I was going to say, I don't know, because this is, uh, maybe use a, uh... Okay, bud. What's wrong? Because this is so, I wonder if, just a grillo pad. You can just rip a corner. Can you refresh that? So, you guys, um, today we're going to be decaling all of our these. Some of them will be a challenge because the molds were on the cheaper side. So there's, like, for example, this one has a dip. And I've never put resin over whatever the quick white Illumilite is. Um, I know it, it wasn't sticking to the paper. So I don't know how well doing my little flood fill will stick or bond. And I don't want it to delaminate. So I'm just scuffing the bottom of the dishes before we get rolling with the decals. Actually, what's essentially going to happen is I'm going to sand and pass it along. Well, it's going to take longer to place the decals than it is for me to scuff the bottoms of these. So I'm just going to sand a stack at a time and then wipe them out with 91% isopropyl alcohol. I am using a 220 grit sandpaper. Um, I could TG, but it's marketed as an epoxy. So I'm sure ultimately this will be fine. The areas that have floating gold like this, I'm not going to sand because floating gold.  It's okay. It's not hot today. You have to stay over here. Okay, right there. So I'm going to sand over here because I don't want him really breathing anything in. What's up everybody? If you're new here, hi, hello.  We go live most every day at 6pm central except for on Tuesdays at 2pm central and on special occasion days. It's a mystery time such as today. Um, there's some interesting weather in town and also we're trying to go watch the hockey game tonight. So we're early. This one has a lot of floating gold in the base. So I'm just going to stand around it.  So for this application, it wouldn't be that detrimental. It would still look classy. But if you're wanting gold to stay gold, then I'm going to go get his fan. So I can blow it the alcohol in another direction. Hi, canvas. So good girl. Get to art. All right. A fan. Yeah. Got to move Austin. I'll move those things back. All right. We're all set up. So we've got his fan. He's up high. Fan is blowing all the smells away and keeping him cool. So what I want to do, I left these two strips to kind of demonstrate how if you have trouble transferring your cut letters, it doesn't really matter what cut it. I use the, what is that? A U.S. cutter. Whatever cutter you use, it doesn't matter. As long as you've got your letters out and your transferred paper. Some people use paper. The others use the roll. I get this stuff at my vinyl plate and it comes in, what, 15 by 36 inches? Three feet. And some, when I first started, I didn't understand how, if you wanted to, if there was a sticker this long, that you would have to take this stuff, pull it off and hopefully press it down and it goes on evenly. I'm sure everybody else does this now, but I just thought, why don't I try to cut a little bit of the protective paper, this stuff here. First of all, let's get however much paper we're going to use. So we have the amount of paper we're going to use.  Get your scissors.  They're, I guess, transferred from, I don't know, I don't know what carrier paper. That's what it is. This is like, you know, so what you're going to do, make sure this sticky side does not touch any of your sticker, on any sticker, on any, whatever you're doing for vinyl like this. So what you're going to do is you're going to hold it. You're going to put this on the, you're going to put this side without the sticky side on the very end of where you're going to be sticking this transfer tape. Now, my letters are just a smidget too big right here, but it's not going to matter because each one of them are going to be individually cut and stuck down. So there's no big deal. Now, if there was a longer part that was sticking out, you'd want to have a little bit more transfer tape. So what you're going to do, you can make sure every side is just perfect. Like I said, it's not going to matter if this is all sticking or if this end isn't sticking because these are just the barely the ends. So we have everything lined up, top, bottom, back, and then the front. So you do just stick that, make sure everything's stuck on there. Pick this up and literally all you're going to do is just take your, take your hand. If you have this type of stuff, if you have more of the paper than you're, you're just going to, you'll probably pull it, but this stuff comes off really easy. And you're just going to pull it in a straight line just like this. And voila, you've got your transfer tape on your stickers. Just like that. And I'll tell you what, I'm not going to do it on these because this is a single one. But let's say you want to put this somewhere. It's basically the same, the same deal. You want to, you want to take this up, make sure everything's stuck.  So that's, you know, you have a little bit of sticky. Wherever you're putting this, that way you can arrange it nice and perfect. And then put this side down, do the same thing. Lift that up and just pull that, pull the, the transfer, the carrier sheet right from under. And it should just go on straight and even. Yeah. A little helpful tip there by, I did all of these at once. Like I cut them out so that I wouldn't be wasting a lot of this transfer tape. I cut them all out and then just kind of lined them up, put that transfer tape and did them all in one time, which was pretty impressive on my behalf, I think. All right. So we're not going to start with any of the S's. I guess we could. It doesn't really matter. There's no order. There's no, like, so what we're going to do is just cut each one of these out. Check it out. Yeah. There's a little scuffy. Some of these are like a little wavy. Oh, I saw my glasses on. I guess some of them are super wavy. So here we go. I guess I should have, I didn't do this on all of it. Do we want to do it like that or like this? Like this? I like that. I like this. People will probably put them like this. Yeah. So try to center it. I don't think it really matters too much. Karen, those color boxes came from our fairy art mother, Judy, but she got them from Sarah Renee Clark.com. And yeah, after I decal them, I'm going to fill them just up to that first rim with resin to seal in the decal. However, I don't know that that's actually necessary. Is she chewing on something in there? No, she's playing with a tree ball. It's just the Cricut Master. Actually, we have a Cricut. Why is there purple on so many of these? I have no clue. I mean, that was, that's the color of that, but I didn't, I don't see it. Okay.  Sand it. I don't know. I shouldn't be any purple on there. I didn't know that sandpaper can transfer. He is the vinyl cutter master, but the cutter we have is a US cutter. Not sponsored, but if you're watching, what's up? So, does anyone have any questions about the decal process? B, do you have any tips, tricks, and otherwise fun facts about using a cutter? I don't. I mean, I wouldn't know what to tell anybody, like, unless, unless they, you know, had a question. All right. It's like, I love my US cutter because of the surface and how big you can cut and how fast and, and all the options. I've learned to use it. I think in a way that it's not like, you know, when you, when you, when you get, when you get something back in the day or now or, and you don't really know how to use it. But you learn, you learn on your own. And then you kind of do stuff where you're like, I wonder if anybody else does this because you've kind of said, you know, you found a cheat code or, or some sort of backdoor little trick that is not like, for instance, when I used to make my shirts or flyers. I used, was it Microsoft Word? No, I use Microsoft Word. I didn't use paint. Like paint was like, I swear you don't need to do this. No, it was, it was, it was word, you know, when they give you the options, all the different colors and the different style of, if you could type it on. I don't know. I just did a bunch of, I know that it had to been unorthodoxed. Whoa. That was a big word. So it's kind of like that. Like you kind of, you just kind of learn on your own and do, do stuff that works for you basically. That's, that's kind of how this big old cutter I have kind of has worked for me. I'm not a very computer savvy person. That's why I never got into Adobe, like Photoshop and whatnot, which I'm sure that's a lot more difficult than a lot of other things, but I just taught myself how to use it. I YouTubeed a couple different things on like how to make a, how to cut out a contour sticker. Which came in real handy. And then you kind of learn to manipulate that.  Look at that. I like this because it looks like it's circled. Did you have to weed that? I did. And I like all these whole things. I just did it all at once. Like I cut them out. It was a 15 by 36 sheet. They were all cut out. And I just, I cut each line out and then weed it that way. Because if I were to try to weed that whole thing at once, it would have been a nightmare. I'm saying they like the V's. The other V's are nice. The R's and the K's are really nice too. What do the letters represent? So these were commissioned for a private party for graduation. And these represent the 90 people that are students that are graduating that are going to be in attendance. And so the letters are the first name of the student. Nope. The letters are the first letter of the first name of each student. It makes me want caramel vanilla ice cream. I don't know how, but this one has a little green dot in it. So if you could do something to cover that would be superb. What would your guess Nancy? So the device should be don't pick a font. The device should be don't pick font. That is really loopy because weeding is a nightmare. That is 100% true. And not only being loopy, but putting it on here is tricky. Because these are kind of like domed a little bit just because of the mold. So you want a thinner letter.  