What's going on everybody? Welcome to Rotoride. It's Djiboy Vortex, and I'm BubbyFPV. And we're back at it again with another community spotlight. This time it's all Bubby's picks. So if you don't like them, you know who to blame. He picked them all. So for the first category, we have Epic Dives Yo. We have Cycle1337. I think that's supposed to be Leet. I think that's like Hackertop. That's pretty sweet. I like that he, unless I'm tripping, he put the cinematic bars. He put the building out of the frame. That's pretty cool. Diving buildings, like super tall buildings, is just always like super cool to me. Like just to see all the still tall buildings from up top, you know. I just love how he gets super close up to the building. And that's a huge one. Yeah, it's a huge, huge building. All that's cool. Do you know where this is? Um, Shanghai, yeah. Okay. That's so sick. That's so sick. I always wanted to dive buildings. Well, I have, I have done them. To go take you for a building dive. We don't have any buildings that tall, though. That's a tall building. Yeah, I went once in New York, but it wasn't nowhere near that tall. There's been nice little B-roll mixed in there. Guys, he only has 47 subscribers, so let's get his sub count up there, because he's doing some awesome stuff. Yeah, that's kind of crazy. Yeah. He only has 47 subscribers and he's already doing stuff like this. Yeah. I like how he keeps doing that, where the building cuts off of the frame. Yeah. That's cool. It gives it almost like a 3D effect. Yeah. Sweet video. Very creative, very good. Who will get next? Sync FPV. Okay, so this, this video I saw this, and I was like, okay, it should go in good old freestyle, but then at the end, there's just this dive, and it's just like, oh my gosh. This guy, I feel like I've heard of before. Oh yeah, definitely. I've seen this guy. Yeah, he's been on a couple community spotlights before, I think. He's got his name on his controller, and he always has stick pants. It's like one of my kind of Detroit spots that I miss. I don't get to fly stuff up this animal. That was sick. Dude, like, I love Powerloop Gap. Still looks so cool. Okay, watch this, watch this, watch this. Oh my gosh. And boom, I didn't, I did not see that coming. I was like, oh my gosh. Because he started like going away from the building, and then going and went, hit that dive. That was solid. Who we got next? JT Wilde. Okay, this guy, I've grown through his channel, actually know him. He does some of the craziest dives that I've ever seen. I don't know. Oh, giving Cricket a round for his money. I don't know if that was luck when he went through the wooden thing behind the structure. That one was super short. I'm not sure about that one. That was super short, yeah. Cricket. Watch out. This guy's on your booty. You better step it up, son. That was it for epic dives. What's the next category? Good old freestyle. Some good old freestyle. A little bit of classic. First up, we have L1 Lithium. Lithium with some ones. Lithium with some ones. Skatepark shredding. Skatepark action. I love how she's just like riding. Oh, this is a girl? Yeah. Oh, cool. I think she's been flying for like four years or something like that. This is so many good pilots now. Yeah, there's everyone. When I scroll through Facebook or YouTube, it's pretty much everybody I see is like, pretty good these days. Just like the pilots. Yeah.  That's different. I think it fits the mood with this one kind of thing. Yeah, it's like shredding. Yeah. Handrail, nice little gap there. Yeah. That's cool. Skatepark too. I like to go there. It's all in video. That was a good old freestyle edit. Good old freestyle edit, for sure. He's done good on picking a good old freestyle edit. PDEVX. This, oh my gosh, like, that was sick. That was sick. And then he does it pretty much again, that backwards thing. All right. Dude, it's fire. That's what I'm saying. There's so many people that are just getting good. Dude, all the backwards stuff, like. This guy, a decent amount of subscribers. You should probably have more or so. Yeah. We're going to give him another another subscriber groove. And you make sure you subscribe too, because this guy's good. Yeah. It's a nice, like, chill music too. Yeah. Which is interesting because he's, like, ripping. Totally ripping. Chill music. Oh, that was sick. That was sick. I'm going to have to watch that again, actually. I do it one more time. He does a power loop, but then he's going backwards, and then a roll backwards. Yeah. That's pretty cool. Pretty rare to see someone do something you haven't already seen nowadays. Nowadays, everything's, like, pretty much everything's done, but this guy's making up some new stuff that we really like. Yeah, that's great. PDEVX. They're killing it, man. JB Photography. That's B-roll in the beginning. Another good music choice. Yeah. And editing, all those on beat. I love when edits are on beat. Yeah. Satisfying. This is, like, a nice fun vibe. Just get out of the woods and just rip it up. Yeah. Like, a lot of his tricks are, like, I guess, landing level with the horizon. This music is so fitting with the song, with his flying style. Well, he's, like, riding on the edge the whole time. Yeah. Strippy spin. Trippy spin. Oh. Oh, yeah, a little combination thing. Yeah, and do another one. And he's still. Oh, and he wanted to do another one. How did he do that? Dang. That's impressive. That's really impressive. Props on that. Yeah. That was, like, three trippy spins linked together. Especially those trees aren't, like, the most tall. Yeah, and there's, like, stuff everywhere to run into. Yeah. I think we're going to subscribe Drew to this channel, too. Yeah. I can't tell if that clip was just reversed. I don't think I've ever seen. We're breaking new ground here with boys and girls. A bunch of new freestyle. I don't think I've ever seen someone do a trippy spin and then just stop it and go back the other way. Like, so immediately. It wasn't, like, a, oh, yeah. Quite impressive. All these videos are, like, blowing me away. Bully, FPV. Really coming at us with some Bando action. This guy has flow. I'm a flow pilot, I guess, and I love this video. Just that one-get-together type of stuff. Feel like this guy's somewhere in Europe. I don't know why. Germany. Germany. Okay, there we go. I didn't see that. I didn't see that. That's cool. That was a good flow. So rewind thing there. Some video, man. That was another good old freestyle video. Keep up that flow, man. Links in the description to his channel, as well as all the other pilots that we're featuring today. Yeah, one left for good old freestyle. Who's it going to be? One left FPV. One left. You know what's funny? What's up? I didn't mean that. What? I was genuinely saying we have one left in this category. Really? Yeah. And he's called also one left. One left FPV. This video, when I first saw it, I was already blown away by the freestyle, but then you'll see his dick camp. Oh, he's kind of like, what's his name? I think Captain Uno. Yeah, Captain Uno. Yeah, I don't know how people, that's crazy. Yeah, that's impressive. One handed flying. And I didn't notice he was one handed flying until like near the end of the video. Oh my gosh. One handed trippy spins. Trippy spins. True. Stuff your game up. Yeah. It looks cool to see someone who, like I have no dexterity to do that. It's a lot of precision for handling the throttle with your pinky. That was cool. That was really cool. I love that move. I don't know, there's no name for it yet, but it's like a half parloop, half yaw spin, half, and then Matty flip through. That was sweet. Awesome video, man. Another solid one. One handed. Yeah, one left. One left FPV, definitely. What's the next category? Cinematic FPV. I am very excited to show you this category because there's some fire videos. Already, I like it. The fog and the violin is definitely cinematic. Got that right. I love just going straight through the trees and just... Like it's like nothing is technically challenging about it, but it's just something about the way the trees are. You're flying a camera around. Yeah. It doesn't always have to be about tricks. Sometimes it's just like filming something cool, just like if you're filming something you wouldn't go flip it. You're filming something cool, so there's so many different uses you can use a drone for, and this is awesome. Yeah. I'm impressed with how good it looks because it's dark. So usually when it's really dark, like a lot of times the footage comes out grainy, but this looks nice even though it's really dark. Yeah, next. Energylandia. I think this is like a video, like an ad commercial for an amusement park. Okay. So I don't know who the pilot is, so I feel bad not knowing who the pilot is. Wow. The editing is really good. It's really good. Like there's transitions are so butter. If you guys know who actually piloted this, hit us up in the comments so we can give them credit. Yeah. Excellent video. The color grading is amazing too. It's like really vivid. I wonder if he edited the video too, whoever piloted it. Oh, was that one shot? Yeah, I think that was. That was, if that was a one shot, that was really well choreographed I guess. Yeah, if that one goes by. And then it just speeds up. Yeah. Yeah, that is sweet. That was... You got two different roller coasters and one shot. And right up on them too. I want to go to the scene part just after watching this video. I know that's what I was just thinking the same thing. I wish we could get some access down here in Orlando. It's like, come on Disney. Yeah, come on Disney. I love that shot at the end. Yeah. I was, he was able to get the whole park in the shot. I'm peanut butter and jealous. Next up, Andre Larson. Like the exotic locations like that. Yeah. I wish we had some kind of elevation here. Ford is this Ford flag. I wonder where he was standing to get like video. Like he had to have been standing on edge to get... That was crazy too. Yeah. I don't know if that was real or not. The lighting in the flag. I love this diving the waterfall because... There's a little rainbow in there. Yeah. And especially it seems like he's like kind of going with the speed of the waterfall. I love how many like... The snow and the green. Yeah. I love how many like different like spots, locations there are in the studio.  