Picture this, amusement insiders, you pull up to Canada's Wonderland for the spring opening weekend. And looming over Splashworks is an absolutely massive RMC, standing in the place of the mighty Canadian minibuster. How do you feel about this? Let's talk about it. So in this video, I want to make a case for RMC minibuster. I know it's probably never going to happen, but let's just ignore that. Let's just put that aside for a second and talk about it because your girl likes to dream. For those of you who don't know me, my name is Jasmine. I used to go as Jasmine on YouTube and my channel these days is called Jasmine Dreams. So you'll notice in my segment here on Amusement Insiders, there's going to be a lot of dreams, opinions, ideas. We're here to just talk. We're here to put our brains off in space and think about what could be, what should be, what we want to be, and then talk about it. So we're going to have some awesome discussions about various topics around Canada's Wonderland as well as other theme parks. But I have some really hot topics around Wonderland and a couple other Canadian parks that I want to start off with. And today, if you guys have seen me in the live streams and the podcast, you will know I have very strong opinions about this particular topic. Okay? Minibuster is a Canadian treasure. All right. Love it or hate it in its current state. That isn't a Canadian icon right there. And I just feel it would be completely wrong for Canada's Wonderland to exist without some iteration of Minibuster. Now, pardon me while I scratch my eye. I also feel it is just wrong to have Minibuster and not have it be a wooden coaster. That is just the legacy of Minibuster, okay? Now, do I think that what I want is what Wonderland and Cedar Fair are going to do? Not really, but we're going to talk about it anyways because we're here to dream with Jasmine Dreams. I don't know what we're going to call this segment. Feel free to throw some suggestions down below. It may be a week or two before your suggestions come into play, though, because I've recorded a few of these in advance. So it is what it is. All right, let's get into it. Minibuster. Here we go. Now, I am not going to get deep into Minibuster's history in this video. For those of you who have been following Amusement Insiders long enough, you'll know that I used to do write history videos and I have done a whole video on Minibuster's history in the past. So maybe Brennan can put a card here somewhere where you guys can click or like a link in the description or something. But go watch that video if you want the full history of Minibuster. Because when you know the full history of Minibuster, I think you'll have a greater understanding for why I don't want it to go away, okay? There's only five coasters that opened with this park and Minibuster is the most iconic of them, if we're being honest. I have a special place in my heart for all of them. But we can't get rid of Minibuster, okay? We can't do that. It's illegal. I just made it illegal. Just now. Anyways, with that said, Minibuster did open May 23rd, 1981, with Canada's Wonderland on its grand opening day. So I don't know how many times I need to say this in one video. I've probably already crossed that limit, but this is an iconic ride. This is a historic ride for this park. But I will be the first to admit this ride is not in the best shape. She's seen better days, you guys. And on top of that, Splashworks really honestly deserves more than what it has been for a long time now. And there's a lot of potential in Splashworks. So there is a general redesign of that whole part of the park necessary as it is. And I think this is just an opportunity for something big, like a ground up RMC. Yes, I am not talking about a conversion here. I think Minibuster deserves a ground up. So it would have to be a new iteration of Minibuster. It obviously would not be the original. Maybe they can use a few planks of wood and be like, the original Minibuster is in there somewhere. But I just don't think it's worth it or cost wise, smart, or given the condition of Minibuster, safety wise, smart to make it a conversion. But I think a ground up RMC would be wild and to follow a similar layout to Minibuster to follow a similar pathway, have it somewhat interact with Splash. All of that could make for a really sweet ride. You guys, come on. I know I can't be the only one thinking this. I know somebody in the comments is going to be on my side here. Okay, ground up RMC Minibuster. It's a solid idea. That's pretty much the video. Not going to lie. That's pretty much the video. Okay, lies. It's not the whole video. Here are some of my more detailed and nuanced aspects to this thought and my justifications. So Splashworks deserves its own entrance. Splashworks, if Cedar Fair was smart, would have a separate ticket price. Do I want this personally? No, I love being able to wander in and just go into Splashworks as well if I feel like it. However, business wise, it's a smart idea.  That said, if Splashworks has its own gate on that side of the park where Splashworks is, and it's now separately ticketed, imagine what kind of incentive it would be.  I want to go on the big roller coaster too. Like, sure, you're going to pull up and you're going to see all the other rides in the, in the, what's the word I'm looking for, in the park skyline. You're going to pull up and you are going to see all those other rides in the park skyline. But a lot of those you've been seeing for years now, a lot of those you've experienced, whatever it is that's going to pull people out of just being a Splashpark's only ticket purchase and into being a two-parks ticket purchase has got to be big. It's got to be worth the money and it's got to be something very different from everything else that you already have written that may make you say, oh, we're just going to get a day ticket one day this year and then we'll get a Splash ticket another day this year. We'll only come twice because then we can, you know, we've done everything. If they put in this coaster that's worth riding because I've been on, I've been on an RMC now. I can actually speak from experience now. I was still advocating for this like a year ago, but now I can advocate for it with experience. You cannot just ride an RMC one time and fully get the experience. Like, you're not even fully in it. You're like, whoa, what's going on for like a big chunk of the ride? So if you want a really re-rideable ride, a really exciting ride, a really thrilling ride that's going to make people feel like I have to spend this money to experience this and then when they go on it one time, they're going to come up and say, I have to go on that again. I only remember the first half because I kind of like blacked out. This is the ride. This is the one. This is this is like a big draw and there's not that many of them out there. Like it will get people to cross the border. And I want to talk about why ground up and how it could still pay homage to Mindbuster. So ground up. I mean, some of you guys have talked to me about this again in the chat during the podcast. It's a great time to argue with me about things. Why a conversion would not be a great idea for Mindbuster? There are many reasons and I would love for those of you who have more information on rides and engineering than I do to sound off on that in the comments. But my reasoning is because it would give them a lot more freedom to play with the layout because currently to try to retrac or redesign Mindbuster in any way along the current layout. There's a lot of challenges. It goes straight through the middle of Splashworks and that may not be what's best for Splashworks in the long run. But we could still have a ride that interacts with Splashworks. Maybe goes around the perimeter of it kind of plays over the entrance to it that goes in between it and the park. There's a lot of options that are opened up when you look at doing a ground up rather than a conversion. But I still think having a ride called Mindbuster, whether it's another iteration of the mighty Canadian Mindbuster or whether it's like Mindbuster's Revenge or some other take on Mindbuster. It's like the ghost of Mindbuster, something. It's a perfect fit for a final addition to Frontier Canada. You cannot deny that. Frontier Canada, like the one thing that would truly cap off Frontier Canada would be a badass version of Mindbuster, like a really rockin' version of Mindbuster. Two, my second justification for a ground up RMC on this case is the wood element. RMCs have come a long way, all right? I know they have a reputation for a lot of things, but they've come a long way and they're these days significantly more reliable than they were in their earlier days. And I think that's going to improve with everyone that we see. And so you have something that is a lot more impressive than the current iteration of Mindbuster that we have. And on top of that, you have something that is able to eat up potentially a lot more people than the current iteration of Mindbuster that we have. Okay, I'm just going to skip to my main thing, which is in a park where you only have a couple of wooden coasters and they're both to the point where it's like not great, like they kind of got to go. And your last major thrilling roller coaster additions have all been kind of same-same in like a few ways. Like they're all BNMs. They all have a very similar feel. There's a lot of people you could, like GP, you could go in and just do like a, you know, a TikTok interview and be like, what's the difference between Behemoth and Leviathan? And they'd be like, one is blue. So you need something that's not, that doesn't feel the same as everything else we've been getting. And you could argue that any steel coaster is not going to have that effect. It's just going to be another steel coaster. It could look a lot different. It could be, you know, inverted or flying style instead of more traditional style, but it's still going to give like another steel coaster. You know what I mean? Wooden coasters are just a very different species of coaster. They're a totally different type of experience and they can be amazing when done well. And even old historic wooden coasters can be amazing when maintained well. I just think ours are past that point. And so we have to do something kind of modern with them that fits Wonderland. And I don't know if going with all wood for both of the two big ones, big ones. I say that like they're big. We're talking about mine buster and wild beast here. Come on. But my, what I'm trying to get at is one of them I think we should keep as a traditional wood coaster and the other one should be a hybrid because a hybrid I just think is a really good fit for Wonderland's crowd. People, if they saw an RMC especially, especially all this talk we've been having about the specific type of crowd that Wonderland gets, how we can't do the chaperone policy because there's so many of that like thrill-seeking age group of teens and young 20-somethings in the park. Those are the people that would want an RMC. Okay, those are the people.  They just, do let it just eat them up. That said, do I think RMC mine buster is what we have coming? Absolutely not. Absolutely not. Me and Wonderland are on two very different pages. We might be in different books altogether but I think if they did redo mine buster and let me just go into, let me just, sorry, let's back up a second. I did not script this video. I used to script my videos, you guys. I didn't script this one. So you're getting unscripted Jasmine right now. This is gonna be a wild series. I don't know where this is going. I wanna talk about why it would be blasphemy for them to get rid of mine buster when we now have Frontier Canada, okay? We finally got Frontier Canada, okay? Mine buster was the only part of the original plan for Frontier Canada that we did get, okay? If they get rid of the only part of the original Frontier Canada when we finally get the full Frontier Canada, I'm gonna lose it, okay?  