from asbestos fiber the pathologic, the phenomenological, biologic concept is very similar, okay? So their conclusion exposing method, method the lining of mice to long multiple carbon nanotubes resulting asbestos like lens dependent pathogenic behavior. That's our main conclusion. So it doesn't matter, it's not a chemistry driven, it's actually a mechanical driven phenomena. If it's long and stiff and you have this toxicity, right? Because it's a, the material, the geometry of the material such that your immune system have not learned how to deal with that, okay? So the second problem I'm gonna share with you is the packaging issue, okay? Imagine nanomaterial such as carbon nanotube intercell, okay, so where do they go? Okay, they go to this important compartment for lysosome. Lysosome is an intracellular vesicle which is responsive for digestion, okay? Our food particle taking to cell and end up lysosome being digested. Lysosome is a highly acidic environment, contain many, many enzymes, okay? It is digestion, degradation, autophagy, metabolism is very critical. The size of lysosome range from 0.1 to 1.2 micrometer. So highly regulated by cell, right? So we are trying to address when a fiber is a problem because a sphere is no problem, right? It's known that if you just feed the cell the sphere, spherical gold nanoparticle, no toxicity. But if you feed them with a lung fiber, then it's a problem arise. So where's the critical transition? How big is big, right? How big is the toxicity starts? So that's a question we address. Here's the key engineering issue we identify as a so-called bio-soft and bio-stiff. So what is bio-soft? Bio-soft we define as the following picture shows, the following movie shows. If the nanomaterial is lung slender and compliant, flexible, then this vesicle, right? It's trying to wrap it around. So it cause, because there's a pinch force, okay? The biological vesicle can apply a pinch force on this confined nanomaterial. If nanomaterial buckles and just nicely package inside the cell, we call it a bio-soft, okay? That means it's in a biological system. It can be nicely packaged, okay? And so this pinch force is about 20 people Newton, right? So we identify. Now, what's a bio-stiff? Bio-stiff if you feed the cell with a lung and stiff nanofibers, okay? You can see this membrane cannot fight against this. This is too rigid, too stiff. It doesn't buckle anymore, okay? It doesn't yield. So instead you have the membrane form a tether around. Deform around this fiber, form a tether, okay? Form a membrane tether. So you have a persistent contact here and here, right? It's like you have a pinch force persistent contact. That persistent contact cause problem, okay? As I will show you here, it's carbon nanotube. Because the fibers material have a high density, high atomic density, okay? There are these material have a high density of atoms here. They have a strong interact with lipid membranes. If you have a persistent contact of a nanomaterial, nanofiber with the membrane, it can extract lipids cause lipid molecule to be extracted along the wall of these fibers. That cause make this lysosome permeable. Make them pores, create pores in the membrane, right? So anyway, so the conclusion here is the persistent contact needs to local membrane damage due to lipid extraction. So we published paper a couple of years ago on this one too. Now we wanted to do experiment. We did my collaboration again. We did experiments to verify this, okay? Again here, we choose the material with the same chemistry. We choose carbon, okay? Same chemistry, all the materials, various geometry, okay? For example, here shows one micrometer long, stiff multi-wall carbon nanotube, okay? In this case, we see that here's a lysosome. We see this fiber is long, right? Cannot be deformed by lysosome actually sticking out just as we, our model shows, okay? This fiber is protruding out, membrane deformed around it, okay? So here shows carbon black, same chemistry.  And they become nicely packaged. 0.5, half micrometer stiff, same stiffness, but it's short. It's also become nicely packaged within the lysosome. A long, a soft carbon, like these are, these are, these are with only a few walls. So the nanotubes are flexible, right? You can see this flexible tube are being bent, okay? Being nicely packaged and bent and packaged within lysosome. It doesn't cause this kind of issue. And tango or soft multi-wall carbon nanotube, even more flexible ones, there's no problem. And carbon nanohorns, these materials like a horn, right? They have sharp corners, but they're pretty short. It's small in dimension, they have no problems. Okay, so the short story is we only find this case toxic, same chemistry, right? For this case, you, the lysosome is not capable of packaging them completely. This can cause problems. This is our definition of bio-stiff. All the other one we call them biosoft, okay? So our experiment, so we further verify this permeability. So we tack on an important protein that's usually contained within the lysosome. It's called Cotapsin B. Okay, Cotapsin B is an important protein that's normally within the lysosome. It doesn't permeable. If the Cotapsin B is released into cytoplasm and the cell die, okay, it's the trigger of apoptosis. It's a very famous signal, upstream signal that calls cell death, okay? So we feed different carbon nanotubes, carbon material here. We find only in this case, stiff multi-wall carbon nanotube involved cause substantial release of Cotapsin B. Okay, this is a clear evidence that lysosome become permeable. A lot of things being leaked out into cytoplasm. Okay, here's a number of cells with Cotapsin B release shows this long and stiff multi-wall carbon nanotube show those dependent release of Cotapsin B. The more such nanotube you have, the more Cotapsin B is being released into the cytoplasm. Okay, just our theory predicts, right? You have the persistent contact, damage cell membrane causing the leaking, causing the leaking, and then we can correlate with cell death. Okay, we can correlate the Cotapsin B release to cell death. Okay, so we monitor the Cotapsin B release. Let me also monitor how many cells are surviving, a surviving cell. Okay, we count down with 24 hours later how many cells are surviving. Okay, so here shows the dose dependent survivability. Okay, for example here, let me show you flexible, for the flexible carbon nanotube, so basically surviving, right? Most everybody is surviving. The only exception of a substantial cell death is this red curve and this green curve. So remember the green curve is the one micron carbon nanotube showing this picture, right? So this red curve is even longer. Okay, red curve is even longer. It's more than one micron long, so-called cell death. Okay, so this again correlates the geometric interaction, the packaging interactions to the cell death. Okay, so based on this insight, based on insight we draw a phase diagram, cytotoxicity phase diagram. Okay, phase diagram. Here we plot two parameters, vertical axis is the length of the fiber, horizontal axis is the diameter of the fiber. Here's the effective diameter because this nanotube is multi-waters, inner hole and outer surface, right? Effective means we're gonna condense with the solid fiber. What's the effective diameter? This curve is the buckling criteria, okay? So this, what is the critical condition for this fiber to buckle? Okay, under this 20-peak-one-second pinching force. Okay, so we did a lot of experiments. Each of these data represent experimental measurement, okay? Red means they're toxic. Cell are dying from the viability, assay. Cell are dying. Blue means the cells are surviving, the non-toxic. Okay, so in this range, this between this buckling and a critical length, this length is given by basically less or some size, okay? Less or some size. It's about one micrometer. And we bounded in this range when the length of nanotube is bigger than one micrometer, but they're stiff, it's in this range, okay? So we did a lot of experiments showing they're all toxic. In this upper corner, they're flexible, okay? They're, this range are slender fibers, they're flexible, they're packaged, they're non-toxic. Okay, there are two data here. Okay, and here, okay? In this range, the fibers are short, right? They're smaller than, the lengths of the fiber are smaller than one micrometer. They can be well packaged, okay? Because you don't have persistent contact. There are thermal fluctuation, the statistical fluctuation, right? The thermal forces would cause this fiber to basically do in brownie motion within the cell membrane. So you don't have persistent contact. You don't have persistent contact, it's no problem. You don't get a kind of a lipid extraction. So it's easy to understand in this range, they're non-toxic, right? Including a little part of the spherical particle, they're non-toxic. So this is a cytotoxicity phase diagram we proposed. Okay, so my last example is I'm gonna talk briefly about energy mapping, okay? So here, the challenge is to develop, try to help our medical school colleagues to develop membrane-active antibiotic, okay? The critical challenge here is these antibiotic molecules, whatever you select, has to be non-toxic to human and animal cell. Has to be specifically talking about bacterial membrane, okay? So there's a lot of choices, right? For example, take a nanoparticle, the dimensionality, whether you take a 1D or take a sheet or take a particle or take a tetrahedral, there's many choices. In terms of shape and size, right? How