School is important, but getting revenge, is more importanter. If you like true revenge stories, you found the best place for your vengeful needs. We start off with a racist teacher who likes to bully kids in his class, but when recorded in during his most inglorious moment, he became infamous.  A teacher falsely claims a student fakes her injury and treats her bad for it, but a protective mom steps in, making sure the teacher receives a fair treatment and return. Lastly, a shallow and cranky dance teacher bullies the wrong girl, for not being as perfect as her own daughter. But her protective dad gets revenge for two. Before we begin, since the like button has an unignorable need for being tickled, tickle it right before it has a toilet emergency. Naturally, viewer discretion is advised. These revenge acts might be disturbing to snowflakes. Some details might be a bit blurry, my story happened around 14 years ago. I'm a 25-year-old male now. Also, stay in for the end, I promise it will be worth it. It all started when I was 12 years old. My grade 7 teacher was a 47-year-old man who was infamous for being intimidating and, in my opinion, abusive to his students. He was the disciplinarian of the school. He was in charge of keeping track of detentions and announcing who will be staying after class every Friday during assembly. We suspected at that time that the reason why he never got fired was either because his students were too scared to report him, or because of the fact that the principal was his brother-in-law. Reasons why I hated him personally, he was constantly making vaguely racist remarks, complaining about the new South Africa and constantly bringing up how his life was better during the apartheid regime, he's a white guy who was raised on a farm. I always felt like he had an issue with me as a person. Because I'm a practicing Muslim. He would make the class laugh at how funny Muslim women looked with their heads wrapped up.  He used to often rant about how the school is no longer a pure Christian institution as it once was back in the day. He would say these things and glance at either me, my twin sister, or the black students in the class who practice their own African religions. When it was his birthday, my mom encouraged me to buy him a gift.  Throughout the day, he would get gifts from students in his class and others. He would get up from his desk to greet and thank them, and then shake their hand. I remember noticing this because I always found it weird when students shook hands with teachers. Because of how small our hands were compared to theirs.  I remember being thankful for not offering my handout for him to shake, because I thought he might have ignored it in front of the entire class.  The experience that made me hate him the most happened just before we wrote second or third term exams. I was walking with my friend David back from the tuck shop during interval. We took a shortcut between the English and Africans kindergarten classes and saw a group of boys huddled together. One of them walked towards us and I saw that he had one of those camping multi-tools with the folding knife out and instantly got a fright. He told us, give me your stuff before I cut your neck and then started laughing and walked back to his friends. It was clearly a joke, but David looked close to tears and I was startled too. I told the guy, Zander, that he's not allowed to have knives at school and that I'm going to tell my teacher. We walked straight to our teacher and when we spoke to him, David burst into tears. We told him what happened and David was sobbing when he said he felt like he was going to die. Our teacher barely looked up from his computer while we were speaking and when he asked for the guy's name, we told him the name and he said he will deal with it in for us to go out for interval again. I went home and told my mom who I felt didn't fully believe me at the time. The next day we saw Zander, we were basically poking fun at him for getting into trouble with our teacher and that he's likely getting expelled at worst or sitting a Saturday detention at best. He laughed back at us and said our teacher just came to his class, asked to speak to him and told him to never bring it to school again. No detention, no suspension. Basically nothing. He still had the knife on him for the rest of the day. We were upset, so we went back to our teacher and I told him that Zander said that he didn't get into trouble for having the knife. He gave me the ugliest look as if I was bothering him and coldly said to me that maybe I should fix my late coming problem before I try to get other people in trouble. I would come late four or five times a month because my mom's car's battery terminals were broken, so the battery would run flat and she couldn't afford to have it fixed. She had to put the neighbor's battery in her car, start it, and then idle it while she took that battery out and put her own battery back into charge up. I felt betrayed by my teacher. The person who was supposed to make us feel safe while we were away from home. When I spoke to my friends about it, they told me that Zander was actually the principal's son, meaning he was my teacher's nephew. I decided to take the opportunity to speak to my friends about getting evidence that our teacher is treating students unfairly. Three of my four close friends had camera phones. I sat in the far left corner, my one friend sat in the opposite corner by the door, our other friend sat in the middle, and the last friend was right at the back of the class by the window on the left. One thing about our teacher, he did not give a flying frick about where we sat as long as we answered him and didn't bother anyone when we swapped seats. We came to an agreement that whenever our teacher would sound like we was going to say something vaguely racist or Islamophobic, we would all discreetly take videos of him. Any private conversation we had with him was voice recorded on our phones. We caught him on camera telling a really racist joke about black people and saying that Hindus must have a lot of problems since they have so many gods. We caught him saying a lot of bad things, but a lot slipped through our fingers because we weren't fast enough. It was extremely difficult to keep our friend group motivated to record him and not tell anyone else about it. It was especially difficult because at the time I had a hand-me-down Samsung which was seen as an expensive phone at the time. My mom prohibited me from taking it to school. She instead bought a cheaper phone for us that could only make calls and send SMSs. This was in case she needed to reach us in an emergency. I got caught several times sneaking my camera phone to school. My biggest mess up at school was when my mom phoned me on my Samsung and I answered it.  After around a month, we decided that we couldn't let it go any further. One of our friends was a black guy named Tendah. Tendah was a problem child. His mom died when he was four and his dad was an alcoholic. He was raised mostly by his uncle who I think was a pimp. He used to act out at school because of undiagnosed ADHD. His dad and uncle didn't believe in learning disabilities and always assumed he was just lazy and badly behaved. Tendah especially got onto our teacher's nerves not only because of his skin color, but because he would bring broken calculators from home and take them apart during class. One day our teacher told him to clear his desk and throw away the bits of plastic and calculator crap. He ignored the teacher. The teacher then started screaming at him and Tendah done the only logical thing in 12, 13 year old would do in such a situation. He mockingly put two pencils into his ears.  The narrow window pane cracked and Tendah's head was bleeding. He told us he was fine during interval afterwards and we put money together to buy him a sprite. I almost cried when my friend, who sat way behind me, said he got the whole thing on camera. We didn't even trust that the whole class's testimony would get him into trouble. We decided that enough was enough. Enter our revenge. First we showed the video to Tendah's uncle who showed it to his father. Then I showed my mom all the other videos and recordings. She lost her mind. One of my friends sent all of it to his older sister who had a Facebook account and she posted it there and tagged the school and as many parents as she knew. It blew up. Parents and people from around the province phoned the school demanding answers as to what is going to happen to our teacher. He was immediately suspended. There were rumors circulating that he had to go into hiding because Tendah's uncle and his friends were looking to get to him. I even met to Tendah's dad for the first time in the weeks after the whole thing exploded. He liked to joke that his dad sobered up especially for this. The principal pulled Xander out of the school. We never saw him again. My mom told me an investigation was launched against the school because of the improper handling of bullying complaints. If I remember correctly, three English kids in my class alone spoke out against teachers dismissing their complaints of bullying by the Afrikaans kids. We were a mostly white Afrikaans speaking school with three Afrikaans classes but only one English class per grade. They called us Saudis, which was short for South Peel which literally translates as Salty Pippi. If you get what I'm saying, it's a derogatory term for English speaking white South Africans. It means you're one leg in a South Africa, your other leg is in England, so your Pippi is hanging in the ocean. We only saw our teacher once after he was suspended. He looked badly beaten up and was accompanied by a policeman and two other male teachers so he could gather the rest of his stuff from his class. But it didn't end there because so many kids needed the evidence that they were being bullied and nothing was done because of it. The CCTV footage was brought up. My friend's mother who was part of the school governing body that time told us a few years ago that when they reviewed the footage, it became apparent that the principal was having an affair with one of the grade two teachers. He could be seen grabbing her booty at the furthest point away from the camera. They slipped up a few times and kissed in clear view of the camera. I guess once you're surrounded by the cameras at work every single day, you kind of forget that they're there. It was very apparent that they thought they couldn't be seen. My aunt sells Tupperware and one of her regular customers and close friends is the principal's ex-wife. Not only did she leave him, but they were not married in community of property due to a prenup agreement. The house they lived in was in her name since before marriage, so she effectively made him homeless because none of his family wanted to take him in. He ran into severe debt from staying in guest houses and burned many bridges from overstaying his welcome at friends. As for my teacher, his reputation was destination fricked. He served jail time, don't know how long, and eventually left the country because it seemed everyone knew his face from the media attention he received. The reason why I made this post, I was never going to tell this story online as I've told it over and over through the years since primary school. But I felt I had to because of what I experienced at the beginning of this year. My family is part of a nonprofit organization that has feeding schemes all over the country. The last Friday feat of February, I'm helping as security as I usually do since we're few volunteers and there's many homeless people who are on drugs and some can get violent. I'm walking down the line to make sure there are no fights or anything that could start a riot and I see somebody. The familiar face in a tattered, yellowish white button down and mud cake shoes working his way down the line. My old principal is standing there waiting for a bowl of stew and bread with absolutely no idea who's standing beside him. Obviously he wouldn't have recognized me but I never forgot his face. I'm not going to lie, I couldn't help but feel all the emotions and a single tear appeared out of the bottom of my left sunglass lens. Seeing him brought back the feelings I had when I was 12 years old in seventh grade, trying absolutely every excuse in the book to not have to go to school and be bullied by my teacher. So yeah, for those of you who are still reading, this is the end of how my friends and I destroyed the lives of my teacher and principal. If you got this far and are feeling depressed, worthless or less than your peers, I love you.  