I'm going to fly from Florida to Vaughn to Mississauga, I guess, and drive to Vaughn and I'm going to pick it until they listen to me because like I said, I did the ride history videos thing. I did the park history thing. Please go back and watch them because I actually spend a significant amount of time on them but it would just be such a mess for them to finally do Frontier Canada justice and then remove the only original part of it that we ever did get before it came to be. The only part of it that even survived the Paramount era. We cannot lose mine buster guys, okay? But we can give it a new version, a new life. We can modernize it. We can give it a backstory. I'm fighting for mine buster, okay? And yes, there is a dragon fire video, okay? If you know me, you're not even a little bit surprised about that, okay?  I'll come for you. Okay, this video is a disaster but closing chapter is what I think Wonderland is actually going to do. One, just keep retracting mine buster until the end of time and like replacing bits and pieces as needed which would make me so sad. You guys went to all this trouble to make Frontier Canada the amazingly themed part of the park that it is, the icon that we always deserved. And then you don't do justice, fix up, re-theme, fully redo the one original part of Frontier Canada that we did get that deserves the second life that she has put in all of these years to earn. All these years, she deserves better. Justice for mine buster. Okay, okay. Now, I don't think Wonderland is going to do what I think they should do but I think they might give us something more like Seven Dwarfs' Mine Train at Disney World which would be really cool but then we'd be down to only one proper wooden coaster like adult-sized wooden coaster at the park and it's a really bad one. So I don't think that's the right solution either and I know that all of the markers that we're seeing for the 2024 and or 2025 edition are all in the Whitewater area so it makes a lot more sense that we're probably going to get that mine themed ride in the form of like a big long... I'm trying to avoid swearing in this video guys. I'm really struggling. I swear a lot. Anyways, I think we're more likely to get a really long and themed log flume ride or I don't know, a water coaster would be really cool. I would be fine with that but regardless of what we get over in that area I just think it would be wrong to do what they've done with Frontier Canada. The amazing work that they've done with Frontier Canada and not do justice to Mindbuster. So what do I think is going to happen more likely? Is they're just going to let Mindbuster limp along for several more years and they're going to give us something else in the Whitewater area. So it's probably going to be like I said, a log flume type ride or I think a coaster like I said that's like Seven Dwarves Mine Train style which I believe that's a Vakoma, right? Yeah, it's a Vakoma. I should have these things memorized but my brain is mush, okay? I'm a millennial. Have pity. Yeah, I think that's more likely something like that or something like what Brendan's been talking about like a mock spinner or something, you know, a little bit more cookie cutter. I really want a super custom ground-up hybrid RMC woody type of Mindbuster. And that's my rant for today. So I hope you enjoy. Please share your comments below on what you think about this, what you think about Frontier Canada, what should be Frontier Canada's final addition, what do we deserve and what do you think Wonderland is actually going to do and what other topics do you want to hear me dream and speculate about in future videos and get you guys all mad and riled up in the comments? Let me know. See you soon. Bye. Also, I just realized I made it through that whole rant video, whatever you want to call this and didn't give any of you context as to who I like really am. So for those of you who haven't been around Amusement Insiders long enough to see my old series and to chat with me back when I used to be on Discord a lot more, I will be on Discord more now that I'm doing this series. I've just been having some social anxiety lately, all right? So be nice and I'll come back. But for those of you who don't know me or remember me, like I said at the beginning, I used to go by, you're interrupting again. Sorry, that was my husband coming home and interrupting my recording. He's going to interrupt again. But for those who don't know slash remember, I used to make ride history videos here on Amusement Insiders and I have my own channel. As I mentioned, Jasmine Dreams is what it is these days. And on that channel, I post a lot of theme park vlogs and other theme park and travel related content mostly captured with vintage and weird camera lenses. So I've been friends with Brendan since 2018 and yeah, started out my my vlogs and stuff at Canada's Wonderland and then have kind of like moved a number of times since then. And these days I'm in Florida. So I do mostly theme park content that's based here in Florida. So largely Disney and Universal to come. I actually went to school for tourism and travel. So I studied theme parks, amusement parks, tourism destinations, like Brendan. I spent a ridiculous amount of time listening to investor calls and doing boring stuff like that for funsies, doing research for funsies, basically millennial things, that's all. Okay, so I hope you enjoyed this video and I will see you next time for another rant to get you all angry in the comments. Okay, bye.


This is Witchspace news for Friday the 3rd of May 2019 I'm CMDR Buur In this weeks news the first interstellar initiative kicks off Distant Worlds 2 comes to an end and distant cake is a thing If you enjoy our content then please remember to hit like and subscribe and click the little bell icon to get a notification next time we upload a new video You can also find us on Twitter, Discord and Instagram and if you want to help support the work of this channel you'll also find us on Patreon Links to all those avenues are in the description below Frontier launched the first of their community goal the next generation interstellar initiatives this week with a forum poll as it turns out a very hard to find forum poll In case you're unaware the plan is that a company the in-game Zenday partners is going to establish a mega ship transport service that will travel to and from the bubble every week to a guardian system outside the bubble that has been chosen by the community The poll for this system, one of five choices started on Monday this week via a forum post on the official forums There is also a post on the in-game Galnet news service that you can access from your ship It mentions that the poll is happening but doesn't tell you where it is The interstellar initiative is also mentioned on two posts on the web version of Galnet and on the game launcher Neither of those stories mention the forum poll and one of the stories just mentions interstellar initiatives as a thing If you click on the story to read it, it opens to a blank page with no text within it whatsoever If you do manage to find the poll then there is some information about the target systems but not only is that in another post and not in the post that contains the poll it also just gives very general information about the systems and is not really anything that might help you make an actual informed decision Fortunately player organisations like the AXI and Canon have stepped into the fray and two prime candidate systems have floated to the top with the other three being quickly disregarded I've included information in the description below taken from the AXI Discord server that details the two prime candidate systems Suffice to say that after the fanfare of the excellent launch video that accompanied the announcement of the interstellar initiative program the actual launch of the initiative itself has been met with a collective sigh from the community The main problems being, it's not in the game, it's very hard to find When you do find it, it doesn't really give any guidance on what you're voting for or why The vote ends on the 6th of May You can find a link to the vote itself in the description below This week the Distant Worlds 2 expedition reached its conclusion at Beagle Point The star widely regarded as being the furthest out easily accessible star on the opposite side of the galaxy to Seoul The expedition left the Polini system on the 13th of January this year with well in excess of 10,000 commander names on its register So huge was the participation from the community that in those early weeks the Sunday night's release of the next waypoint the expedition was hitting was often met with server outages as thousands of commanders all jumped all at the same time, all from the same place, all going to the same place and the frontier servers creaked and groaned under the volume of data being flung about Later though as the expedition spread out things calmed down and they completed a couple of nice community goals that saw the completion of Explorer's Anchorage Give shock at the center of the galaxy Now that the expedition has ended most of the remaining commanders will be winding their way back home via whatever suits them best Most of them More on that in a moment The expedition has seen a massive amount of video and screenshots generated of the things people have found along the way I was about to say I'll miss that element of it but to be honest that has always been a huge part of the elite dangerous experience and I can't see that changing now It'll be nice to see the population of the bubble increase again What's left of the bubble anyway At least the direction of home is now nicely lit for returning commanders with a host of burning stations And finally, as I mentioned previously not all the commanders on the distant Worlds 2 expedition are turning around and heading home just yet It seems that space madness might have infected at least four souls Serial circumnavigator and generally difficult German burpit regular Commander Primetime Casual who you might remember from such efforts as the circumnavigation of Mieter and Hollow which he did on his own and the Quantum World circumnavigation which he did with Commander Greytest has now embarked on what is possibly his most ambitious and difficult challenge to date Primetime Casual has banded together with fellow burpit regulars Commander Sabic Commander Lord Tyvin and Commander Soundo to embark on what the team is calling Operation Distant Cake Setting off as a team this coming Sunday May the 5th the Mad Quartet plans to drive around the equator of the furthest landable moon from Sol in the system known as Sematus Beacon or Body B9F in Ovesi SGY D0 to give it its less than snappy in game name The journey will take a number of stages to complete and after a group drive on Sunday the team will complete the rest of their individual drives at their own pace In total they'll be travelling 4239km in an SRV for some reason each leaving behind a perfectly serviceable spaceship in order to complete the ordeal They did ask me if I wanted to go No, no I don't We wish you Godspeed, gentle goats and rest assured that should you survive there will be a steaming pot of tea some battenberg and a fresh set of tyres waiting for you back at the bubble That's it for now Thank you so much for making it this far We'll be back later this week with more videos Until then, 07 Commanders Follow the Greens on the way out and do keep clear of the toast rack We very much look forward to seeing you next time