I would have flown behind some mountain or something on actually. Yeah, I would have lost all the girls. Stay in the stuff. Yeah. I'm not good at long range because I'm not used to it. I won't fly.  I feel like I'm on my way back. The music almost seems like it was made for the video, not the video made to the music. You know what I mean? Andre Larson, everybody. Yeah. Subscribe to this one too. Yeah. That was really sweet. Ellis Van Jason. This video, I bet a lot of you guys have seen it. It kind of blew up. Like it has like 38,000 views. Kind of like the similar to the last one, like cool mountainy flying. Yeah. Like the speed ramps. Yeah. Makes it more intense. I think you can see the clouds. Yeah, I was going to say that's wild. There's like above the clouds. Yeah. But still in the mountain. I'm pretty sure I watched this video like 20 times before. This one, I just was like. These locations are just insane. Where the heck are these places at? I don't know. No sick. No sick. There's like full package and the audio, the smooth shots. Which category? Young Rippers. That's you. Yeah, that's me. I love this category. Put yourself on the list. You're not supposed to do that. I mean, I didn't. Or who's the first Young Ripper? First up, we have Gunwoo FPV. He is, I believe in South Korea.  I love Bando's and I love Narnia treats. Narnia treats. Yeah, you're definitely a Ripper. He's only 16, I think. You can tell that right away. Got so close to the wall. I like in a video like this for like aggressive freestyle, I guess. I could hear the audio. He keeps getting so close to the walls and stuff. It seems like a lot of building to be abandoned. It seems like apartments or something. Like you could have hundreds of people live in there but it's just empty. Yeah, that was. I like the premise. What are you gonna do on the way down? Oh, you like Maddie flipped her in. That's sweet. Okay, I'm getting that crack. Oh, that was so sick. That was so sick. That was a good Young Ripper. Yeah, epic. Next up, Rooster FPV. Big old backyard or something. Yeah. It must be nice. I'm just open at the back door and just go rip. Dude, Joshua L. Bardwell style. Yeah. The title of this video is Chillin' and it's pretty much exactly what he's just chillin'. Just chillin'. Chillin' some nice chill music. Chillin' out on the way outside. Yeah. I've been watching this guy's videos for, I think like a couple months now, just how good he got like so fast is insane. I think it's gonna be like our talent scout. You're finding all the people we've ever heard of. It's gonna do it for Rooster. Keep it up, Rooster. You're quite the Young Ripper. Yeah. One last Young Ripper. Next up, we have Lee and Vander. What a chill song. Yeah. Dude, this kid's so smooth. That is. He's got some nice smooth flow. Really smooth. He's definitely got a good like constant flow going. It's never kind of a drop to a change in direction, but always sweeping. Oh my dude, he was headed straight for himself. And then he just, I'm always scared about hitting, getting hit by drones. I hit myself with a tiny one, so that kind of hurt. A tiny? Yeah. It was like full speed. I was like, oh. And then we have the unnamed scout. Yeah. So like, that was video, I was like, I have no idea what the heck, what category to put it in.  Yeah. It's only seven seconds. It's only seven seconds. It's only seven seconds. It's crazy. Oh man. I don't know if that's. I'm more impressed with the guy than the flying. Me too. Me too. Do you think that's real? Do you think you actually did the jump? Yeah. Yeah? Yeah, people do stuff like that. Yeah. I want to see more of that, though like drones in parkour go together well. They do. I like how the guy is just so chilled. Yeah, that's what I'm saying too. He just looks casual. He's not like, it's just like, jogging and then like death leap. I think that's the end of the spotlight. Yeah. Are we doing a giveaway? I don't know. Are we doing a giveaway? Drill. Drill. We're doing a giveaway. Yeah. Giveaways. Giveaways something. I need a guidance. Yeah. What's the best motor? Choose carefully. No. Settle the drift motors. Settle the drift motors. I think we should give a set of motors to one of the young rippers. They probably need it the most, right?  Yeah. I think Rooster FBV gets a set of drift motors. Congratulations, Rooster. You got a set of drift motors coming your way. That's been it. That's been it. That's been it. Thank you for the spotlight. Thanks for watching. Hit the subscribe button and then. Smash the notification bell. And the likes and the comments and do all that stuff. Yeah.


from the Sands Convention Center, Las Vegas, Nevada. Extracting the signal from the noise. It's theCUBE, covering AWS re-event 2015. Now your host, John Furrier. Okay, welcome back everyone. You watching theCUBE's SiliconANGLE's flagship program where we go out to the events and extract the signal from the noise. I'm John Furrier, the founder of SiliconANGLE. We are live in Las Vegas for Amazon re-invent. Amazon Web Services re-invent, big conference.   And theCUBE, we love showcasing the hot startups. Albergio, the CEO and founder of IIX and Console, and Paul Gamp, the CTO of IIX and Console. Console A and Console Connect. We just did the event. Welcome back to theCUBE. Thank you for having us. So we did a live broadcast at your event, Console Connect in San Francisco. Small, intimate event, thought leaders. At the same time, Apple had their big event. You guys created more buzz than I've ever seen on a small show because you guys are doing some pretty compelling things at the infrastructure level. Very disruptive, I would say, that enables really secure internet. And I want you to explain to the folks now why the company was formed and why it's doing so well. Because there's a nuance here. The world is evil. DDoS attacks, security. So talk about what you guys have done and what it means. Well, clearly we're seeing enterprise movement to the cloud and we sort of had early visibility into that many years ago. As a result of that, we basically looked at other parallel things that were occurring, growth in issues like DDoS attack, network security and so forth. And recognize that as the enterprise was more and more moving to the cloud and directly connecting to AWS and other cloud infrastructure, SaaS providers and so forth, that there would be the need for a simpler way to directly connect to these business critical applications. And so with Console, we really focused on creating a platform that would extract all of that complexity for the enterprise, for cloud companies, for SaaS companies to really make it simple for them to directly connect. And that's essentially what we've accomplished with Console. So Paul, I want to get your take on this because there's a little, another technical angle. One, the internet's been great. It's lowered the cost of barriers to connect countries, cities, states, worlds. But it's an unsecured internet that's out there. That companies have chosen to lower their costs to use the backhaul of the internet. That's caused a lot of problems. Explain that problem and what you guys do. Yeah, so I think what we've seen is that the mission critical applications have gone from on-premise to on-cloud. Here we are at AWS re-invent. There's no further proof of the success of moving applications onto the cloud environment. But it has exposed that internet, that vehicle that was never designed for mission critical purposes to traffic that it was never intended to do. So what we've done with Console and with IIX is provide a direct and secure path from the enterprise to the SaaS application no matter where it's hosted. So the thing that I like about this business is that the innovation is happening up and down the stack. Right, we all agree. We talked about this a year ago. If I look out amongst the show floor here, I see Juniper, I see New Relic, I see Cisco, I see Red Hat, GitHub, Intel.  