It probably wouldn't have worked as well. Like it definitely would have. The letters would have gapped and folded. They would have been creased or not laying down flush. Is it coed school? I'm sure. There are 90 of these. Reverse weed, I don't even know what that means. That's above my favorite. Taking out the letter and making it square. Leaving a negative space basically. Which? A stencil. Yeah. You wouldn't be able to do that with these. I guess you could do it like this. It's strange that there are so many V's. There are some letters that I would have thought there would be at least one of 90. There's no T's. There's no T's. There was another letter that I was like, no way. Have any of you guys done assembly line artwork? Projects, crafts. Getting ready for an art show festival. Right TG, no T's. Super weird. There was another letter that I was like, are we sure? Two times. It's starting to stall on my side. Was it G? No, there's a couple G's. Wouldn't that have been crazy if T and G were left out? This is a little bit easier because I'm not worried about these things sticking together because some of the transfer paper overlaps. We do cut them out. And it's a little easier. Not being so messy. Must be last names. Could be. Yeah, it would have to be last names because this is just strange. So many N's. There's 23 A's. There's like a little bit of green in this one too. There we go. You can almost not even see it because I singled it. I say because of all the V's. There's a lot of Veronica's at this school. Which is possible. I mean, Mark Cuban is going to be there with his family because his daughter is in the same graduating class. What does that have to do with these? Because Veronica is like a hoity-toity name. Hoity-toity. Yeah, hoity-toity. By the way, I have no problem with the hoity-toity thing. Because all of the V names I can think about in my brain are like picturing somebody with a glass of champagne. Veronica's drink champagne. Vanessa's drink champagne. Vivian's drink champagne. And that's like all the V names I can think of off the top of my head. Victor. Yeah. Victor's drinking. Veronica Val. Oh yeah, Valerie. Vernon. Vernon is not a name anymore. I don't know. I think that's like only grandparents' names. Yes, I graduated high school in a town called Vernon. Hi, Jerry. Susan's drink champagne is the same. Same. Will you put a little sparkle on the top coat? I am not sure. Only because when you add abalone, milky way, diamond dust, it can kind of dull or cloud black. This one's coming up. So the answer is maybe. How long did it take you to make all of them? Actually, it should have taken me a long time. But Illumilite sent me some stuff called Illumilite White. And it was like in the nick of time. Like I didn't even know I needed it. I took this gig not knowing it was a thing. And then I got it and each round of six coasters took like 10 minutes to pour and demold. So it didn't take long. I think it took six hours. The school has an independent secular college preparatory day school for girls. Bam.  She goes to. All right. She does the Google cost 30,000 to 40,000 a year. Yeah. So our kid won't be going there. At least not the first year. I don't know if you guys want to sit here and watch me do. This is all of these. We're so boring. It is boring and tedious. So probably once I get all these sanded, I'll take you guys in the other room and we will pour a couple of the bases so I can show you guys that process and what products I'm using to do that. Yeah, I do. For sure. I heard you mentioned that one was lifting here. My two cents after. Done before resin brush a little poly acrylic on top of the resin. So it was only coming up because he just didn't press it down. But that's a good tip. You're all just trying to add to the. I don't think I have any poly in the house anyways. So what's your favorite letter that we've done so far? Still the V's. We've been on M for a while. So they're probably over the M. How many M's are there? 15, I want to say. Yeah, 15 M's. That's the most letter. Oh, no. There's 23 A's. Is there a trick to getting it center? No. It's just the corners are, you know, I'm hoping that it's centered. But I don't, I think I probably maybe a little bit off a couple, but it's not that noticeable because these letters are so loopy-loo and they're little sticky outies and what not. Brilliant, Karen, thank you. What's up, Lori? Better late than never. I agree. Eyeball. You guys saw an eye in that one? Any time a siren just starts outside of our window, which is not rare because of where we live, makes me jump just a little bit. Lava here coming, no problem. I've only got like 15 more to send and then we'll go start laying out resin. Oh, good eyes, key to center. Yeah. Yeah, yeah, yeah. I'm glad you guys said that because I would have been searching the M's to see which one had an eyeball. Well, these look like T's because they are not T's. I wanted to make the letters a little bit bigger, but some letters are a lot bigger than other letters like the M has that little arch and the K, one of those letters that like the G, the K, the R, they're like, you know, they come down a little bit more and I didn't want to risk making the regular letters as big as I can and then those letters not fit. So I just made them all a little bit smaller than the largest. Yeah, Brenda. It's like getting here. I'm sure it will be before too long. I don't think that it's going to, I think it'll rain. I think it will get some rain, but I don't think it's going to be bad. Like I said, it kind of goes around. It kind of builds and then it goes like southeast and then it gets bad up that way. Looks perfect the way it is. L for my granddaughter, let's see. That's my niece's name. She's my baby niece. Ugh. You had hail in Florida. Wow. I saw that. I saw so many posts like, what is up with Florida? Last act of sand. Lovely. Loving this. What's up, Shannon? Anybody from Minnesota in the room? Well, we're happy you're here, Brenda. Gary the wind was serious. Yes.  Alexis, stage one. Let's go. No. Hey. Alexis and Chevelle. No. She's going to be friends with Chevelle. Yeah. Easy. I have no idea. L for love. I don't know. It's from that movie, The Guy and the Thing. Oh, The Guy and the Thing. Yeah. Dodgeball. They were at some tournament.  Yeah. Because I don't know what that means, love. L for love. Yeah, L for love. I get these wiped out. I'm going to go make sure all the lights are on and everything is prepared. Come get everybody to go over and under. We'll start adding resin.  I don't like all that. I won't be answering any questions.  So we got anybody new? Any first-time watcher? First-time chatter? Long-time watcher? Anybody new from a new state? A new country? Anybody from any country outside of the U.S. or Australia? And high Australia? Yeah, I don't think we got any Minnesotaites in here. TG. And if you guys notice on Facebook, all the people that are coming out now like doing floors, like, hey, we haven't even done a floor. Who's just starting out doing floors? Like, you just can't pour resin on a floor and it'd be good, like, in the long run. People are all trying to be professional now. Like, maybe I'm just jelly because I've never done a floor. But I would love to do a floor. I just don't know how to prep a floor. I know there's a lot that goes in a floor. Like, you know, concrete sweats and doesn't want stuff to stick to it. You know what I'm saying? Other GFs is favorite initials are art. My son's initials are art. And yes, I meant that to happen. Speaking, I wonder if he's going to go watch the hockey game with us tonight. That would be really awesome. The kid's going to be 30 this year. Get out of town. Like, I don't ever see him. He's always up in, like, Plano or past Plano or some crazy town outside of Dallas. I'll epoxy everything. Yeah. I would love to do a floor, man. That would be so much fun. This is a fun one to take a lot of a lot of white and then just a teeny bit of gold coming through there. What if we turned all these into them and they were like, not anyone with a W here? Because that they'd look at the end of the second. Yeah. I'm always like, I'm chilling. I'm going to do the bathroom outdoors. That's cool. Yeah. One lonely eye. Is that a J? Why did I think those were the eyes? This is just a regular hole.  The H's are fun. The G's are fun. I am hungry, y'all. I need some food. G's are nice. My Aunt Geneva would like this G. You know what's crazy?  