All right, I know it's officially after 12 p.m. So the legal tampering period is here But in the move that the Cincinnati Bengals made days ago was placing the franchise tag on wide receiver T Higgins and when you place the franchise tag on a player, then they can start tampering Wow back but with T Higgins With him being franchise tagged this this could go a couple of different ways for the Cincinnati Bengals T Higgins is nice T Higgins is a number one wide receiver T Higgins is a ball I know he dealt with some injuries Recently and I know Jamar Chase he dealt with some injuries recently too but T Higgins is not both of them obviously nice both of them could play some football and With the Bengals, I always felt I think most people always felt like when it came to paying both T Higgins and Jamar Chase Not that it's impossible. It is possible in this ways to work around a salary cap and whatnot But figured a lot of us felt like it was highly unlikely that both of them end up getting their bread from Cincinnati Again, it's still possible. It can still happen, but I just I don't think it's likely and We all kind of all got the vibe that the priority would be on them paying Jamar Chase instead of T Higgins Jamar Chase was I think number four overall picking and just because he was drafted that high doesn't mean that he Would automatically be the priority, but it just seems like He would be the priority in Cincinnati But with T Higgins getting franchise tag like I said, this could go a couple of different ways Him being franchise tag does not mean it's over him, but him requesting a trade. It's like, oh, he requested a trade He was old from Cincinnati. Oh, but that also does not mean that it's over We've seen this happen in the past with so many different players They get franchise tag players don't like being franchise tag They want long-term deals and if they can't get a long-term deal from that current team, then it's a business move That's it. It's just a business move This is letting the bingles know like hey, let's get a long-term deal done If y'all don't want to get a long-term deal done with me Let me go somewhere where somebody will get a long-term deal done with me because I want my bread Staying in the AFC North. We saw this just last year with Lamar Jackson The Baltimore Ravens plays the franchise tag on Lamar Jackson and Lamar Jackson put out that series of tweets that thread like a Request to be traded like 20-something days ago and they haven't on my request I'm requesting a trade again, and it was like, oh my goodness not Lamar Jack So we saw how that worked out a deal was done now with T. Higgins again It's a bit different because of the Baltimore Ravens. There wasn't no other quarterback for them to turn to With the Cincinnati Bengals again I think the priority is Jamar chase as far as deals getting done now You can't rule out because Bengals got some money to play with you can't rule out Him playing on a tag because if the Bengals feel like you know what we don't want to trade him Then he could end up playing on the franchise tag this year and then next year. He could end up being a free agent If the Bengals chose not to tag him two years in a row And then he can go on elsewhere after that and at this point I'm not sure about any other receivers who will be entering free agency at that time, but He would be one of the top free agents available next year if he plays on a franchise tag and they let him walk next year So that's something to think about But you know Bengals like they they don't want to go the comp pick route for T. Higgins And what I mean when I say that is if he were to walk in free agency Bengals would get a compensatory pick the following year You know, they don't want to do that. They don't want to do that for sure So that's what the franchise tag is all about man. It's just a so ugly side of business for the players It does give the franchises the teams a whole lot of control that I know the players just get so frustrated with I'm sure T. Higgins is super frustrated right now with the Cincinnati Bengals because he wants his bread Like any of us will want our bread like think about that put ourselves in in player shoes if we had spent four years with a company and We would do for a big raise and we knew that there were a lot of other companies that would hire us like that And they'll give us a big raise like that But the company that we work for they said, oh, no, oh, you think you're getting ready to go to one of those other companies for free Without us getting anything. I know you ain't going nowhere now You will get this year you will get a significant raise from us But it'll only be for this year and that's it. It won't be for the following And I know when we get raises at our regular jobs It is it goes into effect right then and there so saying that to for us with regular jobs Like, okay, cool. That doesn't sound so bad But in the NFL, we know it's a lot different because it's only for one year and one year only then after that Who knows what it'll be? So we'll see what happens with T Higgins Like I always say subscribe to the channel turn your notifications on because today it's starting off a bit slow But once it picks up y'all already know how it goes with NFL free agency Leave a like on the video because it helps out the channel a lot and it'll allow you to keep on getting recommended to this channel If you like it, of course, I love y'all team keep it clean I appreciate y'all and like T Higgins requested to the Bengals to be we out


You often hear the terms El Niño and La Niña when climate discussions come up. As a skier, you especially appreciate the extra snow that La Niña brings to the northern U.S. La Niña means the little girl in Spanish. La Niña episodes represent periods of below-average sea surface temperatures across the east-central equatorial Pacific. The impacts on worldwide climate of La Niña tend to be opposite of El Niño impacts. During a La Niña year, winter temperatures are usually warmer than normal in the southeast and cooler than normal in the northwest. El Niño and La Niña are opposite phases of what is known as the El Niño Southern Oscillation or the ENSO cycle. La Niña is the cold phase of this cycle. These usually last about 9 to 12 months but could last years. La Niña occurs less frequently than El Niño. So instead of putting all your faith in a snow dance, you just need to know a little bit about El Niño and La Niña and their effects on temperature and precipitation. From the University of Wyoming Extension, I'm Tina Russell, Exploring the Nature of Wyoming.


Hi, welcome to another CTN member highlight. I'm Leslie McVane, and my guest today is Patty Deloise from the Portland Public Library. Hi, Patty. Hi. Well, we're going to be talking about the Civil Rights Film Festival that's going on now for the month of January. And it's an appropriate time because of Martin Luther King Day coming up. Tell me how you got these fabulous films. Well, we've been collecting films from California Newsreel, who produces them. We've been collecting them for several years. And we just thought that they don't circulate enough and people don't know about them. And so we wanted to give the public a chance to see them. And as you say, this is an appropriate time. And to have them at the library where a number of people can come and have discussion after. Does that happen after these films? A lot of people don't want to stay and have a discussion. But we hope that people are stimulated to talk about them with each other, with their friends, to tell other people what they've seen. And if they do want to stay, there's somebody there to facilitate discussion. Yes. I know I've been to the film festivals in the past that you've held. You do these month-long series. And they're wonderful. They're absolutely wonderful. And to have a place where you can watch it on a big screen with other people, it does stimulate thought. It does. And I think when you see films like this in a community setting with other members of the community, as I say, it stimulates discussion. And I hope that it encourages people to be in touch with things that go on in the community, with things that go on in the larger world, and also to connect with the history that's being documented in the film. Yes. And right now with all that's happening in the world, in our country, in France, everywhere, civil justice and civil rights, racial issues, it's really timely that you're showing these films so that people can have some perspective on the history of, in this case, race relations and civil rights in this country. Yes. And I think the films do give a sense of continuity and a sense of history and a sense that this isn't something new that we're just starting these struggles. This has been going on for a long time. Ongoing, yes. And these films, they're an excellent series, and they document how hard the struggle has been and what the sacrifices have been, and really highlight the fact that ordinary people can do extraordinary things. And really make a difference. And these films, at least some of them, are award-winning films that have been recognized for their content. Yes, some of them have won awards at film festivals. The film we watched last week was done by Stanley Nelson. I don't know if you're familiar with him, but he's an excellent filmmaker, has won many awards. And the California Newsreel, where you get these films, has been collecting social justice-related films since the 60s. Yep, since 1968. Yeah, and I think that in itself was quite smart on their part to be collecting our history, a part of our history that maybe other people weren't thinking about so much as far as collecting it. And aren't you lucky that you were able to acquire some of these films? Yes, and we'd like to acquire more. They have a huge catalog, and we'd like to get as many as we can so that people can see them. Because these aren't things that you might see them on PBS occasionally, but they're not things you're going to find in a video store or they're not really accessible. And that's why we are able to buy it from them and make it accessible to everybody. I went to their web page just to see, and it is costly if you want to do something with the film. So we're lucky that we have you to provide these for us. And they do have an amazing catalog of various social justice issues that you can tap into. Do you have any kind of funding specifically for these videos? We don't at this time. But they do give us kind of a break in the price because we are a public library. And they do allow us to show them in community settings. So they are doing their best to get those films out there too. Well, I'm looking forward to attending some of these. And I hope that you'll continue with a series, if not this particular one, another, and let people know more about how they might contribute to this in some way to help you provide this for our community. It's really, really important at this point in history, I think, to remember, as you say, our past, which is our present and hopefully not our future. Hopefully it will make a difference so people can change. I want to do one more plug for your library on my way to work this morning. I stopped in and looked at the fabulous photo exhibit that's there right now. And I just hope all our viewers will take the time to go and see that. It's a little bit about the history of Portland with the old photos from the Press Herald. But it's the current history as well with the new photographers and the new work that the Press Herald's doing. It's a lovely exhibit. You might want to talk with Abraham Schecter. He's our Portland Room librarian, and he put that exhibit together. Well, it's wonderful, and it complements what you're doing with the videos. So the people can come Thursday nights, 6.30, and just take the time to see the photo exhibit as well as the videos. And if people can't come to the showing of the films, they are available to be checked out. That's wonderful. Thank you so much. Thank you.