Dropbox, it's an API services business. So people are exposing their cloud-based services or SaaS to each other in an unsecured backbone called the internet. Al, demystify what that means because the internet is completely secure. I'm just going to have end-to-end encryption. I'm good. What's the real deal with web services? Is that a reality? Is the web secure? And what do you guys say to that? So, I mean, it's a public fact that on average between the point of origination of data and the destination on data over the internet, there's over four organizations between that path. And that path and direction can vary from time to time. To really have a consistent user experience is private and secure. You need to be directly connected. However, historically, it's been complicated to achieve that total cost of ownership is high and so forth. We basically focus on creating the technology that would make that simple to really drive adoption and to support cloud enablement with the console social platform. I think the title reinvent is very fitting, not just for the event, but for many of the organizations that you've mentioned. There's this evolution that's happened, not just with the internet, but with Direct Connector, Direct Interconnection. And it's frankly needed. Businesses want that reliability, they want that security, and they want simplicity. They don't want complicated solutions. So, I'm gonna throw back memory lane back, go back 10, 15 years ago. Oh, B2B exchanges. That was a company-to-company relationship. That wasn't so much a product issue, meaning if Paul and I are, hey, let's work together, our company has to do a deal. Yeah, we'll exchange data on POs and sales or some data exchange happens. Okay, great, secure that up. But now, what if, now today, we're exchanging product services? Completely different, it's no company relationship. The customer uses both of our products in one solution. That's right. That's an integrated stack. Is that the phenomenon of why you guys are growing so fast, or is it something else? No, there's not really one use case. There's quite a number of them. But there is that. I mean, we have the benefit of this many to many ecosystem platform that we've created. It's a non-discriminatory ecosystem platform. It allows any organization to be a member of it and for it to directly connect what's important to its business and other organizations that see them as important to directly connect to them. And so, we are a product, but it's all really about the ecosystem at the end of the day. And in helping empower innovation and supporting our customers and our partners take advantage of the platform. All right, so what are you guys doing here? Okay, we've got a couple minutes left. We're going to talk again on Thursday.  But what's the big deal? You guys, in the ecosystem partner, is it like good sales opportunities at BizDev? Is it, what's going on? Why are you guys here? Are you guys talking? Yeah, we're a titanium sponsor at AWS re-invent. We're also speaking, Paul and I tomorrow, have a 60-minute session on how we've simplified direct connect. And as well, there is a great AWS ecosystem that we're here to put our best foot forward in supporting. And so, it's a great opportunity. It's an amazing event, phenomenal attendance, and we're really happy to be here. So, Paul, I've got to put you in the spot, okay? Yeah, I've been hearing in the hallways that there's been kind of a love fest between Amazon and you guys, company. Why? Is it technical? Is it business model? All the above? Explain that. Yeah, I think what you see with the success of Amazon is the fact that mission-critical applications are moving to the cloud. And Amazon, despite their size and capacity, need partners to help bring the enterprise onto that platform. Amazon Direct Connect, they productize this, the concept of getting direct connectivity. And they need partners like console to make that easy. And we're probably one of the easiest ways to do that. We've also had a cloud router project, our open source project is now available in the Amazon Marketplace, so we're, there was a lot we do in collaboration. So you can explain to someone in the attendee hall here, hey, what do you guys do? Why are you relevant? What would you say to them? I think we're relevant because we provide direct connectivity to SaaS application providers, cloud providers, in a manner that's never been done before. Versus what alternative? Exactly. Yeah. And what's wrong with that? It's complicated.  It requires, the best thing you would see out there is a virtualization of a connection, but it still requires both parties that connection to deal with that configuration complexity to essentially have the packets move. So you can say, I'll pretend my marking out, plumbing's hard and you do it easy. You don't have to become a global network operator by using console. All right, so what booth do you guys add and then how do people find out what's going on with the sessions? So we're a booth 1024 and our session, if you had the AWS app, you can look us up, console or us by name and I hope to see many of the members watching at our session tomorrow. All right, so I got to get a pound of flesh from you guys. I want you both to answer this question to wrap up the segment. Paul, we'll start with you. What's so big about this year with Amazon? Is it a tipping point? What's going on this year for the folks watching who couldn't make it down here? What is going on here? What is happening right now in this moment of history? Yeah, for me I think it's that internet of things. The fact that telemetry, devices, automation, everything's getting onto the internet. And for a company like ours, that's critically important because a lot of the data that you're now getting from those devices is super sensitive and you do not want that traveling over the public internet. So that's what I think is happening here. Al, what's your take? I know, I agree. I mean, the internet of things, I think in many respects it's just the beginning but we're starting to see that at this event. We're starting to see the growing, the growth even with direct interconnection. Many, many organizations, more organizations are starting to ask for that. So a number of things, but I would agree with Paul. The internet of things is a big driver. Okay, thanks so much guys. Engage with us, engage with our hosts during AWS re-invent and all of our shows on CrowdChat.net and the URL for this chat is CrowdChat.net slash re-invent. Join the conversation, keep watching theCUBE. We've got wall-to-wall coverage coming right back all day tomorrow, all day Thursday. Console, check it out, IIX, remember that company. You guys are going to be pretty successful. I think you guys have a really great business model. It's hard to do what you do. You make it easier, congratulations. Good luck to you guys. We'll be right back with more after this short break. theCUBE, live here in Amazon re-invent. I'm John Furrier. We'll be right back with more after this short break.


It's Wednesday and we all know what that day is. It's Hawaii, the state of clean energy day. So I'm really fortunate to have Mark Watt with us from the Hawaii State Energy Office and Mark's the resiliency and energy assurance program manager. So he's a guy that works before the storms hit and when they hit and after they hit to make sure we're all safe and sound and thinks ahead because none of us do. So Mark, welcome to the show. Thank you, Mitch. I really appreciate it. Really am glad I could hook up and be on your show because I think you guys are doing a lot of good work with energy. Thank you very much. So first of all, you have some slides with us and first of all, let's talk a little bit about the kinds of energy disruption issues that we have that we experience here in Hawaii. So would you like here to talk about that? Yeah, sure. I mean, people think about disruption in systems and I kind of look at it, go back to what does energy provide as a service, as an enabler for other functions? So you know that modern society is highly dependent upon energy. You just think about almost everything that you do on a daily basis. You're using some form of energy to accomplish that. And I think over the years, especially here in Hawaii where we have very high reliability of electricity and we have very good weather year round. So we don't get the kind of things that happen in the main line where they're having a fire that takes out the power for hundreds of thousands of people or other disruptions that take out the whole bulk power grid. So in Hawaii, I think we've become very used to having uninterrupted energy. And so people's perception about that it's always going to be there has changed over the years that they become frustrated if the power is out even for a short period of time. So it usually hits doing Super Bowl. For some reason, whenever the Super Bowl game is on, it's been my observation over the last 20 years, we always seem or pretty well always seem to lose the feed. And that's why that's a total disruption. Everybody gets really angry about that. Right, but you know that the consequences of a disruption could be a lot greater. For example, things happen, water supply is interrupted, food spoils in the refrigerator, medical care is impacted, water and sewage systems fail, communication networks, so we rely so much on cell phones, those become disrupted and businesses go dark and if your businesses has to shutter if you don't have power. In addition, I mean, even within the energy sector, you see that gas stations can't sell fuel if they don't have power. And so you get this idea that we have some sort of vulnerability because we're exposed, we have so much reliance upon reliable and resilient power and fuel. And so what happens when you look at the systems, the markets and the resources involved, you realize that everything has to work in sync. There's a lot of coordination that goes on in order to deliver you the fuel and the power that you're using. It's a life-and-death thing too. It is, it is and we see this with how it works even during COVID with the need for a supply chain to be intact, same thing with energy. You have to have a complete supply chain intact from where the energy is produced all the way to the end user. So I kind of look at it like, then you start dissecting it and wear all those pieces in that puzzle. And you'll have to see that most of the pieces are in the private sector because the government really doesn't own a power plant or a refinery or pipelines and all these other infrastructure and systems that deliver those things. So how does this work? Because, you know, private sector government have to work together to make sure that there is this continuous supply of energy. Yeah, so how can we work from home with COVID if we didn't have a very reliable source of energy? But here you and I are talking on ThinkSec Hawaii and our computers are working, everything's working and I can still do my job. I'm one of the lucky ones can do my job from home. But I've lived through, I lived through the ice storm in Quebec in Canada. We're without power for almost two months in