I'm thinking of my son. His name is Austin. He lives just outside of Dallas. He's a sports guy, like big into sports. Anyways, he was born on his aunt's birthday. And the funny story with that, because it's funny, I was born on my aunt's birthday. I know y'all wanted to know that, so that's why I told you. No big deal. That Austin was born on his aunt's birthday and I was born on my birthday? How did they put all of these letters on such a long vinyl piece? That's the good thing about using the U.S. cutter. You can cut up to like, I can cut up. It says 24, 26 inches, 28 inches, but the cutter doesn't go the distance, you know? So it's technically about 24 inches. And up to, you know, 5 feet, 6 feet I think. I haven't cut anything that big, but you can put a roll of paper in there and bada bing. I want to do beach floor so bad. All right, I wish Mike would do one of his bathrooms up there. We come out and do it. That would be so fun in Hawaii. Can you imagine an entire bathroom floor just like a beach? Paint a shark underneath there. Stingray. A turtle coming up. Like, that would be so fun. And I've only seen one beach floor that was worth anything, if we're honest. Like, I've only seen one that I was like, okay, you're awesome. You know what's funny? Every time I start to put these letters on and I know that it's like, I do it like this. I see vamp, like just ready to be like, it's crooked.   She's laughing. What's up Megan? A Thomas Kincaid theme of what? Does anyone in this room? Okay, so there's, oh yeah, that would be good Megan. So if you put 30 people in a room, at least two of them will have the same birthdays. Who in here has a December 2nd birthday? There are 39 people in here. That's true Susan. What is Dev doing with all these bowls? So these are- I like cereal. Dev eats a lot of cereal. I don't anymore. I like totals. Therefore a graduation party and therefore all the girls will drink it dish souvenirs party favor to take with. So apparently it's not that expensive to do floors. I thought it'd be like super expensive, but you can like day rent the prep machinery from like Home Depot. A grinder? Yeah. Or like 60 bucks. I don't even know. So you grind it, then you have to sand it? Then you have to, I don't know, paint it, seal it? Yeah, you have to put a seal coat down. I haven't hand mixed resin in so long. What about that look nice? Pink, light blue, purple, and white. Arlene, it's a student from a preparatory school having graduation party and we were commissioned to do these 90s. These 90 dishes. Or them their students. That will be in attendance. I'm going to stir this over hot water to get it out from. If you ask why she stirs it over hot water. Because hot water thins out. Present. Be careful doing that. Because if you get it too hot and stir it too long in hot water it will definitely activate it. You don't want that. I may just not do the rest of them. I don't know why I don't like it. I'm just saying. I think there might be an extra A. So we'll see. I know we have four or five extras. Sorry for all the movement kids. Ooh, I didn't know you guys were zoomed in that much. I bet that was a real rough ride. My bad. So I can't see what you guys are saying right now. So if you guys have a question or amazing, amazing mods will answer it. And if it's a question that I need to address and just hang out until I get this resin poured and I will get right to it. Also little prayers for round out to feel better and for Clara's mom to feel better. Or if you're not into prayer, like good vibes. Positive thoughts. All right, I went ahead and mixed up. 10 ounces of the. Illumina like quick coat. This is a UV resist. Nope. Yep. It's got UV blockers in it. It's got halls. It's got all things to make it not yellow anytime soon in it. You can get it from illuminlight.com. I have that linked down below and you can use the code YALL10.  I think. I know.  She needs to actually rest. All right. I feel like super specific with how I pour this out. I don't want to fill the bottom of it. I think it'll look fine. So I'm going to go ahead and just pour out some resin in all of them because this is a quick coat. This is a fast setting epoxy and the longer you keep it in a big bucket like this, the faster it's going to heat up and start to cure my bed. I'm trying not to get any drippies anywhere, but it's being quite difficult. Quite difficult. Oh my goodness. I just spilled all over that one. I'll have to wipe it off with a rag in just a moment. I'm going to do these on this side. Why is this so stressful? It just is YALL. I have a little bit left in case anything isn't full coverage, but I still have to get it out as soon as possible. Okay.  I had a bad experience in Alabama with a tornado. It's been a problem ever since. Hate them. Hate them. Hate them. Hate everything about them. Make sure we have all of everything covered, and then it'll just self-level on its own. Oh, it's about to be another... Dang it! Hate it. Hate it all. Hate everything about it. Did you guys hear that? Slash feel it. Slash see it. I'm just pushing all the resin up slightly to the edge. I want it to coat, but not fill. That makes any sense. Don't like that. Don't like any part of that. All that ruckus outside. Not my fave. So just like regular resin, this is going to self-level. It is a one-to-one. So you just mix equal parts of the A and the B. In order to save time, I'm just going to pick these up and check them out, and then move them to the back. I need to heat them. Looks good. But it... Huh? No, not really. The ones that are super domed, I'm just... I'm still just coating them. So I'm hitting it with just a little flame to pop the bubbles. I'm swirling it around just a bit so that I can see and make sure everything is coated. I think that one was not. I need to remind myself that some of these have more resin and then than others. When I pick it up and tilt it around to see if there's anything gapping, it may pour out a little bit. It is starting to set, so I'm trying to pick up the paste some. There are some of these that I haven't touched like this one. Make sure everything is covered. Luckily, though, when I hit it with some heat, it'll thin it out and make it movable for just a little bit longer. It doesn't give me a lot of extra working time, but it'll give me just enough. But if you can get an alcohol rag, I got some resin on some of these, like the lip, and it's starting to cure on me. I'm not going to have time to clean it them off. This one has some right here. Thanks. You want to set it like, all right, it is. It's quick coat. It's curing. That's why I was saying if you could help me get these just here. If I can just get it coated everywhere, then it will self level and fill out. Also, be careful not to use too much heat because it is quick coat.  But in my opinion, it's worse with, except for one of them. Toss them. Will you just help me concentrate on these for right now? I'm just trying to get Bowie to be up here. Perfect. I thought it was a good idea. That one's good. That one's good. This one is not good. This is the bottom where I'm on this side. I got you. I got some on the outside lip of this one right there. There's a hair right there. I got stuff on more of these than not. Let me just go through and make sure I've got everything coated in these and then I will start helping clean them up. Let me start wiping as well. I'm just trying to scoot in and wipe these inside corners or wherever there may be some sticky where it shouldn't be. It probably won't even make a difference. But I'll know and therefore unacceptable. Luckily, this resin gives me just slightly more working time than real quick coat does. And I'm sure it doesn't hurt that I have it poured so thin so it's going to take longer for it to start to cure anyways. So all these need something. Here you go. No. I think so. No. Still no. The downside to doing resin is like flooding resin like this is that now I have to be concerned with little tiny baby hairs landing in it. I'm trying to pick out any little fuzzies as I go while it's still fluid enough for me to do so. As you guys know, no matter how dust-free your area is, it's not just a little fun fact and our new really big dust-free zone isn't quite ready yet, but soon. I think these dishes turned out okay. What do you guys think? I've lost my assistant. It sounds like a fake rainforest, doesn't it? Now that it's starting to cure up some, it's not flowing as much. So I can hold it kind of more vertical and it's not really moving. Wow, that's a perk. It also means running low on time to get this done. Right. These bad boys are done. Oh, thanks, Kathy. Thank you, Jillian. Cardboard, huh? Good idea, mates. They said to put cardboard over it. I happen to be in possession of quite a lot of cardboard. Yeah, but like after you put them on there, I just, I'm going to do this. All right, you guys. Fun times to be had. Honeycomb them then cardboard and put next layer. It did work. So you guys, I'm going to let you go because it's not the most fun to watch me do this. So we're going to stack them a honeycomb style and then cover them with some boxes. I'm about to do that many. And so yeah, that's what I'm about to do. I hope you guys have an awesome evening. Oh, we don't lose power. And yeah, let me know down in the comments below which letter was your favorite. And have an awesome evening. We'll see you guys this day Friday. And we'll see you guys on Monday. Monday. You guys have an awesome night. Be kind to one another because you never know what someone's going through. And always remember, we do the test. So you don't have to, including bowies because they're so cute. You guys are amazing. Have an awesome and safe weekend. We love you guys so much. Please subscribe if you haven't. And leave me a comment after the video posts. See y'all Monday at 6 p.m. Central. Bye. Oh my goodness, look how big she is. She can like reach him now. He's thrilled about it. That's it.


