What I'm going to tell you is that Vermont was the first state to abolish slavery. And over the years, they really polished this reputation. But beneath the surface existed racism that many have chosen to ignore. Madeine Maudine Neal exposed this undercurrent in her talk about the Ku Klux Klan in Vermont. So I hope you enjoy her presentation, which is a very thorough one. And it has many pictures and many relics from the days of the Ku Klux Klan site in the 20s. So without further ado, Madeine Neal of Franklin Street in Montpelier is going to speak. We're going to do a PowerPoint first. And I will get in the process of the pictures. Now, first I want to tell you the cover of the book was designed by Wesley Herwig. Anybody here know Wesley Herwig? Susan Randolph had a little printing shop and he published the book and he designed the cover. And here we go. Did it move? Yes. What was the Ku Klux Klan? Did any of you know that ahead of time? Do you know what it was? Yeah. What was it? The Ku Klux Klan, it was an anti...  Okay. And this one. I don't see very well. Trying to get so I can see it. So people thought they were protecting themselves. People thought they were protecting themselves from them. Hello, hello, hello. That was a turn on and off. Can you hear it? No. Well, I didn't touch it. There, try that. Okay, okay. Can you see that? Okay, it was started by Nathan Bedford Forrest. Not picking her up again. It keeps going out. He was a cavalry officer in the Civil War. They said he never lost a battle. I don't know myself if he did or not. Okay. Can you read that? Is that a yes or a no? Okay. Next picture. That's not the next picture. Because you're going back. You want to go this way? I know. What's happening? That's what there was another. Can you read that? Okay. That is not the clan that came to Vermont. Not that first one. Because it became a terrorist thing. In 1915, another clan was formed by Joseph Simmons. Can you read all that? Born in 1880, died in 1945. He wanted it to be a fraternal organization. And when he saw that it was becoming prejudice, he got away from it. 1924, he sold it to Hiram Evans. Hiram Evans didn't want it to be a fraternal organization. He wanted it to be a money-making thing. And you have read this now. Why was it accepted? Before you go on, would you talk about how you thought it would be a money-making organization? They were charging admission. Not a lot. $10 a person. But that's the first I know. That's the way it was done. The purpose of it is here. You can read that. Okay. Don't change it so fast. Give us a chance to read it. How did I get involved in it? I grew up in Oklahoma. And when I got out of high school, I went down to Texas. And they had an Air Force base there. And my husband was there as an airman. I met him. And we got married 70 years ago. And he was from Vermont. He was born in Berlin. I liked him a lot. And I like Vermont a lot. So here I am. But I was surprised when we were looking at his mother and I was in pictures. And I saw this big, long picture that could expand people in it and say, what is this? And she said, well, that's me right there. Thank you. She said, I'll show a big one. There'll be a big picture of it on the screen in a minute. Okay, good. Okay. So I questioned her about it. She said, oh, it was just a good time. It was a picnic and prayer and singing. Just having a good time and burning a cross. That was fun. But how many of you like a bonfire? They went to schools and gave the kids flags and different things. Unless they were Irish Catholic. In 1970, I took a Vermont history course because I've always been interested in history. And Charlie Morrissey was going to do a history course on Vermont history. So I started in. He said, everybody has to do a project, a history project. And I went to him and I said, how about if I do it on the Ku Klux Klan? And he about fell off his stool. He said, it's never been done. So, okay, I'll do it. So, where would I go to find material on the Ku Klux Klan? Where would you go if you were going to find it? Vermont Historical Society? The newspapers. You got it. Newspapers. The Historical Society had nothing. But every day in the newspaper, and they were easy to find, there was KKK headlines. Almost every day in the newspaper. So that's where I got a lot of my material. And I got a lot of it from personal interviews. A friend about that time, who had bought a new, a different house. And she said, she found in the attic some Ku Klux Klan material. Some robes and some hoods. And if you see the big banner that says Montpelier women here somewhere. It's everywhere on the table. I'll get it. But we'll see it later in the pictures. So I asked Kristen, could I take those to my class to show the teacher? Guess what I got for a grade. Okay. Yeah. Yeah, yeah. He liked it. So, we got all the things and took them to my class. Okay, are you familiar with this house in Montpelier? It's on Liberty Street. And it didn't look like this in 1920. It didn't have that top part on it. But this is where Edwin and Emma Nichols lived. And they were the head of the clan. She was the head of the women. He was the head of the men. I asked her, Charlie Nichols, our former mayor was their child, but she didn't know. I don't think they had children. She was 43 when she married him. But I didn't find any children for them. These are some things that were found in East Montpelier. And when I published my book, people started finding things all over the place. In the Vermont Historical Society, it has a getting a pretty good collection of things. So what did you publish your book what year? In 87. Here are some newspaper items that I found. Cross burning and Barnard. A big gathering in Shaw's Field. 600 to 1,000, I don't know if that's cars or people, but there were a lot of people. In the bottom is later Crawford's Auto World Land. Do you remember that in Bethel? Crawford's Auto. Nice. How were the articles presented? Was it just straight news coverage or was it expose? They were kind of against it, most of them were against it. Here's more items, there were 150 clansmen in Windsor. You can read all that, right? Yeah, give them a minute. Can you see it, Mary? Yeah. That's, okay. The last one is a cross was burned on the Catholic church steps. That brought a lot of attention because two men were arrested that did that. And they were getting ready to have, you have a question? Yes, if they were Protestant, why did they earn crosses? Why not? Crosses. I don't know why, I don't know how that developed. And I couldn't find out. What time period now are we talking about? 1920s. Okay. And that's all. All right. Well, my grandma told me that when she was a little girl living in war town as an Irish Catholic, she was told not to go near the encampment, which was down on the Bad River, right below War Town Village. That's right. Because they were stealing children. Oh, well, okay. We'll have time for comments and questions later. All day if we want to. But I want to tell you about these two men that were arrested at the Catholic church. They were getting ready to have a trial for them, but they never did. They finally had to just not do it. But the lawyer that was going to defend these two guys was named Dean Davis. Oh, wow. Who was that? Governor of the Vermont State of Vermont later. And here's some more, a lot of them. And the last one is Weston's trailer park. Oh, yeah. Do you know where that is? Sure. Well, you know Ken Weston? Well. He lived right there. His wife was a herring. And the head of the women of the clan in Montpilier was Emma Herring. And she was a cousin of Ken Weston's wife. So I expect that Ken Weston was in it too. I don't know. But they did things in his field. And this is a place on the hill where that picture of the big group was made. You familiar with that? What is that in Berlin? In Montpilier, Town Hill. Wow. Yeah. Okay. Anyway, we'll see more about the parade.  Now here's another personal contact I had. Ethel Bailey, any of you know Cliff Bailey? Barry, he's a big businessman over there. His wife told me a personal story. She said, my brother was in the clan, but I didn't know it.  They all drowned because they got swept into deep water. And when they had the funeral, they were in their gowns, their clan gowns. That's the first I knew that my brother was a member. And you're here with Dr. Corson? Corson. Corson and Plainfield? He arranged the funeral for these people. Said when they marched around the grave, they were throwing flowers into the grave. Because flowers, they were throwing flowers into the grave because these were their friends, their clan people. Okay. Another personal contact I had was with a woman that lived downtown, up above Summer's Heartburst or in those apartments. They lived up there and she had a little boy and she made him a robe so he could go with them to the meetings. And she told him one night, there were seven crosses burned around town. And you see the list of them there, she didn't remember all of them, but said they really enjoyed looking out and seeing the people on the street, being really startled by all this. And this is a clan outing of some Northfield clans, more women, and the woman on the right, as you look at it, is my husband's grandmother. And the next one was his mother and two other people from Northfield. They went to Smuggler's Notch for a picnic. Now, doesn't that sound like fun? How many of you been to Smuggler's Notch for a picnic? And here's some other members from Northfield. Mrs. Whitney, her husband ran the Northfield News. You can read the names of these people. So they were all, most of the people who were in the clan were upstanding reputable people like newspaper editors, preachers. They had a lot of meetings all over the state. The big event they had was the parade, and that was in 1927, July the 4th. Why not have a big parade? This paper is printed and it's laying on the table. You might want to read it. I'm not gonna try to read the whole thing to you, but it describes that parade, and this is the area where it was. Can you go back to that for a minute so we can just? Could you go back so people can read it a little bit longer? Oh, the whole thing? Yeah. If you take the paper over there and read it. Well, it's easier for us. Take less time. People like sitting down and seeing it in big letters. I know. I shouldn't have put that in there. It's okay. They say there was 15 to 20,000 people in town that day. And here's the leader of the parade. I would guess that that is Edwin Nichols, the head of the Montpellier Clan, and that is the Montpellier Clan's women. And Louis's aunt is kneeling down in the front row, the second from the right. That's his aunt. And she lived in Montpellier. Kerry Warren? Clara. Clara. Yeah, her name is Clara Warren. I can see it from here. Yeah, but Kerry Warren was her mother. And this is the Barrie Clan. You see their banner? And this is the Montpellier. No, this is the Northfield. And Louis's mother is standing behind the flag on the right end. That's Louis's mother, Laura. And she would have been about 27 years old. And she got married later that year. The Randolph Clan. And these are all in that big long picture, but I chopped them up because I don't know where I could get them into a picture like this. And this is the Boston Tea Party Clan. A group came up from Boston for the big Fourth of July celebration in Montpellier, Vermont. And this is the area where it was