the middle of winter. And actually, you know, we had all our homes were powered by electricity because electricity is so cheap in Quebec. So all the homes were electrical. And one of my neighbors actually froze to death in her living room because she didn't want to leave her dog by himself and they found her the next morning frozen solid on her couch. That's an extreme example of the importance but people on dialysis machines, like you said, communications, what would we do if our cell phone towers didn't work and we can put our text messages? I mean, we'd be like, we couldn't survive on that kind of stuff. Right, so if I may briefly explain to you what the role of the Hawaii State Energy Office is in resilience and emergency response. If we'll go to the slides, please. We talked a little bit about the disruption issues but next one please. And we talked about these kinds of issues that are related to disruptions. And one of the things I wanna highlight here is the last bullet down there is that, there's a lot of vulnerabilities in the way that we manage energy loss and the expectations that people have in restoration and then the public's response. How can we educate people on what the expectations are and how we go about coordinating a unified response in making sure that energy comes back to quickly as possible for critical services. And on the next slide. And we have to plan ahead. That's like when the emergency hits, it's too late to say, well, what do we do first and what do I do second? So that's why we're so fortunate to have you in the Hawaii State Energy Office kind of putting these plans together and I don't wanna steal your thunder but and then trying them out like doing training exercises. So like when it hits, man, we're ready to rock and roll. So. Yeah, yeah. So that's a good segue to the next slide Mitch. Yeah, perfect. So our challenge really is to be prepared as possible to effectively contend with the energy emergencies and threats to our energy security. So this really what our mission is to keep energy flowing in disasters. The reason why we wanna keep it flowing is as you alluded to is there are tremendous numbers of critical services that rely upon energy to deliver things for health, safety and the wellbeing of people and property. So the way we look at it is what is energy resiliency and if you could go to the next slide I'll explain to you what energy resiliency is. So energy resiliency, it's really, we hear about it a lot about, oh, we need to be resilient. The community needs to be resilient and how do we define that in terms of energy? So it's really a focus on people who usually think it's about just infrastructure or systems or something like that but we see that the definition is much more nuance than that. So there's elements of the resiliency that includes such things as robustness. And so when you think of robustness, what is that?  So you have this, like it's hardened. So it could be stronger poles or it could be that you've hardened a pipeline or that you have redundancy in certain systems. So that's that ability to continue operating in a disruption. Another component of resiliency is resourcefulness. And so you think, why is resourcefulness? Because really what you're doing is you're managing that disruption. So this is really about people skills. This is workforce. This is skilled how you respond. And so I think we look at the people who operate the energy systems here in Hawaii be it anywhere from a truck driver, a gasoline tanker truck driver who's moving the fuel. It's not just somebody who's a truck driver. You have to understand all the components about how to handle hazardous materials. What's the best way to deliver that fuel safely? Same thing with linemen and people working in power. Electricity, these are highly skilled and trained people because there's so much danger associated with electricity and explosions if it's gas or other things. So I think about we have to have skilled people that know how to operate the equipment, know what the capabilities of that equipment are and know what the right buttons to press.  Because it's not like, oh, you say, what does highly skilled mean? Well, they have to take annual or buy annual refresher courses and get certified and meet the standards that government has set for their ability to actually operate that truck. For example, I know that from operating hydrogen trucks. I mean, the guy has to be more than just a truck driver. He has to have had the course in dangerous goods transport. He has to know how to fill out all the forms and what the precautions are. He has to have a background check because you don't want anybody just driving that truck. So it's very complicated and it's a challenge and you want your best people doing it and we do. And if I may, another factor of resilience is rapid recovery. So it's the capacity to get things up and running again quickly because you don't want it to take like you were mentioning, like days or weeks to get your power back. That's totally not acceptable here. I understand that the tolerance of people to have outages or lack of things differ depending on where you live. And like I mentioned, is we've become accustomed to having a reliable power and ready supply of fuel and gas at any time, which is a good thing. I really think it really helps to have that certainty. But really to understand how to keep it going, you have to learn lessons from outages or disruptions that have occurred in the past. So what kind of lessons can we learn from Hurricane Lane or Kilauea? There's a lot in every event, there are lessons to be learned about energy supply and distribution. I think it's just learning from them and bringing that knowledge into your organization. And over time, everybody becomes a lot better at managing any disruption once it happens. So as you mentioned, for example, how do you do learning lessons?  It's not just during hurricane season or when there's a tropical storm crossing 140 West that you start worrying about these things. It's a full-time year-round occupation to look at your systems and your people and to strive to make everything work better for contingencies if we have a major disruption here in the islands. And it's your equipment. Things like, when was the last time you ran that backup diesel generator? And has the fuel been filtered or has it got a whole bunch of ickies and algae growing in your filter so that when you really need it, man, it's not there because I didn't learn the lesson. So like Hurricane Sandy, not one backup generator worked during that storm, except the fuel cell systems, they all worked. Yeah, they're great for a short time, but I mean, if you have a prolonged outage, a lot of times these generators fail. They're not designed to operate under load for a prolonged period of time. They're temporary. They're really for short interruptions, short disruptions. So it's your organization that helps us document the lessons learned and manages this whole thing. I mean, it's a huge job to manage all this and you're waiting for the big storm. Sometimes they don't happen. Right, I mean, there's a whole structure. You know, the national response framework, if we could see the next slide. You know, this is doctrine from the federal government down with FEMA and how the emergency management is organized nationally. And so the national guidance is organized around what are considered community lifelines. You'll see here, you probably can't see it, but it's things like, you know, food, water, shelter, health and medical, communications, transportation and energy as being in the center there. And these are all linked. You'll see that there's a lot of interdependencies between the main lifeline systems. And so what a lifeline does is it enables the continuous operation of government functions and private sector operations. So you'll see that if you have power, you can have communications. If you don't have communications, it might disrupt other things down, like health and safety issues. So all of the lifelines work together in order to seamlessly keep a, you know, continuous operation of, you know, even if it's under an emergency situation to keep up human health and safety and economic security. So it's a challenge that is not laid out in every organization's mission statement. So FEMA or in the case of the state, the Hawaii Emergency Management Agency, which are the agencies in charge of the state response, which are the agencies in charge of the state response. Organizes around what are called emergency support functions. And these are just groupings of agencies, whether they're public agencies, private sector companies. And they come together in times of a disaster to coordinate and communicate about situational awareness, resource needs, any, just, you know, any kind of emergency response. And so we have a lot of, you know, emergency response and things like that. So we have what is at the state level called emergency support function, 12 energy. And the energy office, the Hawaii state energy office is the lead primary agency for that emergency support function. Right. So where is, where is civil defense fit into this? Like at the county level, they all have civil defense organizations. So what's the difference between you and the civil defense organizations at the county level. The only county that's called civil defense is Hawaii County. Hawaii County has kept the old name of Hawaii civil defense. But now Hawaii Maui and Oahu, Honolulu city and county are called emergency management agencies because it really gets away from the cold war era about. You know, a nuclear attack and it really looks at. The kind of all hazards approach. And so all hazards is everything from hurricane tsunami floods, fires, pandemics, you know, radiological events, all, all these kinds of things are really like an all hazards approach. And those fall under. Really what's more correctly called emergency management. So. And in real emergency management is a mixture of art and science. Because you have. At the, at the heart of it, it really is about a unifying. Of resources and capabilities coming together. To. To deploy those resources to a disruptive event. Whatever that disruptive event may be. Where's the military fit into this because they're really good at, you know, organizing things like emergencies and jumping in, whether it's the national guard or like the, you know, like, or the regular, regular forces. Right. So the way that it's structured is that.  