okay everybody let me know if you hear me I've got CAH in the middle and I have Kate up as the main one I think Kate's just gonna be a lot easier today I think a lot of things are gonna work today I rated it as well Kate's the main one CAH is in the middle okay 56 we're gonna do 40 by 65 40 by 65 Kate 40 by 65 Kate get the stuff in yeah it hit so fast I got it late but I got it I got it got it got it 65 65 65 be in it you have to be in it it just moves so fast wow better be in it than not be in it I don't think we have to give it any more room though here here I'm moving this my coffee today hold on 18 yeah the first target shoot gosh it looks so good though here I still have it no hesitation for me there just can't hesitate here here here here here here let's see if it breaks 18 if it does I'm taking it ever reset something I'll call it a second time here here here here come on okay this has to break the love for me now I'm just gonna take it cuz it's probably gonna reset up yes beautiful I took it beautiful trade whoa here who got it with me dirty women didn't get it yeah I had my breakfast this morning to the week in earnings season who got it who didn't get it what's the story perfect good everything this didn't do anything yet let's look that looks crappy gosh Goldman Sachs if you feel like getting nuts I could call this today would anyone do it if I called it wow we did the best one and it's still going it's still going is anyone still in it I can't see how you could be though look holy crap wow what a nice move wild weasel got it shower singer got it koala bear didn't get it well I do declare it looks like it's whooshing I predicted a swish today I didn't know it'd be the one we traded 1856 it is this is a swish it's swishing yay and we actually play this whoosh by chance and Netflix wow be interesting to see what this does all right what are we doing next here J&J I'm gonna call this a bit sets up if you didn't if you didn't get the other one 50 by 85 J&J short 50 by 85 J&J short if you want a trade and I would kind of scalp it and don't take it till it hits 50 by 85 J&J short if you want to trade if you didn't do the cape but I'm gonna go back to it 50 by 85 wow this is a really actually very very tight stock for this price for the stock here 50 by 85 if you want something targets low of the day back let's just figure out what it is it's a big move a dollar dollar plus 121 50 low of the day target and J&J if it hits 50 by 85 there it goes there it goes there it goes there it goes I just gave you a trade it just had a big move this is gonna go right down to 122 you could take half off right there low the day though is the target here did anyone just do it another beautiful call forget the other ones this looks crappy this just did it and we got this and it's beautiful and it's gorgeous and now it has to rally and this looks crappy too boy spot on today but how could you possibly have missed that I called it 55 times before it hit your grandkids are distracting you put them in the playpen and lock the door you wanted a fast one today look you wanted the fast one there boom you prayed you got it and you missed it I missed too here I'll watch this I hope somebody did this journey woman didn't get it to please tell me somebody did it quala bear did you do it you said you missed the Kate I would definitely definitely definitely watched 122 though for a bounce big fudge did it watch 122 for a bounce 122 17 it got to well this is very interesting very very interesting I still think this holds I still think this bounces wow can't wait to see what this does 3240 okay this looks fine Kate now I can't call this here again it's too soon for a turnaround but it very well could go I mean look at this let's let's stick to the rules I did the first entry it open and swoosh I think this needs more of a bounce back if it keeps breaking it's gone and I'm not doing anything else today there'd be no reason to that media idiotic good start to the week though for me Bob did the J&J not the Kate Cuda well I told you you could have done it I didn't like it it's a weird looking chart with too many tails and it didn't disappoint it didn't disappoint today where would you have shorted this there's no entry here that makes any sense this looks like a scary monster that tail if you did it you're on your own here this may get a set up here here here if you didn't do this you could still do it it's late put the stop at 122 85 J&J it's 50 cents it's 50 cents this could very well go down and break the low 121 it could go to this is still worth taking or you wait for the Kate those tails look like a dagger like a sword that chops your head off so let's see what this does let's see what this does let's see what they all do that's this Kate if it sets up for a second time for anybody that wants to do it but I'm not cuz it doesn't make any sense and I'm very very happy I think I could call trades my sleeve in fact I will I will without even looking at the candles 89 by 25 there 89 by 25 you could short Kate I'm not even gonna look at anything I'm just gonna call the numbers it's by seeing it with my eyes closed 89 by 25 89 by 25 target in this now is 1775 1750 1760 is realistic big move in this today bigger than it had the other day for the morning that we go look at that we'll go back and look at that and we'll review it feels like that was yesterday didn't hit 89 yet no guy we did the Kate to I also called J&J which worked here this looks good people this looks really really good people 85 to stop you can add under 35 you can get nuts today if you want to everything's working 35 by 85 and the Kate I said 89 by 25 and poor Netflix is suffering suffering suffering let's see what it does 44 43 somewhere in here it's gonna bounce there's just a question