First, the Nigerian Veterinary Medical Association has called on stakeholders both at the national and state levels to collaborate with the association in its campaign on mass vaccination against rabies. The national president of the association, Dr. Olutoni Aditsubiru, gave the chart at the 2023 World Rabies Day, tagged all for one, one health for all, in Lafayette. She explained that, in a bid to tackle the virus, the association had embarked on awareness campaigns across the state of the country on the menace of the disease. Aditsubiru expressed confidence that efforts by her union will in no small measure educate Nigerians about the killer disease. She added that they had also stepped up effort at eradicating the virus by the year 2030, in line with the commitment of the World Health Organization. So we want to all put our hands together to pursue one goal, which is pursue the latest out of the world. It is possible. And I know that as veterinarians, we can do it. People are aware that we have a target and then they have a responsibility. And this responsibility is just to on your animals responsibly. This we can do by making sure that your dogs are on leech. Nobody is supposed to release his dog or cat to be roaming about. So that if it requires, even if it's monthly, it's a transition or weekly, whether it's jingles in our local languages, it will make sure that the message gets right down to the grassroots. We're willing as a government to make sure that this is done so that we can work towards the eradication of this virus as a target.


Thank you for the introduction, and thank you to the selection committee for giving me this opportunity to present a progress report on the FFPE pilot study that we have done through TCGA. For the talk today, I will cover three main things. First will be the co-isolation of nucleic acids from FFPE, some of the preliminary results that we've obtained through the genomic and epigenomic characterization of those analytes, and then wrap up with some conclusions and future plans. In addition to serving primarily as the technical director of the molecular group at the BCR, it's also my great pleasure to serve as a co-chair for the analysis working group leading this effort, and so I wanted to start with this slide to give credit for all of the individuals who are contributing to this. It is truly a classic TCGA approach to new concept discovery. Before I begin, I basically just wanted to frame the context of this pilot and emphasize, or maybe just reiterate as we all know, that massively parallel sequencing has resulted in major advances in our understanding of tumor biology. Many of the presentations that we've seen this morning in fact speak to that, where new drivers of cancer tumor progression are being discovered, new therapeutic targets are identified, and even molecular taxonomies for cancer classification are being identified. I think it is important to note that many of these seminal studies have drawn from frozen biospecimens and the platforms that were used to generate those results were optimized for those frozen tissues. So it is the application of what is being learned through these research projects to the clinical environment that is commonly known as precision medicine, or as Dr. Mongol referred to it, personalized medicine. So one of the challenges in making that translation is that diagnostic specimens that are typically available are preserved as formal and fixed paraffin embedded samples, and it is fairly well known that that preservation method introduces many molecular artifacts. So the goals for this FFP pilot were to first identify and optimize the best practices for the extraction, characterization, and analysis of FFP samples, and then our goal is to define the patterns of artifact introduced by that fixation process, and then hopefully in the end contribute to bridging the gap to this diagnostic material and facilitate the application of the emerging cancer taxonomy to clinical testing environment. So from there I will begin with our results from co-isolation of DNA and RNA from FFP. So this pilot study actually started quite a while ago when the TCGA program office asked the BCR to identify a method of extracting DNA and RNA from FFP tissue, and they asked us to find an optimal method. And so from the BCR's point of view, an optimal method is one that maximizes yield and integrity with the hope that it provides consistent molecular characterization of the platforms outside of our domain. So integrity is measured at the BCR by DNA gel electrophoresis, and what we would expect to see in an intact sample is a high molecular genomic band from DNA and from RNA high molecular weight discrete peaks for the ribosomal subunits on a RNA bioanalyzer trace. So in fact this is what we see from control tissues that we used in this pilot study where we procured fresh tissue, split it in half, immediately froze one portion, and optimally paraffin embedded, formal and fixed and paraffin embedded the other half. It was that FFPE half that we then used to survey different commercially available FFPE extraction methods. So what we very quickly found was that there was no single off-the-shelf method that gave optimal DNA and RNA integrity. So as you can see here from these FFPE control tissues we obtained nice DNA integrity, but the RNA was very much degraded. Conversely we found other methods that didn't perform quite as well from a DNA integrity perspective, but the RNA integrity was much better. Still not up to the standard of frozen, but from FFPE this was perceived to be a pretty strong signal. So from there we undertook a task of customizing the methods that were available commercially to try and improve the integrity of the DNA in the context of improved RNA integrity. And what we ended up with in the end is this TCGA optimized method, where as I mentioned DNA is maximally, the molecular DNA is maximized and the molecular weight distribution of the RNA is also shifted to the trend of less degraded RNA. So here basically just briefly want to lay out what this method looks like, not dwell very much on it other than to say this technique utilizes scrolls, not cores, and we normalize the input surface area for each extraction, not necessarily the number of scrolls that go in. And to point out that after the key point here is that the actual lysis step results in a supernatant that both the DNA and the RNA are further extracted from. So with this extraction method identified then we turned our attention towards patient samples and there were basically three criteria for inclusion in this pilot study. First was the, there needed to be FFPE material available from a TCGA patient and that TCGA patient needed to previously qualified and shipped. And then lastly there needed to be enough residual analyte from those frozen extractions from the previous shipment so that we could send out a set of samples, a cohort that was frozen and FFPE matched patient samples. So after some, after a selection of patients we ended up with 38 patients in the study distributed across six different tumor types and there's quite a bit of information on the slide but I think the only thing I really want to emphasize two things is that the average age of these tissue blocks was fairly uniform just shy of three years old at the time of extraction and that consistent with what we see from frozen extractions the DNA yield from FFPE is also less than RNA and use this point to illustrate the fact that it typically required more than one extraction to meet the yield requirements for distribution and characterization. So all of the data that we have is from multiple pooled analytes prior to shipment. So then if we talk about the distribution of this I'm summarizing here the five platforms that paired tumor and FFPE analytes were sent to and when we distributed these it was DNA and RNA from FFPE tumor DNA and RNA from frozen tumor as well as a germline normal and we sent them to as many places as we had yield to send and there were some that were rate limiting and so that is the explanation for the gap. So at this point then we can start to turn to the results. The first platform that I will show you today is the results from the SNP6 array and we were not very surprised I would say to find that the SNP6 array did not perform very well with FFPE or FFPE did not perform very well on the SNP6 array and in fact zero of the SNP6 arrays passed the QC pipeline in place at the Broad and this was in large part due to the highly over segmented copy number profile and it's illustrated in this figure here where the fresh frozen samples are very tight, excuse me tight low segmentation but the FFPE was hyper segmented and spread out. When put into a cluster analysis FFPE was still able to validate copy number alterations in frozen but if you did not have the frozen reference to map to you found that a there was a high rate of false discovery I guess when you when you mapped it to the frozen you found that FFPE had far more copy number alterations suggesting there's a high rate of false discovery so the conclusion from this was that segmentation artifacts compromise the standalone utility for determining copy number alterations of FFPE through SNP6 array. So the next platform then would be exome sequencing and so I am showing you today results from one center and one of the studies and shown here is a Lego plot that represents the rate of different types of single nucleotide transitions found in a tumor sample relative to the germline control and so essentially what you see on the Z-axis is the rate of transitions per megabase and this is essentially should be viewed as a profile of what the Luad tumor type looks like in this view and from our analysts we heard that this picture is consistent with what was previously found through the marker paper for the Luad study. Now the interesting thing is when we when we compare this picture from the frozen to the FFPE we see a profound enrichment in CDT transitions