on Town Hill. Do you remember what Town Hill is? Yeah, it is. You go out Main Street to the northeast of Montpellier. That used to be the Juliana's house, you know. I'll tell you, it used to be airplane landing field. And in 1927, when they had the flood here, the bakery was running out of yeast and airplanes were dropping yeast up on the hill so they could continue their bakery. Cross funds. Yes, I did make it. My husband worked there for 10 years. The Clan, by this time, is getting a bad reputation. So they were beginning to fold up their things and hide them and deny they were ever in it. By 1930, the Clan was totally disbanded. You can read that. When I was doing my history project, a lot of younger people, which would be my age and years, didn't know that it had ever been here. Would you have known it had been here? Mary Alice did. Not a much by group, but my grandmother was much older, so she must have been in the 1900s. Yeah, well, I would just bet you that a lot of your grandmothers were in it. Well, some of us will have Irish Catholic as a background, so... I don't think that would have mattered. George. Now, this changes the whole subject right here. The witness stump. My husband was working on the neighbor's garage, and he saw this thing and mentioned to her that I had been doing Ku Klux Klan work because down in the corner, it has a KKK-32, and she gave it to him. And up in the corner, in the right-hand corner, with a pencil had been written, and it was so damn I could hardly see it, I had to use a magnifying glass to see it. It said Stan Olin, Chicago, Illinois. Hello? I'm a genealogist, so what do I have to do? I had to find him. So I went through the 1920 census, also the 1910, in the 1930, you can see it better in the 1930, that the fourth person down, Daniel Olin, he was born in Finland. His life's Alma was born in Sweden, and they have two boys, Stanley and Clifford. Here he is in the census in Illinois. I don't know if he came to Vermont and wrote his name on this witness stamp, or if he wrote it there, and then it got brought here. So I didn't find out that much about him, but I did look up a lot of his. What is a witness stamp, yeah? That we did not find out. It wasn't included in the amount of plan research, but because it said KKK down on the bottom, I wanted to put it in my story. Well, I did my research with Charlie Morrissey, and at the end of the course, I just placed my manuscript in the Historical Society, and it stayed there until 1987. It's the end of the slideshow, but I have a lot more stuff to tell you about. Do you want to turn the light off? Yeah. Okay. I have a newspaper article from the Harvard Gazette. February 8th, 1923. So 1923 is when it was starting up in Vermont. And here's what the Gazette said. A Ku Klux Klan organizer, George P. Mason by name, is due to make an attempt to start something in Vermont soon. He is now already talking it up in New Hampshire and has told people in that state that he plans to organize plans all through New England and eventually control the public offices. Last week, he invaded the New Hampshire legislature where it is claimed that he enrolled membership of 20 legislators in this organization. Forewarned is forearmed. If Vermont is on the lookout for him, it may be better prepared to meet the situation when it arrives. So this is kind of warning they're coming. I'm going to tell you some more about Edwin Nichols and Emma Nichols that lived on Liberty Street in Montpeyre. Her maiden name was Herring, and there was a lot of Herrings in there. Any of you related to Herrings? There are a lot of them around. A lot of Berlin. A lot of them in Berlin, and there was a lot in Moretown at that time. And Emma Herring's father was named Charles Herring, and he was from Berlin. He was in the Civil War, and he got wounded. He was shot, and he had a brother's seldom that was with him there, and he put him on a retreating cannon. Then he put him on an ambulance, and then he put him on a baggage wagon, so he managed to save his brother. And Charles had a brother, William. He was the father of Ken Weston's wife. So I prepared a little Herring genealogy here. Okay, another incident that I found was in Southbury. A friend of mine, well, two friends of mine, Don Camp, ever hear of Don Camp? He had a farm in Southbury. Camp Street. Maybe. Well, Camp Street is where all the granite and your fracturing owners live. Yeah. Well, anyway, he was a member of the clan, and another friend was a member of the Grange, and they made a deal. If you join the Grange, I'll join the clan. So they did. Well, this friend of mine that was telling me about it, he said, I was really a joiner, so I got involved and I became an officer. They gave me a special collar to wear when I'm officiating in the office there. But one day I went to a meeting. There was a strange woman there from New Hampshire, and she said, I've got to talk to you. I understand you're harboring Jews. What would that mean?  And said, they followed me to the door trying to beg me to stay.  So he might have appeared to be a Jew, but he wasn't. And when that little boy grew up, he married and had a little boy. Well, neither one of them were Jews. His wife died and he needed someone to take care of the little boy while he worked. And my friend was taken care of him. So that's why they accused her of harboring Jews. But she would have anyway, she was the kind of person that would help anybody. She didn't want to be against anybody. Another case, did you ever hear Cliff Bailey in Barry? He had a wife, Ethel Bailey, and she's the one that said that her brother was in the Klan. I guess I told you all about that, but Dr. Corson prepared the funeral. So at the end of the three month class, I wrote my paper, left it at the Historical Society. And during the time it was there, over 10 years, people were coming to me from all kinds of places. One came from Maine, had a picture just like mine, made the same day. His was number one, mine's number four. So I'm sure there's more of those in somebody's attic around. Pat Shepard is a lady from Topsum. And she wrote to me and said, my mother has a bunch of Klan stuff here. She doesn't really want it, don't know anything about it. Would you like to see it? So we went over to see it, and she ended up giving it to me. And I think there's a picture over there somewhere of all the stuff. It was a little pocket knife that said KKK on it. And there was robes and there was templates. And she gave all those to me. I gave them to the Vermont Historical Society. And you see this, the book standing here on the end is called Vermont Century Book. It's the 200th anniversary of Statehood. And the section in the, and it was kind of laid out every 10 years. The section on the 1920s has the picture of a big Klan picture. Okay, one more character here. Karen Hess was an author. She wrote children's books. And here's what she had to say later on. She said, in 1997, while returning from a speaking engagement, I spent the last minutes of my flight skimming the airline magazine. I came across a short piece about the Ku Klux Klan in Vermont in the 1920s. I read the item, shaking my head in disbelief. Back home, I wasted no time in attempting to disprove the article. But to my surprise, it was correct. I read Maureen Neal's book about the Klan in Vermont. I wrote to her about her research and tried to imagine how I might take this episode in history and craft a compelling story from it. Well, she did write a story about it. And it's over there, a book called Witness. She wrote the book. And shortly after that, in 2003, Vermont had a program. I don't know if they do it every year, but it was called Vermont Reads. You ever hear of that? Yeah. Every town or school, at least, wanted to read the same book that year. In 1923, the book they wanted to read was Witness. 1923? 2000. 2000. Oh, wake me up. 2000. Is that the one called? 2003. Different points of view. Pardon? Is that the book where different people talk from different points of view? Maybe. She wrote it as a play. And in Northfield, she had it performed. And when she chose the people to be the performers, she chose people that had the same background. I have a question. Okay. You talked about your mother-in-law described what the KKK was for her with the social. That's right. So, but they did do all the cross-breeding. So how many didn't evolve from kind of an innocent in Vermont? Did it evolve from kind of an innocent social event to the darker side? Or was that always there? And some people either ignored it or didn't know about it? They didn't know. Like my friend in Southbury, who was accused of harboring Jews, she didn't know she wasn't supposed to harbor a Jew. And really, she wasn't. And you came from Oklahoma. So you actually knew the Ku Klux Klan as a pretty terrible warhead. I knew to stay away from it. My father's people came from Georgia and South Carolina. And they came to Oklahoma in the 1920s. And that's when it was pretty active down there. Do you have any information about when it started over in New Hampshire? Because the people that were honoring my grandmother were coming from New Hampshire and camping out over here in probably the end of the 19th century. I think it would have probably been, Joseph Simmons started this Klan in 1915 in Georgia in his spread all over the country. So it came to- My grandmother was born in the 1800s. So I think it had started in New Hampshire long before. Well, the first Klan might have been, but I don't think it was up here, though. It was more in the south. But 1915 is when Joseph Simmons started this Klan. In Georgia, and then spread all over, right? That's right. Okay. Well, I think- Tell them about finding some of the robes in the houses in Montpelier, the story you told me when we got together. People had them in their attics. I did. I did include that, didn't I? I thought there was another one, because didn't the picture you got of the women of the Clue Cooks Klan? Women? You know that picture of the banner? I mean, didn't you have some that the woman wanted to sell and she took them back? Sorry. I don't know what you're talking about. Okay, but that's the end of my research, but I'm gonna read something to you, okay? This is by Marvin Neumauer, a German theologian. He said, first, they came for the socialists. Then they came for the trade unionists. Then they came for the Jews. In each instance, I did not speak out. As I was not one of them, then they came for me, and there was no one left to speak to me. So people had questions, and I'm gonna help me, Dean, to find- Who has heard this before? Everybody had it up in my classroom. But I think it kind of describes the Klan. Either you know what you're getting into, or you don't. Okay. And the first question is out of the blue. Can you tell me what Ku Klux Klan means? What the words? Ku Klux means circle. Klan was taken from the Scottish clans, and people who had a common interest. So Ku Klux Klan's, and they called it Ku Klux Klan. Thank you. Back to the Cape Cod sweatshirt. So I'm curious, were most people in Vermont in the Klan like your mother-in-law, there for social gathering, and weren't aware of the hatred behind it? That's right. That's exactly right. It was the man that did all the work. Well, even the man didn't know right off what it was. Black and white? In the early 1980s, I worked in Middlebury with a woman who had grown up on Route 7 in East Middlebury. She was