Hello. I'm gonna shave every 1000. Let me see here. Okay but you guys Hey, Carol Guess who? Must have been you that hits the like button. I type slow Yeah, I do too Because it was just when I was coming out the door. I had one like in no views Yeah, I guess slow internet cuz I got a hot spot off her phone But I got her phone right here beside me. Yeah, it said it said one like and zero viewers. I'm like She's like monkey, so how's that possible? I don't know but it says that Where you into today Carol? Had to do this at noon didn't I That's something hot should have put my sombrero on Going in the full lock now. We're just We're just on a stay at home order. I ain't stopping people. Nobody listens. I ain't going anywhere unless I have to And we've got plenty of groofies and stuff I may have to get some more coax, but I won't do that unless we need something else, you know I'm not gonna risk it just to Just to give you some pop got plenty of water in there and tea and stuff like that. I can drink Monkey be out in a minute. She's getting dressed Beautiful day here. It's nice in the shade, but man was hot out there. My son He haven't had rain she's in two months We need it, but we haven't had a hundred things open here It got down I'm going to gas the 60s last night in jack, but it was nice. It wasn't like too cold So we could have the window open So yeah, it was pretty nice, but like I said, we haven't had any rain. We need rain bad I mean I haven't I haven't cut the grass in a month and as you can see It's looks like I just cut it yesterday Well, in fact, it was when I did a video. I think I was on the mower And I did uh, I think I filmed some of that Yeah, I think so that was this it's been a while back and There's a couple right in there. There's a couple longer things, but You can see I mean we haven't I haven't cut the grass in over a month. I think it is Carol's on she was the one that hit the mic, but Of course you can comment it, but it still said zero viewers That's right.  Hey, she's very kind. I should watch them. Say hi monkey Hey, she says rain and then she'll be there. They're going on full lockdown. So Really in texas? Yeah, so She had one got some cigarettes and some coax Oh my god this morning.  Yeah, you gotta get his beer or whatnot But I'm buffering too Are you? I don't think anything's on in the house I shut the tv off my computer's going on My name either.  I can't believe it. What are they going in a lockdown today or tomorrow? I got some meat. I got noodles. I need some spaghetti sauce and we can have spaghetti I made some meatballs too If I can get some ham or some meat Let's see. Everybody went apeshit.  Yeah Yeah Instead of doing what we did and then you only have to go and get a few items here and there All right, you know, right The way Nancy's are in Seattle is pretty much like that too. They say everybody's doing well in Seattle They're they're they're staying at home. They're doing what they're supposed to do up there I don't get these Florida people I don't love like that car just went past, you know, they don't work They ain't at work. It's noon, but they do get off at 11 30 or whatever. No, it's they probably went to the store Nothing else.  Whatever you have at home, you have it I mean, I don't even you know, I'm gonna take her out on the track yesterday, but I'm I'm doing what I'm supposed to do because I mean, even though our closest neighbor on the other side than bushes over there You know, that's our closest neighbor.  I heard a couple were boats yesterday Yeah, you know But they're like, well, I'm out on the water. Yeah, but you're stopping at the store again. Shit. You don't need you know Just to go boating get that beer go home drink your beer, you know Have a fire in the backyard whatever but yeah, we've been just trying to Just trying to stay home and uh, like just get what we need to get like she may have to go get a couple things today, but After that, we'll be home all weekend and they're now they're set now They're saying you have to wear a mask even go into the grocery stores. Definitely now So it's getting worse.  I really don't want to because um, I give you a mask because um You don't want to that well like people with heart conditions Like us like me and older the elderly They're what they this started what two weeks ago To self stay at home, you know to self do whatever So Yeah, the uh So, you know, I don't want to take any chance even with a mask if I don't have to Something dropping on the truck over there The woodpeckers up there And in here and shilly then what her messages are being taken off Mine's on here. Well It says I wonder why what was that? Well, I put that because it's being It was a message retracted. She might have took it off. I don't know I don't I've got all my messages are are approved. So I missed I I got them all Well, so I you know I figure I mean I've read everything I I've got on here I asked her what she was up to she says she been to town and back They said They're going on full lockdown So she wanted to stock up on six and coax Raining here and chili And there's a couple retracted ones I'm removing them because you didn't read them See message above No, I've got two. Yes. I'm watching I type slow stay still your buffering Yeah, hit the like button and comment been to town and back I'm removing them because you didn't read them. I just I just read that. Yeah Yeah, but I'm just saying this is all I've got right here raining here and chili and then Message retracted message retracted and you said I wonder why and then I'm removing them. So I've got you know all that I'm getting Yeah I'll turn this one off. Maybe it will stop buffering. Yeah, and also, you know, remember there's a delay too Yeah, so that could be cozying problem too. So I turned it off Now maybe he won't suffer so much if you'll have to read him off his phone then It's all right Oh I need some salt Yeah, let's go to her old man there. Yeah, you'll see they're just out cruising.  Well, maybe they should have stayed home Or blame trump because he didn't do it. Well, I said to the government I'm not really wanting to get into that too much. But I'm just saying if they get sick they'll blame Everybody but themselves. Well, you were out on your bike. You were out boating.  Yeah, we were going to add some things to do on the corvette and uh, I wanted to do She had a couple things she wanted to do and then we were going to investigate but We were going to stay tonight. Yeah.  We were going to investigate and I just Don't feel like running around Well, let's just stay at home. So I don't want to be out there and then no they just they just want you You know, you can go to the store and get some supplies if you need to Well, I heard that they they haven't done it yet, but I heard that they're going to charge people $500 Or six months in jail if you go out But I had they haven't put that in order yet, but they're talking about Well, that's because nobody's listening. Yeah Like in people on them bikes, they didn't go to the grocery store They're driving all over the place. We're going to grocery store on the boat, you know Like I said, I heard several airboats yesterday over there on the river and it went up when then affected 1205. So, you know Yeah, I don't know But all we can do is what we're supposed to do and You know, so They could they had his texas carol. Is it pretty bad over there? Must be if they're going on full rock down must be pretty bad national guard headed to houston What's that mean? They're bringing in the national guard That's pretty bad They're heading to houston. So Wow Wow Because remember, uh, they did that in Washington. Yeah, right up the national guard. Yeah Well, my mom said they um Those two ships did dock yesterday They are taking the ones who are very sick on those ships to the hospital The ones that are not they're putting them on airplanes and sending them home where they belong So they but they're uh, I guess the airplanes are doing it for free And so I'm like, wow, what if they're carriers, you know So they're probably making them wear a mask on the airplanes and stuff. She says Where's the damn lock down and loaded here? I even carry now You must be getting bad there. You got people down here playing golf. Yeah, the people just listen, you know I don't know I wouldn't think but So it's just like any other day all these cars Then it was a few weeks ago because I think people were listening a few weeks ago But they're getting antsy now and That's what I saw yesterday too and I mean friday is always usually busy, but the week Ago live on friday It was it was like ghost town now Yesterday. Oh my god. You would have thought it was everything was normal.     No I don't figure they're doing anything either No Nobody's gonna be set up If they're far, I'm sure they're gonna be told to leave I'm not gonna sell anything That's why I set up Thursday Because I knew Friday For what, 30 days or whatever I'm gonna be able to do nothing See above Yeah, see, it says see above Yeah, see, Dino cut the grass My little handsome dude Bruno Had to buy a new phone I hate the piece of shit already My old phone was 40 years old And then it says see above So, apparently I'm not getting all of them Had a man walking down the road Yeah, I mean Maybe it's a delay Could be a delay Well, there is a delay Yeah I don't know how long it is But you're getting them Well, I mean The ones I'm getting, I'm reading Oh, no She bought all new stuff for her living room To deliver one piece at a time Oh, no Oh, man That sucks I'd be like, look, man I'm gonna do this on my time I'm gonna be on this day Bring it all One piece at a time Oh, man You got my money and then I gotta wait for my shit Yeah That's just like nice stuff They kept bringing wrong one And I was already paid for And everything It took three months for them to get it right If this one ain't right I'm getting a full refund Because, you know, they take your money And then, well, we'll just give it to you No, when I pay for something I want to be able to use it as soon as I pay for it Well, the thing was It has the long reclining thing So they kept giving me the wrong side Well, I can't use it Because I have a very small living room Yeah, it's a sectional So they had to Eventually, the supervisor Brought two of them And they gave me the right one Finally They could have done that three months before Never did Yeah, but still That's beside the fact When I got the washer and dryer I bought that one at Lowe's I got it, they delivered it Then they didn't bring the right plug They didn't have the right plug for the dryer So I couldn't use the dryer You see, no, that's unacceptable I don't care what time it is You're going back and go get it I'm going to bring Another one this Saturday After Because I was working so much That I could only have them come on Saturday So I ended up going to Lowe's And talking to the manager And getting it that way She said it's easier on her coming one piece at a time But she had bought a new HD Oh, wow 12,000 BTU That's cool Well, remember Get my computer Like what we can deliver for you When?     No, I don't watch South Park Fire went up first It'll be more exciting Yeah The whole video is upside down We talked about it on the last life I've been wanting to do that for a while But I just couldn't think of what to do it on Right I was going to do it upside down But this can really help someone Right I was like Just make it suffer I suppose I'll get a lot of haters on that No I can't see what you're doing That wasn't a point Point is I want to tell how many people want coffee now I don't know I remember this buddy drinking a bottle of cracking last night What's that?  I'm just kidding I know She live it I need to flush my system on Oh Honey where's that bar and chain oil I bought The other day I missed shot her to that No give me the Lucas That's better it'll really coat my insides What I miss I'll flush you out Yeah I'm not going to try it though Can he try it? He can try it Let me know what it's like son New coffee?    