The, the lead agency for coordination of federal government resources, which includes. At certain times, Defend DoD assets and capabilities. Okay. They do not lead the show. It's basically. all those acronyms and they fall under these ESFs. And there's a structure that parallels with the state, for example, ESF 12, at the federal level, that's the US Department of Energy. And at the state level, it's the Hawaii State Energy Office as the primary. But under both of those agencies, there are support agencies that help. And so, you look at how the DOD interacts is, for example, I'll give you a quick example. All disasters are local. So it starts off at the county level. A county says we need help with clearing this road because it's been blocked and there's debris. And so the county looks at their capabilities and their resources and what's on Island to address that issue. If they can't do it with their available resources, they turn around and they make a request for assistance to the state based on the fact that they don't have the capabilities, adequate resources to address that. So the request goes from the county up to the state and then the state looks at it and says, okay, we have some additional trucks, we have personnel, we have some contracts that can help with that restoration. So you see that it's all addressed by at the local level. They're the ones that are really in charge of addressing that emergency. And the state is like a coordinating body at the higher level to address those state resources. If the state says, hey, we don't have that capability and it's a life safety issue and we need assistance, where else are they gonna go? They're gonna send a request up to FEMA. And so you find that it goes to FEMA, FEMA looks at their resources as an agency. FEMA doesn't have very much really in terms of their resources. So they reach out to other federal agencies, for example, with the pandemic, HHS or other sector-specific agencies that have expertise and capabilities in that need, in that lifeline. And then they ask them to help through the state to address the county issue. Again, how DOD gets involved is, for example, I don't wanna get too complicated here, but it is very complicated on how these things operate. There is something called the Defense Support of Civil Authorities and so FEMA can ask DOD, hey, you can help the state. And then they deploy, it could be anything from military, guard assets, it could be a number of capabilities that the federal, the DOD has. I think though the one thing they realize is that there's this structure and these organizational linkages that make emergency management possible in the first place, without this kind of framework and operations plans and people working together in training and exercises and updating plans and doing all this stuff, you cannot expect an effective or timely emergency response. And so for us in energy, that means that we have to stay abreast of what's going on in industry. What's the situation in terms of how many outages? What's the fuel situation? Is there any damage to the energy infrastructure and quickly identify what those needs are, whether it's in the public sector or in the private sector, and then really problem solve on how we go about restoring that capability. So that brings us up to the next slide. How do we keep the energy flowing? I mean, we've touched on it in a way, but this is like a more direct question. Seeing as how- Right, I mean, I just really break it down into responding. And then also what we haven't talked about very much is the prevention and the protection. So it's mitigating risk. So it's the stuff that goes on in pre-planning. How do you mitigate vulnerabilities or how do you, I can mention about resiliency, how do you make the system more robust? And so what we do also is to try to identify and focus resources on prioritizing whether it's infrastructure or if it's policy issues or if it's workforce development to make a more secure, reliable and resilient energy infrastructure. So I'll give you an example. For example, you may have heard that there is a new FEMA program called BRIC, which is Building Resilient Infrastructure and Communities. And so FEMA has just let out a grant period for people to make applications toward getting federal monies to strengthen infrastructure. Right. And so a lot of the focus is on energy issues, and it's not just temporary emergency generators. It's hardening facilities. It's looking at micro grids, options for micro grids, energy storage options. And you mentioned things like talking about resilient communities and critical customer hubs where we're an isolated community. The members could go there and get power. And you talked about hydrogen. If you want to mention that, you know, an option for, yeah, no, there's alternative fuel vehicles, whether they're powered by electricity or LPG or hydrogen that could support emergency response activities. Well, I always got to get my plug in for my hydrogen buses. They're going to have export power units on it. So all your buses become mobile backup power units. So I won't bang on about it because I do it every show, every chance I get. So just looking through some of your slides, let's talk about emergency support functions. And we only have about a minute, about two minutes to go. Okay, then if it's only two minutes to go, why don't we just jump to the end because I really want to plug what we can get out of this about energy resiliency and emergency response. I really think that what the public should know is that the coordination that it takes to keep energy flowing is, it's complicated complex. And though the really critical thing is when we do have a disruption that people act in a responsible fashion. I think in a way, we want to conserve energy for critical services, for fuel for ambulances, fuel for trucks that move debris.  So I just caution people that think about energy usage when there's a disruption and to conserve energy because the more we conserve, the more they'll be available for providing for that emergency response, those emergency response functions. And I think that kind of coordination with the public, private sector and government to help the most vulnerable in the community that are impacted by a disruption, whether it's energy, whether it's a pandemic, whether it's flooding or fire, earthquakes, tsunami, emergency management pools all those resources together to make a more unified and efficient response. Well, Mark, that's, believe it or not, that's the end of our time. So I'd really like to thank you, A, for coming on the show, but more importantly, I'd like to thank you for the services you've provided. You're like one of these unsung heroes that works behind the scenes. And then when the emergency hits, you guys and your organization is there to help us all. And so for that, we're all very, very grateful. Yeah, no, on behalf of the Hawaii State Energy Office, really appreciate you inviting me on to talk this subject. I'll come back if you wanna give me a call. Yeah, let's do that. So with that, it's like Aloha everyone and that completes Hawaii, the State of Clean Energy for this Wednesday. All right, thank you, Mitch.


So, thank you for the introduction and good morning everyone. My name is Ni, I will present our work. This is the new Private Set Intercession Protocol from our SPARS OT Tensions. And this work with Ben Ni and Avishe from Bayer Island University and Mike from Oregon State University. So maybe you attended another PSI talk a few days ago and might know what is PSI, but I also want to go through a simple scenario of PSI. So here we have two parties, Alice and Bob, each has a set of items and now they want to compute the intersection of their set on the way that they don't want to leave any information except for the interactions. So PSI has many applications and my favorite one is private contact recovery. In this scenario you have, we have Alice with the phone and a list of her friends. On the other side we have Bob who is the Skype server provider and he has a list of users. And now Alice wants to find out which of her friends use Skype so that she can chat with them. Obliviously, we want to compute the intersection of their database without revealing anything. So they have many and many PSI applications like DNA testing, online advertising, or botnet detection. So before going into our protocol, I would like to discuss about communication complexity of PSI from General Shumpsons. He one-way function and OT hybrid. So the naive solution, you need to compare every item of X to every item of Y. So clearly the complexity is N squared and later several works use hash scheme to map the item into bin and now compute PSI bin by bin, introduce the cost to N log N. A few years ago with Google hashing, the communication cost is upper linear, little omega N, it's still not linear. And this works is the first linear communication PSI protocol from the General Shumpsons, I mean one-way function and OT hybrid. And the work of four novel and Ostrich keys is the concurrent work. So I will discuss about the comparison in more detail