born in 1921. And one of her earliest memories was seeing a cross burning out her bedroom window on a hill. And we had a conversation about how beautiful she thought it was, because she didn't choose so little. She didn't know what it was about. And having to switch her mind, she didn't learn what it was about, but she said after she got out of high school and someone explained to her what that was about and how she had to switch her mind from this beautiful image in her head of this gold-firing thing to what it really meant. It was a conversation I'll never forget. That's where it was. It was a beautiful thing. I like to see a fire, a bonfire. Probably enjoy seeing a cross. Can you tell about instances of violence that you learned about? Not really. Well, you talked about the fires all over Montpelier. Yeah, the cross burnings, but those weren't violent. They were, like the girl said, greedy. But there was a case in Burlington where two men broke into the Catholic Church and stole some artifacts and they were found and that wasn't really violence, but it was mischief. So I think a lot of them were. I had heard that when they came to Montpelier that it was the mayor or it was somebody high up in the Montpelier City government and that's why they never took them to court. But there is another side of the story, all the victims and the people that had to deal with this hatred. There were Irish Catholics, there were French Catholics, those churches, all that. You didn't do any research with the people who were victims? No, I didn't. I just said what I said was in the newspapers. This was just a less than a year course and she was also raising a family. So she did more research than anybody's done, but it's hardly as exhaustive as it might be, but the resources are limited. They really didn't talk about the victims of the news. No, the newspapers wouldn't talk about it. No, but there must be several people in the community who remember from the Catholic and the Jewish side when it was like living here in those days when you had all the wasps against you. Hey, there's something for you to look up. Yeah. Find that and tell us about it. And time is short, right? All those people are. Well, I'm a Burden, I mean, I have a friend who's great for his wife's great aunt was a member of the Klan and not only was it a social club, according to this person, but it also was a Christian. It had a very strong Christian component to it and she joined and really loved all the festivities and the fire probably, but then other people said, oh, this is a terrible organization. What are you doing being a member and so they quit.  I mean, may Dean at least talk to some people when she did her research in heaven that were in either in it or their parents were in it, their relatives. Okay. Purple. Prejudice didn't just start with a claim. All right. I had ancestors that were Irish Catholic in New York in the 1900s and were signs that they were always on the building. Catholics, you know, Irish. I had a high class of the Catholic. So the Klan came up with something that existed. You can't blame everything on it. The undercurrent of racism as I called it. Do you ever hear the term wasp? Yeah, sure. What is much standoff? White Anglo-Saxon Protestant. White Anglo-Saxon Protestant. That's what the Klan was. Growing up with Yankees that had been here since 1634 on my father's side and my mother was an Irish Catholic. So I got both sides. My father was, my grandfather was Mason and I think the Mason's had something to do with. Well, the Mason's were also a religious organization, Purple Church. Yeah. One of the interesting things about your story is you said that the group here in Montpelier had their picture taken in front of the Catholic Church. And that surprised me because I thought they didn't like Catholics, right? I didn't say they didn't have pictures in front. I said they burned across on the steps at the Catholic Church. She said they did it up on Hill Street, up on what I think it might be at the Giuliani. But they did it right on the steps of the church and nobody would do anything because they were all people who were highly up in the government. Yeah. And they said that the mark on the Catholic Church stayed there and they couldn't get it off. Any other questions from someone other than Mary Alice? No. I would just like to comment that I think we have to be aware that we're living through a time now where there are some groups, crowd voice and some of these others. Oathkeepers, just one on trial. Pardon? Oathkeepers, the president of Oathkeepers. And crowd voice. And we're seeing the same kind of prejudice stirred up that we did then. So I think we all have to be aware that it's around us and speak out against it. Absolutely. That's why I read this for you. Yes. If there are no more questions, I'm gonna try and have somebody help us get this screen off. Yeah, he tried and he wasn't... You can come and look at all of these things as long as you want to. Otherwise we should, let me be on the other side. Whatever. It's not going up there. It doesn't matter. I like to say the more I think, that's why a lot of us don't go to the North Pole. Yay!


Why is the president bragging today about gas prices? Because it's gone down. I love how she responds to that. Like, it was a test question she didn't know the answer to. It's still $1.72 higher than when he took out this. It has come down in a way that we haven't seen. It's trajectory coming down in over a decade. And it is still $4 a gallon. Was that good? You want to talk to a nurse or a teacher or a firefighter? If having that little bit of breathing room doesn't matter to them. Are nurses and teachers and firefighters saying gas prices are only $4 and 11 cents? This is great. I'm saying that even that little bit of breathing room matters to families. This matters. When you think about $0.85 per month for family member, you think that doesn't matter? Oh, yeah. No, $0.85 a month does not matter. I cannot believe how utterly disconnected from reality and the average person this administration is.


So there aren't going to be, we originally hoped that by this time we would have estimates of poverty impacts to present. We don't have that, but we do have a taste, a somewhat simplistic taste of some of the results coming out with regard to sources of yield gains. So the latter part of my presentation, if you hold tight to the end, you will get to see some preliminary findings about what the big winners in terms of yield gains will be. We haven't had time to process the input-output data in order to look at technologies that improve efficiency, that are cost savings, that have other more complicated effects. But there are some interesting results at the end of this presentation. To begin with, I'll start with the process and how we will use these data and how we will get to estimating impacts for the poor and on the environment. So just to review where we are, for those who may have just stepped in or if they weren't here before the coffee break, where we are right now is we have about 55 research solutions that have been assessed by participatory scientist working groups where they've looked at the resource requirements, probabilities of success in terms of having research solutions develop the time frames for those being developed, and then the expected adoption levels and on-farm input-output effects. These 55 solutions are still not all the solutions. Some of them are still being discussed. There are interesting points where we do have particular uncertainty or divergent views where we're still discussing. We also have probably on the order of something like another 20 that we still haven't yet done, that are ones that are particularly more complicated or maybe even areas where we do not have sufficient in-house capacity to make a good assessment and we need more to bring in external expertise. Those assessments of the research solutions have been backed up by analyses of problem prevalence and trends in those problems, and that's been complimented by what was presented in the first presentation, and that's a spatial analysis of trends in actual yield and attainable yields to 2035 so that we identify how the base yields that we should assume for one different technologies come in, what the base yields will be, and so we can look at what that scope for all technologies that help to close the yield gap is collectively over time, how that has evolved. We will use that to basically triangulate the information we get from the scientists. We don't want to be coming up with estimates that say that we are actually exceeding the yield gap through technologies that close the yield gap, so it's a good reality check on what we're doing. We'll take those estimates which Boss gave you a much more detailed picture of. Those are generally in relative terms in terms of effects on input use, different elements of production costs. We have an extensive in-house survey database covering many different Asian countries that we're going to use to get the baseline values in terms of input-output use by operation so that then we can translate those relative changes into absolute changes and we can look at what the unit cost of production is and how that's being shifted by the particular technologies. When we do that, we also will take into account the adoption trajectory over time and the adoption trajectory over time that would be likely for the solution without yield. So that will all be reflected in the effect that's estimated on the unit cost of production. Now why is the unit cost of production important? It's important because A, it's the basis of farm revenue changes and B, there's an important dynamic effect that when the unit cost of production goes down, the general market price will go down and you need to incorporate that in order to see what the farm level benefits are. So the price effects of changes in the unit cost of production are being modeled via our own global rice trade model that's been developed by Sam Mohanty. I won't go into the specifics of the model but I'll just illustrate a few aspects quickly. The model itself then has elements of the cost of production that are used in terms of determining the supply for a given country and basically the changes in the unit cost of production will be shocked into this element of the national rice supply. The national rice supply will then be shocked into the equilibrium of supply and demand and trade for an individual country which then will be equilibrated across the different countries in the model. And so the model includes these 18 countries and most of the countries are sub-nationally disaggregated so we actually get sub-national price effects as a result of the change in the unit cost of production from adoption of the technologies. So once we have the change in the unit cost of production and we have the data on the price effect we can then look at what portion of production is occurring by those under the poverty line and what their general characteristics are compared with average producers, particularly in the area of rice that they have cultivated because that will determine what kinds of farm level benefits they can receive from the technologies considering the equilibrium price response. So we have data sets on the spatial distribution of poverty. We can use that to approximate the number of poor in the different agroecologies and based on that we can look at the proportion