I'm not but they might be But they probably are I got the phone Right here I got one back That's probably what it is You gotta remember a lot of people streaming today App spotlight is running My battery down bad Oh really That sucks Slept enough in my mind Yeah sorry about that I don't think it's your in I think it's us Her phone even though it's right here Like I'm not buffering But by the time it gets to you guys You know Yeah she had to refresh Sorry it's all on my end this time guys Sorry about that What the hell is that I don't know Spotlight I don't know if I had it on my phone Or what but Is that the one with the green Anything you don't use You can take it off But sometimes Sometimes There's some apps you can't take off There's a lot of apps on this phone That I'd like to take off it just won't let you I don't fucking use it Like the Amazon You can disable it In your settings I'm like I don't shop enough On Amazon through my phone to use it So We need to go chip in Get Shae Baron monkey Some Charter Spectrum Yeah I heard they're giving Spectrums giving it out free right now Oh yeah It's for schools and stuff Kids that are online Yeah but they ain't just gonna give it to us No Yeah Well the bills aren't that high They're charging me for the equipment Yeah well once you go cook I'm gonna pick all right off Yeah well we'll get hooked back up It's had some hard times But you know we'll get straight now Uh That was my dream when I'm here During the day you know like I used to Yeah So sorry about that Still you know just gonna keep in touch with you guys See what's shaking Make sure everybody's alright He said he had to let go of his internet for a while He went crazy Well you know And another thing Like you know I'll get a video done Say 10 o'clock in the morning I'll edit it by 11 It's ready to be upload And I can't upload until she gets home Sometimes she won't get home until 6 or 7 And then of course you know For a 15 minute video it takes like 3 hours to upload To where before you know I can just go ahead And upload it and start on my next video So Pam hasn't left the house in a month Good girl I've been out a couple times Yeah I go for work And come home I went to the store the other day with you And I did set up Thursday We go to the store today We'll have to wear a mask You don't have to Yeah but I spent a little time with you But I just don't want you out I'm an essential I think Monkey's going live tonight aren't you No I thought you were doing live stream tonight That's what you said last night You're going to make a video That's why I go to the store Burger King and McDonald's Has awesome wifi I've uploaded videos in the park A lot faster than at home before Yeah they do Spent a while back But yeah they were pretty quick I've done them before But see Like when I had the internet I just run ether My computer is wifi But I ran ethernet cable I could upload a 20 minute video In like a minute and a half And wifi It would do it in 5 minutes See this is I'm not using wifi I'm using in a hot spot So that's really slow Whenever you do that At least we're still in touch At least I can get some videos up Like my job now Yeah All these cars They've been in the house And now they expect everybody to do For another 30 days And people aren't complying now They're going to have to make them Like I said All these people are going to complain if they get sick Well my thing is They'll end up locking Florida Because people don't want to listen I'm being antsy too I'm going to go do stuff I wanted to take a ride up the park yesterday And vlog I didn't I probably could have been alright And not done trouble But it's the point I'm complying I'm knowing what we're supposed to do Oops Spent 2 hours on the phone Getting my net bill reduced I think she gave me a discount So I shut the fuck up Yeah you do that And they'll do that I used to do that all the time With T-Mobile What's the next project? What is my next project? I forget now I don't know What is it? What is your next project?   You don't have to water them Bought me some beer for now That Bought me some beer Now that he can drive If he gets If he gets sick I need my Miller light I lasted Went all that time Listen to those Cactuses Yeah they're out I killed a cactus Yeah I don't have a green thumb She over watered her cactus Maybe that's what I did Yeah these cactuses Because really a cactus you're not supposed to water Yeah these things They're light now but A little bit later this month They'll really be so loud You know it'll be hard to hear out here Through my cactus in the trash It kept poking me Poke it back Carol I threw it in the trash I never ran over that one Yeah I took it out Ran over it with the boat I ran over it with the boat not the lawn mower Yeah the boat But yeah and then you replant it to make it safe Look why I'm to it The thing was like Some things Be well enough alone it's a cactus It'll be fine Well you were guiding me in there Real tight over there Just enough room to Back the boat into Don't let me hit anything I won't Next day I was like you ran over my cactus You were guiding me I couldn't see it was dark I didn't need to be that close to the house anyway Kenny's laughing For dinner since you like cactus That cactus I had was good Yeah you liked that Yeah I didn't really fond of it Too well Well cactus there is no flavor You got a Your flavor in whatever you're cooking it in Like whatever Let's see you fried some potatoes Then you put the cactus in that In the pan That was that restaurant that we went to That we didn't know that was Their buffering Yeah it was a vegan restaurant Yeah I won't get off here Maybe I'll do another live one tonight Okay Can we do a fire?  Yeah let's do it by the fire Hang out Better call them hot dogs though Let them fall off the stick Oh I won't do them No I wish they were something else I could do That's not full of sugar Kenny said hell yeah There you go What's everyone doing? 8 o'clock We'll figure out what we're getting Randy will be 16 on the 22nd Aw It's another year with her Very good You going to have a little party for Like you always do little cake and stuff She makes her cake and stuff It's in June yeah I just don't know the date So we'll just pick a date Make that his birthday Alright Pam you have an awesome day too Be safe Come visit us tonight Yeah we'll have a fire tonight About 8 That's cute 16 years That's old Enough it's a June 22nd Bruno B day Bruno D day Birthday June the 22nd As we know it was in June I just can't remember what day it was on No I'm looking in and I'm looking I get distracted Car went by June 22nd What day is that? It's a different day every year It's a different day every year I think so Alright see you tonight Kenny Have a good one buddy Stay safe and we'll chat at you this evening Carol's going live around 4 or so So we'll check her out Okay Yeah we'll stop and Check you out She's going to go to the store Yeah she's going to go to the store So it's on a Monday she said I don't care I don't care what day it was But we do know he was born in June So I got my glass on But it felt like they were up here I didn't really pull them down You don't feel good? Maybe you should lay down for a while Take a nap I don't watch YouTube and lay down Take it easy for an hour or two I can't believe the traffic Unbelievable She said have a party on the Sunday before Yeah I don't care Don't matter to me if you want to make the 22nd We can Go live with Bruno B-Day party Yeah we can do that That'd be fun Yeah we'll do that I guess she's going to go to the store I'm going to watch some YouTube Check for a little bit And So Carol's going to live about Four We'll check in on you And we'll do hours at Eight By the fire We'll have a fire tonight I'll go out and I'll get that ready About seven I'll get it started real good We'll have a good fire going So We'll do a We'll have to upload a video Okay Carol we'll see you We'll check in with you when you're doing your live stream We'll keep an eye out and make sure we don't miss it Okay So oh man just got three Okay Carol we'll let you know we're on And Then anybody else hopefully we'll see them at Eight Tonight we'll have another fire Yeah so that's what's going on So we'll Back on Eight o'clock tonight by the fire And chat with you guys then So hope everybody enjoys the rest of your day Now I got three on here I'll see you at Monk Monk says she's out for now I'll see you later Alright bye guys Alright good we hit zero


Thanks for checking out this review video. So let's get into season 2 episode 2 of Creepshow on Shudder Once again just so people know no spoilers on this I can't even do a little synopsis because it's forbidden as far as the terms for me to get the screeners and do the reviews And also because the stories in them are so short. It's about 25 minutes ish that doing a little synopsis could just Ruin too many things honestly. So anyway, I will tell you what the titles are and we'll go through these now I will say if you watch my review for season 2 episode 1 This episode I think is a step back from that. I didn't expect that it would stay at that same level There were some real good stuff in that episode this episode there is still some good stuff But it's definitely a step down in my opinion. So now the big thing is where are they gonna go with the episodes coming up I'm hoping they go back up or if it kind of goes like this fluctuates a bit. That's fine That's kind of how episode or a season one was so I would expect that but anyway, this episode comes out Thursday April 8th now the first story is dead and breakfast and I wasn't a huge fan of this one. This one was the Directed by Axel Carolyn Who did film soulmate tales of Halloween one of the stories and tales of Halloween and an episode of the haunting of Bly Manor which I have not watched yet. I need to get caught up on that So that's what that's been on my list so much on my list Written by Michael Russell a who wrote the script for Dubro party massacre three So shout out to that guy because Dubro party massacre three is awesome. It's in my Collection back here very good And also Eric Sandoval was involved in the writing of the script as well This stars the bigger names being Ali Larder who's best known for some stuff like House on Haunted Hill final destination within the horror realm and See Thomas Howell who's best known for the outsiders his roles pony boy than the outsiders But I think he should be best known for his role in the movie the hitcher with Rucker Howard that film is awesome I rewatched it about a year Maybe about a year ago or so. It's so good if you haven't watched that film in a while Just go back it is outstanding and see Thomas Howell did a great job and on that note Ali Larder and see Thomas Howell do a really good job with their roles in this story. My problem is the script is Okay, I mean the story the concept. It's an okay concept, but where they chose to go with it I kind of saw it coming a mile away, and it's just not that compelling in my opinion. It's But the acting is good I enjoyed the acting I do like the initial premise the initial the initial setup of it it's kind of taking kind of a subsection of horror and it's kind of a more like dark serious point Part but it kind of makes it a little bit ridiculous and over the top and has kind of a comedic take on it to a degree which you know That's what a lot of these creep show stories do so you would expect that type of thing But it just it seems kind of boring in my opinion It's kind of boring. There are some things that could have been done with this concept I just don't think it works out. I just really don't think it works out that well But like I said good performances so you can revel in those especially I was excited to see Thomas Howell was in it like very excited because I can't remember the last time I saw him in anything And like I said, he was awesome in the hitcher. I would love to meet him and tell him that in person The integration of comic panels in this is actually worked really well. There's a portion It's kind of used to tell a backstory. That's that's kind of important to tell in this So I like the way they use that They put a I already talked about that my my apologies There is a way they move the story was something a bit implausible and They could have easily actually made it more believable That's one of the things is how they get from one part of the story to another part of the story is not that believable it's more of a kind of Convenience of writing in a sense It's not a huge issue and some people may see it and just be like yeah, whatever. I don't I don't really care that much I'm just accepting whatever is going on But it's just a small thing that I saw and I always