This was the good thing about a dynamic website is that once the site is running, it will keep on running. It does not require a lot of intervention and maintenance because as the data is loaded in the database, or if the data changes in the database that is reflected in the dynamic pages. But still you have to look at certain aspects of maintaining and keeping your website up to date. And what are those things? I will look at this over here. So you see that these are some of the things you have to be looking at, okay? And then of course, testing also, because there is no such thing as an unchanged or constant website in today's domain. The time of the static websites is gone. Even if it's a static website, it requires certain updates. It requires certain changes over the period of time. It is never ever completely static. There may be some of them, but that is not a rule. So let's look at this more detail. So managing your website from afar, from a distance, now usually it is the ISP who is performing their people are accessing the database and maintaining the website along with the database, which is of course, not with reference to the application. Remember that. Few people have full access to their website, few people, okay? And of course, when they maintain their websites, you may move your database back and forth between your computer and the computer of the ISP or the database service provider. Just like the HTML pages, just like the HTML pages, right? So if you are maintaining your own website, now there are two things over here. Number one is that you may be moving your database back and forth between your ISP or the database service provider and your computer. That works for small databases, for example, Microsoft Access. In other cases, what you might be doing is you might be accessing the database at the ISP. When the database is very large, it may not be movable. So you are making changes over their database by accessing your ISP for large databases like Oracle differences are there. So database housekeeping, DSP performs the routine backup, okay? It could be a daily, fine. But the thing is that there may be many, many cases, which I have listed over here in which you might be interested and you might be required and you might need to do the backup on your own, okay? Not leaving it on the ISP or the database service provider, ISP as in internet service provider. Now say, for example, you are a university and it is the registration time and the students are registering for the courses, adding and dropping courses, right? So in that case, it is not wise to wait for a whole day to make a backup while the system might crash for some reason. So looking at hourly, hourly backup, right? Or you may want to place the backup at a certain location. You would not like the backup to be with your ISP, the DSP, but you want the backup taken at your end and then locating that backup somewhere else for reliability reasons. That is one thing more. And of course, if you are taking the backup by, it says the guest book and you take the backup by moving the guest book, okay? And moving an empty database over there, that might create link failure. That might because somebody might access your website while you are performing this task. The best approach, the better approach is to point it to an under maintenance page and once you are through, then you point it back to the actual page. You have to be careful. Why careful?  And it is not good, it is not good to be notified of the errors, omissions and corrections by somebody else. So have a schedule for database maintenance. It could be a minor if it's okay with you or it could be overall. It depends upon your requirements, it depends upon your domain. Remember that this backup is sensitive. By which it involves people from the ISP, it involves people from the administration, from the database, from the web developer team, from the so many different people are there. So you have to be careful. And of course it could be a permanent fixture or it could be routine, it is up to you. So changing, when you make a change, improper minor changes can bring down a major system. There are many, many stories, the system down for hours, the system down for days and so on. Why? Because when the change was made, it was not tested. You have to be careful. And changes are always there. And in case of emergency, streamline the changes because in case of emergency, loop holes are left to be exploited later. You have to be careful. And finally, the person who makes the change is not the person who is testing the changes. That's all I have for you in this world. Thank you very much.


Welcome to the CIS Distinguished Lecture Series. Today we are very fortunate to have Dr. Renu here to give a really exciting talk about the precision in medicine, point of care systems by medical sensing. And Dr. Renu has three affiliations. He's Harvard Medical School, the Children's Hospital in Boston, as well as Northeastern University. So he has a very long resume, so I have to just read most of the highlights here. He received his bachelor's degree from Magistrate University and then did master's program at Columbia University with PhD from State University of New York. And he studied with the Nobel laureate, Professor Harold Urey, and which is a very distinguished scientist. And he also published more than 250 publications and two monographs. He's currently working on the textbook level monograph on the bio-nanoscience. And he's a member of several academies. He was the editor-in-chief for Journal of Bio-Nanoscience and associate editor for Journal of Nanoscience and Nanotechnology, and currently serves on six editorial boards for various journals. He has been interacting with a number of leading researchers in the world from universities like MIT, Harvard, Stanford, Berkeley, et cetera. So he's very well known for the biosensors at the interface of engineering, materials, and biomedical applications. So without further ado, I would like to invite you to join me in welcoming Dr. Renu for this wonderful talk. Thank you. Good afternoon. I'm very pleased to be at the University of Wisconsin in Milwaukee campus. I had been to the Madison campus many years ago. It's quite a surprise waiting for me here that it's expanding. I have been very fortunate in knowing Professor Zhenhong Shen, with whom we share many common scientific interests. And I think our partnership in census will be a tour of the force. Having said that, what I'm going to talk today is on personalized precision, personalized medicine and the various tools that are emerging from nanoscience and nanotechnology, protein engineering, and a number of other areas, and medical areas like nephrology, endocrinology, cardiology, urology goes on and on. If you look at the way this country is operating, we have a health care industry that is escalating. And there's been a lot of discussion in the last 10 years how to reduce the cost. If you look into the breakdown of the cost of the major illnesses, lifestyle diseases, one thing that comes to the attention, redundant, repetitive diagnostic tests, and a somewhat reluctant medical community to share the information. Because of several complications, doctors are not talking to themselves. It is a collaborative effort. It's not a single physician alone can solve the problem. So the repetitive redundant cause or a very large component of the total health care cause upwards, spiraling upwards, past a trillion dollar mark. In fact, a trillion dollar mark would be approximately one-third the GDP of France. Therefore, there is an urgent need to avoid repetitive tests and to make it more accurate. That is where the recent developments in a number of areas, material science, nanoscience, nanofabrication, protein engineering, molecular biology, biotechnology, and engineering all converge. If you do that, the tests become more accurate. Second, why is that health cost increasing? For example, glucose medias have been around for a long time. Measuring cholesterol at home is not easy. Measuring triglycerides is not easy. Now, there are so many analytes, the liver profile, cardiac profile, the nephrology profile. Some of them can be done by an educated patient at home. That will reduce the cost and the information will be automatically relayed either from home or from an ER, from an ambulance or an emergency room. And there is a lot of miscommunication between the two and we are making these tests again and again and again. So, taking advantage of the great developments in material science, nanoscience, it is possible to make these point of care devices home-based physician office, emergency room, or in a continuous dynamic mode where we can measure from a smallest amount of blood, the entire biochemical profile of a person, and from there the diagnosis begins in a collaborative collective effort. In the case of children, this poses neonase, this poses a problem. In the animal science area, this poses a problem because we don't understand so much of, although we are using animal models in biology, there's a lot of gaps in the veterinary science which came to my attention the last four weeks, four months. And I've taken that as a major mission to see that all this, we know in human medicine is transplanted to the animal world as we become more and more civilized towards the end of the century. Now, in the case of neonase, there are special complications. Life is just beginning. At the beginning of the life, there is the biochemical reactions that are taking place. The entire process of developmental biology is not known to us. Now, with all the advances, infant mortality is not insignificant, it's quite high. Now, our groups are working on a fusion of different disciplines of science and technologies. While we say something, say certain types of research we do may seem out of place in a medical school, yet you can see a common thread running through the entire mosaic. Now, we work on energy, photovoltaics, that's mostly and fuel cells at the Northeastern University, sensors, picomolar sensitivity, graphene-based microfluidics. That is done in a collaborative fashion between the Boston Children's Hospital which is teaching affiliate at Harvard Med School and University of California Berkeley, Stanford Medical School and Rice University in Houston, also with MIT and Columbia's partners. So, you can see we've got some of the elite institution, now the University of Wisconsin joins us. Now, we also been known for high density memory. Everything we do is bio-based. We believe in a bio world dominating, entirely replacing the presence inorganics and organic materials and making it more green and energy-friendly. Now, the high density memory storage had been a project that has been kind of hush-hush due to the commercial interest involved in that. All of this, some of it started at MIT and Harvard, then went to the Florida International University in Miami, is a collaboration between NEC, Sony and ourselves. The idea is to create a thin film, protein-based film, which is a photochromic material that can be used for writing and reading and densities can go upwards of 50 terabytes. All idea is to have a thin film embedded in a smartphone or in a tablet. That is quite exciting, but it is taken more than 15 years. Some of the initial ideas were taken from Soviet Union. Naturally, they are very upset with the whole thing and I've not seen a single Russian-made appliance or a