of when let's look at the market market looks good GS flipped over the high glad I didn't call that CAH shut a little bit of a move but basically looks like a piece of crap what happened to hog nope my golly we picked the right thing 89 by 25 if you want to do the Kate again looks fine still but didn't hit yet J&J is going and it's too late now if you're not in it I've called it twice you got to leave it and you will have to wait a little bit for this to make a move does anyone want me to look at anything else or go over anything okay 89 by 25 Kate it's gonna hit during women you should probably do them both you should do them both and scalp them and get some money and get on with your day yes it did cheap alright let's go over it because I'm not doing anything else and I made all the calls I'm gonna make so this was amazing and I mean I was right on top of it very very happy and actually didn't realize it until after I was already in it that I actually did it here which was kind of funny because here here this goes you can do it right now 25 is a stop you take it Kate if you want it there goes it's fine it's good it's beautiful anyways that's that was interesting but I just didn't want to hesitate and it moved into the move so fast and boom boom boom and I said I'm not gonna call this here wait look and I just called it and now this is great so let's just see where the really brothers is gonna go 1760 1750 it's gonna break the low hit on every time frame here looks amazing so if you didn't get the first one there's the second one and yeah 60 65 ish 1750 1750 is a realistic target for this today I wouldn't look for more than that here it goes today everyone do it journey woman did you do it I told you to do both did you so I said 122 is the first target for this be careful because it could bounce there I'm not saying doesn't get back down to the low but it could bounce at 122 it did hit that basically so if you want to be tight with it or get out of half in here you could so if you took this up in here you get out of half in here because it just went to the first number 122 I do think this continues down as well but I don't know when it could bounce back until it does here here here go with it go with it if you're in it everything's working that I'm calling and here this goes and it's gonna get down to the low I could have done this again this is a good this is gonna work koala bears down Kate flat J&J okay all right well if this breaks 122 then ride it down if it doesn't I take it journey woman did I don't think it broke 122 yet if it doesn't break 122 I would take this here and scalp it and it's it's a little late for you to get Kate though I think this is a little too late you only want to do one at a time if this doesn't break don't let this rally over this area here because it could take a good 15 minutes still goes back down again and it just could reset up mean you're at your max limit for for buying power or for size every trade today has worked what I mean your max limit everyone should be up money today I don't know what you mean oh this didn't break 122 I would take it and then I would do the Kate and you can put the stop at 1825 this will be a smoother move because this could have a big bounce I just don't want to see that happen to you okay you have profit good you want to do this you can do it and put the stop at 1825 it's great 35 cents drops down breaks a little take it this is you could do this right in here the stops still good 1825 you do too where was I Kate targets I just said them 1765 1760 1750 low of the days the first target which is 10 cents away so this is very interesting here this did not hold from the morning 4732 4740 people this is this is wild I think this bounces today and then follows through tomorrow I still think this bounces right in here there's no way this is going to go down there's no way this is going to break this area so I don't know what this is doing it makes no sense at all but I think it's going to flip over yep those are targets for Kate looks great congratulations everyone everyone's doing very very well any questions any comments let's see what we have out tonight so we get excited if you're in this there's nothing to do but just sit and watch yourself make money what about J&J nope see I thought I thought of feeling that this would rally because it's just a wilder mover alright let me go back and see what's up for tonight it's Tuesday tomorrow's Wednesday today is the 18th oh yahoo's tonight IBM's tonight let's take a look at this yahoo okay this will be in play be great for us if this gap down great day that you're right you're definitely right about that came in dude that's the show off IBM I mean um the stock is taking a tumble and looked at this in so long does anyone want me to go over anything this day here yeah it was 2013 good call thank you I remember predicting that this talk would fall and break the trend which it hadn't yet but that it would after this day forget if it swished or I don't even know then it really back and I remember arguing with somebody about it some person that was in the finance world saying this chart was lower hadn't looked at it in so long and it did really back after that but it but it did break it did break after that I was right there's something on that day and I don't remember if it swished or there's something I saw that day and I thought this talk is lower and then I did it turned it turned and I haven't looked at it in years we just don't play this it didn't turn immediately after but then it did there was something about that day I think we I think we did it fact I'm sure we did so this is a watch too okay anyone have questions about anything at all everything is fine if you're in this and you want to hold it it's holding the stop 85 is the number but you're gonna have to be a little patient it is going to attempt to break 122 it's got to set up now in the five minute on the 15 minute Kate came back down did not go down to the low is this gonna take some time to go back down and break it probably probably got to be more patient when you take late trades and I'm not saying 945 is late but I'm saying it'd be the second trade in this and you could have gotten out of half down in there who wants to go over what ask me anything you want ask me anything at all