bring these things up when I recognize them There's a coming together of kind of old and new in this that I do find kind of interesting It's kind of like a old version in a sense of this darker subs subset of Horror than I was that I was talking about and a newer version of it And that's kind of interesting to watch in the beginning But once again the whole thing is where they go with it and how that develops is I just think not super compelling Unfortunately from a technical standpoint though, it looks good. Well directed all that stuff Like I said already very well acted and like I said, I saw the end coming a mile away and when it ended I was just kind of like okay. I mean it wasn't bad. It was fine I expect that they're gonna have you know these stories pop up where I'm not that into them now Maybe someone out there really loved this one. I'm sure someone did you can put it in the comments and let me know But overall, I'm gonna give this one story a two-star rating not big on it But Allie Larder and see Thomas how you guys did an excellent job. So good job The story the next story on this one is pesticide, which I liked more than dead and breakfast definitely Directed by Greg Nicotero. We've already talked at length about you know his resume So I'm not gonna go into that written by Frank Dietz who wrote scripts for naked souls mischievous Magic in the mirror just a few that I pulled out nothing super known And it stars Josh McDermott Who's of walking dead fame which I don't watch the walking dead so this is kind of my first time seeing Josh McDermott and he's a great actor and His role is really fun and the way he plays his role is really fun. So I really enjoyed him Ashley Lawrence is in it in a smaller part than than the other two main people in it But she does a solid job with it Obviously, she's best known from Hellraiser films love her met her in person. She's super cool And my favorite in this is Keith David obviously very well known for things like the thing Which is an amazing amazing film and they live to John Carpenter films by the way He's a killer actor the amount of IMDb credits. He has is in frigging sane. I wrote it down 326 acting credits on IMDb He does a lot of like voicing for cartoons and stuff like that, but dude that guy is so prolific. He is Putting in so much work. He's still working. He's a delight in this I mean, I said that McDermott did an excellent job and he really did but every time Keith David is on On screen in this he just steals the whole show. I love him in this. He is so good I feel that way about him kind of every time I see him in a film But in this story in particular, he just strikes you as like chewing scenery like nobody's business His line delivery is epic. I love that guy Keith David is so great So he's one of the best parts about this, but it is a good story It's an interesting story and with the very very beginning you can see very heavy influence through From a film. I'm sorry Very heavy influence from a film that is older that is pretty beloved that has similar a Similar story just just to start this does deviate a lot from that So you can see the parallel with it and I know some people if you're watching this and you've already seen it You know what I'm talking about for sure There are some portions with ADR problems in this now ADR is basically where after the fact that they Rerecord dialogue from the actors in the areas where you know, maybe the dialogue didn't come out that great It sounds a little messed up so they're trying to get like a better quality of the audio there And some of the ADR doesn't really match up all that well and it it's wonky. It's weird So that is kind of an issue another small issue is and some people may not even catch this one, but There's a moment where someone's driving in a car and you can see the scenery behind them The car is turning it's taking a turn But the wheel has not been moved at all like the guys is still just holding his hands Not moving the wheat steering wheel whatsoever and the car is turning That's something that kind of threw me but like I said, that's just probably a small thing most people probably won't Acknowledge that or see that I don't know but let me know if you did Keith David like I said, I'm gonna repeat this Keith David Keith David. He steals the show He's not in it initially but once he shows up and he has a good role in it Well-written character. God. He's great Some minor attention. Oh, no, sorry. I already covered that one with the steering wheel There are some fun practical effects sequences in this especially ones that are in Involving involving. I'm sorry. I don't know what's wrong with me today Involving larger pieces that they made for practical effects So if you've already seen it, you know what I'm talking about those moments look really good practical effects are really good There's some good kind of gruesome stuff to this creepy stuff to this. I like those aspects of it very fun and This one actually plays heavily on kind of morality and socio-economic themes at least initially But then it starts to really kind of wind and you don't really know where it's gonna land in the end So I like the aspect of not really being able to know where it goes Especially after seeing the dead and breakfast one where I was basically like I know Pretty much exactly where this one is going no surprise this one I spent the whole time going through just very engaged because I'm like, I don't know where they're gonna go with this There are so many directions they could go and it was fun. This one is legitimately fun. I did enjoy it It's a good time. So out of five stars with half stars in play I'm giving it Three, I'm gonna have a very solid three. I was between three and three and a half I don't think it's quite at the three and a half, but Three, I'm definitely gonna give it a three. It's a good time. I enjoyed it and it is the Bright spot of episode two So we'll see where we go with episode three, but I'm gonna have the reviews for all of the episodes and Just a reminder. I also have all the episode reviews for season one and the specials And those are available in one playlist on my channel. So there you go Real quick hit the subscribe button if you can for me and you can because it really Literally only takes a second means a lot to me and the growth of my channel and I do appreciate it Every time someone hits the subscribe. I get an email about that I see your profile. I look at it and I say, thank you very much to this person I actually literally do that in my mind when I look at it So I really do appreciate it and just you know, join this nerdy horror community. It's a good time Also hit the notification bell button because that way you will know when I'm putting up more Reviews like this more in-depth ones that have spoilers unboxings haul videos all that stuff and The other thing the big thing is I just have to thank you for taking your time to check this out I really do appreciate it and until next time keep it brutal


My name is Emily. Thank you for joining me for some yoga today. Today we'll get started with our Namaste song. They extend in stretch, sun salutation, some fun movement, our mystery bag, and then our closing. Alright, let's come into a comfortable seated position, take a deep breath in through the nose, and out of the mouth. Hands in front of our heart. Namaste, namaste, this is the way. Light in me, honors the light in you. Namaste. Extend and stretch, extend and stretch, twist and turn, twist and turn. Let's have fun, let's have fun. Can we do that again? Just a little slower. Extend and stretch, extend and stretch, twist and turn with a little breath work and sun salutation. So I'm going to pretend that I have a big balloon in my belly. Have you ever heard of balloon breath? So when we breathe in, we'll breathe in through the nose and just imagine that the balloon is filling with air and then when we exhale or breathe out, we'll imagine that the balloon is losing its air. Can you try with me? One, two, three, hey, breath through the nose. Did you get your balloon to move? Can we try again? How do you feel? Did your balloon expand to get bigger? Last one. We might have to try that after our sun salutation. Let's come to a standing position, and it's kind of rainy. So maybe we can say hi to the clouds. Let's see. Let's breathe, exhale, breathing out. Oh, pretend we're the rain. Just step back and then splash down.  Feel free to make the sound. Let's push back in the downward facing dog and help me out with the sound the dog might make. If you want to shake a tail, shake a tail, shake the other. To a table top position. Now the other day, I was driving down the street and I saw a lot of cows. So let's do our cow pose.  Ooh, what does the cow say? Quack, quack, quack. Now I'm just getting, quack pose, bringing our back up like an angry kitty. Head down, meow. Dropping the belly. Now let's come up on to our hands for some donkey kicks. Feel free to push that. Here's a frog pose. Catch a few flies if you'd like. One, two, three. Ribbit. Last one, ribbit. Let's shake out our legs. We're going to try the sun sanitation one more time. We're just moving through a little more quickly. Let's reach our hands up and down.  So nice. Oh, if you have a dog, please say hi to your dog for me. Coming into tabletop position. Cow pose. Cow pose. Let's do one super duper big frog jump. One. So nice. Feel free to shake it out. I'm going to take a look outside. I'm going to put on whatever I might need. Keep my feet dry. Just in case I get cold or wet. All right. Walk with me. Let's go. Walking, walking, walking, walking, walking. Hop, hop, hop. Hop, hop, hop. Running, running, running. Running, running, running. Now let's stop. Now let's stop. Mmm. Let's put on our good job. Take a look at that pose. There are giant oak trees right out my window. So I'm going to try tree pose. Feel free to follow along with me. And pour my weight into one foot. Oh, and start lifting the other one. So I can either put my foot under my knee, over my knee.  Let's find one spot to gaze at.  Whoa, maybe you flow in the window. Remember, keep that, keep that gaze. Let's try the opposite. Now remember, the foot can come under the knee, or over the knee. Whenever it feels best for you. Feel free to grab hands from your helper's, from your friends. And then if you'd like to grow branches, ooh, let's grow branches up. Little walking. Walking, walking, walking, walking, walking. Hop, hop, hop, hop, hop, hop, hop, hop, hop, hop. Running, running, running, running, running, running. Now let's stop. Now let's stop. Monoculars up. Take a look around. Down pose. But you've got pose too. That's a good one. Alligator pose. I'm going to pretend I'm a big green alligator. Ready? Maybe I'll put my jaws right here. I'm going to put my head on the floor and snap, snap. All right, last round. Let's go for one more walk. That would be fun. Thank you. Walking, walking, walking, walking, walking. Hop, hop, hop, hop, hop, hop, hop, hop. Running, running, running, running, running, running. Now let's stop. Now let's stop. Monoculars on. Did you see anything cool? Did you see anything you can try? I'll tell you what I saw too. I saw a ladybug. So if you'd like to do a ladybug with me, you'd have to pretend I'm a little beetle in my shell. So I'm coming down low. Hands together.  A little ladybug? That's great. Oh, so nice. I wanted to take a look at the mystery bag. Can you sing with me? What's inside the mystery bag? The mystery bag, the mystery bag. What's inside the mystery bag? Tell me do you know? I don't hear anything. Nothing in here. Any ideas? Oh, let's check. Let's check. Let's see if this is what you thought it was. We're talking about balloon breath today. Can you pretend that you have a balloon right here in your belly? Let's breathe in through our noses. Did that balloon in your belly get big? Let's do a few more. I think I need to practice my balloon breath. Now also feel free to ask someone for help, because this is hard to blow up a balloon. I might need some help too. So on the count of three, you might have to take a big breath and blow as hard as you can. One, two. Can you help me again? One, so good. Do you think we can make it bigger? One, two. That was pretty good. All right. So with my balloons, I love just focusing on my balloon. See how long I can keep it up. It's fun. I can do it by myself. Or maybe with my mom, maybe with my dad, my grandparents, my friends, and just try to see how long you can keep it up. Here's something else too. I wonder, if I put my balloon down here, I wonder if I could blow it across the floor. What do you think? Is your breath strong enough to do that? As you count down, that was pretty good. That's a nice way to practice your breath too. All right. Just about that time. So we're going to take our balloons. Ready?      Put your balloon on your toes. Let's see where it goes.  Put your balloon on your belly, on your belly. Put your balloon on your belly. Put your balloon on your... Here we are called Shavasana to have your balloon on your belly and you can even practice a balloon breath here. So let's see. Breathe in through your nose and see if you can make your balloon rise a little bit on your belly. Maybe I need to practice too. Did you see my balloon rising up and down with my breath? I hope so. And just feel free to relax here quietly for a moment.  Push your body up against the wall. Then you can lift your knees.  Ah, this is super relaxing too. So good. Come on down, bring my legs to the ground. I'm in a comfortable seated position. It's just about time. Take a deep breath in and breathe out. Hands to your heart. Namaste. Namaste. Honours the light in you. Namaste. Thank you for joining me today. We'll see you later.