I am a sophomore in high school. We have four assistant principals in our high school for freshmen, sophomores, juniors and seniors. Anyways, my school is a very left-sided school. Most of the teachers are very liberal and honestly don't let you share your own opinion if you are conservative. One day I decided to post my own opinion on my Snapchat story. Bad idea. I posted how it is my own opinion that girls shouldn't wear makeup as I just simply don't find it attractive. Some girls started a rumor that I was talking about a specific girl and she and her friends came at my physical appearance and my nationality. I made a bad decision and called her out for cheating on her boyfriend and called her a hoe. She got really mad, deleted her stories and she made it seem like I was simply just coming at her for no reason. I was already pissed. I have tons of screenshots of what was said by both of us. However, I couldn't stop this now. I had 175 views already and the amount of screenshots I had on that story was 86. People started making memes about me, calling me a weasel, gerbil, said my forehead was big and everything. Many kids made rude memes about me. It really hurt, but I knew I still had some friends. I go to school the next day and I'm getting threats to get beat up and people are spreading rumors about me. I was distraught, but I fought through it. As I was getting my lunch, a group of at least 15 girls circled around me and started yelling at me. The girl I was talking to at the time was on my side surprisingly and she went and yelled at all those girls to back off. Turns out, someone told the sophomore assistant principal. She was pissed that I was just mansplaining and I didn't know anything about girls or women or why they were makeup. Turns out, someone told the principal the wrong story and didn't show the assistant principal's screenshots of everything. Shit escalated quickly and she started yelling at me and I kept saying that wasn't the full story. Turns out, someone made fake screenshots and gave it to her. I was surprised at this and I even showed her everything what I posted, but she said I deleted it. I showed her all the things people made with the name not blurred out and she was now surprised. She called them up and told them not to do it anymore. Then they just got let off with a warning after they left. She told me I was going to have to be suspended. She was a real skank and I don't know why she suspended me for this. I was really mad that I was suspended for at least two days, all because of a Snapchat post that wasn't even that bad. Getting suspended means a lot to me since I'm a good student and I have had perfect attendance since first grade. This is why I was about to get into national honor society. Now, because I'm suspended, my chance became really low. The minute I got home, my parents instantly talked to me and sided with me. They even said I could just treat it like it was a snow day and I didn't even get punished. They were pissed too. This is where the revenge starts. I sat at the desk in my room on my computer and I thought what I could do. I looked up my assistant principal's name on Google to see if I can find any information. I find her Instagram and her Facebook. She's quite young actually, like around 26 to 30. I decided to make a fake account posing as a random person so she could accept my follow request. I had to buy followers and make posts so it doesn't seem to suspicious. I think I spent $30 on followers and likes, I'm not sure. I needed to make sure this would work. I soon got a notification at around 5 or 6 p.m. saying she accepted my follow request. Perfect.  She had around 450 posts, could be a little less, could be a little more, I just assume around 450. I scrolled down and saw pictures of her husband and two year old with her. He wasn't tagged, so he probably doesn't have an Instagram. Not a big deal, so I scrolled down more. Then, I saw a picture of her. Except, she was in a Hooters uniform. I thought to myself that usually people work two jobs, but this picture was very odd. It was one of those pictures you could slide right and you could look at more pictures. The next picture was with a kid that looked really, really familiar. I then saw a bit of his shirt. I couldn't tell what was really on it, but I saw that it had my high school's colors. I then instantly realized who it was. The junior that was the captain of the varsity football team. I was so curious, so I slid to the next picture. He was grabbing her booty while hugging her and she was looking up at him like he was her husband. I was so shook and I had to make a screenshot. I keep scrolling through her Instagram and it's just her and this kid. Then I found the perfect picture. She was kissing him and it was definitely at a house party. I knew what I had to do. I had to screenshot this. Then, I quickly went on Facebook. I found pictures of her husband and he was tagged. I knew his name now, so I looked him up on white pages. He lived at the same address as her, so I had to do this during school. I found out where he worked and so I quickly printed out the pictures that I found. This was the best thing. I also planned to email the pictures to the school.  I went to sleep knowing this was going to work. I woke up between nine and 10 that day and was ready for this. I grabbed my bike and made the six-mile trip to get to their house across the city. I am on the varsity soccer team, so I am quite in shape. This took me about 20 minutes and when I got there I saw his car in the driveway. Great, he didn't leave for work yet. I simply just put the pictures in their mailbox, knocked on the door and biked off as quick as I could. I had to wait until at least seven o'clock to email the school. I made an entirely new email address and emailed it at around 6.30 because I couldn't wait. They emailed back almost right away and they said they will deal with it right away. Since today was my last day of being suspended, I was ready to go back to school tomorrow. I woke up in the morning really happy and excited and I couldn't wait to see what was going to unfold. In our lunch, the assistant principal, along with a few teachers, walk around monitoring the lunch. I then see the principal, along with two police officers, pull the assistant principal and the student out. The whole lunch room gets quiet and so many people are recording. I smirked really openly when I saw this happen. I asked one of the monitors to get a bathroom passed just so I could hear what was going on. He told me I could not go right now as some thing is going on. I was kind of sad about this but also savored Chadenfreude. I knew what was going on but I couldn't tell anyone. Now some stuff happened in the next few days but I'll just sum it up. First day after, the day after this happened she didn't show up, but the student did. I asked him what happened but he wouldn't say. All I knew was that he wasn't in trouble. That made me glad because he didn't have a reason to be in trouble and that wasn't my goal. Five days after, since it was a four-day weekend that week as we had Friday till Monday off, we came to school back on Tuesday. I noticed her office was empty. I was really stunned she got fired and so I looked up on our state's court case search. There was already one case and it was hers. There were charges pressed against her, something like cause child 13 to 18 to view sexual activity. Three weeks later, the school wanted to hide this so of course no news came out but a new case has opened up. Her divorce case. I saw this and I instantly knew I succeeded. She should not have ever been an assistant principal. As she was extremely hateful and biased, you can even add perv to that list. Now, she is serving 10 months in prison and I don't feel bad at all. I'm a 15 year old female and have had a long and elaborate history with injuries, especially breaking my arm. Starting from literally the day I was born, my health has been complicated. I have a few conditions but none serious enough to impact my daily life as I know it. I have a very weak immune system and somewhat easy to break bones.  My mom has obviously known me from birth and knows just the extent of my weird bad luck and health issues.  I don't know why but teachers just love to say that students that get hurt off in our attention seeking fakers and a lot of relationships with authority figures have been harmful because of this. Every time it's the same.  I get bullied for a while and then I have to try and prove that I'm not faking. I could post so many stories about teachers doing this but this is a story about the time my mom actually managed to get revenge. This all took place around seventh grade. I was 12 at the time and I had broken my wrist by climbing on top of something I wasn't supposed to and then falling off like a dumbass. My wrist was in a Velcro cast since I'm allergic to the regular cast material as they figured out through many cases of trial and error. My teacher, let's call her Mrs. Tablet since this took place while we were learning about Egyptian tablets. She taught history and our personalities clashed a bit. I'm more of a creative, adventurous free thinker type and she was very conservative by the book and proper. We hadn't had any real issues before this but we just didn't mesh well. She was absent one day and we had a sub in. I knew the sub is one of my teacher's friends and one of my bully's mom. I didn't like her but I was still respectful in class and just tried to get my work done. She got me in trouble for doodling on my paper once or twice but other than that everything was fine or so I thought. The next day I went into class like usual. We were working on an assignment about Egypt and their tablets when Mrs. Tablet called me over to her desk.  She started asking if I'd know why she called me up to her. I politely told her I didn't. To which she asked how long I've had the cast so far. I responded, about a week now, why? She asked me if I really needed that cast and that the substitute told her she saw me take it off. I told her it's a cast. I couldn't take it off because it would hurt too much. She asked that if she would ask the class they'd say I need that cast. And if she called my parents, they'd say the same thing. I told her yes, since they were the ones who brought me to the hospital. All of this conversation was done right in front of the class. She then dismissed me and people looked at me weird for the rest of class. As I later learned, apparently the substitute had told Mrs. Tablet that I'd taken off my cast as soon as I left class. I didn't, and that I threw it in my locker like, ha, I'm getting away with doing less work. So basically, she pretended I was a bad guy in a crappy high school movie.  