of benefits occurring to those poor households when we take into account their differences particularly in production area. Then on the consumer side it's also very important to look at benefits to the poor. Through the trade model we get these price effects and we know that poor consumers spend very high proportions of their income on rice in Asia. They spend about 50% of their food income on rice since about 25% of their entire household income for the population surviving on less than $1.25 per day in purchasing car parity terms. So that's also an extremely important pathway to impact for the poor. So this was a simple run that we did before looking at an aggregate contribution to productivity and rice supply. It's a very modest set of assumptions we used. We basically said that we will continue productivity contributions at the same level as we've been documented to do historically up until from 1960 to 1998 that we would continue the same level of contribution. With that same level of contribution you end up with these kinds of benefits. It was basically a net contribution of 15 kilograms of rice on average per hectare per year for Asia. With that kind of a contribution it's about 8.5% over 25 years. You end up with a price reduction that ranges from 9% to 18%. The price reduction exceeds the supply increase. So that price reduction then considering how much the poor actually spend on rice it has massive benefits for poor consumers. So if you look here that kind of a price reduction over 25 years would lift 125 million people above the $1.25 a day poverty line if you count the reduction in their expenditure on rice as income. If you look at the proportion of the population that's food insecure if that expenditure savings were used to purchase rice to close the caloric gap potentially 62 million people could be lifted out of hunger. That's a big assumption but it's a possibility. So taking into account the consumer side is very important. Also when you see that these price effects are so pronounced it's important that we consider that when we come back to the producer benefits. So economic and poverty benefits are not the only benefits we're looking at. We also will look at environmental benefits. In particular when you have such a big price response you get an area response. So through the productivity enhancing technologies you actually have less area under rice. And part of that decline in rice area is also an avoidance of expansion. So given that there's a lot of land pressure in Asia and competing land uses some alleviation of that pressure on land we'll save natural land cover such as tropical forests. So taking into account the environmental benefits of that savings of natural ecosystems is very important and something that we're trying to reflect here. The spatial disaggregation of the model in terms of the supply response can help because then we can look at where the contraction in area happens relative to the proportion of safe forest cover. We will also then look at the more direct environmental benefits such as Boss mentioned for AWD in terms of the reduction of greenhouse gas emissions. And we will supplement this with analysis of the health benefits from our biophortification work and potentially some work to reduce the glycemic index of rice. And then we will put together all these different data on expected impact by different impact sources and we'll compare it with our patterns of resource allocation to look at where are areas that are in need of additional investment in order to realize much greater impact. So those are the methodological steps going forward. I didn't want to go into great level detail because this is the last presentation of the day and I want to leave some time for discussion. I'll now present some very preliminary results based on sources of yield gains using the parameter estimates of the sort that Boss was presenting. Now I have to of course give a number of caveats because these are very preliminary. These are not final estimates. We are still in the process of refining and reviewing our assumptions in these sheets. We are though looking at this as an opportunity for feedback so we do want your feedback. Your feedback still can affect actually what we're estimating given that we are still in that process of adjustments. This what I will present is only on the sources of yield gains. So it's a subset of all the research solutions. It only includes those that we have more confidence in our estimates and it doesn't include a whole array of technologies such as those that increase efficiency, those that are quality oriented, post harvest technologies, policy oriented research, etc. Those are incorporated in our priority setting exercise but they will not be incorporated in what's presented right now. This does not take into account the actual effect on the Unicostar production. It's solely based on yield and it's only using the lower bound parameters from scientists on adoption.


All right, thank you. Good morning everyone, a big thank you to the organisers for giving me the opportunity to present some of the work that we've been doing on the self-assembly kinetics of linear virus like particles. I apologise for my voice, I'm a Dutch person so I don't do well at elevated temperatures and under the influence of sunlight. Anyway, so these virus like particles consist of a strand of a double-stranded DNA which have been encapsulated by a tri-block recombinant biosynthetic protein and I'll show you the chemical structure later on in my talk. This biosynthetic protein which actually was designed to mimic the functionality of the code proteins of a tobacco mosaic virus. Many moons ago we investigated theoretically both the stability and the assembly kinetics of a tobacco mosaic virus and that led us to design this protein. But before going on, here are my buddies in crime. There's an experimental group who did all the experiments. There's the group of Renko de Vries at Wageningen University which is in the east of the Netherlands. The actual experiments were done by Amando Hernandez-Cathia who is now at Northwestern. We've actually, I mean, left two years ago. We've since done more experiments. I'll be reporting on the end of my talk hopefully. The theory gang is welcomes us of Willem Kegel who is part of the audience and actually Willem is the chap who drew me into physical virology. So maybe, I don't know, 12 or 13 years ago he sent me a manuscript about the self-assembly of tobacco mosaic virus in fact and I thought it was rubbish. But we wrote a fantastic paper about it. No, no, no, no, no, no, as you mentioned. Anyway, at the time, Daniela Kraft who now is at Leiden University was a PhD student with Willem doing experiments. Her background is theoretical physics. So she turned to the dark side but she wanted to have like a little toy theoretical problem. So the first theory for the self-assembly kinetics was done with Daniela. Very recently, another chap, Mella Punter, who was a master student in my group, he extended the model that Daniela and Willem and I coughed up so many years ago. Okay, so what do we want? Oh, funding. It's rather important. Funding virus-related topic is not that easy in the Netherlands, I think. I mean, so we got some funding from NWO, which is the Dutch equivalent of the NSF and the now defunct Dutch Polymer Institute, which is always extremely generous and I used to have a project with the previous speaker from the HFSP and there's a chap sitting over there who got a salary out of it. Anyway, so our aim was to encapsulate DNA. The reason, I mean, way in the background is gene therapy, double starling DNA, using a synthetic protein. Of course, viruses are the ultimate encapsulation machines. They're made to infect things. So we thought, why not imitate what nature does so well? Here on the left is an image of a CCMV, which is a plant virus. It's very, very small. It's just shy of 30 nanometers across. About half the viruses have this shape. Or, for instance, look at tobacco mosaic virus, which is actually the first virus to be discovered, I think. That one is not longer. This is about 300 nanometers. Now, if you look at the internal structure of CCMV and TMV, it's actually quite different. They're very different. I mean, the shape difference is obvious, but also the way that the genome was encapsulated is very, very different. If you look at CCMV or any other small plant virus, it consists of this shell, you know, of protein. And inside this shell, you've got a cavity. And this cavity holds the genome. This is the genome, where, for CCMV, you've got three different particles that they all have, roughly, 3,000 nucleotides of single-star irony inside. Now, the driving forwards, so actually, I should say, both these viruses have been reconstituted in vitro, which means that you can disassemble the viruses into the proteins and the genome. You can purify them and then mix them into the same test tube. You swirl, I presume. That's what I think that experimentalists do. And, lo and behold, it will self-disable. And the driving forwards, I mean, both, I mean, certainly for CCMV, in the first instance is electrostatics. So the inner surface of the viruses is very highly positively charged. And this is also the reason why you can just yank out the RNA and replace it with anything else that is negatively charged, like particles or droplets or whatever. In fact, I don't know if Adam Slotnik is in the audience yet, no? But actually, Adam Slotnik encapsulated DNA. This is interesting because, you know, this particle is 30 nanometers across. DNA has a persistence length of 50 nanometers. It is no way that you can get DNA in this very, very, very small lumen. So the protein is easy. It adapts to the template that you offer it. But it still has this preferred curvature that is, you know, like 30 nanometers. So what this protein does, it will grab hold of multiple strands of DNA and form a tube with a diameter of 30 nanometers. Our goal was to encapsulate single RNAs, not bundles. And one reason would be that, well, you know, if you have a long thing, long thing object that's more easily absorbed by cells than a fat one. So why not try the trick of tobacco mosaic virus? Well, tobacco mosaic virus doesn't have a lumen. It has a hole which does run through the entire center of the object. But the RNA is not encapsulated in this lumen. It's actually embedded in the capsaid itself. Right? This TMP will not encapsulate DNA, double-stranded DNA. It will do single-stranded if you force it to. So our goal was now to try and combine, you know, the properties of TMV and the properties of CCMV. So the easy, so having physical driving for the assembly and still have used the tricks that TMV use in order to encapsulate something. Because as we shall see, that's not so trivial. Because it's a quasi-linear object.  We tried to encapsulate single-stranded DNA, homopolymeric. So it cannot self-hybridize. So it's a polymer, like a homopolymer. And we tried to do that with a synthetic molecule, which is a naftali derivative, which has three functionalities. It's got a functionality that will allow it to hydrogen bond with the single-stranded DNA. It got this planar group. This planar group will allow it to interact with itself and self-assemble along the single-stranded DNA. And then there is a group attached in order to increase the solubility in water. And now this homopolymeric DNA will not form a helix, which means that you can monitor helicity as a proxy for how much stuff is absorbed onto the DNA.  