Hello everyone, my name is Jack Anderson and Matt to answer your question does my back hurt yet? I'm here to present to you my company Life Changing Foods LLC and more specifically our product Dot E's Life Changing Salsa. Now to answer the question that you're probably asking is the question I get when I do demos in the grocery store right away. Life Changing Salsa, what is that? What's life changing about this? And the simple fact is we donate a portion of our proceeds to build fresh water wells in Ethiopia and that's something that was right from the start. This business began about 14 months ago and when I started it, I had no idea what to expect. Absolutely none. The goal was two fold. I wanted to learn about entrepreneurship. I wanted to see if I could learn something and I wanted to make a positive social impact on the world. A team of students and myself got together and over those 14 months we developed a product, three different flavors as you see in front of you. We built a brand. Believe it or not, we were able to convince some people to give us money to get this thing launched. We started working with reputable retailers and we've seen the screen here. High V, Luns and Byrdies, Amazon are most recently starting next week, Kowalski's Markets and Twin Cities. And from there, our media coverage grew. We've been covered on local media here in Makeda and St. Peter, most notably in the Twin Cities with WCCO Radio and TV. And tomorrow at 12.30, I believe, if you're bored and you want to watch Pioneer Public Television, they're doing a 30 minute special on us. So, channel 10 on RepTV for those that haven't. And finally, in those 14 months we were able to bring in about $30,000 in revenue and most importantly the thing that I am absolutely the most proud of is we built a freshwater well in the tiny village of Decare, Ethiopia and 400 people have access to freshwater well for the first time. And it's an absolutely amazing experience. That well was completed in March and it's really energized the business going forward into the new year. Now, I know this is an entrepreneurship competition. What I've heard is we're being judged on four distinct categories. Number one being progress. Two is feasibility. Three is scalability. And four is innovation. I believe I covered the first two in the opening, so I won't spend much more time with that. But I want to jump to what I think most of you are asking is the innovation piece. I'm not going to lie, I did not invent the salsa. It just happens to be something that I've been working on. It's a billion dollar market across the country. According to different reports, it's the second most popular condiment behind ketchup. And some of you were probably wondering, well Jack, how many salsa drinks are there? I have no idea. There's too many to come. I went down and approached you to our last night phone. I did a little snooping around. 21 different brands of salsa on the shelf, including Harlan. That is about the least innovative, most unimaginative product that's probably up here today. Right? However, bear with me a little bit. The innovation has not come from the product. It does not even necessarily come from the story. There are other groups of businesses that do something like this, including social mission. What comes is when you put it all together and you start thinking about how should this product be distributed. Come back to me for a minute. We're in the grocery store. There's 21 brands of salsa. Most of those brands, almost all of them, are in grocery stores across the country and they're competing for the same target market as we are. You, myself, Montaz, your parents, pushing their carts down the aisle, looking at all the sauces and picking on price, on taste, you name it. And I thought to myself, there's got to be more of a market out there than just that. In a way that I don't have to compete with 20 other brands on the shelves. And so I got back and I looked at the mission of our business, which is to empower people to change lives and the purchasing of our products. And I thought to myself, is there a way that we can reach customers outside of just our local partners in Minnesota? Is there a way that we can work with businesses to connect them? And businesses that are looking more and more to align with social mission-driven companies like ours. And most importantly, is there a way to empower entire communities to get behind a life-changing mission, whether they want to help their local charity or they want to send people to different mission trips and help out in places like Ethiopia? And what we came up with was four sales channels. Three, two of which are fairly original in my opinion and two that have been tried and true. Retail, which I've already talked about, has been our bread and butter for the first year. We're working at Plums and Biodies and Qualsas and things of that nature. The second one to reach those customers beyond the state, we started working with Amazon. I didn't know how that was going to go, but since we started in November sales have increased every single month. And we're selling more and more sauce. I'm actually getting tired of packaging stuff and sending it out to be honest with you. But I like it. People from Kentucky, Oregon, California, everywhere are buying it. We're getting good reviews. And it's fun to see that it's not too hot. Even though it's in the Ministry of Spikes. The third channel is in corporate gift baskets. How can we engage with those businesses that are trying to align their brand with the brand that they want to be a part of? We actually saw on the slides earlier a gift basket. We started selling around Christmas, kind of by accident. And we ended up selling 500 of them. Some of those people have bought it were businesses about 100 at a time. Some bought 20 at a time. But either way, that was a significant portion of our revenue in our first year. It was one of the leading reasons we were able to build it well in March. So we think that's going to grow. We think that's very, very scalable as businesses continue to look in this. We're working with partners that are going to be marketing this in their catalogs. We're going to be working with businesses directly. And finally, what I am most proud of and most excited about, which has not been tested yet, that I have a lot of confidence in, is that fourth channel. Remember I mentioned how can we empower entire communities. I don't know about you, but when I was a kid, whether it was at church or whether it was at school, it seemed like every few months I was out on the streets peddling some sort of product. Whether it was candy, it was salt. I've been told by some of the older people in the audience that some of you may remember Happening's books. I don't know what that is, but it sounds terrible. And I thought to myself, why can't we take our brand, our life changing brand, and equip young people at churches, at schools and say, go out and sell this stuff. Pick something that you want to support. Maybe you want to feed the homeless. Maybe you want to go to a local food shop. Maybe you want to send kids on a mission trip. Go out, sell this.  What's the life changing salsa? Why is it life changing? Well, two reasons actually. I'm going to be selling this salsa to go out and make a point of mission trip to New Orleans or wherever it is. And the company that distributes this is building wells with a percentage of the profits. And they can go online and provide them with all the marketing materials. And I don't know about you, but when a good idea gets around at my church, it starts getting spread all across. People say, you got to do this. And this is something that's totally scalable. When you hook them with the materials, they go out and sell it. They have the orders. I just drop the salsa at their front door and say, thank you very much. I hope that you made a difference and changed some lives today. And finally, I don't want to forget about the retail channel. Those other 20 brands of salsa in the shop, they didn't do these extra things like gift baskets. They didn't do fundraisers. They didn't need to. They worked at it for a long time and they built up their network. They started regionally just like us and they expanded across the country. There's no reason we can't do that. I've sat down with three different distributors. They're very willing and eager to carry this product. I've just told them I can't do it right now. I'm looking for more capital. I want to make sure that we're capital secure and sustainable before I move forward. But they want to get us into Kansas City in the Midwest and they're looking to move us nationally. And that's very, very exciting. But again, I'm not going to do it until I'm comfortable with where the business is at. And finally, to wrap this up, the name of this business is not Life Changing Salsa LLC. It's Life Changing Foods. And I don't know when it's coming, but it's coming soon. There's going to be different sauces and rubs and spices that are going to all complement all four of these revenue channels and they're all going to work together and only going to build a more scalable and sustainable platform for us who at the same time continuing to change lives. And as we move into our second year of business, we'll be moving on to our second one. So with that, I thank you for having me up here today. I wish I'd have brought some chips for you. Maybe we could work on that in the reception. Any questions for me? First, there are a lot of them. One is potentially financing from nonprofit foundations, which would be extraordinarily powerful. So I'd like to think about that. And then also the subscription boxes. This is a real house for a new company. I just want you to be in the box to help students. So with me, you've got to do the boxes and you've started the boxes. So that's what you're going to do. Thank you. I appreciate that. Questions? Questions? Yes, sir. I think you've really thought through the channels. And I've been very thoughtful and really attentive to designing a market. I guess with this first project, I was also curious about the manufacturing of this type of scalability. We had a co-packer called Paul Wisconsin, who has been growing with us. He started with a 25 case order. Now we're doing 250 case orders, and he's waiting for that first. In fact, he doesn't even hit economies of scale until we're doing a 6-9 case order. So I'm feeling pretty good about the number. Excellent question. Other questions, but we're still in time. Who's Doc? Doc E. Oh, the man and the myth, the legend. He happens to be a professor here. He was one of the early starters. He wanted to encourage us and give us a chance to go with this. So these are his recipes. And we just ran into him. I put his name on it for better or worse. But people love the mystery behind the man, Doc E. You can maybe meet him later. Last questions. Last big questions. I love being on the stage. Don't take me on. I'm sorry, Doc. Thank you. Thank you.