I technically could remove my cast since it was Velcro but that would have been extremely painful and it's not like I could just walk around without it. I wasn't going to make a big deal about it at the time but it turns out she was serious about calling my parents. But she made the mistake of calling my mom. Now me and my mother don't have a perfect relationship. She has anger issues so we don't always get along well but she loves me very much and doesn't let people like this mess with me if she can stop it. According to my mom, Mrs. Tablet called her all cocky and said things like, apparently your daughter is wearing a cast, did you know that? And we all know how dramatic teenage girls can be, especially unique kids like her.  I don't think I'm allowed to repeat what was said but by the end, Mrs. Tablet had to retreat with her tail tucked between her legs. She started being even more cold to me after that day and started giving me unfair detentions for not sitting still and doodling on the side of my worksheets. Even then, outside of school, we went to the same art studio sometimes and poisoned the art director against me and got me put into a lower class. When my mom found out about this, she started to feel tingles in her revenge bone. My mom has never been a PTA mom. She honestly can't stand most of the PTA members and their kids are chosen as favorites at my school and use this power to bully others, including me and my friends. My mom is however friends with a very lovely woman on the county school board, which has more power than the PTA. She told her friend everything that had happened and supplied all the doctor's notes needed to prove I needed my cast. Her friend managed to convince the rest of the board that Mrs. Tablet was discriminating against an impaired student, given her comments about me being slower at work and just wearing the cast to slack off, along with detentions related to me not sitting still when she knew I had nervous movements and slight ADHD. At the art studio, my mother pulled me from the classes. I took it upon myself to inform the fellow kids about why. And rumors spread quickly. Soon enough, kids were getting pulled left and right and my art teacher was pulling at strings to try and get the situation under control. I actually think the studio went into temporary closure before reopening again. She was suspended for a week to go under investigation and it turns out she treated a lot of the unique, as she put it, students in a similar way. She was verbally reprimanded by two different departments, and instead of getting her standard raise over the next three years, she'll only get half of that. Not only that, but she had to retake a course on how to properly treat students. And if she's ever caught to cross this line again, she'll lose tenure. She'll just be getting her raise back at the start of next school year. And I just like to say that I hope she learned her lesson. When I was in eighth grade, I tried out for a spot on the drill team. For those unfamiliar with that, it's the group that dances alongside the marching band, and we were separate from the cheerleaders. The reason I tried out was because those girls, you know the ones, pretty, popular, teacher favorites, laughed at me when I showed up for the informational meeting. They said I wasn't right for it and didn't belong. It made me more determined to learn the routines and become more fit so I could show them. I was flexible as much as a 14 year old could be, and at home I practiced, stretched, and I showed up for the rehearsals and tryouts. Only a couple of decent girls would not act like I was poison in the line when we had to link our arms. I passed tryouts because I could do the routine. My dad, who owned his own business doing car repair, wasn't rich, but he managed to pay for my camp fees and uniforms. He dropped me off each morning of summer camp on his way to work. There we learned the routines we would be performing at pep rallies and football games. We had to stand out in the sweltering Louisiana heat and humidity in August and stand in position for minutes at a time. Any move and you had to do kicks. This was a version of pushups or having to run laps where you stood and did sets of 25 high kicks for each infraction. I ended up with blisters on my upper arms from the sun. Oh, let me tell you about the sponsor, the teacher in charge. She was the epitome of a cranky old woman who had an only child who went through these dance teams and did private dance lessons. She was perfect to her mom. At the time, dance moms wasn't a thing but that's exactly like how she was. She would berate me for the smallest things and basically bully me along with the other girls. For example, when I was feeling faint having to do kicks for the third time that day, I was sitting down beside another girl who had pulled a hamstring. She is sitting with us while watching her daughter, who was our choreographer of course, commenting how wonderful her daughter was and said to me to get back out there. She could understand why the other girl was sitting. She was an athlete so she got a pass due to her also playing basketball.  She told me I was rude when I was eating lunch with everyone else around and would join in laughing when someone said they coughed on my sandwich. The other girls would say things to me also and it was as though she encouraged it. Not only is Junior high bad enough but this on top of it was bad. I tried to be strong and show them I could make it work and do what was needed but it made me cry as soon as I got home. Our student newspaper would put out a who's who based on physical traits like best hair and best smile.  The night of our first game performance I was waiting to be picked up with a few others and the sponsor. She went to the bathroom and came back with one girl who was caught smoking in the bathroom with some high school kids. Her punishment was she had to sit out the next game. My dad was running late. He had to work and couldn't just leave his shop unattended and after about 30 minutes she told me you know this means you're off the team. My dad pulled up a few minutes later and as soon as I got in his truck I was sobbing. I told him what happened and he said just a minute I'll be right back and proceeded to rip that woman a new one about her attitude towards me. So the revenge years later this woman brings her car to my dad's shop for repair. He remembers her but she doesn't remember him oddly enough. She was just as condescending as she had ever been treating him like a lowly servant under her because he did manual labor. He was writing her estimate for the insurance and she kept trying to get him to add in stuff that wasn't caused by the accident she was there for. Things like a ding on the back bumper when the damage was in the front, et cetera. He wouldn't do it and she got all huffy and said something like if he didn't she would have his license revoked. Now, since my dad had his shop broken into and tool stolen he had installed cameras. He sent that video to the insurance adjuster when he had a long good relationship within his course of business and she got hit with attempting insurance fraud charges. I don't know the exact extent of it but she should be glad her threats to his livelihood weren't added on there. Thanks for listening to the end.  They really don't matter. Think about who you would feel you disappoint the most whose love you want to keep and who you want to be your best for. Hopefully it's yourself but if you have another person in your life family, friends, et cetera, they matter. Not shallow teachers who think the world should be an ideal version of their own making. Thank you for enjoying this episode which was made with artificial love. Subscribe to receive future episodes and tickle the like button for good karma. Do you have any experiences surrounding this topic? Share yours below. I'll join the conversation and I'll be seeing you in the next one.


Welcome to MOOC course on Introduction to Proteogenomics. We are now moving on to mass spectrometry based data analysis and in this light a new method which involves data independent acquisition or DIA will be discussed today. DIA is an MS technique which involves fragmentation of precursor ions within a selected M by Z range. It differs from data dependent acquisition or DDA where only selected precursor ions based on their abundance are fragmented. In this lecture Dr. David Campbell will introduce you to the concepts of SWATH MS, a type of data independent acquisition or DIA method and the various tools available for analysis of such data. So let us now welcome Mr. David Campbell from the Institute for Systems Biology, Seattle. So I am going to talk about basically SWATH MS and in particular some tools that we use to ensure that the libraries that we use for SWATH MS are robust. That was initially going to be the focus is this one tool but as I sat through the talks and as the schedule changed a little bit I broadened it to include more sort of general SWATH information. I would like to actually before I begin thank some of the people that, there we go. So Muckel has done most of the mass spec work that I will talk about. Dr. Robert Moritz is the lab head and they both of them help prepare some of the slides that I am going to show. So I would like to thank them in advance. So basically this is an overview of the talk. So first of all I am going to try to tell you what SWATH is, DIA or SWATH and why you would want to use it. I am going to talk a little bit about SWATH Atlas and Peptide Atlas and some related resources that are out there that you can use and again you have already heard about some resources. There is a lot of people working and trying to make this data available. Somebody that can use that, somebody that can take what is already available and leverage it for their own research is going to be ahead of their competitors. Then I am going to talk about the specific tool and a little bit about what it might be in an ION library that we would want to make sure is proper so that we can be confident in our SWATH analysis. And finally I am going to give you a short example of a SWATH analysis and to try to illustrate some of the points that I have discussed and I will say that along the way I am going to take a detour that occurred to me during Dr. Mani's talk yesterday. So this is a depiction of SWATH MS and so each of these cylinders represents a proteome. The top of the cylinder is basically more concentrated or basically more concentrated proteins say in the micro molar range and down at the bottom is very dilute proteins say in the femtomolar or atom molar range. And these are the different techniques for mass spec proteomics and I just like to go through them briefly and give an illustration of some of the pros and cons. So the one that we are most familiar with is called DDA. So basically what you do is as the peptides are eluding from the column you pick generally speaking the most intense one you fragment it and then you interpret those fragments that is a spectrum. And that is typical proteomics. That is very good at seeing pretty much everything. So yeah basically it is from most concentrated to least concentrated and from side to side basically means how many of the chemical of the ions at that level you have been able to see. So DDA sees everything that is very concentrated. If you then instead of basically selecting the most abundant precursor if you instead have an inclusion list a set of precursors that you are looking for to begin with you can actually get deeper into the proteome. But it comes at a cost of losing some of the coverage at the higher concentrations. SRM which was explained well this morning carries that trend further still. It is very sensitive but because you have a limited number of precursors you are trying to select you necessarily the mass spectrometer has to spend more time basically finding the fragment ions of interest. And so you don't see nearly as many things but you see them much more sensitively. So that is generally correct. So almost all of these techniques that require sort of precursor information they generally rely on originally on DDA experiments. There is a number of ways you can get deeper into a DDA experiment by doing fractionation by doing longer gradients, by doing different cell types, by doing subcellular fractionation. So basically by really beating this technique to death you come up with inclusion list for this, for this and it turns out for SWAT. So SWAT also in the main way it is analyzed depends on previous information.  