in the FFPE sample and that's a little hard to see but the scale here is actually doubled and so the overall rate or distribution of other transitions is similar between frozen and FFPE it is that we have a significant enrichment in the CDT. So to gain a little more information in about those CDT transitions we bend the transitions by allele frequency and we saw interestingly that the CDT artifact or CDT signature was really confined to the low allele fraction compartment. So when we look at a low allele compartment above 10% we see now the profile of frozen to FFPE is very similar between the two is a comparable signature and then in this low allele fraction is where the CDT artifact segregates. So these results support the use of FFPE for exome sequencing however the additional tools are needed to compensate for this artifact. So the next platform then that I'll present is the mRNA sequencing results and the one of the methods that we use to evaluate the comparability of RNA sequencing between frozen and FFPE samples was through this pairwise Pearson correlation and essentially the takeaway message is there's fairly high correlation between transcript abundance in the frozen and FFPE samples. The lowest average was a 0.85 and the point of reference here is set of technical replicates run on the same platform at UNC. So overall comparable when we do another method of looking at this unsupervised clustering we see that the dominant clustering factor in the RNA-seq data is the tissue type and then within that we see that patients for some of the studies segregate out together suggesting that the effect of FFPE is very low in that in those studies so bladder and endometrial. However in other studies such as Luad and colon rectal we see that the effect of preservation method is much more profound suggesting that there may be some differences between the studies in the influence of FFPE fixation. So if we wanted we wanted to look a little bit more closely at that so the next the way that we did that was to isolate out only the significantly differentially regulated transcripts and so this is a cluster analysis comparing the differences between fresh frozen and FFPE and importantly what we saw was in this cluster analysis there was no dominant effect of tissue type in this we see a reasonable distribution of tissues across but we do see this this fairly profound and somewhat scary if we are approaching a pilot to use FFPE for precision medicine there are some robust differences between between these transcripts but I think the encouraging thing from this is when we drill down and look at the individual patients and then plot the relative transcript abundance of fresh frozen against its paired FFPE we see a fairly linear relationship between the transcription levels or the transcription abundance suggesting that there's a uniform over or under detection within these specimens and it is not a transcript specific effect. So before I get run off the stage then I'll go through the micro RNA sequencing results very quickly and and save with within this platform we used a principal component analysis to evaluate the the effect of FFPE and what we've found in this principal component analysis was that 70% of the variance that could be detected in these samples is restricted to principal component one and that the effect of FFPE through this analysis does not become apparent until principal component five where you can begin to separate out the frozen versus FFPE samples so this suggests in the micro RNA context there's very weak contribution of preservation method on the outcome and just briefly one of the interesting things that we've we've come across so far is that the the number of micro RNA that are significantly regulated in FFPE or at least significantly detected to be different in FFPE is far more diverse than what we find in frozen. So here the micro RNA contributing to cluster formation in frozen here the micro RNA contributing to cluster formation in FFPE so we see greater diversity all micro RNA up that are present here or present down here is highlighted in yellow and then when we map the frozen to the FFPE we see that frozen patient samples map to similar cluster as the FFPE for the most part so the orange and green you see that very easily but then for for other patient samples there is some strain from the original cluster so overall FFPE has a weak effect on micro RNA characterization however there is quite a bit more work we need to do to look into the the cause and effect of the increased micro RNA diversity and so and then just just lastly I can very simply sum up the methylation platform we found very little effect of FFPE on methylation studies and so if we just just go right to the pairwise Pearson correlation coefficients this these are representative of what we see and so with DNA methylation there is very high concordance between frozen and FFPE tumor specimens but we need to declare that this is very much dependent of or at least requires the Illumina FFPE restoration protocol as that was best practice already in use when we ran this pilot study so to to close I've shown you results on our optimization of nucleic acid co-isolation we've found the DNA and RNA extracted from FFPE can be employed for multiple state-of-the-art platforms and then to just rehash the slides that I've kind of zipped through SNP6 array had a high false discovery rate exomes were interpretable with artifact caveats methylation worked very well assuming you're using this Illumina FFPE restoration protocol mRNA seek there was very good correlation between FFPE and frozen samples however there is some differential detection of transcripts that needs to be evaluated further and also that this platform utilize the riboseurochemistry and then lastly the mRNA seek systematic increase in the diversity of micro RNA species from FFPE so in conclusion with 18 seconds blinking red we intend to further this work by analyzing the signature of FFPE through multicenter calling and that work is underway right now we think that will enhance the robustness of our ability to characterize this effect we're also working to delineate the influence of tumor heterogeneity in this comparison that we are performing so again this is frozen specimen compared to FFPE from different areas of the tumor and literally before I walked up here I received the the Lego plots from from that heterogeneity analysis specifically looking for CDT artifact in our luad samples in the frozen adjacent portions and we do not see that artifact suggesting that that the results that I've shown today are more likely to be attributed to the effect of FFPE than a spatial heterogeneity difference and then finally we intend to do deeper analysis of the differences between these two preservation methods so that we may identify bioinformatic mechanisms to correct for these artifacts that's it happy to take any questions and the full-time interest let us give the question for this so if your question can discuss later yeah so so as you know the ps3k kindness is one of the most mutated gene across all different cancer type so our next week is Chris Benz from Bach Institute for Research on Aging he will talk about the domain specific ps3k mutation affects different pathway activity across more than 3,000 pancancer


All righty guys, what's going on? We got Kyle and Sprinkle here with Layton sports cards And we are ripping one box of the tops um zero cool stranger things season four hobby box I mean the first one we get to look at here see if we can get some autos from the cast uh in there Good luck here for Jim. This one is Jim s first stranger things we're getting here. Thank you. Thank you for the order Jim Again guys will have Neil up next than Nathan and then Jeffrey still got time if you guys do want to get something ordered Appreciate everybody hanging out with us so far Let's do it. Let's do it Yeah, they did they did do my boy Eddie dirty for sure man for sure. Let's go out on top though rockin rockin hard Yeah, they got you got like you got like torn apart by birds or whatever that was that's messed up Alrighty good luck good luck stranger things season four See what got here. It's like we got our base take a little slow down a little bit see some from the base here There's a guitar god, there's your boy Eddie Live from the upside down guitar god the insert got in zoe Colonel Sullivan Yeah, pretty much for it's pretty much like once a season like Get somebody you like like um My boy Samwise game G. They killed as well There's I don't know this one is Vecna's curse. That is sick looking man.  It's a cool looking hologram max Base base 11 as well. Yeah, they definitely did my boy Samwise game.  