big should it be? Polarity, whether you put electron, you put polar groups, non-polar groups, and stiffness, right? How stiff these particles? There's a range of choices. So a lot of physical questions you can pose. Each of these physical dimension and parameter probably will influence how they interact, how the nanoparticle interacts with bacterial membrane, okay? I'm gonna show you some really simple experiments showing their specificity, okay? So here I'm showing three different molecules, okay? They are chemical from, they're all 2D sheets, sheet-like particles, okay? Have these benzene rings. We put them interact with the vesicle, right? In the first case, in this case, the membrane stay intact, right? Not much damage, okay? The cell membrane is completely okay. In this case, slide change, O to OH, we see the membrane's been damaged. There's segregation and poor formation on a cell membrane. In this case, let me play this again. In this case, actually, even more extreme, right? We start with the membrane, then we have a huge aggregation here, then the membrane burst, okay, the membrane burst. So that means our nature, bacteria and the membranes, actually very sensitive to all these physical parameters, okay? Yet, we do not understand, we do not understand fully how each physical parameter are influencing this, right? So that's why it's important to study, to map out the energy of interactions. So there, we use a methodology that's been widely used in the chemical engineering, about chemical engineering, look at how molecular interactions is called steered molecular dynamics, okay? Look at how a molecule in the process of going to cell membrane, how the free energy change, okay? Essentially, you map out the energy evolution as this particle go into the membrane, go into the, so I'm not going to go into detail here, given the limited time, but here's our calculated free energy evolution, okay? As a distance from a bilayer center, from a center bilayer, okay? So take an example of adipoline, adipoline is this molecule, the first one, adipoline is this case, okay? Adipoline is this case, we see that it doesn't have much effect on cell membrane, doesn't have much effect, okay? So here that corresponds to this green curve, this green, light green curve, right? Adipoline, you can see that as you bring this molecule toward the cell membrane, the energy is increasing, all right? So there's a strong resistance, strong barrier to penetration, this particle actually doesn't easily penetrate into cell membrane. Take a CD437, CD437 is a second molecule that causes substantial change in structure in the membrane structure, causes aggregation, the membrane aggregation, okay? Make them pour, make them leaking. And in this case, we see that there's a small, as you bring into the cell center, small energy barrier, but then there's a big energy well here, okay? There's a strong motivation for this molecule to be inside the membrane, all right? This red curve is the one I showed cell burst, okay? In this molecule, you can see initial causing aggregation, then it caused the burst, the whole cell, the whole membrane just ruptured, okay? In this case, it corresponds to the red curve. There's a huge energy well, right? That means there's a high density molecule would rush into the cell membrane, oh, it's the membrane interior, okay? So here's some simulation, okay? In this case, NTZ, let me show the movie here. Okay, so this is the red curve, simulation of a molecule interacting, molecular dynamic simulation of interact with cell membrane. This is the corresponding experiments. You can see eventually that the molecule will get into the interior, and okay, the middle one is the CD437, CD437, right? So have this blue curve, it goes to the cell membrane. Okay, both of them actually get into the cell membrane. In this case, you have this segregation, okay? Causing permeable, and that pulling is the green one, corresponding energy curve, okay? This one, there's a huge energy barrier to penetrate into the cell membrane. You can see this one actually doesn't go into the membrane, so the membrane stay intact, okay? Just slight change in chemical. Remember, these two molecules deviates only by a hydrogen route, okay, a proton. Very tiny difference in our chemical structure. Have a huge difference in our interaction with cell membrane. Okay, so we've been interacting with our biochemical colleagues from medical school, Brown Harbor Medical School. So we test these two molecules as new membrane active antibiotic, right? So this was reported in Nature paper a few months ago, we show three things, okay? We show these two molecules, indeed, they're fast killing on persisters. Remember, persisters, they're not synthesizing anything. They're sleeping bacteria, right? The only way to kill them, because they're not, there's no synthesis pathway, they're not making anything. So traditional drug doesn't work. But we say that these new molecules do kill them rapidly, right, giving enough dose. They have low human toxicity, right? They have low resistance probability, right? Remember, we're compared with a drug on the market, Cepro, right? So the traditional drug, develop resistance, substantially, this is 100 days of experiments, along experiments, right? The traditionally drug on the market, develop resistance, show resistance after two weeks, okay? After two weeks, you get bacteria, bacteria become resistant, right? Become resistant, this drug, you have to constantly raise your dosage to be effective. Now, because these drugs are targeted on the membrane directly, right? So within 100 days, there's no indication of a substantial resistance, okay? Anyway, so this is the beginning of this work in this area, so there's a lot to be done. So I think maybe there's an emergence of a simulation-based platform to speed up a drug discovery optimization. Because currently, most people in this field are doing combinatorial chemistry and high-soup drug screening. They're just combinatorial, try, try on air, right? Making this, making that, and cooking just like a cook, right? We're just trying to see whether they have any effect. But simulation does reveal the mechanism, right? Tell us why, tell us the rules, revealing the rules behind this, how they're by mapping on the energy, which we have the computational tool to do that, that will help us greatly reduce the time that's needed for the drug innovation, right? So there's a lot open question, of course, in this area, right? There's a multi-scale from all atom simulation to theoretical modeling, the core screen MD. Actually, this is one of my proposals I'm writing, right? I'm proposing this, a lot of open questions in the field. So modeling and simulation, facility screening, assessment, design optimization of synthetic antibiotic. Anyway, so this is a new thing. Let me, how am I doing in time? So, okay, let me just conclude my talk today by summarizing, okay? So rapid events, experimental computational tools, as well as unprecedented bio-imaging techniques that are facilitating quantitative mechanics-based model of cell nanomaterial interaction. Okay, as I showed this bio-imaging techniques even five years ago, lots of questions we were even possible to begin to explore, right? To study, because not biology so far has been qualitative, not quantitative. But with high resolution bio-imaging techniques, biology started to be more and more quantitative. This is where engineering can make a closer bridge with some of the biological problems. And I show three problems today as a representative examples of study contacting my group. And first example I gave it is the morphology of cell nanomaterial interactions, okay? Here I show that one-D nanomaterial below a critical diameter intercells while tip first entry pathway leading to a pathogenicity mechanism for CNT and asbestos nanofibers, right? This is because there's a critical size in fibers. Below that size cell only interrupts a tip which is very dangerous, okay? Cell only capture tip so this material cell gets stuck, stuck in the endocytosis. Get frustrated. So the second example I gave the study on vesicle packaging of flexible nanofibers. Here the key concept is bio-stew versus bio-soft. So when you say biology is soft but how soft is really soft, right? So we give example, we give a concrete criteria, right? Which is for fiber. For fiber, if the fiber undergoes 20 people Newton it buckles, then it's bio-soft, okay? We are very quantitative here, okay? So shows the persistent tip contact can damage the lipid membrane leading to cytotoxicity. We develop a mechanics-based cytotoxic phase diagram and we validate it by experiments to distinguish between pathogenic and biocompatible behavior for carbon 1D nanomaterial. But anyways, but this one is not limited to carbon 1D material. Can be extended to all the other different type of nanomaterial. The third study I showed is on energy mapping of nanomaterial penetrating the cell membrane, okay? Here the example is how this kind of work help us develop a new antibiotic, right? As our paper just published shows, indeed we were able to identify these two very promising, highly promising compounds that can kill bacteria. Even though we still do not fully understand the physics behind it, right? But so we're working with our colleagues on this. So thank you very much. Let me stop here. Thank you. Thank you. Thank you. And actually I want to make one statement. After the discussion, there's a reception and it's called a mayor. So please, you're welcome to join and have some discussions. Any questions? Okay, any questions? Yes, yes. We're going to make a few... I mentioned his grandfather, the only grandmaker in the back area. Right, right, right, right, right. So far we only have grand positive, oh, grand negative material. We're working on grand positive material. Grand positive material, yeah. So because they have a different membrane structure, they have a different structure, that's obviously very important. How much can the newer patients that you mentioned before...  