My name is Kerry Saunders and my son is Aaron. We were camping down Llangennith last summer with a few friends and family. We went for a walk over to Bluepool and at that time we saw a young boy being washed out to sea. Aaron attempted to go out to rescue him in the first instance and realised that the current was too strong and I went into the water after that. I managed to get to the little boy. Attempted to tow him in various ways. We weren't successful so I realised that the best chance of survival was for us to float on our back until we got separated eventually and then Aaron and I rescued us. We were both lifted to Morrison Hospital by the coast guards where I was treated for hypothermia. We are trying to get signs erected in the area so that people are made aware of the dangers of the current there. We've got a local councillor and MP who are assisting. Hopefully it will be progressing and hopefully we'll see some signage sooner rather than later. I'd like to thank the R&I and the coast guards that came to our rescue. Without them it may well have been a very different ending to our story. I'm sure anybody that was capable of swimming or thought they could attempt the rescue would do the same but they are honoured to be nominated.


This systematic review assesses the characteristics of smartphone-delivered ecological momentary interventions, EMICE, using self-reported ecological momentary assessments, EMACE, in relation to their effects on health behaviors, user engagement, and user perspectives. The results show that EMICE are most popular in the mental health domain, and have been found to be helpful by users. However, there is great heterogeneity in intervention designs and measurements, and further assessment in larger participant cohorts, and well-designed evaluations following reporting checklists is needed to determine their efficacy. The review suggests combining self-reported EMACE of subjective experiences with objective data passively collected via sensors to promote personalization while minimizing user burden, as well as exploring different EMA data collection methods example chatbots. This article was authored by Kim Foong Dao, Katrina Cocker, Hong Laitong, and others.


I've got to be here for at least half of it. So I'm going to talk about results from mainly atomistic molecular dynamics simulations. There's a little bit of coarse-grain molecular dynamics simulations. So the talk is really split in two parts. So the first part I'll be talking about maturation transitions of a bacteriophage that can be driven by pH modulation in vitro. And then the second half of the talk, and so this is a little bit older work that we published a few years ago, and then the second half of the talk is stuff I've been working on more recently. And so I'm interested in non-envelope viral entry and there's kind of two aspects to this problem. One is the pH sensing aspect, which I'm actually not going to talk about today but we've been working on. So I'm going to talk about the result of this pH sensing mechanism, which basically is the release of membrane-disrupting peptides. And so I'll be talking about, in the second half of the talk, the interactions of these membrane-disrupting viral peptides with membranes and then I'll be looking at how they are organizing inside of membrane. So essentially it's the modeling the result of a pH mediated process in the second half of the talk. So to start off with the system we're looking at is the Bacteria Phage HK97, which is a well-studied T-equal-7 icosahedral phage. And so 420 coproteins assemble into the immature Pro-Head-1 particle. There's a cleavage event that leads to the initial metastable Pro-Head-2 particle. In vivo, this is the stage at which DNA would begin to package and it would drive expansion through at least one intermediate state, ultimately expanding and faceting and reaching what's known as the head-2 mature state. In vitro, an analogous transformation can be triggered by modulation of pH. So from Pro-Head-2, if you drop pH, you get a transition to this expansion intermediate and then neutralization is required to reach the final states. I'm actually gonna be talking about, so HK97 is famous for making these cross-links and having this chain mail structure. I'm gonna be talking about a mutant variant which has one of these cross-linking residues mutated and so it doesn't form cross-links, but you still end up at a similar final state what's referred to as head-1. So I'll be talking about the transition between Pro-Head-2 and the head-1 system which is an expansion and a faceting. Do it like this. So the changes that we'll be seeing, right, here's the kind of global changes and then looking at the subunit structure and the capsimere structure. So we have crystal structures of both of these states and when the immature form was solved, the Pro-Head-2 state was solved, there was some interesting structural differences. The title of the paper was an unexpected twist in viral maturation, I think. And so one of the key things that they noticed was this kind of deformation of what's known as the spine helix and then there's also this rotation of this beta-strand e-loop between the states and then there's also a real organization of what's known as the a-loop and that's more obvious when you look at the capsimere structure where you see this skewed hexamer that has kind of a more linear organization of these e-loops whereas when you transition to the mature state you have this symmetric organization of these e-loops. So we'll be coming back to these different structural motifs. So what we did to look at this process was we generated, so we're simulating the asymmetric unit of this capsid and we do that through icosahedral symmetry boundary conditions so we're restricting the virus to remain perfectly icosahedral e-symmetric which maybe is non-ideal but we are able to capture the effects of the full capsid so it's a way for us to kind of mimic simulating a full capsid without the computational cost and so we generate a structural pathway between these two end states and we do that through a fairly simple method of, well it's a somewhat complicated interpolation but that offers, potentially offers some benefits over the more standard thing that people do which is a steered molecular dynamics simulation where you apply force and you drive one confirmation to the other and what can be the problem in steered molecular dynamics is that it's what's known as kind of the low-frequency first problem and that's that the system will move along these softer degrees of freedom initially and then there's all this high-frequency motion that needs to happen at the end of the pathway so you get these barriers that are at the end of the pathway and so what we did provides a more even distribution so here's looking at the low-frequency normal mode projections and the high-frequency normal mode projections and so we see we have a maybe more even distribution of those modes throughout our pathway so from that initial pathway we can try to refine that pathway to a low-energy pathway and we do that through a method called the string method and what the string method does is it essentially tries to relieve forces that are acting orthogonal to your path and so when your string converges all your forces should be directed tangential to your path and we see a good degree of convergence through energetic methods that basically the energy of the pathway is stabilizing as well as structural metrics so the structures along my path ultimately basically their RMSD is flattening out indicating that the path is not gonna be changing more so all I can really say is it's a locally minimum it's a pathway and a local minimum is essentially as far as I can say so we have this locally minimized pathway and we can calculate the free energy change along this pathway and we do that through umbrella sampling so we can simulate all of these snapshots we have along the pathway and we restrain them to remain along their pathway so basically the variable where we're straining along is this expansion coordinate and then we can compute the one-dimensional free energy profile from that and so what we see is we see a quite large uphill barrier which is more or less to be expected and the number we get is very large but maybe not outrageous so 150 kC per mole thermal scanning calorimetry showed that there should be a 250 kC per mole difference in the stability between this mutant pro-head two and the head one state so maybe it's a reasonable change in energy we do, so this pathway is generated without any knowledge of the intermediate and we do pick up that the pathway passes through something close to the intermediate so this intermediate just had a cryoem density I think it was around 12 angstroms and we compare our structures along the pathway to that cryoem density and we do see we reach a point that has a quite high correlation to that known intermediate structure and while it's not sitting in a free energy well it's not sitting in a well it is sitting in a flatter region of the landscape and so that's shown in green so this is the slope of our one-dimensional free energy profile and we see that it's landing in a somewhat flatter region of this landscape okay and I'm mentioning that it's a PMF at pH seven and I'll say why that is it's because we're performing not just standard molecular dynamics but we're performing constant pH molecular dynamics and so without getting into all the details the general idea here is that where we have tight tradable residues that can change the protonation state continuously during the course of the simulation so where this is known as a lambda dynamics method and so there's a lambda variable which would describe the lambda equals zero is a protonated state lambda equals one is the unpronated state and so the general idea here is that we know the pKa of what we call a model compound which would be a single amino acid and we know right so there's a certain pH we would like to simulate at so our standard molecular mechanics force field can give you some of that free energy change but it certainly isn't complete right and so there's some term that we're missing that I'm just terming delta QM and the key assumption in this is that this term that we're missing is basically local effects and we use this essentially correction to then bias or supplement our force field when we simulate the protein so basically we have this pH dependent term and then we have what our molecular mechanics model gives us and so that what gets added to the force field is right our difference between the pKa and the pH that you wanna simulate at what the molecular mechanics model predicts that that protonation change would be and then there's a barrier term which just tries to discourage lambda from existing and kind of intermediate non-physical states and so when you're simulating say an acidic residue at around its pKa so pH