With SWAT basically you are taking chunks and I will go through sort of how we decide to do that but you are taking big chunks of M to Z ranges which contain hundreds and even potentially thousands of peptides and you fragment them all. So you have fragmentation information on pretty much everything in your sample. So right now you are only limited by the sensitivity of the instrument. Obviously in the future we would like to be able to analyze every peptide in the entire proteome plus you know any post translational modifications but it is not clear how we are going to get there and we are certainly not there yet. So yeah so the advantages of SWAT are you get greater depth than DDA methods, you get more consistent breadth, the broadness across seeing all the proteins at a given level and you are able to reanalyze the data pretty much forever. So the idea is that you because you are essentially fragmenting everything you have information about everything. With these techniques you have selected an ion and fragmented it so you only have partial information so this one you have complete information that you can reanalyze forever. So this is a depiction basically this was gone over well this morning but in SRM you essentially have many chemical species eluding at a given time.  In contrast with SWAT basically again you have all these things eluding everything else is the same everything is eluding the same you fragment multiple species at a time actually many more than are depicted here and then you read out all of the fragments and you get a very convoluted or complicated spectrum at the end. So this is a depiction of just how complicated it is and it also shows how SWATs work. So basically the distance between here and here is called one cycle. So initially you do one precursor scan to see what MS1 things you have and then you step across a predefined mass range as quickly as you can and in this case we have depicted 25 Dalton SWAT windows fixed width all the way across and then we go back. So that's one cycle back but you have to consider how peptides elude. Peptides are coming off of the column and they start eluding and there's a maximum and then they come back down to baseline excuse me. So ideally you would get multiple points across this peak and because you're taking all of your time doing all these different SWATs you run the risk of not getting data for across every peak. So these are this basically depicts in time versus MZ space the fragment ions coming from just this one SWAT. So these are not the precursors even though the upper end of the range is the same these are actually the fragment ions and the fragment ions in any one sort of vertical slice are likely to be related with each other. You see this smear across here this is because this fragment width corresponds to the precursor width and this is bleed over of precursor into your fragmentation spectra and for this reason when you make a library you actually exclude any Q3s that are within the SWAT window that you initially used simply because there's so much background. So this is a depiction of the actual SWATs that were used you see it goes from 4 to 425 then 424 to 450 so there's a little bit of overlap in each of these cases as you see and basically there are 32 identical windows and then you go back again but it turns out that that was the initial way things worked but the instrumentation companies came up with improvements in the software and the way the instruments worked and in fact it turns out that if you look at sort of normalized M to Z space there are a lot more chemical species that have M to Z ranges between four and four hundred and a thousand then out beyond here so really what you want to do is have a variable SWAT windows and so you see down here where there's you know it's still going up you have a width of about six or seven units but by the time you get into this regime it's just six and so the minimum width would be right at this peak and then when you get out here your width might be you know a hundred or something like that and the idea is to try to make sure that there's a same number of precursors in every window so that you can give your machine the best chance to analyze everything and basically so our SWAT setup uses a hundred different windows and so you can imagine that the machine has to be pretty fast to step over that so because the output is so complicated you need a high resolution mass spectrometer so you need high resolution MSMS or else you'll have no way to really distinguish what you're seeing you need a fast cycle time because you're going over all these SWATs you need to be able to get up and down so that you get reasonable sampling over any one peak and so this is an idealized peptide elution profile and of course in reality these things are all overlapped I mean at any one time many things are alluding so if you take human cells there's something like 800,000 different or something on that order different possible peptides and you think you're alluding them over 90 minutes off this column even if you figure you're not going to have half of them there that's a lot of things alluding at any one time and also a rational selection of these Q1 windows as I described these variable with windows goes a long way towards successful outcome so how do we analyze SWAT data so most of the data that is used it's depicted in the top row and that's library based analysis so we take DDA data we make a spectral library and then we use a spectral library to put into format usable by all these different software tools skyline has its own methodology for doing this as does peak view so a number of tools can can do this conversion for you but as was pointed out that requires that you do DDA runs so ideally when you want to go to a SWAT experiment you wouldn't have to go and run 50 or 100 DDA runs just so that you can make a library to analyze your samples I mean what if you're looking at a thousand human samples or or a whole bunch of different animals perturbed in different ways you know it really wouldn't be reasonable to go and make all these libraries just for this one time use and so we advocate the use of these repository libraries that can be used to do these analyses so so our software can read and write any of these formats there's also another way a library free method and this one is from ISP it's in the trans proteomic pipeline it's called disco and it's the data independent signal correlator so basically what it does is at any given time it simply looks at the fragmentation pattern and it it looks for signals that are rising and falling in concert in unison and it assumes that those belong to the same chemical species the same the same peptide it also uses these precursor scans to watch the MS the precursors come up and down and it essentially makes a pseudo mzml I don't know so mzml is a is a common file format that is used for encoding proteomics data so everybody has different instrumentation some people have thermo instruments some have abcyx so mzml is an attempt to to make take all these independent and different vendor formats and put it in a single format that people developing software can use and so what disco does is it basically it mimics a dda run and then you do a normal search and you come up with results so I'm going to talk a little bit about peptide atlas and swath atlas so peptide atlas is something that's hoping hosted at the ISP and basically what it does is it takes mass spectrometry experiments from all over the world and imports them and analyzes them in a consistent way and comes up with highly highly curated sets of of peptides and highly confident sets of peptides it's it's actually very we strive for very low error rate as a consequence we throw out a lot of data but but the data that's in peptide atlas is actually very good and it's broken down by tissue type by organism and it's searchable and it's it's a reasonable first place to start if you're embarking on a proteomics experiment if you if you want to do proteogenomics and you don't have the tech the know how to basically do analysis of certain cell types you can actually get some expression data directly from the peptide atlas that may be useful so peptide atlas is sort of the umbrella the umbrella property or the umbrella service here at ISP there's also something called SRM atlas where we've basically similar vein we've taken SRM data from various labs and we've compiled it and made it available for people to use and built an interface so that people can look at it and swath atlas again is yet another data type so the the first one peptide atlas that's DDA data SRM atlas is SRM data and swath atlas is swath data peptide atlas is by far the most developed SRM atlas and swath atlas provides some searching interface so that you can get the data that you want and hopefully render it in some way that's useful for you to do an experiment down the road both of these two sort of generate the inputs for experimentation on your own and we use a common code base and a common back-end database to to make these so I would be remiss if I don't mention the human SRM atlas so this was an effort in Robert's lab to basically cover all the all the proteins in the proteome the then known proteome and to do that we basically took unoprote and made at least five for five peptides for every protein based on things that have been seen in the past and we actually used synthetic peptides so these were were made so that we had a high confidence of what we were we were looking at and then for longer peptides we took up to 10 and basically this this gives us an ability to look in theory across the entire proteome in addition to that we did two other things that I think are useful in the context of proteogenomics so one of which is we looked at all the different variable spliced protein forms and picked peptides for those proteoforms and we also looked at about 5,000 well conserved snips so we have we have peptides for a lot of different things and it's possible that that would be useful in the context of proteogenomics so SRM Atlas this represents what you would happen if you had done a search for say this protein each one of these lines represents an SRM or MRM assay an assay is basically a Q1 value a Q3 value so a precursor M2Z a fragment M2Z and retention time as well as a relative intensity so that you can rank it relative to its its neighbors we have all these links you can look at spectrum chromatograms so this is we did all these different collision energy so for this Qtuff so that for any given protein fragmentation there's sort of an optimal collision energy we actually looked at all these different collision energies and you can see that the the behavior of the fragment ions as a collision energy changes this is an example of a chromatogram this is following one of these links and also of course you can see spectra in here the swath Atlas right now has has really three functionalities so first of all it's a repository there's we have several several different libraries from different organisms rat human number of bacterial organisms the