So we mix them together. And what happens is we see a CD signal coming up, and the CD signal tells you that there is a co-assembly into a helical structure of the DNA and that let's call that a code protein. And so this is what happens. So on the vertical axis, we've got the normalized a CD signal as a function of the concentration of this object relative to the concentration of the template for three different template lengths. So this is the longest, so 40 nucleotides, 20, 10, and 5. So we're adding here more of the code protein. And what we're seeing is that the CD signal increases, so it gets packaged. The DNA gets packaged. Actually, it doesn't really get packaged at all. It's really difficult to get full 100% coverage of the DNA. And actually with hindsight, that makes sense, of course, because it's like a one-dimensional absorption process.  I will create vacancies. So DMV is smarter than this. At the time, we're not as smart as DMV. By fitting the experimental data to like an eising-like model, we can extract stacking energies and binding energies. By doing the same experiments as a function of temperature, we can even extract and apply the model. We can extract enthalpies as well. And so here again, the CD signal versus now temperature for one concentration of 0.25 millimolars at this stoichiometric ratio. By fitting the theory to the experimental data, we get enthalpies of, let's say, about 20 KBT. Now, you can see that there is a reasonably well-defined elongation temperature. So what you can now do is plot this elongation temperature as a function of the concentration of the code protein, the quotation marks code protein. And that gives us these straight lines, which you expect, actually, Vanthoff's law. And now the interesting thing is what you can do is you can also not add the template. And these objects will also self-assemble without the template. And but it does that at much, much, much lower temperatures, except for this one. So this is G1. That's this one. At large enough concentrations, actually what happens is that the self-assembly takes over from the templated assembly. So what you have is a competition between templated assembly and self-assembly. This is something that if we want to design a virus, a linear virus, we have to make sure that does not happen. Also, okay, so how does TMV do it? So TMV doesn't follow this route. It doesn't rely on linear type absorption. What TMV has is what is known as an origin of assembly sequence, which is the same thing as rating the packaging signal. And assembly actually happens bi-directional. It starts at this packaging signal and then runs that way. And then later on, in the last part of the assembly, the assembly goes in this direction. Let us forget that this is about 900 nucleotides upstream. The RNA cell is slightly over 6000 nucleotides. So we can forget about the last part. The last part is boring. Now looking at images like this one, what we're seeing is indeed linear object growing as a function of time. And if you focus onto one of those images, you see a rod and what you see is two strands of RNA coming out. This shorter strand is this bit over here. The longest strand that's all of this that has not been encapsulated yet. So this is one of the views that people think is how assembly happens. Under conditions of neutral pH and physiological salt, the protein itself is not in monomeric form. It's in this 34 CP bilayer disc. Actually, if you reduce the pH a little bit, this bilayer disc morphs into what they call a lock washer. At low pH, this lock washer will then self-assemble into helices of various lengths. But we are at neutral pH, so this is a stable form. This means that this form is a high energy structure. Now what the original assembly does, it pokes a little finger. It's a stem loop type thing, interestingly enough. It pokes a little finger into that hole and it uses a conformational change in the protein. And it's this conformational change in the protein that actually causes this bilayer disc to morph into this lock washer. What next happens is that either monomers attached to this lock washer or complete bilayer discs, which then also morph induced by the first one that has already morphed. So this is a luster. So actually turning this into this costs a lot of energy. Turning this one into that costs less energy because of the presence of the first one. And what happens is that the remaining part of the RNA strand is drawn in through the hole in the middle and finds itself lodged in the core of the protein, which has undergone this conformational change. The question now, of course, is why so difficult? And actually, I should say, is this not just TMV that does this? This whole bunch of viruses? Yeah. Papaya mosaic virus, clover yellow mosaic virus, papaya ringspot virus and tobacco. I forgot what the R means, a virus. So apparently this is the way to do it. Okay, so a teeny weeny bit of theory, not too much because this helps us to design the protein. Let us just look at a simple language adsorption. So you have template, this is our DNA now because our goal is to template DNA. And so if you have an attractive interaction between the code proteins and the template, then you'll get language adsorption, which is not very cooperative. It is really difficult to get the all of the binding sites covered by code protein. So you could say, well, these proteins might interact. That reduces the interaction energy further, because more negative. So that drives further assembly. So far so good, but still it's a one dimensional process and an entropy will make holes in this. Making holes in a capsid is not so good for the virus because nucleases might just think something to nibble at. That's the end of the virus. Right. So worse, if you allow the proteins also to interact, then they will also self assemble a solution. Oh, not good either. Right. So, so binding will give you on this is good interactions are good and interaction between proteins are bad as well because you get self assembling. Now let's look at this at this red. This is also well in physics land, everything is fair. This is a protein, a code protein, and it's not assembly active. So it's in this in this low energy state. Let us presume that a template now has an origin of assembly where these proteins would preferentially absorb onto and then change the conformational structure of their protein. Right. So more from red to green. It has to pay something for that. But now the protein can recruit other proteins which are of the wrong color, turn them into the right color and gain, whoops. Okay. And gain a free energy of binding, a free energy of interaction, lateral interactions, and then there is a little bit you have to invest to get the process started. But this is a nucleating process. Right. The statistical mechanics of this is really easy. It's called the zipper model. You have in essence three knobs that you can turn. The first knob is what we call the mass action variable. This mass action variable is a product of the concentration of free proteins, 5p in a solution times a binding constant. Or you could say, well, it's equal to the volume fraction, 5p divided by some critical concentration of proteins, like a critical aggregation concentration. The second thing which is important is the stoichiometry. So the stoichiometry tells you what the ratio is of the number of binding sites in your solution and the number of proteins. And stoichiometry turns out to be extremely important to the success of this process. And finally, because this is a nucleated process, it's a cooperativity plays an important role and it's captured by a parameter that we call sigma. And that's again a Boltzmann factor involving this free energy that you have to pay, H, any free energy epsilon of the binding because the very first protein that binds doesn't have a neighbor. So it actually in essence loses that. Okay, so this allows you to calculate the size distribution. So how many proteins are on average assembled onto the templates? So G is the binding energy of the protein to the template and epsilon is the binding energy of two code proteins that are in contact. And then H again is this free energy that you have to pay in order to morph the protein into its wrong shape. It's the stacking energy, yes. Yeah, it's a stacking energy. And then if you look at the stacks in the free solution, that might have a slightly different stacking free energy because the template will position the planar groups in such a way that you're not losing as much entropy. As you would in free solution, but never mind. So you can also write down your rate equations. I'm not going to show them. You know, you have an empty one, and then you have one, you have two, you have three, one can fall off. And in essence, you've got so forward rates, which depends on the number of proteins on the template. This is the backward rate, this is the forward rate. And these two are connected through microscopic reversibility, meaning that they're connected to these energy scales that we have in the problem. And then you stick this into Mathematica or you write a program and you can calculate the non-equilibrium distribution of the number of proteins absorbed onto a template, right? Let's first have a look at the equilibrium distribution because that tells us why it is advantageous to zipper. And the reason is as follows. So on the vertical axis, we've got the base 10 log of the probability of having n proteins on the template relative to having no proteins on the template as a function of the number of binding sites. And so these lines are what we find depending on the ratio of the concentration relative to the critical concentration, right? So this blue line gives you this ratio, we're just below the critical concentration, 0.98. This red line gives you the distribution exactly at the critical concentration 1.02, just above, 1 to almost, just above. So whether you're below or above, you always have an exponentially decaying function, but it depends on from what side of the assembly. So if you're below the critical concentration of your exponential decay makes assemblies even less stable than the monomers, I mean than the free templates which sit over here. Exactly at the critical concentration, we have got a flat liner. We've got a flat liner which is below the critical one, the empty one. So you still don't see templated assemblies. You have to go slightly above it and only slightly which means that this process is highly cooperative. You have to go a little bit above it. What we see is that we have complete ones, so an exponential distribution now starting at the complete ones and going down. And this means that intermediates are suppressed. To suppress intermediates is crucial. Why is it crucial? It's crucial because you do not want to waste protein on incomplete viruses. Right? So this is a way of having either no templates covered by protein of only ones that are complete, more or less. And this means also that if your stoichiometry is not right, which means that if you do not have a lot of protein, you're not wasting that protein or putting one or two proteins on a whole bunch of templates. Those few that you have, you put on one, that one survives. So it's a highly nucleated process. There's a lag time. I'll show if I'm not going over time, I might be showing you some graphs as well. And so first of all, the transition is really sharp. So there is a really critical concentration.  