So these two common eye shows seem to meet that criteria. But so far it's still, lots of things are not fully understood, a lot of physics, even though we're doing a combination of a combinatorial chemist to try different things and see how it works, right? So there's a lot of work to be done. It's not fully understood yet. Yeah. What I showed simulation today mostly are full-atom. Mostly full-atom simulation. Oh, coarse-grain, yeah, yeah, yeah. Depends on, right. Because a full-atom simulation is very expensive, right? Very expensive. But for example, one of the things is coarse-grain I showed today. Yeah, these things are all full-atom. But one thing that coarse-grain, this is coarse-grain, right? We're trying to simulate the interaction between a bilayer and a carbon nanotube. So the deformation is actually large-scale, large-scale. So it depends on the scale of interest because in order to model all the atoms, then it's too expensive to do this. So the coarse-grain depends on the problem, I would say. If you're interested in problems looking at the morphology, looking at a configuration from a physical point of view, some coarse-grain is necessary. But if you look at the molecular interactions, if you're focused on the energy mapping, how a single molecule goes into cell membrane, then the full-atom simulation is useful. Right, right, right, right. There's a various level of coarse-grain. Essentially, it's your condensed a cluster of atoms into one particle, right? Then you'll try to capture the energetic evolution. From energy point of view, you're trying to make them more or less equivalent, right? From free energy point of view. Just like our model, you can see it's ultimate coarse-grain model, right? We condense into one parameter. This is actually an awfully complex problem if you do an atomic simulation. So we just condense into energy functions, right? Giving a single parameter. This is like ultimate coarse-grain, right? You reduce the degree from into one. We're talking about the length. What about the geometry of the point at the end? Oh, yeah, yeah, yeah. Right, so you're giving a tip here, right? The tip here, so in this particular example that I calculated here, we're showing tip is always rect tip. So as the fiber changed in different orientation, that tip is always being fully wrapped, right? So they don't come in to this free energy function tip because they're always covered, right? But in the initial stage, it will play a very important role. Like let's say a particle just first touched the cell and it started becoming important role. I also published a paper in those, saw the initial stage of interaction. So the geometry, the key geometry are important at different stages. Once the fibers in or they're already, they don't influence energy. But the initial stages actually create dominant. I just didn't have time to touch this question. I gave a, it's not in my one example, right there. But they're very important, very important, but not important in this question. Yeah, you mentioned that there was gonna be a product research involving bacteria and let's see how three new interbiotics, but I was wondering if this could also be important to other areas, like other kind of sicknesses, like for example cancer or even maybe viruses. It's also part of that. Yes, yes, right now a lot of people actually use nanoparticles to do cancer diagnostic example, right? Because the cancer cell membrane also have different characteristics compared to normal membrane. There's a lot of receptors. They have different electromagnetic properties. So you have the nanoparticle, you can code in this cancer cell. It's very important for diagnostic. So nanoparticle actually, so I just didn't give an example here because there's a lot of people in the community who does that. Cancer treatment is one of the very important application areas for nanoparticles. I was just wondering, so are you sure there's no receptors involved? I wonder why. Sorry, sorry. I wonder why the end of biotics which are really non-toxic or less much less toxic to human membranes? Oh yeah, yeah, yeah, yeah, yeah, yeah. Very good question. Yeah, I didn't have time to discuss it here. Human bacteria, human membrane, the bacterial membrane, they have different compositions, right? So bacterial membranes have different lipids. Human membrane, for example, have cholesterol, have many other. So they're competing different. So we did the same calculation for human membrane. Okay. And this free energy mapping, right? If you consider human membrane composition, they're completely changed. Thank you guys for the experiment. We don't take an actual living, we just make the vesicles which have the composition, you know, the polyglycerin present in the leaves so we can see what's in the back. We have done a summer work but not complete. Because I want to do this, this is a separate version of the stuff, right? That's right, that's right, that's right. No, we have not completed this at this beginning. We have, there's so much open questions, so many open questions. So it's amazing, yeah. Right, right, right, right. Yes, yes, yes. A lot of things worth looking into for this. Yes. So you were talking about this one traditional antivirus, what do you find? Oh, Wendy. Right, right. So what we call one-dimensional material is that one dimension is dominant in your lung, fiber-disciplinary.  So like a plane, right, you look at plane. The thickness given, thickness prescribed. But the other two dimensions you can design. Like one that you can design with lens. The cross-section pretty much, it can still be contuned to some extent, but pretty much it's clearly, right? But the other dimension you can play with, right? So this is, this is like jargon use in the animal-material community. See the blood in that, they work out in material. Oh yeah, yeah. They're just a sub-family of 1D material, right? Well, but they're very important in terms of flexibility. Single or multi-platform, very different flexibility. Thank you, brother, for your talk. I have a question on this slide. I mean, when this cell, so the red curve is for the cell with bars, right? Yeah. Okay, so point goes to minimum and then goes back up. Like when it hits the minimum, is that the point when the cell bars? Right. This name on here corresponds to, this is from center, right? It corresponds to some place like here. That means when this part goes in, this is the minimum energy state, not right in the center. Right in the center, because this is bilate, it's bilate. Right in the center, it's a hydrophobic region, but that's the high energy point. The lowest energy is right in the middle here. Okay, why the burst is a very good question. I didn't quite address that, right? So amazing that the energy map in this interpathic already reflected the consequence, right? With the burst, I mean, that's totally rational kind of investigate. That requires some more bigger scale model. For example, we use free energy based modeling to model the aggregation and deformation of how the membrane developed force, right? So that's, this is a completely open field. Completely open field. Where we do not fully understood how this vessel burst, okay? Even without anti-matter, you know what I'm talking about. How the force formation, the rupture in the membrane, it's a lot of people are working on this. But here we say they're highly sensitive to nanomaterials. If you put different materials, you can cause the membrane to, you can destroy the membrane, you can keep the impact, you can cause the leakage. There's a lot of things, there's a lot of biophysics that needs to be explored. Do you think bacteria will actually form a chemical resistance to this kind of like the chemical resistance to other active gases? Yeah, so thing we see here, we put them together in 100 days, we do not see a clear increase in resistance. While we do see, you can see the slightly, of course on this scale, it's almost flat. But there's a slightly increase in resistance. What we find amazingly, the bacterial cell membrane become more negative charged over time, okay? So I think the bacteria is trying to, trying to develop some resistance, but not so successful yet. So we don't know if it's long enough, maybe there's, it completely changed your composition in cookie, right? Bacteria learn how to put some other molecule in and change this energy mapping. But so far, we don't know you, this is a good question. But they do become more electronegative, more negative charged, right?  So they're trying to do something. Yeah, yeah, yeah. Anyway, evolution is a strong force. It's a very strong force, yeah. We don't know what eventually it leads to, but it's certainly a lot of interesting open questions. Sometimes when cells absorb some of those nanomaterials that they ask for the help of other immune cells in order to take care of that nanomaterial, is it possible to actually trigger an immune response by using nanomaterials that get stuck in the membrane of the cell membrane? Right, so in this case, right, the frustrated endocytosis is a trigger to immune response, where it should not, right? Because there's no, because what cell, what an immune cell does is trying to clean all the dead cells, bacteria, all the foreign substance away from the body, clean them up, right?  