biggest one that we have is this so-called pan human library basically it was an exhaustive effort done in Rudy Abersol's lab to to sequence some 60 different samples a number of cell lines a number of primary tissues a number of fractionation techniques that came up with with basically assays for well in excess of 10,000 human proteins and so we also allow people to basically using one of these libraries it turns out that different people have different experimental setups and so using one of these libraries you can provide certain information is in which proteins you want or which mass ranges you want to use and you can basically make a subset library that is more amenable to your research and we also have this tool which i'll talk a little bit about which is which is an assessment tool for an iron library so you can take anything so you can make your own iron library in skyline and maybe maybe you're not getting the results you wanted well you can take it and upload it here and it will give you some feedback on exactly what's in the library and you can see if there's something that that indicates why it wouldn't be working properly and once you do this you get this very complicated output which i'll break down a little bit so it doesn't look so daunting so yeah i'll talk about this assessment tool so this is sort of the the workflow of creation of libraries and then swath analysis so this here the simple sample prep digestion is is what was talked about at length yesterday i mean you have to get the samples you have to figure out how much protein is there you have to know you know what kind of contaminants you have you may do fractionation so there's there's an awful lot of work i mean it's it's easy to put it on a flowchart but there's an awful lot of work that goes into this then at some point somebody does dda and uses a search engine and you come up with a validated set of identified spectra those spectra are then combined into a spectral library a consensus spectral library and what you do is you basically take all the observations of a particular peptide ion so peptide ion is a peptide plus any modifications plus a charge so if you have the same peptide in plus two and plus three that would be two entries in the spectral library and it turns out when you're doing mass spec you often have noise in your peaks and by combining all the different occurrences of a given spectrum in your data you actually attenuate the noise because peaks that are real will tend to add up and peaks that are are just background noise will tend to attenuate and so this consensus library is really the basis for all of the all of this sort of inclusion list all the library based techniques so this can be taken and made into an srm library directly with spectras or it can be made into an ion library and so an ion library is what we call the libraries that are used with swathms and then we can basically take this ion library and and generally we make the ion libraries as complete as possible and then we apply our own particular criteria so every time you do mass spectrometry you have you have a mass range you're looking at you may have a preferred sort of b or y ion composition so you can apply these kind of filters and make an appropriate subset for your data and then of course you do your din analysis with one of the tools that i mentioned so one thing that i decided during dr monnie's talk it seemed like a lot of people were a little confused and i think that for this audience so so it turns out that the spectras tpp spectras ms proteomics tools as well as this uh this data set are available on docker so docker i think for people that don't have a lot of software background i mean by all means if there is if there is a platform that you can use that like galaxy that has everything built in use it but if you want to download something onto your computer and run it docker is an excellent way to do it because basically all of the maintenance is is handled for you and all you have to do is run it and i thought i'd explain a little bit more what what it is so you can see that this is meant to be sort of a container ship and these are containers but really the the whale is a computer and it could be your computer it could be a server in your department but it's just a computer but it's what it's doing is it's running these little mini computers on it and and these running mini computers are called containers um so basically it runs a guest on a guest computer um on a host computer so it runs one or more little guest it's almost like a little computer on a host computer so what you there are two concepts in docker so the first is images it's like it's like a hard drive so basically it has an operating system like linux it has some software so it knows how to do one thing really well but it's just a file it's just you know it's handled by the docker software but it's just a file when you actually start the docker that's when you get a container and that's what dr mani was talking about oh so i lost a couple so basically uh so so you could you could have a little hard drive icon if it was showing up for for sort of all of these things so you could have on your computer you could have a protigy container or an image you could have a tpp image and you could have a fire cloud image but the neat thing is on any given computer you can actually run many of these things and so the images is just sort of this set of instructions it's like a little computer that you can run on your computer but you can you know you can distribute it uh you we're talking about fire cloud basically they can take and they can put their software this little container on as many nodes as they want um and so or you could you could run multiple jobs on your own computer so the and uh so anyway it's but why is it so neat and so so i think from a from a non-software expert perspective i think that the neatest thing about it is that the developer takes care of it so you guys tried to install our studio and protigy and you had issues with the versions of r so i'm actually responsible for for maintaining different versions of r and python back at our at our lab and it's really a headache because basically some really smart postdoc in your lab makes some software that you want to use but he's gone now or she's gone now and it depends on python 2.6 and then somebody else comes along and says oh i have this new software i want to run they download it they say oh i have to upgrade python and now all of a sudden the old thing doesn't work so what docker does is is everything is is separate and so it basically it's there's no contamination of the environment and it may seem like a trivial thing but that really is one of the biggest headaches is the fact that all these software things have their own dependencies and by having the developer maintain the environment it's it's makes it much easier so there's no conflicting versions so basically each docker knows what it does and it does it well the updates are generally lightweight so the first time you download something maybe it's a gigabyte it takes you five minutes to download but the next time somebody just made a little tweak to it you just say oh i want to get the latest thing it looks and it does it by what is called layers and says oh i already have that i already have that oh i have to download this one new thing it takes 20 seconds and boom you're running the latest and greatest and so it's really convenient way to update and keep your software updated so it can run virtually anywhere it can run on your computer department server or as we heard yesterday on on the google cloud or microsoft azure or amazon web services and was also what was also asked yesterday was can i modify it so it turns out yes you can download any docker well almost any docker you can start it up interactively and you can make changes you can put your own software on you can add a little pipeline and then you can save it as an image and then next time you go in you can basically run that image you have that container you don't have to update it again so the one thing that i should point out is you can run these on your computer but it's not a miracle if your computer is slow the docker on your computer is going to be slow it doesn't make you suddenly have a super computer but if you have a reasonable computer you can run some pretty complicated software and there's not a lot of overhead running the container so it would run just about as fast as if you installed it yourself and and took care of the headache so anyway so what what runs on docker well as we heard yesterday the fire cloud uh you guys also heard a presentation about base space well it turns out base space these are all all docker images that each one of these little apps is a docker image that runs on amazon web services and in fact they have a thing for proteomics called one omix um Illumina does and basically we've developed a couple apps and they're basically docker containers and the reason they do that is it's very easy everything is all self-contained they don't have to worry about collisions of all these different software packages that they're they're offering um the tpp runs on docker open ms runs on docker that's one of the things we used to analyze swath python r you guys were having trouble with r you can download any version of r you want and then boom it's it's running um there's also this thing called uh biocontainers and the biocontainers folks uh basically maintain all these different tools uh sam tools is is a big uh genomics tool for doing alignments and um basically it's one of the tools available in galaxy uh and this is uh this is an easy way so these are basically biosoftware that are maintained that you don't have to do anything you don't have to go search and form you can just go to this one place and look and see is it does it have anything that i want anyway so that's the end of the sermon i think docker would be neat i think i think it's worth checking out in today's lecture you will introduce to the greater depth and consistency of data independent acquisition or di a method over data dependent acquisition or dda method softwares like skyline the spectronaut peak view and open swath they require a library for di data analysis softwares like disco and di empire they do not require the library for di data analysis the softwares disco from isb mimics di a runs like a dda run it performs normal search and interprets the results resources like peptide atlas asarum atlas and swath atlas they contain information from dda experiments asarum experiments and di experiments respectively these resources are freely accessible to the research community and contains a lot of valuable information the dockards let the users analyze data from multiple places and offers an easy interface and workflow for data analysis in the next lecture we will look at some key features of swath atlas and how it can be used to accelerate di data analysis thank you