And finally, even if your protein concentration is just a little bit above the critical one, if your template is long enough, it will actually encapsulate it. So the cooperativity increases with the length of the template, which is why you can increase the length of your RNA of TMV and still have complete viruses. And in fact, there are zillions of patents out there where people put genes on the genome of TMV to produce in tobacco plants. Okay, so first of all, the slope of this line depends on the binding affinity. So the larger the binding affinity, the more steeper the slope. But at the same time, stoichiometry does the opposite way. So if you starve the solution of protein, then the slope goes down and down and down, which means at some point you're going to lose complete coverage. Actually, you will not get any coverage at all. Okay, dynamic. So let us do an experiment. And let's take a bit of DNA, which is considered 51 binding sites. The cooperativity has a value of 0.007, which is a typical level of cooperativity, that you also find in supramilocular polymers or in organic chemistry. It's ethrop... I'll show. So it's this Boltzmann factor involving this free energy barrier that you have. And then the absence of a bond, which also acts like a barrier. Yeah, not so. All right, la-de-da-de-da, whoops. Q is 51, segment 0.007. Sorry, possible, finding an one-dimensional object like that. Do you remember the bigger... Yes, yes. ...in what they mentioned, or the same distribution? Yes. And the level of size of the slope... Yes, I agree, but we're actually making use of the fact that you have to turn the protein into something that is able to absorb. So you're killing entropy through energy. I forgot to mention that, yes. Entropy bad, energy good. So we're killing... In essence, why you don't have a phase transition in 1DE if your interaction range is finite. Right? So by having this... Yes, yeah. It's a different type. I can talk about it after my talk, if you want, because it's a little bit technical. Actually, it's not that technical. Okay, so we start off with only empty templates. And that's here, this axis here. As a function of the log of time, I've scaled the log to this level of cooperativity, because the level of cooperativity dictates your lag time. So that's taken out now. Now, for two different stoichiometry. Stoichiometry is zero, means an infinite amount of protein out there. So you're never depleted of protein. And stoichiometry is 1.5, which means that you haven't got enough protein to cover all of the assemblies. Now, for short times, the probability of finding empty proteins, empty templates starts at one, and then it goes down, and down, and down, and down, it's a song. And it's only at the later stages that you start to see a difference between excess protein and not excess protein, so depletion of protein. It takes a while for the proteins to register that they're not enough of them around. And then if you have excess protein, in the end, you end up with almost no empty templates. If, however, the number of proteins is not sufficient to cover all of the templates, then at some point you start to level off, because some of them will remain empty. This is exactly what we want. Notice there is a little bit of an undershoot. If we look at, on the right-hand side, so in blue, that's the probability you have a complete capsid. So for the stoichiometry of zero, so an infinite amount of protein, it goes like that, and it's really sharp, and whoop, goes like that. However, if I don't have enough protein, I will still be able to make complete viruses. This is the cool thing about this. And it goes up. I overshoot a little bit, and then I go down again. Right? So we have undershoots and overshoots, and here these undershoots and overshoots, they only in the theory happen only if you do not have enough protein in the solution, never when you have an excess protein, never. And this will, I'll show you later if I have enough time, anyway. Well, you can also look at the average coverage, theta, as a function of time, for two different concentrations now. So the red curves are for an excess protein, and the blue ones are for not enough protein for two different concentrations. So the concentration is here, e to the power of three, that's about 20, I think, 20 times the critical one, and e is about three times the critical concentration. So you see that the larger the concentration, the quicker the assembly goes. And this is also anyway what you expect, because the thermodynamic driving force is larger. You can't see it, but there's a little bit of an overshoot over there. So again, if you don't have enough protein, you get an overshoot. It is very, very small here, but the largest overshoot we can get is about 100%, and they're very, very long lasted. So the relaxation time, it takes, I mean, it takes a very long time for this maximum to go away again. And I think this, you know, crawls back a little bit to what some people have called the pseudo law of mass action, because if you think that maximum that I can see and you cannot see, because I believe it's there, you know, it takes a very long time to even out. So if you take that as your fraction occupied size, and you calculate back your binding free energy, then the binding free energy that you get is wrong. Okay, so there are overshoots, undershoots, depending on the concentration, depending on the stored chemistry, overshoots in theta, and that's the thing that we're going to measure, we only see up here, so for large values of lambda, so in the conditions of excess protein. Okay, I'll come back to that hopefully in about five minutes time, if I have that time. Okay, let's go back to our TMV protein. So if TMV protein has functionalities, one functionality is one part of the protein has to wrap itself around the RNA and do the death now as it were, it undergoes a conformational change. There's a part of the protein that is involved in the interaction with neighboring co-proteins and then there is a part that imparts colloidal stability, we call this shielding colloidal stability to the particles, because you don't want them to drop out of solution. So this is our well, I mean if you make something in the lab it doesn't look as nice as when nature makes something, but it does a trick, so we have a binding sequence, it's basic, it's like 12 lysines, it's positively charged. There is a silk-like block which consists of a certain number and times a silk-like sequence going in adenine, blah, blah, blah, and we can vary the number of those sequences, one, so zero, one, two, all the way up to 18. And then we've got our shielding block which is a collagen-like block of a 400 amino acids which imparts this colloidal stability to the complexes, because you're neutralizing something that is charged, so you need your colloidal stability. Now it turns out that this silk-like block is able to form either a beta sheet or a beta barrel, depending on how big this N is and depending also on the concentration. If we for instance take four of those units, then this object will not undergo this conformational change into a beta sheet and we know this because we do circular dichrism. And what happens is here, so this is a two and a half kilobase pair of double-stranded DNA, this is an AFM image, and essentially what happens is you have, because this is positively charged, it will just adsorb like in the usual way, in the Langmuir way.  If you take 10 of those sequences or 12 or 14 or whatever, then what happens is that this is the same length of DNA. Now it's packaged into something that is a lot shorter than the original length, so somehow it's been packaged in this object. If you expose this to nucleases, it's protected. And so it shows that having this conformational change is kind of important in order to be able to protect the RNA, in this case the DNA. Okay, dynamics, because we're interested in dynamics, so this is what happens in, if you do an experiment, so you mix the artificial co-protein with the DNA, and every so many minutes you take a little bit of a volume, when you deposit it on the surface, and you do your AFM, you clean it, etc. And so this is a short time, somewhat longer time, somewhat longer times. So we're actually seeing growth from one end. It starts to grow at one end. Hang on, we didn't put in this origin of assembly. I'm good, thank you. We didn't put in the origin of assembly. So why does it start at one of the ends and not somewhere in the middle? Well, look at its tail. This is a big, it's a polymer. It's a coil, it's a random coil. Radiative generation, 50 nanometers. So you're absorbing, actually this goes back to a question that Robin was asking about an hour ago. You know, if you take a polymer, you try to put it onto the surface, this polymer senses the presence of the surface. So the surface in a way repels it because of entropy. So if it absorbs towards the end, you don't have that yet. So even though we did not build in this origin of assembly, by accident we did. But hey, who cares? Okay, now I'm almost done. So we did, as a function of time, measure the length of this growing virus-like particle. This is for the CS10B, so we have 10 of those sequences as a function of time. These bars are the experimental data, and in red is our theory. Now, I will not bore you with how we did the curve fitting, because we had many parameters. The curve fitting is extremely difficult, exactly because you have so many parameters. You have to just believe me that we didn't do anything wrong or fibbed or whatever. I'm not going to tell you that. It's less than what string theory is, I think. Anyway, the cool thing that you already see is this exponential distribution for late times. That's this exponential distribution, this inverted exponential distribution I showed you that came out of the equilibrium model. So for late times we were able to extract the energies that we need to put in. We did the same thing for a protein with 14 of those sequences, and again, so these are the experimental data, and these are our curve fits, which are not brilliant, but they seem to follow what we see in experiments. And I have to say, so these are AFM-based measurements, your statistics are not that good. So one of those bars here could represent maybe 10 viruses, excuse me, virus-like particles. So your statistics are not that good. Anyway, so we can, from our curve fit, we can extract the energies. So the binding energy to the surface, the protein-protein binding energy combined gives us minus 17 kT for both. The free energy barrier to assembly is 1, 6 kT, and 3 for the other one. And we can also back out to the forward rate, which is in minutes of four times the center power of nine. Now, what these numbers mean, well, we have to do more experiments to see changes in these parameters to be able to interpret what we are seeing. What we can do, of course, is compare with our curve fitting, what we did ages ago, to the experimental data on TMV. And interestingly enough, so the binding energies are very similar. So we get 17 kT for TMV, an energy barrier of 7 kT, and a forward rate, which is actually a factor of 10 smaller than for our artificial volume. Okay. I mentioned statistics, and this, I have a little, how much more time do I have? I have two more slides, this one, and