So the immune cell always working very hard, right? So, but if it gets stuck like this, it triggers a great immune response for no reason, for no good purpose. Because our immune response cannot kill this, no matter what you do, right? So this immune response is unnecessary, and this leads to problems, right? You've got a, like a permanent inflammation somewhere in your body, right? So this is what the problem triggers, a lot of mutation, a lot of other things, problems, right? So the fibers are pretty dangerous to our body, in that sense. We've got to be very careful. Oh. No, the trigger-ready immune response is trying to get rid of it. Oh, yeah. You can't swallow the whole cancer cells. Oh, that's a great idea. That's a great idea. You know, the immunotherapy, just one Nobel Prize two days ago, right? This is a great idea. Yeah, yeah, yeah. This is a great idea. Maybe nanomedicine can further the messed up view, as well. That's a great idea. Yeah, I haven't thought about that so far, yeah. It's a great idea. Yes. In the industry, occupational exposure to nanofibers, especially those biologically long biologically set points, has there been an increase in the fragmentation of the disease? Sorry, I didn't capture your beginning of your speech. There has been an increase in disease and occupational exposures using nanofibers. Right, right, right. Oh, in occupation, asbestos is one prominent example. Asbestos is one, but there is a lot of new ones. Right, a lot of new ones. I don't think there's enough data. FDA regulation on nanomaterial currently focuses on chemistry. They haven't focused on the mechanical aspect because there's not enough data there, right? So far, they say, this is toxic. This is not toxic. Mostly, almost always based on chemistry, based on the chemical composition. But the mechanism effect, people start to recognize that. But haven't been to the FDA regulation yet. It's important work then. Yeah, yeah, right, right. Thank you, thank you, thank you very much.


The sound you hear is that of a grave being dug on a hillside in a lonely section of England in the 17th century. And that was English thunder of the era. And that, a soft English drizzle. Water, earth, sky, elementals. Surrounding James Alsop. Money clipper, high woman, father-in-law, grave digger. It'll take him about another hour to finish and about another hour to give it a corpse. For in that time, James Alsop will add murder to his other attributes. And tonight, my report to you on the Alsop family, how it diminished and grew again. Crime classics, a new series of true crime stories from the records and newspapers of every land from every time. Your host each week, Mr. Thomas Highland, connoisseur of crime, student of violence, and teller of murders. Now once again, Mr. Thomas Highland. The year, 1673, and the place, Ham, a small village about four miles from London situated in the county of Essex. Now, the month is March, at the tail end of a wild winter. A winter of great snows, much poverty, and much returning to the earth of the folk of Ham, young and old. Never was there such a winter within the memory of what villages were left alive. And now that the snows had melted and the first warmth blew across the heath and into cold stone rooms, there was stirring among the hamlings who had survived. Man and wife smiled on each other where yesterday had been chill. Life must go on. But what of those whom the ravages of the season and starvation had made lonely? What of, say, a widow? What of, say, widow Ursula, who lost her husband to the wild dogs two days after Christmas? What of her? Well, let's see. You be lonely, are you widow? Aye. Ah, it was a cruel thing as happened to your husband. Cruel. Then now it is the springtime. Aye, and March is soft on the moors. April is a good time for marrying, widow. When I married last, it was in April. It was April after a soft winter when my husband built this house that we married. It was a good marriage. April is soon and you should have another husband.  You make a young man a cumbly bundle. Daddy? Aye, you're welcome, widow Ursula, a cumbly bundle indeed. How long has it been since your wife passed on, James, also? Ah, six years, come next blooms every day. I heard she was a shrew. A witch, a shrew, a gollygog. But a woman of fervor and she fruited me with two sons. Aye. I have seen the younger one. He said to me the widow Ursula had looked upon him. Aye, a robust lad, sinewy. Will you marry my son? What do you call him? Christopher. Christopher. A likely name for April. Your nurseula will make a likely bride. Aye. Marlon. Oh, Father, what did the widow say? Will she marry my brother? She will. To us, William. Aye. So, my brother Christopher will take a bride. Your brother is not like us. He needs a woman to take care of him. He is a douch. Perhaps. And cowardly. Maybe. For instance, he needs his father to propose for him. He has a shy matter. Cowardly. For instance, down there, where the carriage will roll soon. Would he have the courage to stop it and point a knife and aim on money or life and mean it? No. Not Christopher. There. Around the ill's curve. The carriage. Father. Aye. So, the two of them, Father and son William, galloped down the hillside, donned masks and stopped the carriage. And with a welladay and cocked pistols, they robbed the driver and the passenger. History records that the passenger, as coincidence would have it, was none other than Dame Maggie Chavez. It may be assumed, was even then on the first leg of her mad dash to Lisbon. Galants that they were, the high women, took from the lady only her gold coins. Her jewelry, the famous mementos from the thy count of Gluster, was not touched. This might have been, however, because of the hiding places for gems that the Dame is famous for. And with another welladay, James Alsopp and William rode down to the town of Ham, and to home, and to their workshop. Here, they divested themselves of garments necessary to robbing on the road, placed them in a trunk, took out a pair of shears and went to work. This year's mintage is harder than last year's. Work, meaning clipping off small edges of the gold coins, decreasing the diameter, in other words, without decreasing the face value. So, when you were finished, you had the original gold coins and gold clippings as a bonus. Else the shears are doled. Else you, Father, are growing older and weaker. Aye. Aye. Aye. I said it. Did you? Now! What do you say now?  Aye. I meant a gist. I asked your partner. Get back to work. There's a pointless sovereigns on the table to be clipped down. Clippings taken to London to be sold. Aye. Oh, be it. Christopher. Come in, son. Hello, Father. Hello, William. Did the day go well for you, son? I dreamed how it would be to take a wife. You like the taste of blood, William? I dreamed how it would be to take a wife. And it was a fine dream. Yeah, I spoke to the widow Ursula. And asked her what I told you, Father. Aye. Will she have me? In April. I'm grateful to you. Christopher, you make your father proud. Did you have a good day on the road, Father? No, that's thoughtful, you asked. Are you a good day indeed? A carriage with a rich lady. Ah, and much gold, I see. One thing, son. When you're with wife, no talk of this and what happens on the road. Not a whisper on it. Not a whisper? Yeah. We have to do much clipping, you brother. Take our handful of coins, lad. Buy some things for April. When it came, it was the mildest April in years and the good town's folk of Ham wore homespun linen of most feathery weight for the nuptials. The widow Ursula to Christopher all saw. There was much gaiety. Laughter spilled as did the cooling spiced mead. And as sometimes happens at such festivals, a group of the younger set thought they spied a druid and ran off into the woods to give a chase. But the older set, the ones who had already chased their druids, stayed and they smiled kindly upon the bride Ursula. She took the hand of Christopher and led him into the warm stone cottage. My husband. My bride. You dance well, the Saraband husband. Do you not have a good word for my jig? In the Saraband you well and lifted me high. You are comely, bride Ursula. You are young and handsome. I love thee. And you will gentle me and be ardent. Aye. And I to you. Christopher. Aye. Your father is rich. He is. He has told me I do not need to employ myself at anything for a year as a wedding gift if I wish it. He must be very rich then. What does he do for his riches? I cannot say. But you are husband. Then come to me. Yes. Which was the talk in April and in May when the bloom was on the sedge. Come, dearie. Tell your wife what your daddy dear doubts for his riches. And in June when the drone bee danced for its queen. If you do not tell me, dearie, I shall be unhappy and you shall be sorry. And for the rest of the summer, love and curiosity and a Christopher who wouldn't talk about his father. But in September when the first leaf dropped. Now what does he do, your father? Come here. I will not. Bride. Wife now. And I ask you, what does your father do for his riches? What curiosity is yours, Ursula? And who would not be curious? The way your father is hardly home, nor your brother. The way they ride the road so often and after lock themselves in that room. What do they there? And why am I not permitted there in their work room? And why does your brother journey to London so often? Come here. When you will answer me not before. I will not answer you. I'll tell you this only. It is September and you have become a shrew.  