Welcome to Center Stage. I'm Crystal, not Donna. Unfortunately, Donna had to go and do something from Family Matters urgently, so Donna, I hope you're okay, and I hope you're watching this. Happy to be here in replacement of you. Today, we're going to be talking about something very, very interesting for me, Passionist Theater, so it's always lovely to talk about the magic behind the scenes. Now, what magic do we have today? Shakespeare magic. So, why don't we introduce my wonderful guest today, who is going to talk about being the director and a professor at UH. How about that double bill, huh? He can talk and act and direct. So, let's introduce Professor Paul Metri from the University of UH, I give you well, Theater and Dance Department, Professor in Voice, Movement, Shakespeare. Stage combat. Ooh, I like that one. I get all the fun stuff. Come on, I'll give you more. When you're an actor yourself? Auditioning, yeah.  You created the Seattle Shakespeare Festival? Yeah, I was the principal founder of Seattle Shakespeare long ago. I actually started it right out of grad school. I was at the University of Washington, and I still had money left over from a slightly illegal student loan. Who has money left over in theater? I didn't eat that year. It was worth it, right? It was good. Yeah, so Seattle Shakespeare Festival started in 1991 and still going strong. It's kind of the premier classical theater north of Ashland. How do you start a festival? That's fascinating. Well, that's another topic. But it's kind of indicative of how do you create art? So I saw a need. I saw a niche. There really wasn't a lot of Shakespeare being done in Seattle at that time. Really? The Seattle Rep would do a show maybe every other year or so. There was a theater called Intiman that was the classical theater, but they had actually started to do more check off and more contemporary things. Right. So I just kind of saw that and coming out of grad school, I had just done a season at Utah Shakespeare and said, hey, we should be working on that. And I know a lot of actors that they want to work on Shakespeare. Yeah, who doesn't want to. So you could get them to work cheaply. Get the hungry starving actors. Yeah, because Shakespeare is great to work with. Shakespeare is so rich. It's wonderful. So for two years, I actually just produced on my own. I just kind of tried to do almost everything on my own. I remember driving Shuttle Express to go to the airport. I would drive that all night long, then basically get two hours of sleep, come back, rehearse, try and get one more hour of sleep, drive for another 10 to 12 hours and do that. What was your first Shakespeare play? That I ever did. Well, for that, when I produced on my own, we did Romeo and Juliet. Okay. And it's actually very similar to the Baz Luhrmann one, but it was 10 years before. We set it in modern times. All right. I love that. And kind of changed it up to kind of have this, it was kind of more of a yuppies and punk type of. Cool. I really love that type of modern interpretation. It was a lot of fun. It gave us a lot of freedom to do whatever. I remember cleaning blood off the back wall that we had to do because Mercutio gets shot by Tybalt instead of Stab. So we had that kind of stuff. And it was actually Romeo and Juliet and violence is actually an interesting thing because later on during Seattle Shakespeare, we would use that play to bring anti-violence programs to the high schools. Right. You know, it isn't just, oh great, let's do a show of a lot of violence, but it's also being able to use Shakespeare as a tool to teach certain things that society needs to. So have you always combined the concept of education and theater and performance or is it just something innate that you'd like to carry out? I think it kind of has to do it, especially with Shakespeare because you have to learn how to do it. It doesn't tend to come naturally. How do the younger generations take that play and what do they get out of it? That's very difficult in translation. I mean, Shakespeare said the whole idea is to hold the mirror up to nature. So in any time that I think about a play is trying to say, well, what do we want to say with it specifically? Not just, oh, let's set it on the moon because that would be fun. But if you're going to make a concept to try to go all out with it and try to say, if this is what it means, how does that change all other choices within it? 12 Nights is kind of a good example. That was my perfect segue. So why 12 Nights and why? What's your concept of this one? 12 Nights. See, this is actually a production that I did way back in 1996 for Seattle Shakespeare. And we toured it up to Whitby Island and did it in Seattle. And I later did the same concept in Cairo, Egypt when I was teaching at the American University in Cairo. Is that in Egypt? No, no, said in the way that we did this. So the way we do it is I was heavily influenced by the movie Strictly Ballroom.  I was, I was a dance major. You can tango? Really? When I first went to the U-Dub, yeah. And then I changed over to theaters. Me too, I did the same thing. I love it! It's great.  Yeah, yeah, yeah, of course. And U.H. is theater and dance. And dance, yeah. It's all related. It's one of the reasons I wanted to come here is you had the combination of dance. So Strictly Ballroom, yeah. So we said Illyria in Twelfth Night is this mystical, I mean, Shakespeare's geography was not so good. Can you recap for people who aren't as familiar with Shakespeare's stories in a nutshell, what is the long line for Twelfth Night? Well, for Twelfth Night, like the plot line, you could kind of say like many of his twins get separated and come back together at the end. So that's kind of what happens. In the meantime, they all find true love. Right, but then there's a lot of gender crossing and humor in falling in love with the wrong person and all that kind of stuff, right? Well, for me and for this play and for this particular concept of the production, to me it had to do with what is Illyria. And Illyria to me is this glitzy world, kind of looks like a 40s MGM musical type of thing, where it's all about the glitz of it, you know, the glamour or whatever. It's people that are in love with the idea of being in love. They don't really know what real love is. Until Viola gets shipwrecked into this place for us, shipwrecked. She just kind of lands in it a little bit like Wizard of Oz. But she's the catalyst. She's the instrument that kind of allows Orsino and allows Illyria to say, oh, that's what love is. That's what it really is about and they really start to feel something. Whereas all before they're just professing to be in love or Illyria is kind of professing to be in mourning. You just don't really believe it until these guys come along, Sebastian and Viola, the twins.   And, you know, and it's just the most beautiful thing in the world. And Viola dressed as a boy says this and Olivia goes, oh, that's love, you know, falls in love with him, her. What's that context to modern day youth? I mean, do you think the younger generation now also have kind of a misconception of love?  It sounds corny, but how do we take that with us today? What I would say is that plays into why is Shakespeare important? Shakespeare is about words and it's about language. Shakespeare's working vocabulary was 30,000 words. In modern, you know, normal American vocabulary is 8, maybe 15. Exactly, exactly. And that's what the, there's a linguistic theory that, you know, basically says if you don't have the words for it, you can't really experience it. Well, what are we going to do with Trump? He's got less. Sorry. I heard that knock. Yeah, I mean that, I think, if you look at how President Obama speaks and the amount of, you know, words that he uses. Articulation. And it is, it just kind of dumps everything down and simplifies everything. And I think that that's really true, as you're saying, with the younger generation and the idea of being in love. I think Shakespeare, you can look back and look at these words and go, oh, that's what love is.  But instead, it's these, this deepness, you know, read the sonnets, you know. Yeah, it's a lost art, isn't it? Yes. Did you ever see Captain Fantastic? No. Oh, please do. I will. I mean, I'm sorry, I don't want to offside track here. But really, because they go off and they, they are so well read, living in the forest growing up, not going into a normal social environment. But the teenage boy falls in love with a girl and he starts ranting these, these philosophical sayings. A little girl, this young modern girl is like, what the, you know. And it's really quite interesting what words can mean. Yeah. Or the lack thereof. The words are powerful, which is as we're finding the whole, you know, Twittergate thing. It's just, you know, how you use words, they can wound or they can heal your choice. And is it a loaded word or is it oversimplified where it's lacking so much depth? Mm-hmm. Yeah. Yeah. Do you have a favorite part of Twelfth Night? Oh, I love it. Probably the... Without giving away the story? Probably when the scene with Olivia and Viola is really, really just beautiful, I think. I think the poetry is fabulous. How our actors are doing it is a lot of fun. And then later on, I think part of that fun too, is you get to see them later on too after Olivia falls in love with Viola.  And he's like, no, no, no. All right. I'm a servant.  No, you know. But even the cross-gender thing, I think I find that very, very relevant today, you know, does it or maybe not. Because Shakespeare in times, what's seen before is times in terms of talking about, you know, men and women and your identities and who you think you're attracted to. But today, this whole neutral gender thing going on is a very, very different concept. So you say you think you're in love with someone. You think you're in love with the opposite sex, but you're not. So are you in fact, I don't know if those issues come up in it? They are. And if you want to stress them, you certainly can. I think Hawaii Shakespeare did Twelfth Night last summer. And I think that they dealt with it more from that kind of modern standpoint. For us in setting it in this time period, I think that begs other certain choices to be made. But we do talk, I mean, in a way we have Festi, the clown, and we've chosen it to be more androgynous. In a way, Festi is almost like, this is what Viola could become if she ends up just staying in Illyria. If Illyria is almost a trap and just, you know, she just can't get out of it. So Viola's kind of whole struggle is, why don't I just tell people I'm a girl? Right. Why don't I just do that? So the photo is that Viola? No, that's Olivia, played by Emily Hodley. And that is Festi played by Rachel Lueno. Oh, okay, all right. Yeah, so you can see the clown, yeah, we're going much more with androgynous. But the Festi kind of then also has the ability to kind of see what's underneath all the glitz as well. Right. Festi as the clown is able to say, this is what's really going on, you know, and sees through Viola's disguise, and it's kind of going, I know what you are. And the brilliance of Shakespeare, though, you always have one character who's going to spill it if you can read between the lines and not. We're going to take a quick break, and a lot of interesting stuff with 12th Night. So I'm going to pick your brain a little bit more after this, and don't go away because we are going to talk and dig more into 12th Night at the University of Hawaii. Aloha, and happy New Year. It's 2017. Please keep up with me on Power Up Hawaii, where Hawaii comes together to talk about a clean and just energy future. Please join me on Tuesdays at one o'clock. Mahalo. Hey, has your signal just been taken over, or am I supposed to be here? This is Andrew, the security guy, your co-host on Hibachi Talk. Please join us every Friday on Think Tech Hawaii. Hi, I'm Marianne Sasaki from Life in the Law on Think Tech Hawaii. I'm delighted to tell everybody. I'm so excited. I'm going to Washington to march with women on Saturday, January 21st. It's going to be a huge, huge event. And I think we're doing something in Hawaii, too, aren't we? Yes, we are here on O'ahu. We're going to be at the Capitol starting at 8 a.m. Back here on Center Stage with Professor Paul Mitri, talking about his directorial tarot performance, upcoming performance of 12th Night at the University of Hawaii Theatre. Now, 12th Night, if everybody knows this is a Shakespearean play, but Paul, your take on this is to combine ballroom dancing, which I think