Let's see an auto man. Yeah, those hollers are cool, man I didn't know they had those in here. I like that. There's a Wayne There's a massacre at the Hawkins lab insert No, Vecna Murray. Come on. Let's see an auto Bob the brain There's an insert for Robin. Oh, we got something going here. Well thick one That's a good thing. Thanks. They blurred that out for us. We're we're family-friendly stream. Yeah, and we've got a bada boom That one is um sick. Well, I don't know what that is. I don't even know what we call that Bada boom there on the Yeah, I don't even know what to call that but that's a sick one there It's upside down like on purpose. I think the wording it like turns upside down when you turn the cart around That's kind of cool the Bada boom looking at it from the upside down. There's Lucas We've got Hail Lord Vecna. That's a sick one there That one looks like a little lava design Hail Lord Vecna Cool-looking design there not numbered Susie Chrissy and Argyle Dylan and Argyle would have been friends They'd have been booling. There's a will insert car little Um, it's like a quote card Yeah My boy will with the bowl Bull cut this one is the Nina projects. I think these are like episode posters the Nina project. Come on. Let's see an auto See an auto I don't know. I can't say that B star at Upscore chin chin and this one is it's a 99. We got our first numbered card It is seven miles. So these are like little upside down little portal cards. You can look into the upside down That is kind of cool seven. That one is seven miles to 99 there. Well, Steve First numbered card for us. Yeah, I like those. Those are kind of cool to looking into the portal kind of thing This one is another Another hologram here. This one is Vecna's curse Fred Bunsen getting lifted up in the air. That's kind of cool, too, man. I Like the holograms the lenticulars Hey, what's up, MOOC morning sir morning? How we doing today? Once your Sunday morning going so far got another one seven miles on the base this time Eddie on the base right about halfway Going there's Dustin. We got for Chrissy about to do my boy dirty there There's Eddie on the for Chrissy insert It's another Hail Lord Vecna insert there looks like a lava. Those are cool gold RPA Ben Matherin. Let's go Yeah, that's awesome No, no guaranteed auto we'll take that for sure for sure There's a nobody deviates from the plan little quote card for Nancy So one is a satanic cult bringing evil to your home little hellfire club It's another one of the the episode name episode name cards On the hellfire club episode. Yeah, that's awesome. You congrats Give me eighty one dollars for every ghost Spencer Spencer's doing business got goodbye Papa on the quote another lenticular coming. This one is Vecna's curse this time It's Nancy lenticulars Robin and Max on the base. All right. Well looks like the next high bid would be 82 dollars. Who's in the one once going twice Nancy This one is oh, I thought that was backwards that is a he's been watching you insert there. I thought it was backwards. It's a It's not he's been watching you a little tv card there There's Eddie on the quote There's a destined that one's Eddie in the upside down Jamming out Is that master of puppets he was singing? Yeah, there we go Oh, we're up to 85 for puka There we go the move we got a we got an auction going for you We're currently auctioning your card. You were not trying to auction. There's a uh Vecna's curse This one is Patrick McKinney. Oh wow that is a uh Wild looking little one there McKinney on the base This one is pink number to 86. It's gonna be can you hear me? On the pink number to 86 This one is it's time On the quote card Mike base and last pack here for you. Good luck Jim I'm a fun uh Fun first look at these man. I I'm I'm interested and I like it And a see you on the other side on the quote card and base jonathan on the back Alrighty there Jim that'll do it for your first box of um stranger things hobby. Thank you so much my friend That was a fun one. We'll get that right out to you


Okay, yeah, it was good. Okay, thanks. And she will give us a presentation on interpretable embeddings from molecular simulations using caution make sure variation of all the encoders. The floor is yours. Thank you very much. Hello everybody, I'm going to ask you while Ganesh from maximum Institute for polymer research in mice and I'm a post up there. So as let me try again. Yes, it works now. As we have seen, molecular dynamic simulations is one of the areas that can most benefit from the developments in the unsupervised machine learning I will say, and I would like to motivate with the classical time scale problem here. We are okay. We are here ish nowadays and we always want to sample the longer time scales and there are a bunch of methods that allow something this region more effectively, and one of the methods is the collective variable biasing. So the idea is picking some direction and applying a potential along this direction and accelerate the sampling this way. So this requires good selection of CVs beforehand. And what's considered as a good CV is that that allows crossing over the free energy barriers and also separates metastable states and therefore characterizes the slow motions and hence have common models and the common choices are some internal angles, pairwise distances as coordination numbers are and also non generic highly system dependent and highly complex descriptions of the system. And a good CV is not only helpful in altering the dynamics in a control manner but also for interpretation purposes. For instance, like when building kinetic models, such as Marco models. So in this free energy plot that you're seeing this darker colors are low free energy regions and sense regions, and our aim is to properly locate this high energy barriers, also in the low dimensional space, so that we can preserve this metastability structure. And then we take one step further and build a hopefully build a kinetic model and define or describe the rate of the transition between these metastable states. But this, as I said this requires picking a good CV beforehand, and which is a difficult task and maybe data driven techniques can help us to choose one beforehand. As we have seen auto encoders are along with other dimensionality reduction techniques auto encoders, which are special type of neural networks offer an easier route for such tasks. And they're special both type shape, fewer notes in the bottleneck layer forces the network to learn the essence of the data and discard the irrelevant in more information basically and the loss function is the discrepancy between the input and reconstructed version or decoded version of the input basically. So, here, and there's another flavor of auto encoders, which are called the variation of the encoders which is just a probabilistic spin to standard auto encoders. And the idea is instead of just learning and I think from high dimensional space to low dimensional space. Now the idea is enforced and also inferred the parameters of distribution of the data in the latent space. So we learn to model the data fit structure, in a sense. So, now we're interested in finding the posterior distribution meaning the posterior distribution meaning the probability distribution of the latent variable that given the input x. But that this calculation is not easy because of the interactivity of the evidence and what is done is what's called variational inference hence the name variation will take orders. So instead of calculating the true posterior, approximating the posterior we can easy to sample or let's say tractable distributions. So this makes the problem. Actually an optimization problem where now we are trying to minimize the kale divergence between the true posterior and the approximate posteriors and with some manipulations this can be written as a difference of two terms. Where the first one is the one that pushes the latent space to the chosen prior and acts as a regularization and the other one is trying to maximize the reconstruction. So the likelihood of the decoder or minimize the reconstruction error in the sense, and this nicely follows the neural network view as well. So these components corresponds to correspond to each other and that's nice thing about the variation will take orders is that when the prior distribution is chosen as a Gaussian there is an analytical solution, which makes things a lot easier. That's what you say. But, as I showed there are two different terms in the last function now the first one the left one you see is when the this hand written digits example is strange with the reconstruction also only which is just the standard auto encoder and the plot plot in the middle shows that when we only take into consideration the regularization term, which does what supposed to do acts or pushes everything into each other and mixes everything. And when the both terms are considered in the last part. So we have some sort of separability and also some structure, but if our aim is to further cluster these data points in this latent space this landscape is still not very good for clustering. So the Gaussian prior actually has an anti clustering effect. So therefore we suggest replacing the mini model Gaussian prior with multimodal Gaussian or Gaussian mixture in the latent space so that we can give a space to expand the latent space a little bit more. And so for that we introduced the categorical variable why, which can be considered as the cluster ID. So it allows dimensional reduction and clustering together basically that does it simultaneously. I should say that the number of clusters here is a hyper parameter but as I will be showing it acts like an upper one so so this will come but let's start easy and this is an example of one day for well potential without any dimensional reduction, concentrating just on the clustering. So this is the latent space that we obtained from the Gaussian mixture model which follows closely to the probability distribution, coming from the potential. So this is the accuracy metrics that we obtained by taking the cluster IDs of from the GMBAE method which is pretty high. And as a comparison we're also looking at the variation low tank order case, and what it does what was expected from the variation low tank order pushes things a little bit close to each other and making the clustering difficult actually. So it seems like a good improvement I would say. So the next example is iron type of type which is a benchmark system for conformational dynamics and it's very well known that the metastable states are lie in this emotion run plot so they have these special names for the metastable states. So, and we use the, the pair wise distances augmented with the language as feature as input to the auto encoder and we got this to the landscape for interpretation purposes we always kept the landscape as to the, and, and these are the cluster IDs. So all at this point we noticed an interesting property of the method although we trained the method with 10 clusters we only got six from zero to five, and the other clusters were empty, so it can be said that the method can explore the inherent cluster clusters in the system I would say. So, next we looked at the cluster where these clusters are in the emotion run plot. So