You will rue it. Take a fresh horse. After we'll clip the coins and you will go down to London. What hails you, father? There's no humor in you anymore. You worry about Christopher? He's not happy with his wife. As I said it. He says she is a shrew. And what woman is not? That she's curious. As the woman Alice is. That she needs to know what we do. You and I. So she would gossip about it. We are all women. And money clippers. She would gossip and the law would hear of it. And we would jig for my gullows. It grieves me. It would be a terrible thing. Hello father-in-law. Brother-in-law. I had never been here before. In the workshop before. I thought to clean it. To dust a bit. What are these clippers for? Get out of here.  She will gossip father. I. And the law will hear of it. I. The gullows. And death. Not for us. To work William. They clipped a pile of sovereigns. And William took the clippings to London town. And the rest of the night James Alsop sat with his thoughts. In the false dawn he came to a conclusion. His daughter-in-law had to go. Good. You are listening to Crime Classics and your host Thomas Highland. All America is in CBS Radio's 21st Precinct. Tuesday nights every week the stars address brings you gripping and suspenseful stories of behind-the-scenes police work in the world's biggest city. Here 21st Precinct on most of the same stations tomorrow night. And now once again Thomas Highland and the second act of Crime Classics and his report to you on the Alsop family how it diminished and grew again. England in the 1670s. It was a mess. First of all the English didn't like the Dutch and the Dutch felt the same about the English and they were constantly sending each other's frigates to the bottom. The King of England was Charles II. Fain for being a member of the Stuart family and marrying with Catherine Burganza and for appealing the oranges of one Nell Gwyn. It was as I have indicated a yeasty time for meant everywhere. Colonists were going off in every which direction to America, to India and to the Caribbean. It was the decade two of the great Feathergill hoax and it was the decade of plague and fire. Let's look again at a corner of it. 1673. The place, the village of Ham and the woman named Ursula walking through a meadow at dusk. I think only Midland. What are you talking about? I saw the woman Ursula walking through the meadow as you said she would be. And did you kill her as I said you should do? I am not sure. Therefore? Therefore what? I am willing to take with half the amount of money which you promised me. I promised you a fee, assassin, for the killing of Ursula. I hired your sword for a death. No half payments for work not completed. Well, no fault of mine. I stuck her with my sword which she moved quickly away. Then I saw some people who were coming attracted by her cries. And my son, William, who sent you to me from London, said you were the finest of assassin. Another day and I will try again. Go back to London. My pay.  What you deserve.  Your wife is a fortunate one. That the wounds are not grey. The villagers who saw the attack set a rider in a cloak and mask chased her about the meadow with a sword. For what reason? For what reason I cannot conceive? A mystery. Who would want to do murder on such a fine woman? Huh? Yesterday you called her. I even worse. But I did not mean it. As I said, what has happened? A mystery. Who would want to kill her?  She walks alone in the meadow at dusk. If she ever recovers, I will tell her to do so no more. While I'm gone for the surgeon in Yorkshire, you will take care of my wife, father. I that I will. I'll watch over her. I will. I will. I will. I'll watch over your wife. One young Christopher rode off to Yorkshire and what with William in London selling coin clippings, James Alsot was left in his house with Ursula, his daughter-in-law. September nights and a gathering storm and this... What? Ursula, thirsty as a body is wont to get when it has been stabbed. And the man who could slaker's thirst moves but not toward Ursula. Toward the tool shed for a shovel then toward the hill. You know why. That's right. To dig a grave. And while he was digging, you remember it started to rain and James Alsot lifted his mouth to it because the body is wont to get thirsty when it is digging a grave. But, Alsot finished his task, took shovel back to tool house and did what quite a few grave diggers do even to this day. Marky! Another. Good evening, Judy. By a hollyhock by one.  Wear it yourself.  I have only two more to sell and I have nothing to do and no one to sing my song to. Oh. Tonight I'm busy, Judy. And what could you be busy with in such a storm? To help my daughter along a journey. A journey to where? To her cousin in Scotland. If I had a drink, Mr. Alsot, I would drink to her safe return. Ah, Marky. Three drinks. One for me, one for the lady and for yourself, Marky. Yeah. Well, now we will drink to the safe journey and the safe return of my daughter in law, Ursula, who goes to visit her cousin in Scotland and who probably will not return for a year or so. You had to go and find out I was a thief, did you not? I can't stand to see you suffer, girly. You'll never suffer again. Yeah. There's a burying to be done. Pastoral scene in England under Charles II. Cutthroat carrying victim through big storm to grave. A few more details. Darkness torn apart by lightning. Silences shattered by thunder a sea of mud. It was perhaps 300 yards to the hillside and when James Alsot got there, the graveyard dug was gone, washed away. So what we've got now is the same pastoral scene in reverse. Cutthroat carrying victim through big storm back to scene of crime. Sure. Cutthroat in a quandary. The plan was a very unique rehearsal. Now what? Proving that things gang after glee. Yeah. I told everyone you were going for a visit. The question. What shall I do with you? Seeking the answer. The answer? Not quite. This time the answer. Change everybody into warm dry clothes and having done this, the stroke of genius. Remove the glass from a window frame into the room. Light it. And set up a cry. Murder! Murder most foul! Help! Thieves of common aid murder! Help! Yesterday a man tried to do it while she walked the meadow. I've heard. And now you succeeded. Are you to cry for merciless? I came quick to her, but only in time to see her attack her flea. Flea? How? Through the window, sir. Which window? That one. That one? With no glass in it? Of course that one. You're sure of it? Aye. Come here. If the thief and murderer escaped through this window, how did he get through the cobweb which covers the opening? Sir. It's a cobweb, as you see. From side to side. So? From top to bottom. So. Sir, then how did this thief of yours get through it? Sir. You're under arrest, James Alsop. Arrest. For the murder of your daughter-in-law. Charlie? That's James. Alsop was committed at Manchester Jail. His sons William and Christopher were not to be located. Justice was swift and sure. James Alsop was sentenced and led to the gallows. And, as he mounted them, a thing happened. A messenger with good news, especially if your name was James Alsop of Ham in the year 1673, and the hangman had just indicated that you stick your neck in a noose. New evidence from London. A man who calls himself Topham says he can prove that James Alsop is no murder. He saw thief lead the place of murder through the door. That's the way it was. I forgot. Bring down the accused from the jeep. But our story still has a Dolores end. Mr. Topham from London turned out to be none other than the missing William Alsop, son of James. He could have stayed in London safe and alive, so he must have loved his father dearly and loved his friends. But the only way he changed history was this. Father, son, William. What happened to Christopher? Well, he had heard what happened to his wife, so asked himself why return to Ham. He stayed in Yorkshire, where he met the widow Patricia. She was comely, and the next April they were wed. She made puddings and was an uncurious woman. And gave to the world eight Alsops. In just a moment, Thomas Highland will tell you about next week's crime classic. The Alsop family, tonight's crime classic was adapted from the original court reports and newspaper accounts by Morton Fine and David Friedkin. The music was composed and conducted by Bernard Herman, and the program is produced and directed by Elliot Lewis. In tonight's story, Ben Wright was heard as James. Thomas Highland is portrayed on radio featured in the cast were her Butterfield, Ellen Morgan, Betty Harford, Terry Kilburn, Richard Peel and Raymond Lawrence. Roy Rowan speaking. And here again is Thomas Highland. Next week, Rome, Italy in the year 62. My report to you will concern a young chap who wanted to murder his mom. It's listed in my files as your loving son, Nero. Thank you. Good night. Later tonight, CBS Radio brings you to an hour of sprightly romantic comedy as the Lux Summer Theatre stars Anne Baxter in The Affairs of Susan. Lovely Anne has the most difficult time making up her mind over her very assortment of suitors until... But suppose you're here at all for yourself later tonight on most of these same stations when CBS Radio presents the Lux Summer Theatre. America's 45 million radio families listen most to the CBS Radio Network.