Don't like it. Please don't use it. Don't bother and if you don't use it We promise we won't hate you or just just a little bit The digit Genesis of it. I wanted to write things. I wanted to write novels, you know It's it's some kind of pick-up lines. Yeah, you know, I'm a writer and I was looking for tools to help me writing efficiently and I Stumbled up on right room. What is right room right room is a full-screen monochrome text editor Yeah full-screen monochrome that is black and white or green on on black, etc text editor, but I Feel myself. What's the point? Yeah, you know, it's text just like a normal text editor But you can edit any kind of text with a text editor But the fact that it's full screen and it's completely full screen You have absolutely no menu bar at all Focuses you on writing you have absolutely no distraction when you're full screen and when it's only text and text only You don't have to bother about is it bold? Is it emphasis? Is it a title? Is it we don't care? We don't have to care. You just write Formatting will be later on Right room, it's a full-screen monochrome text editor and it costs 24.95 US dollars and it only works for the Mac That's not I wanted to have I wanted to have it for Linux, you know I want to go into four years as I wanted it to have it on Linux. I thought myself well a full-screen text editor That that must happen. I must Be already there So I searched the friendly web and I stumbled upon this post on the open to forums where a new woman and Nicolape Rougier Did a pattern that pies single Python script with 500 line that code only 18 kilobytes including comments That just worked It's full screen text editor. It's nothing That's my fault. I just changed the image. It's just having a screenshot of a text editor. It's not that interesting It was still a full-screen monochrome text editor and it costs nothing because it was GPL version 3 and it runs in new Linux You may think now My favorite text editor does exactly the same or I have to do is to Hit the control alt F1 key and run viral remax or whatever I don't care, but you know real programmers. They use Emax now. They use them now You use add a cat or the butterfly that Changes the ripples of the atmosphere of the upper-down atmosphere Parallel is not for geeks. Parallel is not for coders hackers Hackers have tons of tools in order to edit text because text is a basis That's that's their basic work. So it's not for you It's for usual normal people normal Joe who wants to edit text in his own environment except that instead of having many Windows opened it just has One single frame black and white by the way XKCD rocks The pilot of pie There are many pros in my own opinion. It just works first It was lightweight just an 18 kilobyte Python script is nice. It's in written in Python. So I can rock the code understanding it and eventually Modify it and I can modify it and publish the modifications because it's JPL version 3 the cons were there are no actual really handy Theme management if you have if you want to have green on black instead of a white on black, etc It was hard-coded in the in the script. So it was not that nice. It was not localizable I'm French want to have all of my software in my own language and I wish that everyone can have Software in Spanish in in in German in whatever. I don't care Yeah, and it was a little bit rough at the edges. A lot of things were missing in fact So I decided to fork it For car interesting Because you can do it. It's JPL version 3. So I started up a project on on launch pad Though the advantages of launch pad is that you have the code management You can have any as in as many branches as possible. You want it's easier for the translators If you want to use Rosetta, there is a bug manager that I'm using usually as a task manager And there are mailing lists so you can discuss topics Started up a org domain name very simple website. So just a Few few lines of information about the project analysis each channel on 3.0 And then a very nice bunch of developers just join in So when the project started to become a little bit bigger, we had to Organize it in teams. So there is a parent team anyone can can join in Piran bus quad if you want to get spammed by bug mates, you can subscribe it Piran dev is Only for developers hackers. It's on invitation only and Piran that means that's the basic administration releasing etc and There's in fact, there is one Key developer on this project, which is Florian Heiler Unfortunately, it couldn't make it to the first day. I'm so sorry about that There are plenty of hours of people who contributed it and you might note that I'm not on this list because I've become really too busy to to contribute to this project and I'm also I'm just supervising it from from from a distance Still following it very closely in order to keep it as simple as possible as usable as possible, etc But for young Heiler rocks What are the main features now it still edit text and text only no formatting You can edit multiple documents at the same time if you want it has an autosave function, which is Much more interesting that you have autosaves that have experimented. I've Often lost my data because of autosave. In fact, Piran saves Your current