later. But for now here is the outline of my talk. First our protocol is based on a P version first. So I will present its functionality and how we use it for PSI. And then I will present our past OT PSI constructions and its experimental numbers. So for simplicity I want to present the random OT where Alice input is just one bit and she receives the OT value which corresponds to her choice. On the other side we have Bob, he receives two OT values from the protocol. And the important thing is that if Bob doesn't know Alice, Bob doesn't know Alice choice bits and Alice doesn't know another OT that Bob has. So this is the functionality of random OT. And now I want to show you how we use it for PSI but it's not a correct protocol that we want to use. So this one is for the PSI with a small domain and the basic idea is that the party will run as many OT instances at the size of the domain. So it means that here we have domain A so we have eight random OTs. And for example if Alice has number one, number three and number eight and indeed her input to the OT depends on her set. So with the domain, so look at first random OT, you can see that Alice has number one. So her input to this first OT is a big one and she receives the M11 corresponding to her choice. On the other side Bob receives two random value OT. So now for the second OT because number two isn't in her set. So her choice bit to the second OT is zero and she got M20. The same thing for number three with the choice bits is one so on and so forth. So now if Bob has number two, number three and number four. So Bob only look at the blue values M21, M31 and M41. And the next step if Bob will permute the value and then send this value to Alice. So Alice also only look at her red value and now there's the comparisons. And you can see that M31 show up in two sets. So number three is the interactions. So this is the just example for PSI using random OT but they have two problems. Only each OT needs communication. And the second one you need to use the OT for every item in the domain. And this one is impractical for the big domain. So how to handle these problems? So I will go to our, I will tell you our solutions using our SPARS OT. But before that I want to like tell you about OT tensions, which is first proposed by Beaver already two decades ago. So the OT, OT is very expensive if you've seen public keys. But now you can have a small number of public keys OT and then extended it to get many OTs as you want using symmetric keys. So that one is very efficient in terms of computations. But you still need to pay 128 bit per each OT. And I also want to go a little bit more detail about how OT tension works. So first party run a small number of base random OT and this one you put the key. And each party gets some random output that they use as a seed. And the next step the party will locally extend their seed to many values through PRG. And the length of the total length of the expansion is the total number of OT instances that they want. So now looking at the OT instant I, Alice somehow can compute her OT output. However to help Bob, our book hits OT values. Alice has to send the OT correction values, which is the value PI here, which looks random to Bob. So having PI Bob now can compute MI0 and MI1. So this is how OT tension works. And now for example, if Alice has number 3, number 7 and number i, she only cares about the red value here, number 3 and number 7 and number i. Similarly, if Bob has number 5, number 7 and i, he only cares about the blue values. So if you use the current OT tensions, you have to send all the PI values for every item i in the domain. And this is expensive, so we want to avoid it. So we have two questions right now. The first question is how to send PI without revealing i. And the second question is we need to generate many OT, many number of OT, but we only care some of them. So we have a new idea to avoid sending the HEAL metric. So Alice puts all of her PI values into the polynomial. So for example, this polynomial goes through the point 3 and P3, where 3 is the in her set. Similarly, 7 and P7 and i and PI. Alice sends the polynomial to Bob. So it's easy to see that if Alice has n items, this polynomial has n coefficients. So it means that sending the polynomial is equivalent to sending n values. So now having the polynomial Bob interpolate that one at the point P to get P5, at the point 7 to get the P7, the OT correction value and i and PI. And the main observation is that if both parties had the same items, first number number 7. So Bob will get the correct correction OT values that Alice has. So with P5, Bob can compute his OT values, M51 and M50 and M51. With P7, he can compute M71 and the same thing for MI. And now on the other side, Alice can compute M31, M71 and MI1. So the parties compare the OT values in the clear. And you can see that both parties have M71 and MY1. So it means they are in the intersections. So we are done with the first questions using the polynomial. And now we are going to the second questions. So recall that both parties have to use PRGs to send their seed to many values. However, we only care some of them. So very simple solution is we replace PRG with the PIF. And we have several tricks to compute this PIF efficiently. But you can see the people how to do it. So we are done with our protocol. For the security, the view of Bob is the polynomial over the correction OT values. And Alice, you heard inputs to interpolate this polynomial. You can see S1, S2, and S3 here. However, because all the PXE randoms. So this polynomial looks random to Bob. Therefore it hides the Alice input set. For the view of Bob, also for the view of Alice. I think the item Y isn't in her set. So Alice never interpolates over this point. You can see Y and PY, the correction OT value PY here. But the point she interpolates is something like here. So we show that if the capital P and small p e4 in the meaning of hamming distance. So the size of the view size should be around 3.5 security parameters. And now the OT magic and Y1 looks random to Alice. So the proof is a bit complicated. So just see the paper for more detail. And the bottleneck of our construction is the polynomial operations. The reason is, you can see, it has the heel degree and the heel size is very big. So similar to previous works, we use hash to be scheme to map the items into the bin. So we have many small degrees polynomials and one per bin. So it's very efficient to interpolation. However, we need to pet the dummy item to hide the actual bin size. So this one increase the communication cost. So we use a specific new hash scheme so that the number of dummy items is only 2% of the set size. So for the comparison. So our protocol, we have two protocol variants. The first one have the big polynomial and the second one is the small polynomial with the hash scheme. So for the first one, we send the big polynomial, which is equivalent to sending n value. So here we have n value to send and n value to receive. So the communication cost is about 500 bits per item. For the second variant, because we use the hash scheme, so they have 2% in number of dummy items. And we use two hash functions. So we have 1.0n sent and 2n received. And the communication cost is about 600 bits per item. Compared to OT-based PSI, their protocol somehow needs to use about 1.5n of private membership. And in which 50% is the dummy items. And they need to use 3 hash functions. So it means they need 1.5n sent and 3n received. Their communication cost is about 500, or about 1,100 per item, which is 2 times more than our protocol. So for Delfi-Helman-based PSI, they need n sent and 2n received. You can see the protocol finger. And their communication cost is about 576 bits per item, which is still more than our protocol. And we are the best, our protocol has the best communication cost. So in summarize, compared to Delfi-Helman-based PSI, our protocol is 10 to 20 times less communications. And we have 2 to 7 times faster in the term of training time. Compared to OT-based PSI, our protocol is 4 to 40 to 50 times less communications. 6 times slower in LAN setting, but 2 times faster in one setting. But our protocol is the cheapest in the measurement of computation and communication cost. And we use Amazon Web Service price for this one. And our protocol is the first linear communication protocol in OT-hybrid and one-way half functions. And the work of four, Noble and Ostrowski is the concurrent work. So if we put our two variants in the computation and communication graph, one variant is here with the loud communications. And another variant with the fast in term of training time. And compared to our previous work, our protocol is cheapest one. So for the LAN setting, we are in this zone. And for the warm setting, we are in the dark zone. So for the future work, you can see that the bottleneck of our protocol is the polynomial operational term. So any alternative solution for that one? And this talk is for semi-honest setting. So can we have a linear communication PSI, but for malicious setting? And the last question pointed out by a reviewer that can we use our SPA OT as the multi-query OPF? Yeah, it's done with my talk. Thank you. Any questions? If you want to further compute on the intersection, is it easy to generalize? Yeah, so right now we send all the OT value in the clear. So this one allows the receiver check whether the, that's one review the intersections, right? So I don't know, I mean, maybe you have another way to send the polynomial. So the party can check the intersection, intersection in some circuit or whatever. So the party will learn the share of the bit, whether they are in the intersection or not. Yeah, thank you. Any other questions? Okay, let's thank the speaker again. Yeah, thank you.