Explaining the 17 goals for the 2030 Agenda for Sustainable Development. Let's explain how do vegans get protein? Meet Scott. Scott is a wrestler at his high school, but lately he has been feeling sluggish and fatigued. After doing some research, he decides to cut processed food and dairy out of his diet and becomes a vegan. He starts to feel a lot better and cutting weight becomes easier for him too. Scott's teammates start to worry about his weight loss and fear that his new diet is going to make him weaker and ruin his chances of winning any matches, ultimately bringing the team down. Scott notices his muscle loss but doesn't want to give up the way his new healthy lifestyle makes him feel. He has to figure out where to get protein and still keep his vegan diet. Luckily for Scott, he can have his protein and eat it vegan style too. By consuming a variety of whole plants, Scott can receive all the essential amino acids needed to maintain a proper metabolism and function naturally. Plant-based foods such as lentils, rice, beans, and quinoa can all provide healthy alternatives to red meat and other animal-based proteins.  Also, natural animal protein is wrapped with unhealthy saturated fat and dietary cholesterol. While plant protein is packed with loads of vitamins, antioxidants, minerals, and fiber, all essential components of preventing diseases and living an optimal healthy life. Switching to a vegan diet to get your protein also cuts back on the cruel acts of raising animals to be slaughtered and the toxic growth hormones that find their way into our bodies. Decreasing the amounts of toxic chemicals that are ingested through animal house and farm spraying can help lead to healthier people around the world. You don't have to sacrifice healthy eating in order to get all your essential proteins. It's possible to live an active life like Scott while not eating meat, dairy, and other processed food that can make you sick. Your body is a machine. Make sure that it's running on the right fuel.


All champions have discipline. When others choose to stop, you don't stop for anything, because it's the discipline to work hard. Even in times when all seems hopeless, you gotta stay strong and remind yourself who you are. Remain disciplined and your time will come. What is up smart phone filmmakers and welcome back to my channel. This is the best time of year to start filming with your phone, because the video quality you get out of it is just very good. We almost always see at least some level of camera improvements every year and it's only getting better and better. I'm always looking for the best possible ways to achieve a cinematic look in our videos when it comes to using a phone. With the iPhone 13 lineup, we have come a step closer to achieving that look. If you haven't heard of it already, it's called cinematic mode, which by the way, the video you saw was shot with it. So what does cinematic mode do? Well, it's basically port your video on your iPhone, which blurs out the background. That creates what is called a bokeh or depth of field effect. This makes your video look more dynamic and helps isolate the subject, allowing the viewer to focus their attention on what's important. By the way, this talking head video you're seeing right now is shot on the iPhone 13 Pro Max using cinematic mode. For those that don't have the iPhone 13 Pro Max, don't worry. You can still achieve a blurry background with your iPhone. I will leave links in the video description below for you to check out. Before the launch of cinematic mode, the process of achieving a blurry background wasn't easy. You either had the method to download a third-party app like Focus Live or Protake App, but these come with a lot of imperfections or use the method of buying a DOF adapter, which can be quite costly and you aren't as flexible when shooting. I'm not saying that cinematic mode is perfect, but from my user experience, it's more advanced and overall gives you a better quality of depth of field. The footage looks convincing, it does a great job of processing the image in real time and you can create some cool focus transitions. So how can we use cinematic mode in our videos? First, launch the camera app on your iPhone 13 and swipe to select cinematic mode. Once selected, you have a live preview of what the cinematic footage will look like. You can record in 1080p 30 frames per second and it works with the front and selfie camera. When a person or an object is detected, which it does quickly, it automatically sets them in focus, making the background blurry. By tapping on the screen, you can set your focus as you record. Tap again to track the focus. This is great for moving subjects. Tap holding on the screen will lock the focus. If you have two subjects in the frame, cinematic mode will place the focus on the subject that is closer to the camera. If the subject then turns their head, the focus shift towards the subject that is further away, creating a focus pool and it does it quite organically. If you want to change the amount of blur in the background, you can tap one of the symbols in the top right hand corner. A slider will appear at the bottom where you can change the blur amount between two for very blurry and 16 for less blurry. I suggest you keep the blur amount between F5 and F8 to get a more natural looking shallow depth of field. There is also a hidden menu when tapping the arrow to the left.  I would generally turn down the exposure to a minus one because the cinematic mode tends to overexpose the image most of the time. The lighting bolt allows you to turn on or off the flashlight for additional light. And keep in mind that cinematic mode works well in good lit environments. You can also use the wide or telephoto lens by tapping on the one time button. However, the ultra wide lens is not available in cinematic mode. A good way to save time is to preserve the camera settings when using a cinematic mode. This way you can set your values such as the exposure and f-stop. And when you open cinematic mode again, the settings will stay the way how you left it. This also works with the normal video mode on the iPhone. Now, the beauty about recording your videos in cinematic mode is that you can change the focus and blur even after you shot your videos using the camera app's editor. So let me show you how to edit your videos that was taken in cinematic mode. In the photo app, tap the video that you would like to edit. Select the edit and the video timeline appears below your video. The white dots are the points where the focus has changed. You can move the indicator to scrub through your footage. What's great is that you can see what's currently in focus indicated by the yellow square. And it also recognizes people or objects that are in the background, which are indicated with a white square. This is where people get a little bit confused. Now, below the timeline, you have these white dots, which represent the automatic focus pool made by the camera app. These can't be deleted. The yellow dots represent the manual focus pools that were made by me, for example, or by you. These can be deleted. Like I mentioned, a focus pool is basically the change of focus in a scene. On the top left, you can tap the focus button to switch between automatic focus tracking or your manually selected focus point. Since I want to manually adjust my focus, I keep that enabled all the time. You can tap on the white square to change the focus point to that person or object. You can also tap anywhere else to set your focus point. You can double tap on the subject, which will enable automatic focus tracking, which will track the subject throughout the video. To lock the focus, just tap hold on the screen. To change the blur amount, just tap the F button and set a different value, which will then be applied to the entire clip. And this is really awesome because if you find the blur amount too unnatural, you can always change it afterwards. You can also remove the bokeh completely by tapping on the yellow cinematic button at the top of the screen. So guys, as you can see, the editing process with cinematic mode is user-friendly and it is fun to work with. By the way, you can also edit your focus point and change the depth of field using other editing tools such as iMovie or Final Cut Pro, which is really exciting. Cinematic mode isn't perfect, but the quality of depth of field you get out of it is truly amazing and it is getting closer to taking over the jobs of DSLRs. Yes! Yes!  