it matches pretty well to the metastable state definitions, and then we took next step and build the Markov state model using only these six cluster this course six cluster descriptions, and then the model satisfies the Markovianity and such so for this example at least this clusters were appropriate for kinetic purposes I would say. And other example is now we don't have for this system, nor reference kinetic model or we don't know the number of clusters actually that would be needed in the system but again we trained with 10 clusters. And this system is 15 residual long peptides, and this is also a representative system for helix coil transition and in our simulations coil status sample pretty well, and helical and herping structures were rare in the system. So what we obtained is again this is we kept the dimensional test to the and we got this landscape with this cluster IDs. And as a qualitative analysis we looked at these structures around the cluster centers, and this suggests that the structures become more and more extended as we go along this direction basically. And we further validated with more called quantitative analysis. This shows the average fraction of helical helicity I would say, and as the color goes from blue to red, then the structures become more and more extended, which is in agreement with the quantitative analysis before. And as a further step we also looked at the distance RMSTs from the reference structure which we chose as this helical structure, which is also in agreement with our previous conclusion. We made another test case on polystyrene which is this material that we are seeing every day from computer packaging to food packaging basically, and it has an interesting property. And polymorphs meaning that this molecules have that exist to at these two structures with different arrangements with hands different properties. Experimentally these are the known polymorphs and simulating this transition is extremely difficult. Actually, a PhD student in our group spent a substantial amount of time iterating over the off the shelf methods to simulate this transition and finally could obtain a good landscape that characterizes all the crystalline phases and we instead we decided to follow a purely data driven approach. And we use the slatum description as the input and try to get different crystalline phases with our method and it seems like it does a pretty good job. Differentiating different crystalline phases. And as the next step, although my examples were all from molecular dynamic simulations. I haven't really exploited the fact that our data is a time series data basically and connected in time. So a trick that can be done is just instead of trying to predict the original input. I the idea is predicting a time like version of the input which brings sort of smoothness to the latent space and some dynamical connectivity. So we tried that with this. We shaped potential system. And when we don't add any time like to the system, the division boundaries around this line. So it perfectly when we add the time lag. So it perfectly identifies the different clusters. So at least for the system time lag helps separate the clusters. So this, this is something that one can always incorporate when working with the time series data, I would say. So overall, I introduced a method. Gaussian mixture variational to encoder, which helps promoting the separation of metastable states for our applications at least and it does it simultaneously dimensional reduction and clustering. So it is a generic method, so feel free to adapt to your cases as well. Although I just focused on the molecular dynamics data. So, the paper is here and also the our implementation is there and I would like to thank my collaborators Tristan Barrow and Joseph Rutsunski and Adrian Banerjee from Max Planck Institute for Polymer Research. And thank you for your attention and I'm happy to take questions. Thank you very much. We have time for a very quick question over there. Thank you very nice work. And so from what I understand the to get the collective variables in this way, you need a very long and the trajectory to begin with. You can also kind of somehow iteratively explore, like use that collective variable to sample more, or do you always need to have exhaustively sampled this before you can get the variable. We haven't specifically tried that but that's a good idea one can, of course. Okay, thank you very much. Thank you very much.


From the heart of Silicon Valley, it's theCUBE. Covering CloudNOW's seventh annual Top Women Entrepreneurs in Cloud Innovation Awards. Hi Lisa Martin with theCUBE on the ground at Facebook headquarters. We're here for the seventh annual CloudNOW Top Women Entrepreneurs in Cloud Innovation Awards. Excited to welcome from the at scale team to theCUBE for the first time, Sarah Gorick co-founder and chief architect and award winner. Sir, it's so great to have you on the program. Thanks so much, it's great to be here. So I mentioned you joined a number of your peers at at scale who've been on theCUBE, so we're glad to have you on. So co-founder of a tech company, we're here celebrating women who have not only founded companies, technical entrepreneurs, venture about companies, really hard things to achieve. Give us a little bit of the backstory about the co-founding opportunity that you and your co-founders had about what five or so years ago at at scale. Yeah, so a number of us, a number of the founding team came out of Yahoo where in the analytics group, we were seeing that the scale of data that companies were operating with was changing and the operational environment was changing with both public and private clouds. And even at Yahoo as a technology company, we found we were struggling both internally to develop the tools that we needed and also to find tools on the market that serve the needs of our business users and our executives and our accounts and the sales people. So we realized that this was just a sort of pivotal moment in the change of the way business was being done in the valley and that there was this great opportunity to really help companies connect to their data wherever that data might be and whatever types of data they might have. So about five years or so ago, from your perspective, you are a STEM kid, you've got background in math and physics. So you knew from the time you were a kid, I love this is what I wanna do. What were some of the things that inspired you as being in many industries is challenging but tech is as well that inspired you to not only continue doing what you love but to actually step out of maybe you'd say a comfort zone of a large company like a Yahoo with your co-founders and start something brand new. Sure, I think it's the key thing to me is you have to sort of just believe in yourself and be your own champion because really everybody out there who accomplishes these things takes these steps and says this is gonna be my moment, this is gonna be my thing and I think whether you're a woman or a minority or a man or anybody, you just have to be confident in yourself. Look for the things that you really enjoy. For me that was math and science and technology and just sort of find here's my niche, here's something I can be really good at and become an expert in that area and that's sort of something you walk into over time and sort of develop that confidence to sort of strike out and do some amazing things in your life. How do you find that for those next generation or even those that are in tech now to go, I don't feel confident enough or they might feel some pressure. Do you have a recommendation aligning themselves with a mentor whether it be male, female, whatever, to help them just kind of take stock of what's really important to them? Yeah, I think finding people who have been through it and talking to them, whether it could be a boss or a coworker who's done interesting things in their lives or an old teacher or a present teacher, I think finding somebody who can sort of give you that story of like here's how I felt because you have to have confidence in yourself but really nobody feels confident all the time. Everybody gets anxious or fearful before doing something new, that's part of it and sort of learning to look back and say, well, I've been successful in the past and I've overcome these obstacles and I'm ready for another one. I always think something that's so interesting is the concept of imposter syndrome and how many people suffer from it and I didn't even know it existed until a few years ago and it levels, okay, I'm not the only one and so I think that's something that, some people are kind of born with that regardless, like you love math and physics and you're gonna keep going but for those I think even sometimes acknowledging, oh, this is something that a lot of people that have confidence now didn't have back in the day, I can overcome this as well. Right, I think everybody has a little bit of imposter syndrome at times. I just think that the world can seem like such a big and challenging place but really it's all made of people and they all have the same sort of interests and desires, not necessarily all the same skills but deep down we're all people. Exactly, so tonight though in the last minute or so, you have the opportunity to present in front of one of the most influential females of our time in technology, Cheryl Sandberg. Can you think back to yourself in college and if you'd known this opportunity was gonna happen, would you just said, yeah, that's about right? You know, I would have been a surprise but I think with work and with sort of following your passion, I think anything's possible. I love that. Well Sarah, congratulations on the award and for all of your success at AtScale. We congratulate you and we thank you for stopping by theCUBE. Thank you. We have a great night as well. Thanks. We want to thank you for watching theCUBE. I am Lisa Martin on the ground at Facebook headquarters. We'll see you next time.