Good morning, so we were discussing protein ligand and protein-protein interactions using NMR spectroscopy. So in the last class we started to understand what are the NMR parameters that can be used for understanding the protein-protein or protein-ligand interaction. We started with simple analysis of line width, the line shape, the intensity and the resonance frequency which can gives the idea about the protein-ligand interaction. Then we went ahead and we dissected the protein-protein-protein-ligand interaction into two parts where we are detecting first on ligand and then we can go and understand the detecting on proteins. So on the ligand detected experiment I started with STDNMR, saturation transfer difference where basically you can find it out binders it is a high throughput NMR techniques for fishing out the binders of a particular protein and then I discussed the line width how the ligand can have a sharper line upon binding the line width can increase so that itself can tell about which are the binders if they are binding to a protein or not. Then also like we discussed what NMR can be like STDNMR can be used for so it is a high throughput techniques. So we are going to discuss little more detail how STD can use for finding it out the binders and quantitative parameters of the binders. So here we are directly observing the ligand that is binding to membrane protein in a living cell that is what we are going to do is advanced version of STDNMR called STDD. Saturation transfer double difference right so double difference we are going to do. Discuss now STD we understand to just to make you more familiar in STDNMR what we are doing. So here is a binder here is a protein they are binding binder like a small molecule ligand has its own resonance protein will have its own resonance. We are going to saturate the signal exclusively from protein if this guy is binding the small molecule is binding the effect of that saturation will be transferred to the ligand and when we take a difference spectrum when it is not saturated or saturated far away and saturated on protein when we take a difference the epitopes that binds to the protein will be illuminated that is the STDNMR saturation transfer difference. Here we are going to now discuss the saturation transfer double difference right so what are the double difference and how you can be used for understanding the intact cell protein. So like you know like as a biologist you know the all protein cannot be purified right and purified to high concentration and all those. There are various intrinsic problem with protein purification and instability. Now still there are some proteins which cannot be taken in solution but they can be taken in a in a in a membranic environment but again the reconstitution and all those are going to be really really tough right. But those are main targets like one of the example is GPCR G protein coupled receptor they are the like a major target for drugs because they are receptors for various molecules. So about 30 percent of protein are membrane protein membrane bound protein or integral membrane protein one of them is GPCR and these are important drug targets. So if you cannot understand their binder then we are losing out lots of things so can we come up can NMR can come up with a technique where we do not need to purify protein but still you can investigate what is the binder how we can investigate that was again offered by the mayor group and that was published about 15, 17 years ago in Jax this technique called STDD and STDD saturation transfer double difference right. So now we are not going to isolate protein we are taking them in their natural membrane environment and try to understand how they are binding. So NMR has to characterize at an atomic level the binding interaction of a cell surface protein in a living cell and that is implemented as this double difference technique. So how we are going to do? Here is our whole cell right and here is a receptor and in this receptor some binders are there right. So they are going to bind here so binding reaction is K on versus K off right. So these are the binders here is a free ligand and upon it is binding. Now we are going to selectively saturate the receptor so receptor concentration of course will be very less so we have to choose a region typically methyl region where these receptors only gives the signal and we can even increase the saturation duration such by increasing the duration of this pulse or strength of the pulse and then when they bind the effect of that binding like effect of this binding can be eliminated by saturating this receptor pulse. So if receptor signal are saturated it will transfer to the binder which is small molecules and that we are going to elucidate it in the STDD NMR. So typically experiment is done like this we have a cell suspension and we have cell suspension we are taking a append of sample A will contain cells, cells and ligand and in sample B we will have only cell right. So this is the spectrum we are recording the spectrum of A where we have a protein signal like our cell signal and the ligand signal so cell and ligand is here a spectrum of D where we are doing STD of cell only and then we are like a then we are going to take a difference STDD. So here we have saturated the cell look at the effect of those cells saturation on ligand here just we saturated the cell and then we took the difference of these two. So now if you look at there are some things getting eliminated if that is getting eliminated it says that there is a binder which is which has bound and the signal the sharp signal that is coming here are from those binders the ligand that bound. So this is the saturation transfer difference to emphasize again we have taken a cell suspension split it into two in one we have added the ligand in another we have not added the ligand. In the first sample we did a STDD NMR like saturated the protein cell signal or protein signal and then without saturation we also did that we recorded the STD NMR for first one similarly we recorded the STD NMR for the second one we took the difference of those two so spectrum first minus spectrum two the illuminated signals are coming from this double difference STDD and that is telling that ok these are here as the ligands that is binding. So even without purifying protein taking from cell suspension we can find it out the ligand that can bind to particular receptor right. So that is the STDD NMR. Now I will briefly discuss to you group epitope mapping by saturation transfer difference. So group epitope like I was talking about the epitopes that binds. So basically what we are going to do is identify a segment of ligand that directly interact with the protein or receptor right. So the same concept little bit trick we have to do same concept of the STD suppose this is my ligand and this is the receptor. Now receptor is a bigger molecule ligand is a smaller molecule we are again saturating on the receptor molecules and we are looking at the effect of that saturation on the small molecules. So receptor and protein are in exchange KON and KOFF right. So here is a free ligand and here is a receptor ligand so receptor protein complexes. So we saturated essentially the receptor. Now in this experiment we are going to increase the saturation time. So the closest one will first enumerated once we increase the saturation time the now the second one the second one which is closer here the third one this is the third this is second and this is the first. So now not only we find it out which is actually interacting but also find it out how far they are from the receptor and that is called actually the group epitope mapping. So by increasing the saturation time recording the various sets of STD enumer we can find it out how where they are positioned where they are positioned in the receptor binding site. So typically these are the pulse sequence so to have a simple explanation of these for like a here you have a for D2O sample you can have pulse of saturation.  So during this like all the signal coming from the other interactions will be refaced and essentially we are seeing the saturation effect for D2O if you are doing in water we need to also saturate the kill the water signal so this is done by one of the water suppression technique that we have discussed earlier. So and remaining will be suppressed by the gradient so use of this selective water saturation pulse with gradient we can kill the water and rest remains as it is like STD. Saturating on protein signal detecting on the ligand signal and we have to do on resonance and off resonance in the interleaved manner right. So what is on resonance like when we are saturating the protein signal exclusively off resonance when we are not saturating protein signal so our irradiation is somewhere far right 20 degree 20 ppm or so the T1 row here we have added again to remind you it is for filtering out or removing the protein signal that might come in this. So T1 filter ensures that we are only detecting the ligand and effect of the saturation on the ligand that is what actually it ensures. So this is typical pulse sequence that we have for D2O sample and for water clean is important so we kill water using this 3919 pulse right. So here like for an example here we have only protein you can see really really broad signal so we irradiated the protein at minus 2 ppm that only protein can have. Now if you add this T1 filter you can see now the lines becomes really sharper right so you only we are getting few peaks and one day with T1 row filter you can see the lines are really coming out. Therefore if we are studying the complex system like this just like here we probably need to put this T1 row filter that defages the protein signal just filters out the protein signal and we can get a sharp line which is shown here. So protein here irradiating protein at minus 2 t 1 D with T1 row filter you can see already we are seeing sharp signal and 1 D with T1 row filter you can really get a beautiful sharp spectrum coming from the interactions right. So next is like we find it out epitope we also find it out the position of different epitope how closer or how far they are this the how high position is important because now as we discussed your medicinal chemistry friend can come and start modifying it doing the molecular like a jugglery just to increase the binders increase the size of binding so that we can tune it tune the binding property. So that is helped by STD and STDD and group epitope mapping. Now next is can we find it out because that is what we are saying can we find it out KD of the binding yes we can do how this is the simple experiment. So now this is the simple experiment that we are going we are increasing now ligand concentration and ligand concentration can be increased and then STD amplification factor how many times we can get the more STD signal that can be detected. So with concentration ligand and STD amplification factor we can find it out the KD of this so how STD amplification factor helps so we know the I0 that without anything when I sat when we saturated and we divide by the I0 multiply with the ligand excess how many times ligand was excess using these we can fit this curve and that gives us the KD dissociation constant. So STD amplification factor it is possible to quantify the active ligand concentration which allow to estimation amount of protein needed for STD experiment and it is also a specific of the ligand which has STD amplification factor of 10 or so so using some of these techniques we can probably get kind of the STD amplification factor how many times STD and this also opens an avenue on finding it out the binding strength. So here one can find it out with STD amplification factor like we discussed these guys we discussed this binding so how far or how close