then the next one, and then I'll shut up. So I will not show you some more new theory, unfortunately, even though that's the more interesting. No, no, no. Experiments are cool. So what we did is twice, okay, so how can we get better statistics? You know what, I mean, doing AFM is not going to help you. So what we did, we replaced the DNA by a proxy genome. And this proxy genome is optomechanically active, which means that if you stretch it, its luminescence changes, which is really cool. And this is what we find, right? So by looking at the luminescence, by looking at two peaks and taking the ratio, we call this the fraction of a torte structure. Torte is like, do you stretch on it? So this is at a concentration of 0.06 micromolars for stoichiometry. So F plus here means the number of positive charges relative to the total number of charges. So if F plus is a half, I have perfect stoichiometric matching of the total number of positive charges and negative charges. Let's look at 0.1. So I don't have enough positive charge. I don't have enough protein, so my system starts the protein, time equals zero, goes up, the signal goes up, and I goes up a little bit more. At exact stoichiometric ratio of 0.5, that's the green curve, it goes up, it sort of seems to level off a little bit, and whoop, it takes off. And the same is true if we have too much protein there. Now, this now we interpret as initially as a Langmuir-type adsorption. So the first thing that these objects do is they just indeed adsorb onto the DNA without doing anything to the DNA. You have to wait a little bit. This is the nucleation process. After a while, they start to think, hmm, maybe I want to package it. And that's when the zippering happens. So there is a nucleation process, and then zipping and packaging of the DNA. And so we did the same experiments at a concentration 10 times higher, and then where things started to happen. And this is also, I think, where our model protein starts to diverge from the actual TMV code protein. Because for instance, if we look at the blue curve here, that's at a stoichiometric ratio of 0.7, so we have 20% more charges positive charges in your solution that you actually need. You know, you've got your lag time, and then it goes up, starts to zip, and then, oh dear, I have an overshoot. And my first reaction was, I predicted this. Except this happens at excess protein, where my theory tells me, no, Paul, there's no overshoot. Okay, so how do I weasel myself out of this? Because I will. First of all, we know that if we only have protein in the solution at high enough concentrations, it will form assemblies. So there's parasitic self-assembly. And second of all, we know, and I don't have time to explain this, by using a different optomechanical probe that is able to measure to probe whether or not I have one template encapsulated, two or three or four. So we know that what happens here is also a co-assembly of templates. So instead of encapsulating just one proxy genome, and we're encapsulating many more. Which is also what happened with CCMV, remember? The early experiments of Adam Slotnik. Okay, with this, okay, I'm not going to show this. No, no, that's, no, no, no, no. We have a theory, blah, blah, blah. Does the theory explain the data? Yeah. Okay, so finally, so protein polymers can be designed to mimic co-proteins of linear viruses, but you're cool. Our model tri-block protein co-polymer successfully encapsulates DNA. And in fact, it transfects, which is kind of, I didn't mention that. I think that allosteria and directional assembly are crucial if you want to encapsulate a linear object. So you have to beat entropy. So the kinetic zipper model describes the time evolution of encapsulating DNA reasonably well. We predict over and undershooting the conditions of excess DNA, oh, I should have, excess DNA, not excess protein, and overshooting under conditions of excess protein may occur in competition when you have, for instance, micellization. That's what a model actually tells you that it didn't have time to explain. And with that, I would like to thank you for your attention. And is there any questions? Just let me know.


n't let it wait too long but we also don't have enough knowledge to know that'll probably be some time in october once you have the strategic plans at your priorities shortly thereafter we have a conversation with you about next year's budget the fy 25 budget on what how you see those priorities moving forward and what your thoughts are what parameters you might want to put on as far as budget limits or tax rates or whatever because then we will be starting in november with our own staff presentation to give you your budget in december so going to be a lot of uh thought processes coming up in october so just enjoy your september and when you're not looking at assessments on the board of civil authority still as we're talking about budgets are we still on track to have a fiscal 2023 i believe so for the next meeting which would be just one meeting later than normal thank you okay come check in with the boss back yes yep she's nodding uh and then one other thing i didn't want to add it to tonight because we just discovered this so late but um you may recall it right after the flood you issued me some authority to take actions and flood related and that actually expired on august 23rd so we might need to re-up that so um you won't probably don't need a million dollars now but it would be uh but there are still some FEMA related things it would only be those kind of things to move along so we might ask you to do a call-in meeting sometimes just to re-up that you're the only agenda on the item there's some more period of time at a lower level so great because we've been kind of operating as though we're still in place so i i missed that is that it that's it okay so we can adjourn at 9 22 p.m thank you all thank you


Ramila, at the heart of all the argument, the violence, the discourse, both rational and irrational that's been going on, is the notion of citizenship. Shall we talk about citizenship historically speaking? When did this notion come into being? The thing we forget is that citizenship actually is a very modern notion. He goes back to ancient Athens and Greece and whatnot, but let's not forget that citizenship in Athens was extremely limited. It was about a third of the people living in the city of Athens that were citizens. The rest were slaves and other categories of people who didn't have the freedom of citizenship. That's a different story altogether. That's an older society with its own background of tribal structures and this, that and the other. Modern citizenship is something that comes very specifically at historical moments. What is this historical moment at which it comes? In the 18th century, you find a change going on where various things happen. Industrialization begins, capitalism comes in as providing the economic base of this new form of society. The middle class emerges. The middle class is very aware of the fact that the older authorities, the feudal aristocracy, basing themselves on the labor of the subject people, were declining. The middle class was taking over. You get a historical change, very fundamental historical change, where the old relationship of the Lord and the subject, the King and the subject, gradually changes to a new relationship between the state and the citizen. Now both the state and the citizen are new because the state under this kind of system is very different from the monarchy of earlier times. The citizen is no longer a subject. The citizen has rights and that is the meaning of the rights of citizenship. The citizen has rights and duties and these rights and duties, obligations, whatever name you choose to give them, are in a sense what is spoken of in the Constitution. Before we get to the Constitution, suppose I introduce the Indian context here. Yes, there is a new contract between the institution of the state and the individual citizen and the citizenry as a collective. But in the case of India and many other countries, this is the result of a struggle for freedom from colonialism. So there is an additional edge there in the sense that a certain contract has been envisioned and that is what, when we say the idea of India, is that what we are talking about? Where the colonial system interferes, let's not forget that in a sense citizenship didn't really exist under colonialism. India was still the subject of the British rule, the British monarch. So that old idea of the king and the subject continued under colonialism in the case of colonialism in India. The real change really only comes when India ceases to be a colony and becomes a nation. Now my point is that that change in the relationship and the understanding of what is meant by the citizenship or by the citizen was not fully discussed, explored, understood in these years of independence. There's been a little hangover of the older subject idea. And the realization that the citizen is the independent thinking Indian who has views and has the right to express those views, that didn't quite establish itself. So I think that one of the problems that we have today is making those in authority, anyone who's in authority, understand that in the change to what we call the nation modernization, whatever it may be, the notion, if you're going to use the notion of the citizen, you have to understand that it's a changed relationship.  And then the setting of this particular contract, which is why I refer to the idea of India during the freedom struggle, is taking cognizance of the fact that this is and will always be an extremely diverse citizenry. Absolutely, absolutely, that is understood. And that is understood in the sense that citizenship where it has succeeded, and you know, this is a change that practically every part of the world has been undergoing in the last 200 years. In some parts actually seeing it now, I think, in contemporary Europe as well. And in some ways also questioning the basis of citizenship and so on. What is very, very clear is that if you are going to continue with democracy and the notion of citizenship, you have to understand that the definition of a citizen is not by indicating one item of identity. It has to be multiple. It has to take in every citizen no matter what their thinking may be and what their pattern of culture may be. It does have to include everybody. And that's something which I think is causing a lot of problems because there is an assumption that, you know, the whole question of majoritarianism comes in here. And you can't have democracy if you are going to base your citizenship on majoritarianism. Chief Justice has made special mention recently of the duties of the citizen and of course immediately one wants to ask about the relationship between the rights of citizens and their duties. So if you were to sum up some of these rights on the one hand and some of the duties, what would you? I would say to treat it from the basics going up, the rights would be the rights to at least a minimum reasonable livelihood, which means food, water, housing, etc., employment, education, healthcare, and very important, once you've got these, the freedom to say what you wish to say, free speech, free thinking, and social justice. That is the only way in which you can then begin to talk about the possible equality of every citizen. Unless every citizen has these rights, equality is not possible.