Alright, I'm gonna keep this short and simple, just like the MHA intro. I have no voice right now. I am chocked. I was sick for a week straight in watching last week's episode, made it even worse. But I am NOT going to miss an episode of Chainsaw Man. Episode 10, bruised and battered. Last week, I can't even recap last week, I don't have the capacity to. Two things, Akama has the power of the Death Note. And two, Kobani actually popped off saving Denji. Well, alongside Makama, I guess. But still, we're gonna hop into this one if you're new to the channel. Drop a sub and hit that big boy bell and let's get into it. I'm guessing this is gonna be a lot of, like, fallout from everything that happened last episode. We're gonna find out a lot more, hopefully. Is that Aki? Oh yeah. And now Denji's the one sitting bedside with power. Eating an apple. Wasn't Aki eating an apple? Are they just reading together? This is so funny. Relinquish the apple. There's more! Did she just realize there were apples? Did she just realize there were apples? And Aki's just laying there. He's like, my gosh, Himeno's dead. I love Denji and power so much, man. The glasses, dude, I think. Pauwee. Me and Pauwee. He sees how much is getting to Aki. I think it's mostly because of Himeno, and they're the cigarettes, too. They were so close. I knew the sort of scene was coming, man. And Himeno, lots of her, too. There's no chance she can be alive, right? I think the only reason I hope that there is or think there might be is because, like, her body just straight up vanished. But it's because she, like, sacrificed herself. Like, sacrificed her physical form. How much time did they take off by using it? He only has two years to live? How much did it shred off? How much time he already had? Oh my gosh. No wonder she didn't want him to use it whenever they were in the elevator. I don't know how much he had originally, but that is crazy. I'm just gonna think of her. Dude, taken so quickly, too, just out of nowhere. That episode had to be one of the biggest pop-offs in any show I've ever watched, in terms of just, like, bloodshed. I don't know where. Go comfort him. Nah, he doesn't really get it. Aw, she was now his buddy, too, technically. They agreed to be friends. It's like he had to, like, manually connect that line. So why isn't he sad? It's like he's gonna manually realize he should be sad. Damn. Probably not. Yeah, probably not. So who does he really care about? I think he'd be pretty sad if Makama died, yeah. I don't know for the right reasons, but... Three days. He's so bizarre, man. Ah, like if he had been like this, if this had happened before, which, uh, was part of him, then he actually would be really sad. Yeah, part of his soul's gone. These were the two that were with Makama. Makama. Duh. Coach. Bro, he's still in the hospital. Oh, my God. My coach. I don't know. Oh, my gosh, I totally forgot there was even an intro. How far in are we? Six minutes. God dang! So, like, a turtle around Engie. Who is it? I mean, no one? Also, the graveyard is this? Is this, like, Himeno's grave? It's a Chigens voice actor? One of these three questions. He's just figuring out who you are right away. No, we don't answer like that. Oh, shoot. Wait, isn't this the guy that talked to Himeno? Whenever Aki first became her partner, I'm pretty sure. I'm pretty sure that's who this is. Because he talked to her about how the successful ones, the ones that don't care as much, or they have, like, a screw loose. He's the one that said that, I'm pretty sure. He seems a bit odd as well, so it might work out perfectly. Okay. Simple guy, just like them. Why did it sound like he broke their backs? This is a wild initiation. Those regular humans? Oh, my gosh. He broke their necks. Why does he just have a bag of blood? Oh, to heal them. That is so psychotic. Wow, that's crazy. Was he testing them? He was just confirming what they are? We're in danger. He's coming up with game plans just for them. Custom build strategies. The best Devil Hunter. Wow, they got a pretty good trainer on their hands then. What? Are they actually gonna fight him? She just gave him a blood hammer. I feel like he's gonna roll them, like, instantly. Oh, this guy is badass. I can already tell. It's always those boost-up dudes. He just does it with such grace too. Smoked them. Both of them instantly. Immortal. What? They literally can't die? I don't know, man.  They're crazy. Wait, why is he doing that in bed? Oh, he's trying to show them what his double contract is? Oh, yeah, because it said, what have you put in my mouth? Wow. Boy, none. Just negging him. Dang. Just asking if he wants to retire two more years of just living. Oh, man. Saying he can barely even pull his own weight. Oh, this is so sad. And him and O's gone. If he quits, he has nothing to live for. I thought he doesn't have anything to live for, but I don't think he would feel like motivated to even exist without that goal. Yeah, gotta have a screw loose. Have they said that one guy's name yet? That's training, dingy and power? I don't think so. Makama, I'm guessing. He looks so sad, man. This is so depressing. Who is this? No way. Do you not show us who this is? Is it a relative? Oh, my.  They're still here. They look absolutely traumatized. Can we train somewhere a little less depressing? Dengie? You good buddy? What? Was he like seizing? What's happening? She's just knocking it back in the place. My gosh, dude. They've both lost it. I mean, Powers never had it, but then she's lost it. 20 times? Oh, my gosh. This training is psychotic. Perfect. Realistically, he does want them to be stronger. It's just crazy methods by spirits. Oh, like being drunk. It's going to motivate them to defeat him so they can stop doing this training. Those smart characters in a manga.  All right. I'd love to see it. 100 plans. I'm sure they're all very credible. This is such a cute, like, power duo, dynamic duo. They have glasses on. Oh, my gosh. Are they about to fight right here? He dodged that? This guy's an animal. What? Oh, he is that? Are those to refill? Like health pots? Oh, no for her to use. He fights so calmly. It's crazy. Like, no hesitation. Rip the glasses. Oh, that plan didn't work. One out of 100. I got to commend them. It was actually a pretty good plan. Yeah. Putting the blood bottles and then shooting them down. That's pretty sick. They'll get them eventually, guys. Maybe. I don't know. Probably not. Leaves her anemic from using all that blood. He's out of booze. Good work, team. That's it. That's their entire training session. Was that ambush? Now we... Back to the... Back to... Dude, now it's just getting mean, man. It's just getting mean. Okay, now we really know Dengi's limits. Oh, they're not done. Oh, all right. Damn. This is going to be some really rough training, man, as if it isn't already. What is this elevator? Wait, we're not going to see who's in the hospital? Eh? He's making a deal with a stronger devil? I mean, if you have two years, you know... Where are they going, though? Are there devils down here? Oh, my gosh. He just gets his pick of the litter. Like an animal shelter, kind of. Not really, no. Is he talking about Himeno or whoever visited him in the hospital? Why don't we even get to see? Oh, Himeno's sister. Himeno always dealt with people being mad at her for losing her. Oh, wait, there's multiple letters. It's dad feeling any better.  Don't worry about saying food. That's a pretty serious concern, but I'm worried about him. Is he taking his meds? She wrote that to her sister. Forced Aki to get his ears pierced. Nothing else we could go private instead of saying. How do I get Aki to quit? Brought it up, but he brushed it off. Oh, man. That is so sad. I think she wanted both of them to quit so they could live a life together. Oh, my kill me, man. The future devil? Future? What does he have to give up? Oh, my God, wait. It's got to be a really powerful devil for those kind of deals. My goodness. That is absurd. The future devil. Can you see the future? That makes sense for something that powerful. Is it okay for him to be in there with the future devil? I'm sure they're not very happy after being stored here. Is that an eye? Oh, it is an eye. Oh, that is so creepy, man. God dang, dude. That entire thing with him, Mano, is so sad because I don't think she ever really made it clear to Aki that she also had a crush on him as well. And that's why she wanted to work with Denji to slowly try to get to that point. And now Aki knows, I mean, Aki doesn't know that she had a crush on him, but how she was serious about them both quitting. And now he's locked in for two years, two years of life he has left, supposedly, because of this deal with the devil. Oh, man. Now, I've been introduced to this crazy mentor that's going to train Denji in power, who I actually am a pretty big fan of. I love whenever the sensei or like the trainer, the master, he's like, it is seemingly impossible to defeat in a battle like this because it shows how far they're going to have to go to eventually defeat him and pass this grand test of his. But, man, things are just not looking good for my boy Aki. It is so sad. But, guys, that has been it for episode 10 of our chain saw, man, watch this. There's only two episodes left in season one. I've been enjoying the hell out of it, and I hope you guys have as well. With that being said, I'll catch you guys in the next one. Peace, peace.


A Minnesota judge is considering whether Governor Mark Dayton's veto of the legislature's funding was unconstitutional. The bitter legal dispute between the Democratic Governor and Republican-controlled legislature had its first hearing in Ramsey County Court. Chief Judge John Goothman is also considering an agreement between the two sides to temporarily fund the legislature through October while the case plays out. It's unclear when a decision may come. Dayton zeroed out the legislature's funding last month to force lawmakers to rework a $650 million tax bill and other measures. An attorney for the legislature argued it was a blatant violation of separation of powers, but Dayton's attorney says that the governor has the broad authority to veto any appropriations. If you've enjoyed this segment of Lakeland News, please consider making a tax-deductible contribution to Lakeland Public Television.