is a really interesting combo. Now, I know you have a little dance background, but why did you choose specifically ballroom dancing and how do you think that works with this play specifically? I think especially like setting it in a ballroom and using the ballroom dance, there's so much etiquette to it. There's so much glamour to that, elegance to it, too. And at the same time, though, is what is underneath it? Where's the passion underneath it? If you think of a tango, it's technically brilliant, but it also has the reality of the feeling under it, too. That, to me, is that dichotomy that I think Shakespeare should have. Absolutely. You've got to blend technique with passion. Sure. But my interpretation of 12th Night often is because it's shipwrecked, so it's on an island, and people are in shreds, and they're kind of like not really in the best threads. So how did you turn that into...? That, to me, is tempest. Okay, true. Everybody's like that. In this one, it's really only viola, and let's face it, it's a pretty flimsy plot point. She goes, oh, I'll dress as a guy and serve the Duke, you know? Okay. So it doesn't really hold up, you know? So for us, also, it's kind of, well, Ilyria is this strange world, you know, where you can kind of be like, didn't I just see you as the captain, and now you're doing this over here? So it's a little bit like Oz, I think, in those... Do you think this is a particularly musical piece? Is that why you brought this into it? Yeah, I think... Twelfth Night's kind of the most musical of all of Shakespeare's plays. Is this the one where they have the famous line, is music, the food of love, play on? That's the first line, very first line of music, be it a food of love, play on. That's one of my favorites. Yeah, and Festi sings three songs in it. Okay. It could have been four. So there's plenty of music in there, and that for us, also, is kind of indicative of this world. So the way that we're using it, too, so we use music and dance as kind of transitions between the scenes, either to help the characters get out of a scene or to show the characters coming back into a scene is to use dance as a metaphor. Did you find it difficult to use your cast and their talents with dance and music and voice in that combination now? You know, we're in educational institutions, so that's our job is to educate. Right. And I absolutely am indebted to Mike Romney, who's the choreographer. So he's an MFA student in the dance program, and he teaches ballroom for us at the school. So I said, hey, Mike, would you want to choreograph that? And he leapt at it, and he has been working so hard. Many of the actors had had some dance, but not a lot in many cases. And we have at least one dancer who doesn't have a lot of theater background, and she has grown herself totally into that. So that's also why I love doing the show right now, is it allows different sides of our department to come together and work together. But some of our actors, you know, they are so versatile. They're absolutely triple threats. So watch them and enjoy their work. What do you think? I know it's hard to speak on behalf of them, but what do you think their biggest challenge was during this process with you in particular? Dealing with me. Yeah. Oh, no. I'm sorry. That too. You know, our students are really, really busy, and especially, you know, if they're good, they're in high demand. So they are really constantly trying to balance, well, okay, Rachel, who plays Festi, she just did Elephant Man at Minoa Valley. She teaches two classes for us, beginning acting. She's carrying a full load of classwork also. Right. And if they're my classes, that means you're doing a lot of other scenes and a lot of memorization. Right.  That's all right? That's Rachel. Right. So she's also being asked to be in, you know, X number of other scenes for directing students and all of that. Isn't that wonderful?  If you're good, you're going to be busy. That's, ideally, you do. And that's kind of why... That's part of her job, is to learn to balance all of this. Right. And that's... In theater, a lot of times when it rains, it pours. Yes.  Yeah. And in that meantime, you've got to still be training. Right. You can't just, you know, say, oh, I didn't get any work. So keep learning new skills. Keep learning new skills. And so for this one, the dancing, especially, Mike is really, you know, taking her through her paces for the dance part of it. And it also gives her a chance to show off her vocal work, which actually, an elephant man, we had used her as a vocalist in that too. Okay. So, again, it's like you learn some skills in this show. You apply them to the next one. Yeah. You learn some skills here. You apply them to the next one. And isn't it wonderful to be able to see such talent in Hawaii? We have an amazingly talented stable of actors here. And the question is just, how do we let the world know about it? Yeah. How do we get people to understand that, to see them? You know, we've rejoined the Kennedy Center American College Theatre Festival, and we co-hosted it last year with Shamanad. Okay. And so we were able to show our Hawaiian Theatre program. We were able to show musicals, some scenes that we had done from Sunday in the Park with George. Our students competed. Our student, Nathiel Nimi, won the National Directing Competition at the Kennedy Center. Wow. We have great, great students. And people don't know about it. No. That's the crazy thing. I mean, I really think, if I can get on the soapbox for a second. Yes, please. I can compare the arts to theater. You know, the arts to sports. You know, you pick up the paper, and you see about student athletes in that. And they get a lot of great coverage to it too. And money. And sponsorships. These students that you'll see on stage, you might very well see them in a movie. They might become the next Matt Damon.  They're that talented. That's so wonderful. And that's just so inspiring too to know that, you know, you're in a small community, a small island, but you've got such versatility and talent here. Wonderful. Wonderful. It's so underestimated. Yeah. You know, they don't have the kind of head start that their peers do if their peers are studying in New York or L.A. or Chicago. So our students have to work doubly hard. Yeah. Because once they go there, they kind of have to start from scratch. Right. And we've got some students there doing some wonderful stuff. And I'm glad you, as a professor or as a director, as a creative person, is continually being inspired by people and works that you want to work on, right? So what inspires you to keep going and creating new projects? I like a challenge. Yeah. We just did Elephant Man at Manoa Valley. Right. And that had been on my bucket list, because it's one of those things you look at and you go, that looks hard. I should do that. You think? I should probably try that. Okay. If it's hard, it's worth doing. Yeah, sure. They struggle itself, right? Yeah. What are some other things on your bucket list that we should know about? Well, there's... Give me one. Give me one you really want to do that you haven't done yet. As a director or as an actor? Oh, two questions. That's what we've got. Yeah, about that. Or both. Okay. Sooner de Bergerac. Oh, okay. That hasn't been done in Hawaii, huh? None since I've been here. And my colleague, the late Glenn Cannon, he was so wonderful. And after seeing the company that we started, All the Worlds of Stage, one of the first things he said to me was, you need to do Serino. And so just to honor him, I would like it somehow to be... As an actor or director for that one? Well, I'd probably do both. How do you feel about self-directing and acting in the same production? Do you see it in films? And it doesn't work often. Yeah, I think it depends. I don't know why it doesn't work in film. They have a lot more money. Because they have huge heads. They have a ton more money. They're totally obsessed with themselves. And that's a thing. Too much money. It gives you too much room. Absolutely. It can. It can. It's dangerous. I think in theater, it depends. And there's certainly some productions that I'd say, yeah, there's no way that I would want to direct myself in. But some productions, the character that I play, like there's one called Inventing Van Gogh. Okay. And I played Patrick, the artist, not Van Gogh. So I was able... Patrick spends most of his time watching. Okay. So in a way, you're going, well, I'm already watching. I'm just watching from a slightly different angle. I've also found that with many of some of the people in town that I keep working with, Rob Duvall especially, that we have a wonderful working relationship in a way that we can just kind of direct each other. It's not comfortable. Absolutely. We share pretty much the same vocabulary. And certainly as I'm training students then to come up and be my colleagues, for instance, we did proof for all the worlds of stage. And there were four people in the show, myself and Rachel and Alex Monroe and Hannah Schauer-Galley. And we basically said, we're going to direct ourselves. So we called it Collabber Acting. So how do you come to constructively criticize each other? You have to leave the ego at the door. You just have to check it. Easier said than done though, right? You know, if there are people that you admire and there are people that you respect, and it's something that I try to teach my students anyway is that never be a diva. The world is way too many divas. Yeah, I totally agree. So there's just no room for that in the rehearsal hall. And theater doesn't breed divas anyway, usually. I've seen some. I had to tell some stories about some, not here, but yeah, my work at other theaters, I told some stories to my auditioning class recently about some of those. Oh, okay. But there's, yeah. And I said, please, never be a diva. There's just no reason for it. It's about the work, you know? Yes. Stanislavski said, you know, love the art in yourself, not yourself in the art. Yeah. So if you can do that, then I should be able to go, yeah, tell me what you think. I may not agree. Right. That's certainly fine. But you can welcome that feedback. But yeah, and I might go, I didn't think of it that way. Let me try that. Let me see what happens. Oh, that's important. That's a person too, because you're always growing and you're always morphing so fun in your career. So if there are some young people, well, not that young.  What's the way to go about it? Well, what I tell my students, it's about you have to become aware of yourself first, you know? You got to know exactly how you come across. You have to be willing to use your own feelings to make it real for yourself. And I think a lot of actors, partially because what we see on TV, we do see a lot of fake stuff, you know? Oh, God, yeah. And a lot of that gets held up as props. You know, it's like, oh, this is really good. So you have to kind of have your own work ethic and your own bar, you know, that you say, I'm going to work to that. I'm not going to try to bring that down here. Good point. And be real to yourself. But Paul, before we forget, let's remind everybody when this upcoming performance of 12th Night is. 12th Night opens January 27th on Friday night. It will be at 7 o'clock that Friday, and then two shows the following, that next Saturday, the 28th at 2 and at 7. And then the following Friday, February 3rd at 7, and then the 4th at 2 and at 7 also. This is not at the theater. This is at the campus ballroom because it takes place in a ballroom. Yay, so lots of dancing and singing. Yeah. And if food be the love, no. If music be the food of love. Lay on. Lay on. Thank you so much, professor, for being here, and good luck. Thank you. Thanks so much. God, I screwed up on that line.