consumer product in any department store, anywhere in the world, including Moscow. Now, we were also working and imaging fluorescence tomography, but that project is not moving anywhere. Then we have the bio-nanorabotics, which is funded by the Gates Foundation. The whole idea is to import more human-like characteristics to robots by using biosensors. Then we also have a last area, smell receptor arrays. It started with the founder of the Bose Electronics. The whole idea is to create sensors, smell sensors. How do we digitize smell? That is science fiction at this moment. So, we are supported by many laboratories, protein structure, dynamics, protein engineering, protein-based device fabrication, which is challenging. Many, many device fabrication technologies cannot be used when biomaterials come in. They are very sensitive. Carbon nanotube, which is based out. Graphene, which is mainly with Rice University, Professor P. M. Ajayan. And now with Professor Jan Hong Chen here. And University of North Texas was also a collaborator and the Vanderbilt material characterization center for nanoscale systems, solar cell assembly and testing at the National Renewable Energy Laboratory. All the clinical part, the medical part is all done at the Boston Children's Hospital, Brigham and Omen's Hospital and some at NIH and UCSF. Microfluidics is in the laboratory of Professor Dorian Leapman at University of California, Berkeley in California. There are many numerous collaborations and network of collaborations across Europe, Japan, some in China, Australia and Canada too. So, point of care system for ultra-sensitive quantitative detection of blood analyze relevant to diabetes and coronary diseases. If you look at the chronic illness in the USA, there will be a percent of the US healthcare dollars spent in chronic illnesses. Now, if you look at the two lifestyles, one of them is a lifestyle, this is diabetes. Diabetes is a silent killer, where the diabetes type one, type two and there must be some way of measuring the glucose. Glucometas have been around since 1975. We can also measure the HVA1C, that's fairly constant over a period of time. There is no need to measure them continuously because the dynamics of glucose is very different from the glycosylated hemoglobulins. Then we have the heart disease, the deadliest disease, the cholesterol, high density lipoprotein, low density lipoprotein and triglyceride. So, we need a point of care device that in a single platform can measure glucose, cholesterol, total, break it into HDL and HDL and look at the ratio as well as the triglyceride and an HVA1C channel two. If you look at the glucose excursions in type two diabetes, if you look at the variation into the day, it kind of fluctuates depending of what time you take the meal, it goes up and then down and up and down. Now, a stage has come and the glucose measures we are using are not sensitive. 15% or even 10% or 9% accuracy is not acceptable to the FDA because more original cases of diabetes will be treated as diabetes and you start giving the medications, whatever it is, glucophage, glucomet and genuvia or many other and metformin as well. So, the side effects of some of these is enormous. So, we got to really pin down who is really diabetic or pre-diabetic. That means we need an accurate measurement of glucose. Pancreas is the most sensitive sensor for monitoring glycemic dynamics. The pancreas detects the change in blood glucose concentration and releases the appropriate hormone by a complicated interconnected signaling pathway. So, we have a project organ and a chip, pancreas and a chip. That project is still in its infancy. There are quite a few groups working on it. We were in collaboration with a Wies Institute at the Boston Children's Hospital and also with the School of Engineering and Applied Sciences in the main campus. There are many biosensors, sensors and biosensors. You have an analyte and a response. There's an analysis, signal detection, sample handing and preparation. There are many ways by which there are many different techniques for biosensors. Fluorescence, DNA microarray, surface plasmond resonance, impedance spectroscopy, scanning probe microscopy, atomic force microscopy as quartz crystal microbalance, surface enhanced Raman, electrochemical, amperometer. The last one field effect transistor is the mode we have chosen because it's very sensitive and very accurate. Amperometric lab and a chip consists of at least four microchannels. Flow to micro chambers, one consists of glucose for the second one is for cholesterol, third for triglyceride and the last one can be turned on and off. That is for the glycofumoglobin. Now, this is a flow chart of the device. We work with the tiniest amount of blood that you can think of. There is no point in getting large amounts of blood in certain cases. In the emergency room, it's got to be the smallest, tiniest of tiniest. Then it undergoes a separation into four channels, as I mentioned.  The basic processes involved in using for example, glucose oxidase, cholesterol oxidase, cholesterol esterase or lipases for the detections, ultra sensitive detections depends not only on a platform, that platform I will come to is a graphene, single layer. It can, the basic processes are electrochemical in nature. Redox reactions, transfer of electrons, occurs at working electrode, produces current, electrons flow to the counter electrode and the current is purported to the concentration. They are very simple things. So, there is an oxidation. The loss of electron and the reduction again of electrons is the redox reaction. So, there is a movement of electrons all across. In the case of glucose determination, we use glucose oxidase as a probe. Glucose oxidase undergoes a chemical reaction, releases hydrogen peroxide and the electrons flow from the glucose oxidase from the center of it, called the flavin adenine dinucleotide on to the electrode. Either carbon nanotubes or graphene, now graphene, they are completely abandoned carbon nanotubes and direct transfer is inevitable. Now, the fact which is the electrochemical hot of this enzyme is a cofactor, very important cofactor. Without it, glucose oxidase is not active. It's oxidized, electron acceptor, FAD cofactor is involved in several important reactions. Not only in glucose oxidase, it can exist in two different redox states and its biochemical role usually involves changing between these two states. The complete oxidation reduction cycle involves two protons and two electrons. Cholesterol determination, LDL and HDL concentrations are the two different enzymes. One is cholestrolox esterase, the other one is cholestroloxidase. Both of them contain the same cofactor, flavin adenine dinucleotide. LDL determination will block the LDL. We use the anti-human beta-lifer protein to block it. So, we are looking only at LDL and the total. Convert to cholesterol using cholesterol esterase, oxidize the LDL cholesterol, we use cholesterol oxidase and finally, it can go into an amperometric detection or interfere with the FET sensor. Total cholesterol determination extracts the total cholesterol ester, convert to cholesterol, oxidize and then it goes into FET or amperometric. Amperometric, we are not doing anymore. Triglyceride, we use a different enzyme. That enzyme has been cloned in our laboratory. We make all of our proteins. They are protein kitchens. Lyserol-3-posphate oxidase called GPO.  Conversion of triglyceride oxidize the glycerol-3-posphate and eventually goes into FET. Now, in the case of the glycosylated hemoglobin HVA1C, it undergoes the glycosylation to fructosyl peptide, then proteolysis, then fructosyl valine is actually fructosyl valine. Aminoxidase is the enzyme, releases hydrogen peroxide and the electron. Now, we were earlier using carbon nanotubes. We are decorating the carbon nanotube by covalential linking various types of proteins and lately microRNAs, too. Immobilize the protein. Three-dimensional electrode improves sensitivity. Immobilization is done by introducing carboxylic acid groups by acid oxidation. Activate using EDAC, which is 1-ethyl-3-diamethyl amino-prothyl-carbodiamide. Stable active ester, N-hydroxy succinamide. There is amide bond formation, nucleophilic substitution reaction. Now, all the proteins are expressed in yeast system. We are using E. coli. Problem with E. coli, highly glycosylated, phosphorylated, proteins cannot be expressed. So, they will not fold correctly. They are folding in the protein. When you express it, it is very important. So, we have switched to the yeast system very successfully. Now, expression and purification in pro-proteins transform into p-care-pastore stream. Play transformments. There are several steps involved here. Now, the general strategy is whether you look at a solar cell, photovoltaic cell, or a biosensor to design a point of care, or you look at any other area of topic that you are working on, like memory project, for example, photovoltaic. There is an organic, inorganic material, a synthesis cloning by molecular biology in the case of proteins. It forms nanostructures and there is a nanointerface. That nanointerface is a gateway. So, then there is an organic-organic interface, inorganic-organic interface, inorganic-biointerface, metal oxide, organic-metal oxide, bio-nanointerface. So, if somebody were to ask me why in the children's hospital we are not shining light on the patients to generate electricity, but there is a common element here. Unless we understand how the electrons flow, there is no way we can design the optimal, efficient biosensor or point of care device. Now, graphene functionalization has been problematic. We use a linker molecule. We work on single layers. All that is done in the laboratory of Professor P. M. I. J. M. at the Rice University in Houston. And then we functionalize it using a linker, organic-linker molecule. So, very simply in organic compound, we should synthesize in large amounts and that links to the enzyme or the protein. Now, the functionalization protocol, you incubate the glucose oxidase with a fly millimolar linker molecule, one pyrin butanoic acid succeeding in the middle ester in dimethyl formamide for two hours at room temperature. Then it is washed with pure DMF and deionized water. The linker modified graphene was then incubated with 10 ml. Now, glucose oxidase is again originally obtained from the fungus. We can commercially buy them but you got to purify them extensively. Protein purification is an odd. There is nothing called a 100% pure protein. We can call it 99% plus. Now, therefore, we have cloned the glucose oxidase and so we use the cloned recombinant glucose oxidase. We also have developed by protein engineering method very large number of mutants that can interface with the single layer graphene very effectively. Then we use the Fourier transform in Roman in the signature region, amide one and amide two in infrared and amide one and amide three. They call Boxer stretching region which show perturbations that will indicate that the enzyme has been successfully bound. It is retained in that state.  