four what you see is you can see a lot of titration events so this lambda variable jumping between zero and one whereas if you're simulating relatively far away from its pKa you would see that it's basically fixed in the protonated state around lambda equals one. So with that we can come up with, we can get a fraction of the unprotonated and protonated forms of the different residues that we're titrating during the simulation and that allows us to estimate a pKa of those residues. From that we can use something called the Wyman-Tanford linkage equation which relates the free energy change, the gradient of the free energy change with respect to pH to the change in charge between these two states and ultimately what the way we use it is that if we have a, we know the free energy change or we have an estimate of the free energy change at a given pH using these pKa's we can try to estimate how the free energy would change at a different pH and so rather than having to run simulations at all these different pH's we can try to make some estimates or predictions about how just based on knowledge of these pKa's how that would modify the free energy changes. And so when we do that, when we look at our state, so looking at the initial transition from pro-head two to the expansion intermediate which should be favorable under acidic conditions, we see that the trend is correctly predicted that basically as you decrease pH the transition becomes favorable and then as I said you need to re-neutralize to or you need to raise pH to go from the expansion intermediate to the head one state and though it doesn't, I don't think it gets quite favorable the general trend is correct in that the free energy is decreasing as you are raising pH. And so we can basically map these changes onto our original PMF computed at pH seven. And so what we can see is as you decrease pH right you're essentially increasing this well for the expansion intermediate and then as you are increasing pH you're flattening out this landscape as you go from expansion intermediate to the mature or this head one state. And so it doesn't actually, we would expect it to get negative and it never quite does, but it does flatten out. So we're happy with that but I think what's maybe a little more interesting is that we're able to identify residues that we think are key in controlling the dynamics between these states. And so what we can do is we can look for residues that have shifts in their pKa's between the different states. And so we're just looking at this acid induced initial part of the pathway between pro head two and the expansion intermediate and picking out the things that have the highest pKa either the most positive pKa shifts or the most negative pKa shifts. And our kind of thinking or hypothesis about this would be that since this is an acid driven conformational change that things that are increasing their pH are gonna want to change a protonation state and would be related to the structural reorganization of the capsid and things that are maybe, while they're resisting protonation and maybe you could say they're resisting the transition have decreased pH's and one thing that jumped out at me was that you find that these things that are resisting the transition are at the interface of the capsid at the interface of the capsimere and HK97 assembles from capsimere. So it would indicate that these are residues that are important for stability, right? So we kind of are relating that these things with downshift pKa's are important for stability and things with upshifted pKa's are important for mediating conformational dynamics. And so we can go through and I'll give a couple examples. So of our top most shifted residues one that has one of the most downshifted pKa's that we think is important for stability indeed form salt bridges at the capsimere interfaces and if you mutate this residue you don't get assembly. So it prevents assembly. So it is critical for stability and assembly. A couple more residues. So these glutamic acid residues are located at this three-fold axi and they form what's been termed the three-fold staple and so they form a series of salt bridges and right. So it's between each of the pairs at this three-fold axi, each of the protomers at the three-fold axi. And again, if you make mutations to these residues you again prevent assembly. So another example of residues important for stability that we're picking out from our pKa shifts. Okay, so now let me talk about the ones that have that we think are important for mediating structural changes and we'll be looking specifically at these three structural motifs that have been highlighted in the literature and that I highlighted at the beginning of the talk, the spine helix, this elope rotation and the kind of hexamer reorganization. So the first one is the spine helix and this one was interesting. So this glue 153 is located on the spine helix and it forms a salt bridging interaction with arginine 210. But that salt bridge is there both in the pro head two and the head two two state. So in blue is our salt bridging distance so we have that there, both there in the beginning and the end but what we see is that and then in green is the rotation of this helix so essentially the straightening out of this kink. So what we see is that essentially you need to break that salt bridge and it allows the helix to rotate and then that salt bridge comes back in and locks it in place. And so by allowing that glue to change its protonation state we think that facilitates that conformational change. We also pick up several residues on the elope that are right in the elope is known to undergo this rotation. We focused in on E149 which is down at the base of this elope and it forms an interaction with Q195 on a neighboring subunit and essentially we think that as that interaction breaks as well it allows for mobility of that elope helix and we see that the distance between these residues correlates well with the elope rotation that it undergoes during the maturation. And then the last structural motif was the aloops at the center of the hexamer and we pick up two residues that are on those aloops and so when you're in this asymmetric configuration in the skewed hexamer there's salt bridging interactions that these acidic residues are involved in and so essentially you need to disrupt those salt bridges to allow for this symmetric structure to get adopted and the other thing I'm showing here is there was some HD exchange that showed that most of the changes in the elope HD exchange profiles were occurring between the ProHead 2 and the expansion intermediate state and there was very little change between the expansion intermediate and the head one and all I'm showing here are contact maps between the different aloops and essentially it's showing it may not be terribly obvious but it's showing that there's more change between the ProHead 2 and the EI state and then there's pretty minimal changes between the EI state and the head two so indicating that the structures we're seeing are consistent with to some degree those HD exchange experiments. So the last thing we did for HK97 was right so I'm talking about in right phages don't really need to be pH sensitive since they inject their DNA and are probably so we would like to see if we can make a connection to what drives packaging or drives this maturation in vivo which is the packaging of DNA and so can we relate what DNA would be doing to what we think pH is doing and so the way we set up this calculation I'll try to take a minute to explain it is that so right ultimately we would like to know what the effect of DNA is on, so essentially how is DNA changing the free energy to cause this maturation transition so what we know are, so we have experimental measurements so we have predictions for what this pH driven maturation changes and so we can formulate a thermodynamic cycle and try to get at the DNA driven free energy change as a function of what the pH driven maturation would be and then minus the effect of DNA packaging on the ProHead 2 state versus the effect of packaging on the Head state and what we're talking about here is really just, so essentially we're just assuming that DNA is gonna cause an upshifting of the pKa's of the acidic residues on packing and negatively charged DNA in there and that that would raise the pKa's to some degree of these acidic residues and so we don't really know how much DNA gets packaged in the ProHead state so the packaging starts in the ProHead state and so our assumption here is that so basically there's gonna be some perturbation to these acidic group pKa's and we're just gonna look at and say, well you're gonna have some, potentially you have some fractional change in the ProHead state versus the change you get when DNA is fully packaged and so we can look as a function of different ratios of what the pKa perturbations are and see how much perturbation is needed to make this transition favorable given that you're starting at a highly unfavorable state and so what we see is if there's very low perturbation to the ProHead state which would be a ratio of zero you can achieve, this would be favorable just based on pKa shifts at somewhere around I guess it's 0.3 or 0.4 pH unit shifts but even if you have a fairly sizable change in the ProHead state too so say a 0.6 you still are only requiring somewhere around a half of pKa 0.5 or 0.6 pKa shift so to conclude on the first part on the HK97 section so we use basically pathway refinement methods to and constant pH to calculate pH dependent free energy change well and from the pKa's we're able to map these free energy differences onto our our PMF or one dimensional free energy change and we identified critical residues that we think are either controlling or connected to both kind of stability and then structural reorganization of this capsid. And then in the end we use this thermodynamic analysis to try to understand how the DNA could be perturbing these pKa's to drive conformational changes as well. Okay, so that was work I had actually started quite a few years ago when I was a postdoc with Charlie Brooks and continued on for a few more years. Stuff I've been working on more recently and what I've been quite interested in is trying to understand non-envelope virus entry mechanisms and particularly what's happening in the endosome and how viruses are sensing environmental changes and then how are they getting out of the endosome and this is the aspect I'll focus on today. And I would say just generally right so our understanding of envelope viruses is somewhat better I think than the non-envelope viruses so for some viruses anyways this fusion peptides have been well studied. Influenza is probably the most well studied example whereas in non-envelope virus entry there's considerably less is known. So our model system that we look at here is called flock house virus so it's a small t equal three icosahedra