Definitely knowledge supersedes marks. But do you think that marks do not have any importance in our individual's life? No, definitely marks are important. Why? Because see, when you go for an interview, the interviewer doesn't know you have knowledge. In 5 minutes no one can assist your full potential. But then the marks reflected in your CV is going to show what kind of person you are.  They are not going to check how much knowledgeable you are. They are going to see if you have the potential to learn. Are you eager to learn? See, you always need to keep your mental elbow on. When we drive and when we are learning to drive, there is this elbow sign. Now what that L shows that the learner, the driver sitting inside the car is learning how to drive. So you as students at this point need to keep your mental elbow on. Obviously, whenever you are reading something new, just try to look at it from a different perspective. Marks do play a very important role and you need to do smart work. That is very important. See, hard work is good. Smart work is better. Why? Many people think smart work is shortcut. No. Smart work is smartly designed hard work. You have to do hard work like this to get the right outcome. I have seen a lot of students who fail in the net again and again. They come to me and they say, ma'am, we have so many marks. Tell me where you made a mistake. This is a very difficult situation for a teacher. Your marks, you told me. You did not tell me what the answer to which question you answered. Where did you make mistakes? What topics did you leave? We are teachers. We are not experts of hypnosis. We cannot hypnotize you and we cannot tell you where you made mistakes. Students make mistakes here that when they are given their answer sheets, they never analyze their mistakes. You are doing the same and getting the same results. If you keep doing the same, you will get the same results. If you want to change the result, you need to change your strategy. It is as simple as that.  Check where the mistakes are. You need to play smart. If you are writing about a writer or suppose there is a text, Virginia Moles to the Lighthouse. You are drafting an answer from a 20 mark. The problem comes here to the students that the guide books market is very easily available. We all are aware about that. I have seen a lot of friends of mine who read the guide books and don't know the numbers. Then they say, we have learned so much. Nothing happened in this quantity. Look, quantity is never important. Quality is important. What do you have to write in 6-7 pages? If you quote some lines of Virginia Moles, Virginia Moles has more plays and novels. If you are an Afra Benka or if you are talking about Virginia Moles' work, then Mrs. Dalloway would be there. When you quote lines from them, that showcases that you have read beyond the text. And that is how the examiner is going to see that you stand apart from all the other students. Everyone is writing from guides. Even you will get the same marks that everyone is getting. So, play smart. That is important. Make your notes. There is so much material available on the internet and very good material. I am telling you, in guide books, you get such a weird material. Sometimes, you know, my friends used to tell me that is guide. When I used to read those articles, I used to feel, oh my God. They are insulting this writer. What is he writing? Read good scholarly articles on the internet. You just need to copy few good lines, paste it, make a word document. That is important. On my YouTube channel, I have posted a few videos on how to secure good marks on your bachelors and masters. So, you can refer to those videos also because a lot of students constantly used to ask me how to read and what to do. And I have been a topper in my graduation as well as in my post-graduation. So, I had made a strategy. I used to write answers and I used to always get good marks. Where everyone is getting 40-50, I used to score 70. It was not like that. I was working like 24-7. I was doing a lot more. But I was using smart strategy. We all have read the story of the crores in our childhood. What is that? That was the first story I think we would talk. And that story teaches our smart work. When you take up a career and you start working day in and day out, you can please have a seat. So, you start working day in and day out. There would be times when writers don't block. Writer loves to write. But there are phases in his life when he doesn't feel like picking up the pen. My favorite writer is Amit Rupati. And he was writing a book on Ravan, the third series in the Ramchandra series, the third book in the series. So, I was following him on twitter and I saw that he said I cannot write for 6 months. I am like I wake up in the morning and words flow. But for this 6 months, I was not able to write at all. So, this is going to come in everyone's life. But the case is that when you do something that you like, when a student asks me that I am Dryden said this about Plateau. But Plateau said that this is confusing. And when I research more on Dryden, I try to understand what he was actually talking about in his essay. I feel good that wow, I have discovered something new. This same thing helps me in breaking up. Because life as teachers also. You can ask your fellow teachers, you can ask your seniors that teachers also get to learn something new everyday. And when they discover new things, that is how they break the monotone. And every time you teach someone and that student appreciates you and says that ma'am, I fell in love with this character I fell in love with King Lear. I still remember Ranjit ma'am was teaching us Ranjh Feel Park. And we are having so much fun in the class. I mean, I fell in love with Fanny. I mean, I was a girl who never liked female characters. I was like, what is this? She keeps crying. Look at all the female characters of Victorian age. There are very few who have been shown that they have given up their stand. I was always like, they cry and then say that they have done this. So, you are standing for yourself. But then when you look at characters like Fanny Price and when you have teachers like Ritu Maan who teaches you, Ritu Maan had taught King Lear. Oh my God!  I mean, he is just trying to raise his own good daughter. But then when Ritu Maan was teaching us how to understand that character, how he is standing in the rain, in the storm, in the school, what is his position and how he is making his mistakes.  So, when you discover and read a new text every time, your good insight makes your day. You feel so good. When students call me and say Ma'am, you have studied literary theory, you have had a lot of fun, and you have come to understand as if you have not come yet, that simple message from a student, that is the best gift a teacher can have. So, I think that can answer because a CA or an engineer, when he creates something new, he is an architect. When he makes his building, he sees it, he sees creation. He feels like, wow, man, I have been something really good. That same thing happens with a teacher and that happens with every profession. Yeah. Any more questions? You can sit down please. So, what happened was just the way you said, my father and my grandfather, they both are like book lovers. So, my childhood memories are that my father used to come from the office and used to have that whole collection of stories and used to read that. And I was like very small, fourth standard or third standard. So, I used to try to read it, but it was all Greek to me. Okay, what is it? I don't understand anything because it was an abridged version. Now, what happened was that gradually, I met some fantastic English teachers who inspired me. I still remember when I changed my school, the first class was of English teacher and she was teaching us daffodils. And I don't know, I started crying after reading that poetry that my God, if this man would have been alive, I would have married this man. Such beauty, my God, he's talking about a daffodil site in such a beautiful manner. Just like Sir said that, you know, heard memories are sweet, unheard are sweeter. All these lines from literature, happy families are all alike, unhappy families are unhappy in its own way, they are tall story. When you read these lines, you're like, oh my God, in these five, six words, they put so much wisdom. So, this happened. Then in the tenth, I became the president of Literary Club. So, I got the opportunity to conduct a few group discussions and book readings and all that stuff. Then in the eleventh, I became the captain of my house. Even after becoming my captain, my whole theme board was made up of all the literature. All the writers were going on.  This was the situation. In the twelfth, I became the head girl. Then I took part in a lot of debates. When I took part in debates, I used to research in any debate map. You have to quote these good writers so that your debate looks impressive. Then when I was reading these lines, child is the father of man, oh my God, I'm like, what, what are you talking about? So, all these things connected. Then finally, I decided that I am not interested in commerce. Let me choose English. That time, I was not sure. The dilemma that you are going through, even I was going through. Then what happened was that I watched a speech of Steve Jobs. He gave a very good speech on the graduation day at Stanford University. He was freely available on YouTube. In that speech, Steve Jobs said that when I watch my life in flashback, I can connect to all the dots. And I can know that a failure in every life, every good thing, has made me to this position that I am in today. We all know about Steve Jobs. He was snatched from them. Then again, he developed another company which purchased the apple. Okay, so if you look at this journey, you will find out that connecting the dots is important. You also see in your life that all the things that you have with you are still bad. Have they turned you towards English? That is very important. That all the things that you are getting in this way, that you are getting closer to English. That you are in love with the writers. That is how, see trial and error. Why are we having so many dating apps these days? Tinder and everything.  Then from the one who is coming, you take your relationship forward. Career is like that. That you explore different career options and then see where you are having fun, what work do you feel like this boy who is playing violin. You can see that he is passionate about it. That is why he came out with such a fantastic piece. Give me a violin today. I will tell you what I will do with this violin. Something will happen. I will tell you what will happen. Because I don't have that love for music. But then give me a good book. I can read it and I can analyze it in a very different way and I can put forward in front of you in a way that you start loving that book. That is the job of teacher. If you think that you explain a book to someone and that person is into that book totally. Like in the season of Game of Thrones, you saw an episode and you can't stop. So I think that is the right choice.


Yannis, one of five, four color patch auto. Dude, that's gross. Big one for Patrick Z. Dude. This is a 1920 flawless card. That is awesome. No way. Yannis, baby, one of five. No way. We got Yannis in it. It has been a long time. That's awesome.


The wait is finally over as tonight the puck dropped on the beavers men's hockey season and what a better way to do it than with one of your biggest rivals as North Dakota came to town. A packed house to see the beavers and they did not disappoint. In the first period the beavers on the power play. Brendan Harris hits Adam Brady. He snipes one in from the slot. The beavers take a one to nothing lead but UND dominated the second period. Jordan Kawaguchi speeds ahead for a breakaway chance. Hank Johnson makes the save but tries to cover but he doesn't have it. Gabe Bass ends up tipping in the rebound. They tie it up at one but in the third period BSU working in the corner. J. Dickman finds Owen Sillinger wide open for the one-timer. He beats the goalie. That would be your game winner. BSU gets their fifth straight season opening win. If you enjoyed this segment of Lakeland news please consider making a tax deductible contribution to Lakeland PBS.