If you are evidence of corboxyl functionality. Then doping leads to the discovery of new layered materials. One of the common doping that we found very effective is the fluorine doping. There is lightly fluorinated graphene, moderately fluorinated graphene and highly fluorinated graphene. Size specific fluorination or doping is still a black box. So, we are trying to use some enzymatic methods by which the fluorine atom can be placed in the center of the graphene or the size of the graphene or the edges. We were using carbon nanotubes up until about five, six years ago. The discovery of graphene completely overshadowed the carbon nanotube. It's a very simple discovery. Theoretically, it was possible to take a carbon nanotube, rip it apart, lay it on a table, on a flat table. You basically end up with a graphene. When I visited the laboratory of Sir Konstantin Novosilov and Henry Guine at the University of Manchester, about approximately 10 years ago, I came back not very impressed. Much earlier to that, a postdoc of mine came to my laboratory, to my office, at about one in the night. He was a little unusual person and he said, I did something bad. I said, what happened? I broke, broke what? I took a carbon nanotube and made that into a flat shape. My next question is, is it stable? Theoretically. He came back and said, it's low and behold, it's stable. And I asked him, where are we going with it? He said, I don't know. There must be some experimentalists who can create this. The meanwhile, graphene was born in different laboratories. It's simply not one laboratory alone. There was a lot of controversy on the water of 2010 Nobel to Henry Guine and Konstantinovacero. They were taken by surprise too. Scientific community was taken by surprise. Today, we've gotten to a stage. There's no need to go back to carbon nanotube. We never go back in science and technology. Sometimes we revisit or quote some classical papers in physics, for example, in mathematics or even in chemistry. So the question is not what we can do with graphene, the question is what we cannot do with graphene. Therefore, we are looking beyond graphene. We are looking into silicene, organosilicene. We are looking into molybdenum sulfide, dope graphene, fluorine dope graphene, boron dope graphene, which is suddenly assumed. Tremendous importance in the last two weeks. The rate at which you see news items and graphene is coming is astounding. Nothing like this has happened before. What's the because we have the internet? Any information from any place goes to another place very quickly and people are hungry for information. Now, having said that, we were using the carbon nanotubes as a platform long time ago. No matter where you use graphene or carbon nanotube, the first step is immobilizing the protein. When proteins are coupled to carbon nanotubes or graphene, there's a charge transfer. I talked about charge transfer before. From the protein to CNT or graphene, which is the fundamental principle, foundation for the construction of any biomolecular sensing device. The nano dimensions of single wall carbon nanotubes and graphene, of course, their electronic properties make them an ideal substrate candidate for anchoring the proteins for biochemical sensing. Integration of carbon nanotubes proteins by covalent attachment influences the conductance of CNT. The three steps, carboxylic acid groups are introduced by acid oxidation. Activation is done by EDAC, 1-ethyl-3-diamethyl amino-proxyl-carbodiamide, then stable active ester using N hydroxyl succinamide. Then there is an amide bond formation between biological macromolecules, protein for example, or an enzyme and the single layer gases. All these proteins are expressed in our laboratory. Expression of glucose oxidase was being done from fungus. Isolation was done. Now, we have cloned it. They created a library of mutants. What is subjective in using protein engineering here? Number one, to be able to produce them in large amounts. Number two, to be able to site specifically mutate them to make the interaction between the enzyme and surface, one side of it and the single layer graphene. So, they switched to yeast system, picaplastores, which is an ideal system for those proteins that are highly post-translational remodified. Now, the expression and purification consists of several steps. The project strategy is basically either CBD, I mean graphene is produced with CBD, a dope graphene or it could be an organic molecule or it could be an inorganic surface. Making a nanostructure, there is a nano interface through which electrons have to go through the gateway. No matter whether you look at biosimpsons or point of cat devices, photovoltaics, memory devices, there is an organic, organic or inorganic, organic or inorganic biointerface. We have got to understand this interface very clearly. Now, graphene discovery goes back to Manchester according to the Nobel. Today, there are probably about 18 or 20,000 papers that are flowing in. The rate at which we are reviewing papers, all of us, on graphene is astounding. I wonder that I am sure there is a lot of repetition, a lot of incremental things being reported. So, the CVs are bloating. When you see numbers 200, 300, 500 papers, I think if you finally it looks like a kale in the supermarket, huge. When you bring it home and boil it, it shrinks. So, there is a lot of redundant, repetitive papers that are coming up too. Now, graphene synthesis is straightforward. That is a by CVD chemical vapor deposition. I will not go into it. Transfer the graphene for a flexible electrode. Then, the functionalization to create corboxal functionality. The same thing we did with the carbon nanostructures. Now, the functionalization protocol, GOX was incubated with a 5 mm linker molecule, a simple organic molecule in dimetal form of mine for 2 hours at room temperature and washed with pure DMF and deionized water. The linker modified graphene was then incubated with 10 milliliters of the rest of the buffers and we used the Fourier transform and infrared. You can see the infrared here. You can see a shift in the carbon illustration region, indicating that the attached, covalently attached protein or enzyme is really bound to the graphene single layer surface. You also use Raman. Raman is more conclusive. You take the Raman and infrared. Basically, your evidence that the functionalization is gone through and the proteins are attached are quite intact. You have been looking at graphene forms. This was in collaboration with Vanderbilt University. We are not doing it at this moment. Now, the doping leads to the discovery of new layered materials. Fluorine dope, hydrogen dope, nitrogen dope. There are several dope. We are an European dope graphene. Fluorographene, graphene, the problem is a zero bandwidth.  If you open the bandgap by means of doping, slight specific doping is still stopped. You need some enzymatic methods by taking the fluorine, putting it in selective spots that is still much more a conjecture at this point. Now, boron doping has been looked at experimentally and theoretically. We use a density functional theory. We do a lot of things. Physical chemistry is the same time. So, to do this all under one roof is impossible. So, we collaborate with many institutions and laboratories on the campus and out of the campus and out of the country too. Fluorographene comes from Professor Ajay's laboratory and in future it will be coming from the University of Wisconsin-Milwaukee from the laboratory of our GenFoN channel. Now, we also use the fluorine 19 and the carbon 13 magic angle spinning to characterize these graphene and right now we have done a lot of atomic force from studies at the highest resolution possible. We also do small angle neutron scattering as well as inelastic neutron scattering. Now, cryo electron microscopy is done in Oxford an instrument that can go up to 2.7, 2.8 angstrom resolution. Cryo electron microscopy is getting to age, it is catching up. But ideally if you can crystallize the material with very poor long range order, it is very tough. If we can do it, then we can use X-ray crystallography. So, X-ray crystallography will not work. We have got to look at small angle neutron scattering, small angle X-ray scattering, cryo electron microscopy as well as molecular dynamics which use X-ray crystallography. Now, the protein engineering at glucose oxidase, these are some snapshots at 200 nanosecond simulations. Now, we have gone into microseconds. Microsecond simulation of a graphene complex for protein sitting and graphene surface requires petaflop computer. We have gone to the highest level possible sphere. This is a 100 nanosecond snapshot from molecular dynamics simulation and glucose oxidase interaction with graphene. What are we trying to do here? We are trying to make the interface between single layer graphene and the protein as efficient for optimal transfer of electrons. Now, there is a cross talk between the enzyme, the probe and the graphene surface. Electrons get quite lost, although the electrons can travel up to 30, 40 angstroms quite easily. So, the current that you observe is directly proportional. That is where the FET sensor becomes very sensitive. Optimizing the electrical communication between enzymes and the electrode is critical in the development of biosensors. That is, in fact, a major, major challenge. Enzymatic biofuel cell and other bio-electro-catalytic apparatus. One approach to address this limitation is the attachment of the redox mediators or relays to the enzyme. Here, we report a simple genetic modification of a glucose oxidase that is being published in materials today to display a free thiol group near its active site. This facility is the size-specific attachment of a nullumized modified gold nanoparticles enzyme which enables direct electrical communication between the conjugated enzyme and say graphene surface. Glucose oxidase, cholesterol oxidase, cholesterol estuaries, lipases, in the case of HbA1c, fructosylvalene, amino oxidase, and you are going to hear the ammonia sensing in plasma, which is one of the most complex projects coming up in a short while. So, glucose oxidase is a particular interest in biofuel cell and biosensor application and the approach of pre-wiring enzyme conjugates in a size-specific manner will be valuable in the continued development of these systems. Now, if you look at this, these are some amperometric results. They are not very satisfactory before we switch to FET. We switched to FET approximately 2012, about five years ago, four years ago. Before that, we were using amperometric acrochemical methods as sluggish. They are not very sensitive and you want to reduce them to a handheld device to measure glucose, cholesterol triglyceride, HbA1c and several others. You need to have only FET type of sensors. This is a cycling ultramograph for cholesterol bio probe with different cholesterol concentrations. It is sensitive, but not as sensitive as you would like it to be. This is a graphene-based sensor response to glucose concentration, the resistance on this axis and the glucose concentration on the x-axis as a function of glucose for a single sensor is shown for three sequential experimental measurements. Now, if we reduce all the data that we obtain on the glucose FET sensors, cholesterol, triglyceride, HbA1c and now with the liver enzymes, with the nephrology, for example, kidney injury molecule, creatinine, blood urea nitrogen, the whole variety of things. In fact, the other day somebody was telling me we can have a couple of blocks of people. We can line them up in a cube. Neurologists, obstetrician, gynecologists, nephrologists, urologists, cardiologists, it goes into several areas. All of them are interested in the sensitive point-of-cat devices. Now, microfluidics has been around for