Say goodbye, Sony a7. You know what? I'm probably gonna wait a couple of years. I don't think it's that quite ready yet. Now it does take practice using cinematic mode and getting to know when to use it because for certain situations, it doesn't always look natural and you get that sort of halo effect. So play around with the f-stop and see what works best. I hope cinematic mode will be available in 24 or 25 frames per second since that is the cinematic frame rate to achieving a realistic motion blur and is the standard for movies. The great thing about cinematic mode is that you don't need additional equipment to achieve a blurry background and it does it pretty good, especially that you can adjust the focus even after you shot the video allows for a lot of flexibility in post. With that said guys, cinematic mode brings a new level of filmmaking on an iPhone and makes shooting videos even more enjoyable. Now I would love to hear your thoughts on the cinematic mode. Would you use it in your daily videos? Let me know in the comment section below. Now if you're new to my channel, make sure to subscribe as this will help the both of us. If you're new to smartphone filmmaking, I got a free guide that will help you get started making quality videos with your phone. Now one of the best ways to learn is to join the smartphone filmmaking community on Facebook that I created where you can share your work and get feedback from others. Thank you guys for watching, stay mobile and I will see you in the next video.


Howdy, my name is William Rummy, I'm the Fort Worth Municipal Court Director. Today I want to spend a few minutes to talk about a part of the court that many people may not think about, at least initially, and that's our administration team. So when generally you think about court, and specifically municipal court, you think about the judges and we think about the clerks and the marshals and the prosecutors, defense attorneys, and the defendants. But we may not think about the administration team, which really is the center of the hub of municipal court in order to that to function. So today I just wanted to spend a few minutes to talk about the great team work that we have here in municipal court and administration. So we do have administrative services manager who generally oversees our admin team. And then we also have a senior accountant. We have several account techs and a senior administrative tech. So let's walk through kind of the different things that they do every day to empower the rest of our staff in order for the court to function on a daily basis. The first thing that kind of process-wise we'll talk about is from the financial aspect. So obviously as a court, municipal court, we are a fine-only court. In addition to Class C criminal offenses, we also have civil parking cases. And so as a result of either a liable finding under a parking or a judgment of guilt in a criminal offense, there is a fine collection aspect to that. And so when we collect those fines and record those in that moneys, we have individuals to ensure that A, whatever we are collecting is done in an appropriate process. And then that information goes to two locations technically. So there's the moneys that we collect, and of course there's financial payments online, your credit card or whatever it is. And so we do reconcile those on a daily basis and ensure that whatever we're saying is collected has made it to two locations. The first is to the bank. And the second is to what we call RGL or General Ledger to ensure that those match. So on a daily basis, staff is managing that, ensuring that. There's also payments that go out from the city to the state, whether it's state court cost or also can go to maybe the local jurisdiction related to those cost and fines collections. So that's one component that we have staff who help support that. The next piece is really what you would think of is every organization is going to have both the finance collection, but as well as HR. So we have staff who's assigned to ensure that for instance, time sheets and HR processes, whether it's a medical documentation or those types of things. So on a daily basis, we have staff who supports the rest of the court in order to ensure that those things are happening. People are getting their paychecks on time. And then the next step would be our accounts payable, accounts receivable. So obviously we need supplies, here where we're standing in one of our courtrooms and there's chairs and tables and all types of different things, pens, pencils, information. So on a daily basis, they're ensuring that staff has the tools and equipment that they need to be successful, whether it's a city marshal and a vehicle and an MDC or even the uniforms that they have or whether it's a clerk in the courtroom or a judge and making sure that we have robes. So even in the facility itself, ensuring that we have janitorial services, we're paying the water and electric bill just like everybody else does, we have to do that. So we have a great team that does that on a daily basis. And then in addition, there's another piece to the admin team is called our budget. So we do or are required to on a yearly basis prepare and submit a budget to the city manager's office. Ultimately that goes to the city council for approval. So we have a team who works on that throughout the year, it's just not a one time event but throughout the year. They help to ensure that we are maintaining our budget, whether it's revenues or expenses and then also that we are managing and preparing for each year to submit a new budget. And then one final piece I just want to mention is our from an IT technology standpoint. So again, as I stand in this courtroom and it's, you know, we're up to 21st century. Then, you know, we do have computers and we have scanners, copiers and we have TV monitors. So there's microphones. So there's all types of technology that's involved both here in the courtroom but also for every staff member. And then when you think in addition to with COVID, we've really moved to a virtual court venue where we work with WebEx and lots of other venues where we can virtually see folks they can see a judge and have their case process. So we have a technology group who team who comes in and works both with the hardware and the software to ensure the staff have the training they need the equipment they need. And for that we work in partnership with our IT department and again, so just wanted to spend a few minutes to talk about a group that maybe not everybody would think of which is our administration team and the great work that they do every day to help us to be successful, to help serve you as the citizens. So thank you and have a great day.