Baby vanished during car crash. Police found her in the most unexpected place. Driving is always contained in element of danger. No matter how careful a driver is, there are plenty of other factors to consider in ensuring the safety of everyone in the car. From road rage to distracted drivers to those who hit the road under the influence, it only takes one wrong move for a tragic accident to occur. On Friday night, five people from Arkansas were on their way home from attending a family event at a friend's house. The night was chilly and the car was running smoothly until suddenly, out of nowhere, a massive truck sideswiped the vehicle. In an instant, the riders were thrown away. The four adults were hurt but conscious. Suddenly, they realized that the fifth member of the family, an eight-month-old, was nowhere in sight. It was as if she just vanished into thin air. What happened in the next hour was something that will make everyone believe that miracles do happen. Trina Johnson and Jacizia Coulson were good neighbors from Hope, Arkansas. Living close to each other for years, the two women had become best friends. Both of them were living with their respective families who had grown close to each other as well. Whenever there is an event to attend, Trina would always call Jacizia. They were the best buddies. From cooking new recipes to shopping the latest stuff in the market, the two women were almost inseparable. When Jacizia got pregnant, Trina had been incredibly supportive. She used to accompany her to the clinic whenever the baby's father was not available. Her daughter, Justice, had been excited as well. Whenever food Jacizia asked for during her pregnancy, she delivered. In terms of love, concern, and cooperation, it was easy to say that the two families had been merged into one. Their happiness was doubled when Jacizia gave birth to an adorable baby girl. They named her Bryce. Trina's family just loved the outdoors, and whenever they had the chance to travel, they would always include Jacizia's family. However, when Bryce was born, there were lots of escapades that Jacizia missed since the baby was still very young. One night, Trina received an invitation from one of her friends who had moved out to another neighborhood. It was a dinner party, and naturally, her first thought was to invite Jacizia and her baby. At first, the new mother was quite reluctant, as her eight-month-old baby was still vulnerable to the weather. But after some time, she also said yes, having heard the promise that the party won't take long. Bryce was not used to traveling, so Jacizia decided to have his sister, Shamira Coulson, to go with them. It wasn't a problem, as Trina's car was able to accommodate them, as well as the baby's belongings. Before long, Trina's car was on the highway. She was with her daughter on the front seat. Jacizia, Shamira, and Bryce were seated comfortably in the back seat. The baby was evidently curious about the evening lights, and she kept looking out the window. The four adults were amused at the baby's reactions to her surroundings. Trina, on the other hand, was in the mood to plan for the next getaway. She was saying something about an out-of-town trip, when suddenly they heard a deafening noise. Before they even realized what was happening, their vehicle flipped over into the night. Trina Johnson had always been a calm and smooth driver. That night, she was traveling on Interstate 30 in Texarkana, Arkansas. They were not in a hurry to get home. That's why Trina was doing it cool on the highway. Moreover, she had four passengers and one of them was a baby. Despite talking with her friends and family, Trina was confident that her main focus was on the road in front of her. Her car was alone in the lane when suddenly, in a split second, she felt the unfathomable force of a gigantic vehicle from her left. In the blink of an eye, her head was spinning and she was being thrown out of her car. Trina was sure her car had collided with another vehicle. The massive force of the collision had activated the vehicle's airbag and in an instant, she'd flung into the air. However, before she could even process the damage in her body, another passing car hit her again. Justice had seen everything. She was screaming all the time but was too late to pull her mom to the side. When she looked down again, she saw that blood was all over her mother's body. Jekiza and Shamira couldn't believe that within seconds, everything in their lives could change. A moment ago, they were at the back with baby Bryce. The next thing they knew, they were both on the road with minor cuts and their heads were spinning like crazy. When the realization that they'd been involved in an accident hit Jekiza, her mind instantly went to her baby. But alas, the eight-month-old baby was nowhere near her. Despite feeling weak, Jekiza crawled and looked inside the flipped car. She wasn't there. Jekiza saw that her best friend Trina was badly hurt. However, Justice was motioning that she was still alive. There was already a crowd of people tending to her. The other motorists had stopped to help. Some even called the authorities to report the accident. Seeing that her sister was safe too, Jekiza had only one thing to check, her dear baby. She was shocked and temporarily not able to move. However, not knowing where her baby was was worse than anything. Within a few minutes, Jekiza heard the siren of the police cars. She also saw an ambulance. She whispered a prayer of thanks because she knew that Trina badly needed help. But while the authorities were certainly close to putting the situation under control, Jekiza couldn't afford to stay seated on the road without seeing Bryce. Her sister Shamira began to search the car as well. But there were no signs of the month-old baby. Jekiza began to scream. The police officials heard Jekiza's hysterical scream. The woman was frantically searching in the car. At first, they thought she was just traumatized by the collision. But when they got close, they realized that the woman was looking for her baby. When the officials found out that there were five passengers, they began to help the two women to look for the missing baby. All of them were shouting the baby's name. The cops had surveyed the road and were also able to keep the crowd from interfering further. But try as they might, they couldn't find Bryce. Jekiza knew she had acquired some cuts and bruises. The emergency team approached her to stabilize her condition. However, she knew her wounds could wait. At first, she was thinking that Bryce had been trapped inside the car. But the cops had already checked the whole vehicle from inside and out. Still, no one knew where she was. The officials were feeling positive that they would find the baby nearby. However, they've checked the road and had widened their radius. It was like they were actors in a nightmare movie. How was it possible to lose a baby in a highway with a blink of an eye? The police and the emergency team who responded to the scene knew their prime responsibility. They'd quickly scanned the scene of the accident and had already transported the badly hurt driver to the hospital. They'd also confirmed that the other three passengers, Jekiza, Justine and Shamira were in safe condition. But one thing also baffled them. Where was the eight-month-old baby? If she'd been thrown out of the car, they couldn't find her nearby.  The other officials feared that their rescue mission would soon turn to a recovery mission, but then they heard a sound. Jekiza had already recovered from the shock. However, she felt as if her heart was not beating at all. Her baby was still nowhere in sight. It's as if she'd vanished into thin air. She'd circled the car and even chucked the trailer truck that had sideswiped her vehicle. Several negative thoughts were running in her mind. What if some traveler had taken advantage of the accident and it snatched her baby? Jekiza was trying to block the thoughts, but her stubborn mind just couldn't stop thinking about morbid things. Every second that passed by felt like a knife plunging deeper and deeper into her heart. Then suddenly she saw some officials running down the hill beside the highway. The seasoned responders were used to seeing hysterical people during an accident. However, that night they completely understood why the woman was acting so crazy. She couldn't find her eight-month-old baby. It was just possible that Bryce had walked away from the scene of the accident. Then amidst the siren of the police team in the emergency car, a soft sound was heard. The bewildered officials sprang to their feet and followed the source of the sound. 25 feet off the road was a grassy man-made hill with a storm drain. At first, the officials thought it would be quite impossible to find Bryce in there, but as they were approaching, they were hearing some sounds. But it was not a sound of crying or screaming. Hopes began to rise. To their wildest amazement, the eight-month-old baby was indeed inside the storm drain, sitting in there as if nothing had happened. For a while, none of the cops dared to speak or move. A sudden feeling of calmness swept them all. After the momentary shock, the cops got their wits back and began to manage the situation. The storm drain was located under the bars and they were deeply puzzled by how the baby ended up under there, clearly unscathed. Bryce was staring at them, looking amazed. She wasn't crying at all, not even moving. It's as if she was waiting for the cops to find her and get her out of there. She only showed a different reaction when she saw her mom, Jekesia, who was crying from relief. The storm drain was a crowded space and the metal bars that covered made it not easy to remove the baby. The rescuers had to think of a way to pull the baby out as soon as possible without hurting her. The panic was slowly settling in and it was evident on the baby's eyes. Three firefighters were assigned to do the job. Even though more officials were willing to help, the limited space didn't allow everyone to lend a helping hand. They just relied on the logic. She accidentally got inside so they could also get her out. The firefighters were amazed enough of what happened to the baby. However, their admiration for Bryce had doubled when the rescue mission began. She was just eight months old, but it seemed that she completely understood what was happening around her. Bryce had been extremely cooperative all throughout the whole process.  Bryce didn't cry at all. She was making the task easier for the rescuers. Rescuing little Bryce made everyone stop in their tracks. Jekesia was extremely relieved to know that her baby was alive. It was only then that her heart seemed to beat again. The firefighters told her to wait as they would do everything to save her baby. When Bryce was finally out, Jekesia wept and laughed at the same time. She hugged and kissed her little sweetheart like there was no tomorrow. Her little angel was alive and it was like a miracle. Everyone in the scene that night was cheery-eyed at the reunion of the mother and her daughter. Earlier, Jekesia and the rest of the group refused to get medical attention. Trina was in the hospital already and they'd heard the word that she was going to make it. When the authorities found out that everything had been put under control, they ordered the rest of the group to seek medical attention. While Bryce seemed to be okay, Jekesia couldn't just rest at all. She needed to make sure that her baby was fine, both from the inside and the outside. After making sure that the driver of the trailer truck was already under the custody of police, all of them were taken to the hospital. The next morning, what happened to Jekesia's family, particularly to the eight-month-old baby, had grabbed the public's attention. News channels and other media outlets just couldn't believe that Bryce had survived the accident unscathed. According to the hospital report, Bryce sustained a scratch on her forehead and nothing more.  For them, it was an amazing story of survival. It was not documented by the authorities. One would surely find it hard to believe how Bryce managed to stay safe and sound. Many believed that what had happened was a kind of divine intervention. Others claimed that guardian angels were indeed real. Bryce was found 25 feet from the car and was found safe and sound. No one could argue that what happened was a miracle. The four passengers were treated at a hospital and were immediately released. Trina was left as she still needed a few weeks to recover from her injuries. However, the doctors ensured that she would soon be able to resume her normal lifestyle and be fully recovered. The truck driver that caused the family's vehicle to flip was already sighted. He was charged for an improper and unsafe lane change. Up this day, Jekesia and the rest of the family firmly believed that what happened to Bryce on that fateful Friday night was no doubt a miracle. The accident had brought them a kind of life lesson and a memory that they would forever carry in their hearts.