a long time. The flow of a viscous fluid like blood requires understanding and manipulating the microfluidics to ensure the efficient functioning of the design biosensor. It provides an understanding of the behavior, the size, control and manipulation of fluids that are geometrically constrained to a small, typically sub-millimeter cave. Microfluidics goes back to 1950s. What is happening now? I hope this trend becomes solid. The discovery of something in the laboratory, a prototype sitting in the laboratory, going into some kind of a pre-commercial prototype and then various phases of it before it ends up in the market. It's a long chain. It's kind of getting reduced now. That trend is good. If you look at the history of science, everything that was discovered in the last century, some of them took a long, long time. There was no communication at the time between scientists. The left image shows a microfluidic test bed made from polycarbonate using hot embossing. The fluid channels are 100 micrometers wide, 75 micrometers deep. In the middle of the device is a graphene FET sensor with gold contacts that are accessed through the holes in the plastic cover. The right image shows a magnetic sensor chip with micro-fabricated electrodes integrated into the polycarbonate. All this is done at the University of California, Berkeley in the laboratory across the Dorian River. Now, initial experiments with the device, large range of glucose concentration is got to work in low glucose, mid-level glucose and high glucose. High sensitivity at low concentration work repeatedly with somewhat reduced amplitude. This is the figure at the bottom. It shows a microfluidic channel in the plastic polycarbonate shielding, gold pads opening for electrical connection. Sample volume in a microfluidic channel is approximately two microliters. The device fabrication, I am not an expert in microfluidics. All of them is done in Berkeley in the laboratory across the Dorian River. As I said earlier, it is in thermoplastic devices, polycarbonate or acrylic. Hot emboss devices. There are several steps involved in this. In the sensor fabrication, a gasket is created in a plastic sheet via hot embossing. A functionalised graphene sensor is placed inside the gasket. Using a mask, two gold pads are spreaded for electrical connection. Now, the IGN ships the entire device with a gold contact to us. Then we make sure they are alright. Functionalization is done in Boston. Microfluidics are there. And MIT comes in in a variety of stochroscopic techniques like surface environment in Norman. With a group in Columbia, we collaborate with also. Microfluidic channels are printed on a double sided tape and placed on top of the graphene sensor. The device is covered with a plastic sheet leaving openings for the electrical connection and inlet and outlet of the microfluidic channel. This is a schematic of the biosensors chip. There is the inlet for the blood here. The plasma separation is also a pump a micro pump and there are several channels that you see that in a little better. The prototype micro scale device that we have created single platform again I repeat is a multiplex sensor. One of the very few multiplex sensors that may become available in the market. So you don't have to be running around with different point of care devices. One for color store, one for glucose you know that will act to the cost. And the cost of this device is relatively it's affordable. If the glucose meter runs in about 18-20 dollars somewhat subsidized with glucose and blue shield this may come to maybe another 10 dollars more. Provided the patient is educated and he doesn't scared looking at the values. Another problem, a fear complex for example in micro RNA detection let us say a patient wants to test his blood sample for micro RNA for any type of carcinoma and he sees a value that is abnormal. What should we say? He gets scared so we want to avoid that shock and so some of these devices may still may have to be used in a physician's office or in an emergency room or an ambulance by trained paramedics but others can be done at home. Now since there are several field effect transistors the channel of which a pre-functionalized graphene and operated back gated mode. Each covalently anchored protein and graphene would be sensitive only to a particular targeted species this is a very important point. Emerging two-dimensional nanomaterials alone cannot are not expected to show any specificity. Specificity is something that is germane to the world of biology. A single enzyme can detect only one probe at a time sometimes multiple probes. So the specificity you want to call it two-dimensional nanomaterials comes only when you bring in the world of biology and life sciences into two-dimensional nanomaterials. Now one of the most complex projects we have is the high-sensitivity multifunctional biosensor for microRNA quantitation this entire project except for some parts of it in Chilin's hospital MIT, Berkeley is located at the Stanford Medical School of Radiology and the PI of this project is the person with a long name not as long as mine. Gene expression regulation by microRNAs in cell mechanism. MicroRNAs interact with the 3 prime UTR of messenger RNAs. Low microRNA, mRNA-based specificity can block translation each microRNA can potentially interact with several hundreds of mRNAs. So gene expression by mRNA degradation of translation the role of microRNA and as a biomarker we are all after biomarkers has been emerging in the last five years. The whole thing started with a crisis in the year 2013 2012 when a patient had a rare type of cancer bile duct cancer called Cholanguocosinoma then the oncologist in Danufarba Mayo Clinic also suggested let us measure the microRNA see we cannot be making experiments with a patient who is sick finally he died at that point there were only two or three laboratories Mayo Clinic University of Tokyo Danufarba no one else can measure, quantitate the microRNA now when we lost a few line species at home presumably due to carcinometrosis there is we try to measure the microRNA even the veterinarians will not agree that microRNA is biomarker for a few line stations but we did send the samples to University of Tokyo Mayo Clinic Stanford as well as Danufarba all of them confirmed the absence of microRNA later unfortunately after the passing of this line we found by an autopsis there was carcinometrosis which caused peritoneal fluid to accumulate and histona was bulging some of the veterinarians were so comical they thought giving him a four course dinner and making him eat because he has been starving for a week was an absolutely preposterous every crisis, every challenge is utilized whether human or non-human towards some new scientific discovery and this goes on endlessly all the 24 hours so the point here is the microRNAs are biomarkers before the the present surgeon general of the United States who is in Washington at this moment I have no idea at all he was proposing let us have a screening at the neonatal stage for microRNA can we determine this is also catastrophic if a child for example can be abused is diagnosed as a proclivity to its carcinoma in the later life what do we do it can be abused medical abuses are quite dangerous and they are going to be there for a long time to come so the idea is to have a microRNA screening in the life what do you do with it that is another question that is where the cancer therapeutics comes the role of microRNAs has been very well established in breast cancer, certain types of cancer and they are all 22 male polynucleotides so we make a sense and they are conjugated hybridized on the surface it is a very complicated concept the multifunctional PLGA nanoparticles for antisense microRNA corgidine peptide there are several steps involved in that strategy is currently used for microRNA quantitation quantitative RT-PCR persists the copy numbers from microRNAs TACMAN probes the quantitatively measured microRNAs molecular beacons microRNA hybrids next generation sequencing microarrays the scheme of graphene based sensing so microRNA consists of a synthetic polynucleotide 22 residue long synths and antisense are hybridized we look at the fluorescence properties for example graphene concentration dependent quenching for the 165 antisense microRNA as you increase the concentration so we use the fluorescence here fluorescence is a very sensitive type you also use impedance spectroscopy surface enhanced roman now to get the microRNA to a POC a point of care device it is very complicated there are many microRNAs they are in a sea of microRNAs I am not sure how we are going to solve the problem but the role of microRNA in carcinoma is getting firmer and firmer and firmer day by day like breast cancer lymphoma I think it is pretty well established we still have the pancreatic cancer liver, hepatococcinoma it is very tough we are working with transplant surgeons to get biopsy samples from outside the country I had to be able to extract the microRNA and look at them so here comes the transplant surgeons if you are complaining about one brand that was left out see this multi-channel the schematic illustration I propose microfluidic sensor functionalized with multiple microRNAs targets for evaluating a panel of microRNAs from blood samples now comes the most challenging project they were taken up ammonia gas sensors have been there for a long time they are quite sensitive but to be able to sense ammonia NH4 plus not NH3 in plasma it generates at the bedside in a static mode and a continuous mode we use a family of proteins called ammonia transport protein AMTD ammonia transporter from AMT or orange family structurally and biochemically well characterized under lab as extensive experience in purifying and handling these proteins XA crystallography and ammonia transport protein has been done fortunately in order to visualize the conformational rearrangements there is a channel there so this is a channel protein that is embedded in a liposome NH4 goes plus 4 goes in finally there is a release of electron and on to the graphene suffer but the problem of getting a protein a proteolyposome to bind graphene is not yes where we are still experimenting with it now in order to test the AMTD activity we reconstitute the protein into liposomes and the activity is measured using surface supported membrane electrocosiology so there is a channel protein this protein is so big there are 11 trans membrane channels in it this is the early molecular dynamic simulation of the ammonia transport protein inside the bilay this is average at 10 dummy atom models of the ammonia transport protein from cryo electron microscopy not from our laboratory from our collaborators at the university statclad in Glasgow in a partnership with Oxford University and Grenoble in France so it is a multi national effort that is required here the future perspectives we are trying to use a mediated transfer a mediator is usually a ferrocenium the ferrocy nanoplasmonics for the detection of glucose now the challenge comes in continuous glucose monitoring what do we need that we all saw the variation of the glucose at the time of the day therefore you need to have a bedside glucose continuous glucose monitoring I think this is still a far cry I was tossing into some of the drug companies none of them are sure that they can come up with a continuous monitoring we need one for cholesterol so it is a lot of work here but