least symmetric capsid. It's a member of the notavirus family which infects insects is an RNA genome and so here's the capsid structure and here's the asymmetric unit structure. It has an autocatalytic maturation event which cleaves the 44 C terminal residues to form what's known as the gamma peptide. The crystal structure resolved 21 residues the first N terminal 21 residues and this peptide is termed gamma one and the first things I'll talk about are related to gamma one which has been the subject of some older in vitro experiments and so the gamma one peptide forms an amphipathic helix and like I said it was resolved in the crystal structure. These phenylalanines are highlighted and I'll probably briefly touch on those toward the end of the talk. And so there's a fair amount of structural conservation at least certainly among notaviruses so I'm showing a view from the inside. So maybe I should go back when I... This is a key point that, so you don't see the gamma peptides on the exterior of this capsid. So they're sequestered on the inside of the capsid and they need to get externalized. And so here's a view basically from the inside of these various capsids looking down a five-fold axi and you see a similar structural motif between notaviruses and tetraviruses and also this one bernie virus that you have these peptides aligned along the five-fold symmetry axi and the general thinking is that this would be the point of externalization that they could, there would be some dynamics at that five-fold, a pore could be created at the five-fold symmetry axi and these peptides could exit through that symmetry axi. Lytic peptides and non-enveloped viruses are quite prevalent. We keep learning, finding more systems that have either a lytic peptide or some component of their capsid that are able to disrupt membranes. So for many different virus families and many of these are triggered by, well there's some stimuli that triggers them in some cases receptor binding. Low pH is also a common stimulus for activating these peptides. So what do we know about gamma? So gamma is critical for infection. So you need cleavage of gamma for both infection and also for in vitro liposome disruption.  And a key point to showing their importance for infection was what are known these intrans experiments where a non-cleaving virus, so a virus that doesn't cleave its peptide will have very low infectivity but if you supply a wild-type VLP with that non-cleaving virus, you get robust infection. So having those cleave gammas is critical. So there is a pH dependent process to this but as far as we understand it's the release of these peptides that is pH dependent. So you get a maximal, this is die leakage, you get a maximum around pH six which would be consistent with kind of endosomal conditions.  And so the interpretation is that the pH six condition is exposing or externalizing these peptides and then they will disrupt. Working just with peptides alone some experiences have shown that very similar die leakage profiles occur at different pHs as well. So essentially to get out you need this pH change but once they're out they have pH independent activity and also raising endosomal pH also inhibits flockhouse virus infection. Okay, so the first thing we were interested in looking at is trying to characterize these Gamble One peptides in membrane environments and there's some data showing that the composition of the membrane will affect how well they disrupt membranes. So on a neutral zwitterionic POPC membrane ratios, lipid to peptide ratios of around 30 to one are effective at disrupting these membranes whereas against a negatively charged POPG membrane you need much higher peptide concentrations to achieve a high degree of leakage. And so we started out trying to see if we could get some insights into why that could be. All right, so let me speed up here. So here are some of our questions. So the first thing we looked at was membrane binding and we used the Martini coarse-grain model for doing this and Martini suffers in its ability to model protein conformational changes. So what one needs to do is supply what you want your secondary structure to be. So we can run simulations where we, what I'm saying is turn secondary structure on which means I'm forcing the peptide to be helical or I can turn it off and I can let it be something that resembles a random coil. And what we see is that for PC we get, it requires helical formation to get a stable binding whereas if it's not helical we only see kind of transient contacts. Whenever we have some degree of negative charged lipids in the bilayer we always get rapid binding and stable binding. I'm gonna move quickly past this because I think I'm getting a little behind schedule. So from those coarse-grain simulations we can convert those two atomistic simulations and we can run molecular dynamic simulations to see folding or see the stability of these different configurations. And so on PC it's initially helical and we find that it remains helical and on PG we have a disordered structure that on the time scale of a microsecond remains basically disordered and surface bound. So that's not all that interesting or maybe all that fair of a calculation. Okay, so what we did was maybe a little bit in fair calculation was to say, okay, let's start at the same initial structure and we'll just swap out the lipids. So we do what we call lipid substitution simulations where we take a snapshot from the peptide bound to a mix of PG and PC and then we swap out the lipid type and allow the simulation to fold potentially and we see that on a PEC lipid we recover that folded state and on a PG lipid we get relatively low degrees of folding and we also get deeper insertion into the membrane with against PC. And then the last thing we did here was we said, okay, well if it's really unstable on PG then if we start as a helix and we run on PG we should see some destabilization and the result was that basically we didn't so we still see the same degree of insertion and helicity and that led us to think that what the charge, introducing the charge might be doing is raising the folding barrier and so that ultimately the folded state is more stable but having some charge in the membrane may prevent or slow down the folding of these peptides which would relate back to why, so the presumption is that helical structures are needed for lysis and so if you can achieve helices quicker or more favorable on PC then that may provide some explanation for the difference in peptide concentrations required to lice these different membranes. So since I'm running short basically what, so Gamma 1 there's been biophysical studies, it's small, it's easy for us to work with but it's really not very infective so the full length peptide is much more infective than this would be Gamma 1 so its biological relevance is somewhat limited I would say and so we really wanted to move towards understanding how the full length peptide behaves differently than the Gamma 1 and so I'll go through this quickly, we did martini simulations again with Gamma 1 in transmembrane configurations and basically they're all unstable, when we do martini simulations in with the full length peptide and essentially what we do is we put the Gamma 1 region in the helix and we allow the C-terminal region to extend, we find that the C-terminal region always inserts into the membrane and all these different oligomeric states are fairly stable but okay that's martini, it's not necessarily the best for modeling proteins and so I got an allocation on this Anton supercomputer that David Shaw built and we were able to run some longer time scale atomistic simulations and I'll just finish up by going through what we've seen and we've just been getting this data recently so when I run Gamma 1 at a surface bound configuration essentially it stays at that surface bound configuration when I run the full Gamma, now I'll come back to this in my last minute is we see that aggregation is happening and these peptides are basically coming off the membrane so we were thinking we may see insertion and we're seeing the opposite, we're seeing aggregation and essentially them coming off of the membrane when we, so then we can just insert them in a transmembrane configuration and so the Gamma 1, the shorter Gamma 1 peptide has, essentially there's nine peptides in here and for them transition to the surface on the time scale that we simulated for and maybe more of them will if we run that longer so they don't seem terribly stable even in all atom simulations whereas when we start the Gamma 1 in a transmembrane configuration they do remain stable and so the last point is just looking at water flux and pore formation so we were quite happy to see quite a bit of pore formation so we see a little bit of water when Gamma 1 is there and then we see quite a lot of water when the full Gammas are there and so we see increasing amounts of water density in the interior of that hydrophobic slab as we run these simulations and these are over eight microseconds here's just looking at different kind of pore formation so we see trimmers and we also see a a hexamer structure and I think with that I'll make my conclusions well actually I had one more thing to say so the aggregation thing is it really kind of has changed my thinking on this a little bit and it's made me wonder so I had always viewed that these peptides, you're in the endosome the peptides externalize they find their way to the membrane they create a hole in the membrane and maybe that's not how it works and so maybe they're being tethered within the membrane and so either in a VLP that may be due to this propensity for aggregation in a virus it may be due to interactions with RNA so the termini of this peptide is GF, GF which I don't know a ton about RNA binding but apparently that's a consensus RNA binding motif and so I've written a new anti-allocation and I wanna look at basically the effect if I kind of restrict these peptides if they were remaining the termini were remaining in the capsid and then also try to map RNA onto these peptides by doing single nucleobase simulations and this model has some attractions it provides you a high local concentration there's structural organization there and also you have the virus is very close to I mean the virus is right there and so if you're creating a hole it seems that it would be easier for the virus to get right through there and so with that I'll just thank my funding from NIH and computing resources Shivangi Nanjia did the work on the Flockhouse virus and then I collaborate we're doing experiments now on these peptides with Nathan Alder at UConn and then the HK97 stuff was stuff I had started with Charlie Brooks and Jack Johnson has kind of encouraged me on both of these projects because he's done much of the structural biology work on both of these systems so with that thank you for the opportunity and I'll take any questions.