they are what is the STD amplification factor like this molecule was binding so we did various ligand concentration of STD ligand to protein ratio we put it the how the signal is getting saturated of different atoms H1, H7, H5, H4 and methyl we plotted the signal intensity with respect to ligand concentration and we calculated the STD amplification factor while they binding to HSA you can fit it STD amplification factor alpha times of the STD ligand concentration and one can determine KD. So essentially for each of these moiety using STD amplification factor we can find it out one of the idea we can have the idea of a KD how strongly or how weakly they are binding. So this simple experiment helps us in getting the signal. Now can we also go advance version of the STD right what I mean STD were just basically the 1D experiment where we were recording 1D can we combine this with a 2D we can get even more information. So yes STD can be combined with Cycose or even Toxy like we can get even in 2D fashion so that even our ligand becomes bigger we can get some information. So here is one example when we have combined one can combine STD with a Toxy. Now you can see here different peaks you can identify in Toxy region and since we have done STD so you have to record 2 experiment Toxy without irradiation Toxy with irradiation or Toxy on resonance irradiation Toxy of resonance irradiation and take the spectrum you plot with a various ligand concentration calculate the STD amplification factor and one can see for various epitopes the STD amplification factor can be different and you can again the map the group epitope very nicely and also possibly you can get some idea about the KD of this right great. So that is the extension of STD NMR saturation transfer difference NMR. Now just to again revise you a little bit here is my receptor molecule here is ligand molecule that binds this is non-ligand which is not binding they are like in equilibrium K of K on binding non-binding so binder binds and non-binder does not bind. So we have a KD ratio K of by K on or you can write this K on and this you can write as a K of this will be K on by K of right and we have selectively saturated the protein in the STD fashion we took the difference where there is no saturation here there is a saturation so some peaks have lower intensity so this is no saturation called off resonance and saturation called on resonance took the difference here is a difference spectrum. So now we are getting the signal from the epitopes that binds so that is a STD NMR. So now to sum up a ligand based NMR experiment screen to determine in a quantitative manner which compound binds to protein in the context of drug discovery we can map the ligand a more advanced example to use the NMR for direct characterization of protein inter like a protein ligand interactions at the molecular level through the identification of important ligand commodity one can find it out and one can determine the dissociation constant between the protein and ligand where we are taking the ligand at a various ratio we are plotting the STD amplification factor by fitting it that one can find it out the KD dissociation constant between the protein and ligand that is what we can achieve using various STD experiment. The next technique that can be used is called like a transfer NOE protein ligand complexes that is called transfer NOE. Here again we are mostly detecting on the ligand side so two techniques already I explained you looking at the line width which is determined by the relaxation property the next we talked about STD and it is a variations STDD group epitope mining and how we can use for KD determination. Now the third techniques is transfer NOE this is applied to a system where it is in fast exchange so the ligand and protein is in fast exchange KD is typically 10 to power minus 7 mole and K of rate is like a the T1 inverse here we are going to absorb the NOE because the NOE between the ligand and protein that is a transfer NOE. So we have to collect standard 2D NOG experiment like we can do 2D NOG experiment where ligand is added to the protein molecule and if there is a transfer of signal coming from the protein that is transfer ligand we can detect it. So ligand shows a signal set regions averaged over bound and free state here also ligand should be more like access relative to protein we record a series of NOG experiment and we can find it out. So a strong NOE developed in a complex is transfer to free ligand state and one can measure the free ligand resonances it is a it is even applicable to a mega like a molecular weight which is large kilo Dalton several kilo Dalton. Observe NOE can be used to determine the bound confirmation of ligand like if suppose this is ligand this is protein and we are getting the NOE from the ligand to protein we know that distance. So distance is kind of measured here and that distance fix the orientation of the ligand in the binding pocket and here the sign of NOE cross peak like will be opposite and you can get looking at that this is coming from the transfer NOE. So here we have a ligand which is small molecule protein big molecule we have a long tau strong NOE and when it is in free it will have short tau big NOE. Now when we record the spectrum of protein ligand complex one can see that these peaks which are coming here it is of negative sign and when the positives like when there is a color change positive peak sign negative peak in green. So the sign change of the cross peak indicate the binder and if they are not binding they will not bind. So here one can see here for the these guys the peak sign is changing. So here a protein ligand is happening some of these you can see these are not changing. So if the sign of the peak has changed that means it is binder they are coming from the transfer NOE experiment. So now since we find it out what are the binders you can use this for getting a protein ligand complex. So protein ligand complex can be determined by transfer NOE. Since now we have established the NOE connection between protein and ligand we can fix the orientation of these ligand in the protein complexes and using few of the experiment we can dock it and create a dock model which will tell that how these proteins are binding. So one can illuminate some of these residue that are in context residue from protein that are in context with a ligand that is how you can create a protein ligand complex using transfer NOE experiment. Right. So next experiment that I will be discussing is a water loxie experiment. Water loxie as name suggest it is a water ligand absorbed by a gradient spectroscopy. So here the concept is like there is a protein and there is a ligand but in between there is a lots of water. What kind of water? Some bulk water, some biological water. Biological water which is in vicinity of the protein molecule. So water loxie is used extensively for screening again the protein ligands. It is again a high throughput techniques and it is a reliable method for identifying small molecule. The requirement is that the binding affinity should be from micro molar to millimolar. So what we are going to do here? So assumption is there is a ligand which is binding to protein and in between there are lots of water. So and those are in exchanging. So what are exchanging? Bulk water, bound water is exchanging and then protein when it is binding that is also exchanging with the water present here. So exchange of water that is what its name is. Water ligand exchange that is happening and we are observing using gradient spectroscopy water loxie experiment. So actually it is ligand absorbed and like we are observing the ligand. It is NMR method that absorbs the ligand. It is widely used for study protein small molecule interactions. Here in this case we are irradiating now water. So this irradiated water, what is, so there is a like a nuclear overhouser effect and chemical exchange that is happening with the bound water, the protein and the ligand. So here transfer of magnetism between water molecule with protein and small molecules happens by a nuclear overhouser effect because they are in close proximity. So if they are in exchange and they are in close proximity when we irradiate on water molecule the irradiation can be transferred to protein molecule and then eventually can be transferred to ligand molecule. That is what essentially becomes active. Irradiate water gets transferred to protein, gets transferred to ligand if they are binding and that is how you essentially understand using water loxie experiment. So pulse sequence is essentially simple. So here you excite water and then mix so that the magnetization of irradiation is transferred and finally you suppress water, look at the signal that is coming from the ligand molecule and effect we are seeing on the ligand molecule. Typically we look at the small molecule. So few things you have to take care that pulse length is correct. So if you have a correct pulse length you are basically getting to get a good line shape. If you start with incorrect pulse length you see the distortion phase. So now you can see the sharp line. You can assume that sharp line is coming from the ligand molecule. Traditionally it is like a plotted negative for like when there is no water loxie signal when there is no transfer. So that does not mean that this is giving negative peak. It is plotted like that. Now we do water loxie here. Now you can see that in presence of the protein molecule you see few of these peaks. So in case of BSA you see the C and D. So these are basically with correct and incorrect pulse length again you can see it. So essentially you have to compare these two and you can see now the peaks are coming here and these peaks are coming because the transfer happened. So water molecule in water loxie a small molecule that don't bind to protein molecule does not give a positive NOE signal with water while protein will display the negative NOE because that is how it is plotted. And population that are bound to protein they will display the negative NOE signal. So that is what we are seeing. So now the experiment was done with one molar of tryptophan and yeah in presence of BSA 100 micro molar 1 millimolar of ligand signal 100 micro molar of protein signal experiment was done and NOE was observed on the ligand and this says that they are interacting and that is how you can find using water loxie. So here I will end it with water loxie. So these three four techniques that I discussed where ligand observed the line width detection the effect of T2 the STD NMR and some variation of STD NMR STDD then we looked at the transfer NOE and finally water loxie. So these are the four commonly used techniques that are there when we observe ligand. Now the next class onwards I am going to discuss the protein detected experiment which can be used for understanding the protein ligand protein protein interactions. So here I sum up and I close here I look forward to you with lots of questions if you have any doubt do not hesitate to write to us. I will be happy to answer all your questions for protein ligand interaction where we are detecting on the ligand molecule. Thank you very much see you in the next class.


Lookout World, Lindsay Lohan's back in a brand new film straight to Netflix. Oh. Called Irish Wish.   You can like shit, that's fine, but I didn't. I didn't like this movie at all. It's Irish Wish. Check it out on Netflix, or better yet, don't. I'm doing the service of watching this trash so you can hopefully avoid it, unless this sounds like something right up your alley. No jokes that land. No real action, no real comedy even at all. The romance is entirely superficial and half-baked. The plot, the concept, it's just wash, rinse, repeat, recycle from a million other Hallmark movies. And there you have it, Irish Wish.