This is stock number 11506A, we are here at Summit Automotive in Fond du Lac, Wisconsin, you're new and used sports car headquarters and today we are checking out this Ultra Clean 2019 Ford Mustang GT Premium. This vehicle has the 5 liter V8 motor which pumps out 460 horsepower. This car has been fully safe and inspected by our service shop, has a fresh oil and filter change. All the fluids have been checked and topped off and this car is 100% ready to go. Shadow black is the color we shoot all of our videos in 1080p, 60 frames per second. So if you have HD capabilities on your computer tablet or smartphone device, turn them on right now because it is definitely your best way to check out the quality and condition of the car before seeing it in person. And if you'd like to check out all the photos on this car in the upper right hand part of your screen is a link to our website, click that and check us out there. This one comes with the 19 inch painted alloy rims and it has the Brembo brakes on it because it has the GT performance package on this car. This one has Michelin Pilot Sport 25540ZR19 tires and these tires I'd say are probably about 60 to 70% of the tread left on them. Front fenders in excellent condition, I didn't see any dents or dings on that. Has the LED running lights, the LED fog lamps and the projector lamp headlamps. Front bumper and lower valance are in excellent shape, I didn't see any dents, dings or cracks or scuffs on it and the hood is in excellent shape as well. Notice you get the little pony inside the light there. The passenger side front fender is in excellent shape as well, no dents or dings on there and the passenger side rim, no scuffs or scrapes. As you go down the passenger side of the car, take note of how clean the body is, how reflective and mirror like that paint is. We take these HD videos so if you are far away or even if you're close by and you just cannot make the trip down but you're still interested in purchasing the car, you can see the car, hear the car and of confidence in the vehicle that you're looking at before you even get here so that when you do get here there's absolutely no surprises. That's why we offer the Summit Auto Fastpass option in the upper right hand part of your screen, a link right to our website where you can apply for financing, get pre-approved, see a payment quote, basically make the car yours before you even get here so that when you do get here all you got to do is drive the car, love it, you'll be all set to go. Back rim is in nice shape as well and those back tires have just as much to try to do as the front tires. And around to the back of the vehicle rear bumper is in great shape, you do get the quad tipped dual rear exhaust and it does have backup parking sensors, LED sequential tail lamps, nice deck lid spoiler and the trunk is in nice shape as well. We'll take a look inside in just a second. Down the driver side just as clean as that passenger side, no dense ding scuffs or scrapes on that rear quarter. For full disclosure this back rim is in excellent condition as well. And it does have the capitalist fuel fill which is a really nice feature, never get gas on your hands again. And down the rest of the side of the car looks really good, didn't see any dense ordains. You get the heated mirrors with the line spot mirrors in it. Side directional signals, this one has the intelligent access, you just have the key fob in your pocket, it'll automatically unlock. Inside you get the GT premium package, gives you the black leather interior, side curtain airbags, no scuffs, scrapes, rips or tears on those seats. Both of these front seats are heated and cooled, power driver seat, weather tech floor mats, does have the auto headlamps, power windows, power locks and power mirrors. Back seats are in excellent shape as well, it does have latch child safety system if you want to try and get a car seat back there. Good luck, and then those seats do fold down for extra storage. We'll hop inside here and take a look at the instrument cluster and the radio. See this one has 11,647 miles, the instrument cluster is very nice and clean. You get the leather wrapped steering wheel, Bluetooth and information center controls on the right, cruise controls and audio controls on the left. This one has the sync, 8 inch touchscreen radio, AM, FM and Sirius XM radio capabilities as well as Bluetooth. You have your climate control here as well including your dual climate control and this one does have the factory navigation system on it. The GT performance package also gives you this oil pressure PSI and vac gauge right there and you get that machine metal finish to the dash, looks really good. On this radio as well you have your backup camera and then you have all your different apps and stuff including Android Auto and Apple CarPlay where you can project your cell phone to the screen so if you have a different navigation system such as Waze or Google Maps that you like better than the factory you can project it to the screen, it's a pretty cool system there. Then you have your heated and cooled seat buttons, your push button start, stability control, this is your different drive settings. You have sport, comfort and normal and that comes with the GT performance package as well. Also your drive modes is what comes with the, so you have sport, track, drag, snow, wet. So you get those there, six speed manual transmission has the nice big keyless entry, weather tech formats and passenger seat are in excellent shape, no rips or tears on that and the headliner is absolutely perfect. You do get home link buttons for your garage door security systems and lighting systems and we'll start it up and take a quick look under the hood. These Mustangs always like having the door open when we start them they sound great. I personally like to thank you for checking out the video today and hopefully from this HD video you'll be able to tell just how clean this car is all the way around inside and out. Under the hood we have the 5 liter V8 motor, engine bay is very clean, it's very smooth, 160 horsepower, once again this car has been fully safty to inspect by our service shop as a fresh oil and filter change, all the fluids have been checked and topped off and this car is 100% ready to go. See more pictures of this car, one of other 450 new and used cars, trucks, SUVs, minivans, Wranglers, sports cars, Mustangs, Challengers, Chargers, Camaros, Corvettes, you name it we got it GT500, go to that website right there www.SummitAuto.com full pictures and descriptions of every single vehicle from two locations all at SummitAuto.com if you'd like to check out more HD videos you can go to YouTube.com slash SummitAuto. Remember to like, subscribe and share on this video and all the videos that you see there. In fact any second you'll see a link to subscribe to our YouTube channel on your left, a link to more Mustang videos like this one in the upper right, a link to this vehicle on our website in the lower left and a link to our YouTube, latest YouTube video in the lower right. Click those check us out and we're super excited to help you with this ultra clean 2019 Ford Mustang GT Premium with the GT performance package in shadow black. Thanks again for checking out the video.


So here's some 3D printing here with the Outworks. So who are you? Outworks is a small startup. We started our business maybe eight months ago. It's still start-up stages. We are trying to provide a new Regency 3D printer for individual and so-called entrepreneurs. So you make larger, easier, faster? You make large? What's going on here? Actually, this size is the largest build volume. Can you bring it up? This one? This is the largest what? Largest build volume desktop Regency 3D printer ever. It's very detailed, huh? Yup, it's in-plane resolution, it's 1 tebakin micron, and G-height resolution is 25 to 200 microns. This looks really nice. Which one is that? Which church is this? It's a cathedral in Notre Dame. Notre Dame, yeah? Yeah. And how long does it take to create one? Did you make this today? Actually, yesterday afternoon we started and we get this printout today morning. How does it work? Okay, let me introduce a structure. This is a resin tank. Pour the resin as a low material. We are using a maker juice and you can use any kind of resin. You'll be curable for the resin. Pour it and then down the plate to sink out the resin and then turn on the inside. Inside the stuff? Yeah, inside. Turn on the LED light. Yeah, then put the image to the LCD between the LED light and the LCD tank. The resin tank. Nice. Then the image can be projected onto the resin. After some time, after explosion, then it is cured, solidified. Then it's one process to make one layer. And then pulled up and pulled down again to make some gap, to make another layer. So it's the same step. Alright, how many layers per one church? Actually, we set 100 microns to make it. And this might be 20 centimeters, then maybe, how can I say, 2,000 layers. And how long does it take one layer? Normally 20 seconds. 20 seconds per layer? Yeah. And after that they change color or you spray it or how do you make the different colors? Of course, it's kind of post treatment. So you have all these different demos you have shown here? Right. Different demos, different designs, different wedding. And complexity can be very high? We think so. So where are you based? We are based in South Korea, but we have a small office in San Jose. And what's going to happen in the future? Are you selling these? Yeah. We went on the Kickstarter two months ago and we got some funding from the backers. How many backers? 100 people buy it? Yeah. Now 100 backers are paying us and 40 people are buying it. So this machine will be shipped next month to our backers. How much? Because they already paid around 4,000 US dollars. And some people would like to get this one, visit our website this weekend. And we will also review the site with limited quantity that price will be US 5,000, around US 5,000. And we will start our commercial production sometime between March and April next year. Then the price will be higher than might be 6,000 US dollars. So what do you think about the ID Tech Act show here? That people come and it's very interesting? Because we are invited to the launch pad by supporting Qualcomm, thanks to Qualcomm and ID Tech. We've met enormous feedback from the old attendees and some new debaters. So a lot of business in the 3D printing in the future is going to be a lot of things happening? What's going to happen in the future for 3D printing? I'm not sure but we believe. I and my staffs are believing 3D printing can make better work because 3D printing is a tool to release individual creativity. And combining different materials will be possible? Of course not for this machine but we have a complete roadmap to combine different materials, different colors. Electronics? Of course. Maybe? Should be. Of course. Cool. So please keep an eye on. We are on a small start-up at the initial stage but we have a very big vision and we have technology. We have very good development steps. So please keep an eye on.