10 English words ending in LESS. In English, the suffix LESS means without. For example, something that is colorless is white or transparent. It has no color. So in today's lesson, you'll learn 10 English words that end in LESS with example sentences so you can see how they're used. Number 1 CARELESS means without care. For example, my sister is very careless with her possessions. She left her laptop on the sofa and later someone sat on it. The opposite of careless is careful. Number 2 DOUBTLESS This means without a doubt. Something that is definitely true. For example, the prices will doubtless go down around Christmas time. There are always holiday discounts. The opposite of doubtless is doubtful. Number 3 ENDLESS Endless means without end or very long.  We often use the word endless to exaggerate how long something is. As in the example, it doesn't necessarily mean that there is no end. It just means that something is very, very long. Number 4 FEARLESS The word fearless means brave completely without fear. For example, John loves radical sports like rock climbing and skydiving. He's absolutely fearless. The opposite of fearless is fearful. Number 5 HELPLESS Helpless means unable to help oneself. For example, little babies are so helpless, their mothers need to do everything for them. The opposite of helpless is something like self-sufficient, someone who can take care of themselves. Number 6 HOMELESS The word homeless describes someone who has no place to live. For example, in the city center there are a lot of homeless people who sleep on the streets. Number 7 HOPELESS Hopeless means without hope, without possibility of success. For example, the doctors say it's hopeless, his cancer can't be cured. We usually describe a situation as hopeless. For your own feelings you should say, I have no hope, not I'm hopeless. For the opposite, however, you can say I'm hopeful if you do have hope. Number 8 RESTLESS Restless means agitated, not relaxed. For example, I've been sitting at a computer for the past six hours and I'm feeling restless, let's go for a walk. The word restless is not the same as tired. The word tired means you have no energy and you want to go to sleep. The word restless describes a situation where you don't want to stay still, you want to move or change something. You can be restless due to emotions, worries or simply after a long period of inactivity. Number 9 TASTELESS Without taste or flavor. For example, the food at that restaurant is cheap but it's pretty tasteless. The opposite of tasteless is flavorful or tasty. There is the word tasteful but this is very different. The word tasteful refers to decorations or words that are good and appropriate for the situation. But the opposite of tasteful is also tasteless but used in a different way. In addition to meaning without taste or flavor, the word tasteless can mean not appropriate. For example, it would have been a good movie for kids except it had a completely tasteless sex scene in the middle. Things that are tasteless in the sense of being not appropriate may be vulgar or visually ugly or simply not appropriate for the situation. Number 10 USELESS Describe something that cannot be used. For example, this brochure is useless. All the information is out of date. We'll have to call to get the current information. The opposite of useless is useful. So let's review our 10 words ending in less, careless, doubtless, endless, fearless, helpless, homeless, hopeless, restless, tasteless, and useless. All of these words end in L-E-S-S. You're learning a lot of vocabulary at Espresso English, but how can you remember the new words? Well, one of the best ways to remember new words is to put them into practice. This means using them in practice exercises as well as trying to create your own example sentences with the new vocabulary words. In the vocabulary builder course, each lesson comes with a quiz as well as writing exercises that will help you put the new words into practice. Visit EspressoEnglish.net, click on courses, and look for the vocabulary builder.


21 Savage has been arrested by U.S. immigration and customs, aka ICE, for allegedly being unlawfully present in the U.S. Reportedly, he's originally from the U.K. and has overstayed his visa. A CNN reporter quoted an ICE spokesperson saying that his whole public persona is fake, but a lot of people are saying that's a bit of a stretch and also that ICE is a truly awful institution. Fata senior editor Lawrence Burnie tweeted, Demi Lovato got caught up in the controversy after tweeting that 21 Savage memes were her favorite part of the Super Bowl. Many people are arguing that the jokes and memes take away from the real issue, though. As one person tweeted, A lot of other celebrities are showing their support for 21 Savage. And when right-wing political commentator Tommy Larran tweeted, I got one, two, three, four, five, six, seven, eight ICE agents ready to deport you, Cardi B responded, Don't make me get my leash. A reference to an earlier kind offer from Cardi to dog-walk Tommy. And as media guru Karen Sybil aptly surmised, the 21 Savage arrest is beyond sad. He's been doing such incredible things in the community and changing the stigma of how people perceive rappers to think slash be. Ariana Grande just received the best gift in the world. When Ari tweeted at Tyra Banks asking what she would have done with her hair on the make-over episodes of America's Next Stop Model, Tyra tweeted back, Cool that we got the answer to Ariana's pressing question and that it is the correct answer. But what we really need to know is, oh my God, what happens at Tyra Banks' personal branding course at Stanford? Turns out Tyra started co-teaching the course in 2017 to Stanford MBA students. It's called Project U, building and extending your personal brand. According to the glass description, it will teach students all about personal branding and how it can be unleashed as a valuable competitive advantage. Say what you want about the somewhat terrifying prospect that these days, everything we say and do online has to feed into a strategic personal brand representative of your desired personality and analyzed by everyone from prospective bosses to tender dates. Tyra's smart. She's transferred from model to very successful presenter slash businesswoman slash author slash producer with a reported net worth of 90 million US dollars. She's acutely aware of the power of a strong visual identity, whether that's a smile or a ponytail. I think I need a new haircut. Other fun facts, this class will be a highly interactive learning environment with wait for it, wait for it, image transformations. You can literally get a Tyra makeover as part of your uni course. Presentations take place through YouTube recordings, TV broadcasts and Facebook lives and cover everything from pop star here to ice cream. And you can be a new ice cream company and say, say 20 years ago, I'm going to come up with the best vanilla in the world. Ain't nobody going to be paying attention to that. However, there are two guys, one named Ben and one named Jerry that said, you know what? We're not going to be the best vanilla. We're going to do some decent vanilla ice cream, but we're going to throw so much stuff in there. We're going to be different and that made them stand out. The only question left is, do they offer international scholarships to struggling presenters who base their entire personality on the fact that they just spent six months on crutches? Please tell me what to do with my hair, Tyra. Tomorrow by Together aka TXT, a K-pop's hottest new boy band. They've got 1.2 million Twitter followers and counting, an official community website for fans to connect on and countless YouTube videos of people analyzing small but very necessary details about each member like their star sign or the fact that one of their nicknames is apparently Cucumber, because he is tall. All before they've released any music. Well, they have released a handful of videos for all five of the members, which kind of resemble ads for baby oil. All that one direction fragrance ad where they smelled flowers and focused a 16mm film viewfinder that isn't even attached to a camera. Eager fans have spotted clues in the videos, including more scope messages which spell out dreaming or secret and that each member has an animal to represent them, like a butterfly or a gecko. So why are they so popular when we barely know anything about them, except for one crucial point, that they're formed by an entertainment company called Big Hit Entertainment, who have one other boy band which they made six years ago and that is BTS. So are you a TXT fan? And so please tweet us why we want to know what you like about them, who your favorite is, what's their music going to sound like, what's their music going to be about. Let us know everything that you want to see in TXT.


Welcome to the Top 10 NBA Playoffs Fantasy Performances in Fandall History, which dates back to 2009. Just a warning for all you haters out there, you're going to see a lot of LeBron James on this list. He owns the majority of these spots since, well, he has played in 266 playoff games in his career. Let's get right to it. Starting at number 10, LeBron put up 81.1 Fandall Points against the Hawks in Game 3 of the 2015 Eastern Conference Finals to put the Cavs up three games to zero. King James finished with an insane triple double, 37 points, 18 boards, 13 assists. The Cavs went on to lose to the Golden State Warriors that year in the NBA Finals, but LeBron was an absolute tear during those playoffs. At number 9, it's another Cavs LeBron game, this time in 2009 Fandall's first year in existence. James scored 49 points in a one-point loss to the Magic in Game 1 of the Eastern Conference Finals. The Cavs went on to drop this series, but LeBron did help fantasy players with 81.2 Fandall Points in that first game. LeBron also holds the number 8 spot with 81.7 Fandall Points as a member of the Cavs in 2018. The King dropped 46 points and grabbed 11 boards to help Cleveland force a Game 6 in the Eastern Conference Finals. That year LeBron and the Cavs lost to the Warriors in the NBA Finals again. Coming in at number 7 with 82.1 Fandall Points is, you guessed it, LeBron James. This time we go to 2012 in South Beach. The Heat beat the Pacers in Game 4 of the Eastern Conference Semifinals as LeBron posted 40 points, 18 boards and 9 assists. LeBron and the Miami Heat went on to win a ring that season, the first championship of LeBron's legendary career. At number 6, we finally see a new face for this top 10 list. The Dallas Mavericks, Luka Dončić. Luka led the Mavs to an overtime victory over the Clippers with 43.17 boards, 13 assists for 84.9 Fandall Points. Luka's triple-double in the bubble was fantasy gold for DFS players who rostered him. We now return to our regularly scheduled programming for spots 5 and 4 on the list. Yes, LeBron James. The fifth best Fandall performance in playoff history came during their 2016 NBA Finals as LeBron dropped 41 points to win Game 5 against the Warriors, good for 86.7 Fandall Points. At number 4 on our top 10 countdown is LeBron's 87.1 Fandall Point performance. It came when he dropped 41 points again in Game 6, that same final series against the Dubs. You may remember LeBron and the Cavs came back from a 3-1 deficit to win the finals in 7 games. Okay, I can now promise you LeBron is done on this Fandall Top 10 list. Haters, welcome back. At number 3 is Portland Trailblazers superstar Damian Lillard who scored 55 points, pulled down 11 boards and dished out 6 assists for 88.2 Fandall Points against Denver in the 2021 playoffs. What's crazy is that wasn't even enough for a Game 5 win over the Nuggets. Despite it being quote, Dame time, the Trailblazers were knocked out in the first round by the short-handed Nuggets. At number 2, Russell Westbrook in 2017 with OKC. He posted 51 points, 10 boards, 13 assists and an incredible 93.5 Fandall Points against the Rockets in 2017. Cracking 90 Fandall Points is the stuff only legends do. It's only fitting that this legendary performance came from Mr. Triple Double himself. And coming in at number 1, the greatest fantasy basketball performance in NBA Playoff history since Fandall came into the world, Kevin Durant in a Brooklyn Nets uniform in the 2021 playoff against the eventual champion Milwaukee Bucks in the Eastern Conference semifinals. You remember that game, right? When Katie posted a Triple Double of 49.17 rebounds, 10 assists, worth a whopping 96.4 Fandall Points and one of the greatest playoff performances you'll ever see. And there you have it, the top 10 NBA playoff performances in Fandall history.