when we do research we are concerned about something very important here simply we do not want to make a device we want to understand the science behind it would that science open up new avenues for example a merger between material science and enzyme catalytic as I mentioned for site specific doping, fluorination enzymatic processes these are some areas which will bring in new science the science as it is developing now we are more going into application science but while forgetting fundamental seminal advances in chemistry and physics is what prevails the whole that is what I would like to see any project this lab and a chip has been around for a long time but a multiplex sensor is a technological engineering challenge there are emerging platforms boron nanotubes molybdenum single layer is very promising our team should have my name at the very end Dorian Leapman at the University of California Berkeley P. M. H. M Ph. T. Rice University of Houston, Texas another long name from Stanford Medical School J. R. T. Berry M. D. is an expert in periodic diseases Michael August is a medical intensive care unit Joseph Bonaventure renal division Harvard Medical School urology Harvard Medical School or not J. Well at the University of Stathclades Flavon-Mathfilipec at the University of Warsaw and Dr. Saunia Bishwanathan at the Newton University Hospital she is an internist those three people inspiring us are not humans at the bottom of the list so this is an overview of the Harvard Medical School the quadrangle of course this place had been very historic children's hospital itself had two or three Nobel laureates three Nobel laureates the way you count on Harvard is a bit funny because people are everywhere every hospital tries to put their name so totally it is generated maybe a couple of dozen Nobel Prizes in Physiology and Medicine one way or another and quite a few of them still are around you can see them in the cafeteria sometimes colliding with them but I don't think there are any any special facts about them they're just like that the Children's Hospital in Boston this is my lab, this is the 14th floor my lab has two neighborhoods Harvard Yard great inspiration as I said this is a partnership not only between humans as well as non-humans right now as I told you we're extending this to an important problem renal collapse right now creatinine and blood urea nitrogen are two biomarkers we have a kidney injury molecule CHIM-1 and CHIM-2 came out of the beginning of the hospital we are doing a lot of structural biology too at one time I was only doing structural drugs but nobody buys it now unless it is tied to a disease or to a device or something that the consumer best buys otherwise you don't get it even with all that we don't get money scientist and money are two folds apart they are not being good entrepreneurs only a fraction of the intellectual property generated right in this building probably finds his way to the master's place unfortunately so working in all kinds of directions I want to thank all of you for listening to me, thank you Thank you very much Dr. Manu for the wonderful talk we have time for questions so many questions for Dr. Manu actually I have a question I know we have many students here so what they would like to know probably is what are the most opportunities for the biosensor research in particular development for engineering and real applications which we are focusing here on so maybe you could give some advice to them some advice Sensors are there to stay with us no matter where you look all kinds of sensors have been around for a long time biosensors that is the procedure in your laboratory for example here and bios and several others have become very important in medical diagnostics right you may also find applications in security for example in homeland security right when a hidden explosive or some thing is in the suitcase you got to go to parts per billion some of them may emanate or nitrogen oxides may come out of it ammonia may come out of it or it might be some other substances abuse so it has applications in those areas as well and these are getting so small that we do not know we may not be even aware that a sensor is around us therefore in terms of job opportunities first of all training in this area as I say it we got to create a new workforce that workforce will be very different from the workforce we need the training at one time maybe 10-20 years ago we talk of biosensors we need a strong background not only in biology but in biochemistry as well right and the ability to express these proteins is a biotechnology concept so the type of courses that a student should take to prepare himself for the future job market is quite complex we are not sure we got the faculty in all the universities to be able to teach this we may have to create a new generation of faculties we will not have any mind sets or open mind like all of you the younger generation here so job opportunities are growing if you look at the number of medical companies that are working in diagnostics take Minnesota now we have got the electronics we have got the Boston scientific and Maple Grove Decton Dickinson then you have got the Thermo Fisher right there are several companies that are coming up worldwide market for this is increasing I have seen some of these figures being put out when you ever start the company like you the statistics I wonder they are true they are probably true and also there are other companies competing with us this is a politically contentious issue right now companies in China in India in Taiwan and just about everywhere and therefore job opportunities should be opening up I am also intrigued how much of this workforce will remain here you know the eastward migration may occur too that is already happening in areas like information technology and software and so on and so forth therefore I think the future is very bright the question how we train them and also the background of the student assistance 4 year colleges are not doing a good job students end up in the 4 year college from a school that is lackluster has not paid any attention to training all these things worry me so there must be a very robust teaching program jointly for example the medical college of Wisconsin should join Heinz medicine campus your has been well established I know whenever you have a huge towering institution like Harvard and MIT Boston University and others feel a little not so happy and they try to compete I don't think competition is there the collaboration is the way to do it even the magnetization I start talking about the material modification you mentioned to functionalize the big protein or enzymes on the carbon nanotubule but there are also records saying easy to peer all those big functional groups do you have any suggestions for utilizing the boundary between the protein and two ways of looking native enzymes what we call as wild type can do the function but certain residues in say for example an ammonia transfer protein might be binding to the graphene so if you mutate those residues electro statics you may be charged there lysine, oxygenine mutamic acid you can modify them so that you can make the attachment much stronger that is where protein engineering comes but I have not seen we have done a lot of glucose oxidase mutation ammonia transfer protein is a beast if anything can happen to mess up a protein in nature ammonia transfer protein would be the most notorious member of that so that is hard so you can genetically modify them using biotechnology that is what I have studied I do not think it is any rule of thumb question related to the glucose sensor so we know for the home testing now the glucose has stretched across about one dollar or less a piece so I consider that as pretty affordable so what do you think about detection at home are you trying to make it cheaper or are you trying to make it more robust cheaper see there was a time when we were working on the memory project a questioner has supposing we need one ton for making let us say protein based memory devices thin film to be put into the tablets and smart phones at that time the price was about 600-700 dollars per gram today thanks to China the prices come to something like 10 or 20 or 30 dollars per gram so the cost of the one component of the material is coming down now the question of graphene mass production is being addressed again there are Chinese companies that seems to be ahead of just about everyone in the world so the material cost is less fabrication cost and testing and other things will be there so what will be the cost of the glucometer using the new technology using graphene microfluidics where not made any accurate calculation I don't think it's going to be bad but cost again is dependent on the demand so if the cost is comparable or in two times the cost of the glucometer as the volume of the sales increase it is going to come down and the lifetime is another question there is a time when people are saying I will only buy devices that will last lifetime I don't think it makes sense people who want to throw away they throw away a culture I think that this country has been singularly blamed will continue for some time many of us go back to an old device and take it to repair it who will repair it for what telephone breaks it breaks what we do with the components inside it or the green or the benign that's another question that we have to begin to work if the cost will come down eventually it's comparable it's not too folk maybe too folk now the question is now how many different analyzed in the blood you want to measure continue glucose is one now what happens in the continuous monitoring wirelessly the information is sent to a central desk in a critical carry on it there are the human elements to the concern she has to pass it on to the physician right wherever he is and if you are going to for example I tell you this outsourcing business in healthcare one block number one sensitive information is going on by the time I get a number to ring in India and pick up I want to pick up are we going to a lunatic asylum nobody picks up the phone send me a text message what's that somethings don't work by this so there is a human element involved so how many analyzed we want to measure continuously glucose maybe ammonia plasma and if you are talking of really very large scale multiplex system that can measure everything all you need is let us say 5 microliters of blood that information integration of all that information requires a physician's intervention that's where I think we believe in the medical community believes artificial intelligence for pain and fucking growth it will take a large amount of data collate and say these are the problem 3 4 5 6 7 8 9 even before the patient arrives in the emergency room so it has got to happen with such great speeds right go back to artificial intelligence will become very common algorithms are getting all the time better there is a number of interesting challenges and questions what will be the healthcare like in 2050 2100 we can make some educated guess about what will happen or at the same time the human life span is going up too not noticed by many people are living longer posing a major problem for the social security administration, Medicare, Medicaid and whatever that plan that we had being revamped into whatever that it may be revamped we do not know collaborating not one single person can do it thank you thank you