document when you manually saves it it saves it on for example example dot text and When it's autosave, it's on a temporary file. So any automatic Save is on this temporary file and when you manually want to actually currently record your data It recalls on the current file or the actual current file. So that saves you a lot of time Trying to get your Backward your backup Full screen works on your head Which was pretty cool that I have a dual head at work and and Unfortunately, it didn't work now. It works and we have several available Themes they can you can send themes by the way via launchpad Any kind of color you want for text for background just works. That's fine. And the themes are customizable that is to say if you Import one theme and you want to change it. You can do it very very easily via the preference style It's translated in a lot of languages. We got Welsh These are the complete translation for the the the earlier releases now The next release is not yet translated in as many languages as of this Plant features Spell check that would be very handy for writers in order to have their Spell check the actual spell check in their own language We want to drop JTK source view because it prevents us from being completely fully compatible with Windows and macro sx. We want to port Pyram to Windows and macro sx and using JTK source view doesn't help Many other themes. Maybe what you want your wish here go down on the website on on launchpad File a work report saying I would like this to happen and we'll plan this or not In a more distant future We had the idea of a theme bizarre when anyone can send themes for pyroom and With a screen capture of the theme and people could download it and install it on pyroom. Why not? It's a web 2.0 system Maybe a plugin system because a lot of people are asking for very simple syntax highlighting such as a markdown or Restructured text etc. And we don't want to do it right now currently But for example, we would like to say you select the text and you hit Control B in order to have a build and it just adds the stars at the end and at the beginning of the text So that would be quite handy, but we need a plugin system in order to do that And it's not yet ready. And if you have a genius idea, go ahead, please dive in Please don't ask for syntax highlight. I told you this is not for color. This is not an idea. You see, it's just a text editor We won't do Python syntax highlighting. We won't do C++ Java Baguette and snails syntax highlighting and if you want specific market markup highlight Just wait for the plugin system and we won't have any IRC client integrated into pyroom. Never If you want to try it, it's just it's just a Python setup usual normal Operation or you're installed pyroom on Fedora. It's been packaged for Fedora very quickly It's packaged in DBN seeds. So it's packaged for Ubuntu 8.10. Oh It's not 8.10. It's 9.0. Yes, it's packaged in the in the PPI's for intrapid and it will be packaged in multiverse for Junty Which is about to be released in April And if you want to package it for your favorite distro, please dive in go ahead to get in touch the launchpad website Which is a main website or pyroom.org in order to have the download very easily very quickly and a hash pyroom and free node So that's it. That's the end. Thank you. I think we have five minutes for questions. That's a long time Yeah, yeah, that's that's an interesting question In fact the first time that I just try to launch pyroom on a dual head it just split it through through the two screens and I had the the actual Window the actual text window right in the middle of it. That was really wrong So now when you use it on a dual heads when you when you launch it on one for example, you're right inside screen The the text is On the right hand side and the other side is completely blank. It's completely black if you have a black background for example So it works that way and that's much better and much more interesting. Yes, please. I use g-edit But we're floating higher. I don't know and I don't care One more. Yeah, sorry. Yeah. Yeah, the question was about fonts You can use the font you want It's a mono space. It's not a mono space. It's Whatever it's in your thing a theme is its colors and and the font and with of texts It's in percentage per your It's percentage With with with the width of the of your screen 45 60 percent, whatever. Yes There is help On the bottom of the of the of the screen of the of the text window There is text say saying that this document has been saved and it disappears and for example control Control H its help. So this help is localized. It's very simple Now this it's not a large amount of Of Strings but you need them of course in order to if someone doesn't absolutely Speak English. You definitely need to translate even the smallest the smallest string Yeah, yes, it's there is a debian package in seed Seed it's the instable debian Well, I don't actually currently know if it's going to be in the next debian release in a February One week hopefully anyone else No No, now try to use it. Thank you very much


Hello, my name is Tim Griffin. I am going to give the introduction to our workshop for GCC 2021 on pandemics research using mass spectrometry. So, real quickly, overview and objectives of our workshop, we are going to introduce Galaxy as a framework for a solution for data analysis across the omics domains, focusing on MS-based proteomics. We hope to provide some hands-on experience in using Galaxy in these applications, demonstrate how Galaxy can be used for mass spectrometry-based proteomics, as well as a little on multi-omic analysis using COVID-19 studies as representative data, and lay the foundation that we hope you may take this away and use Galaxy in your own work and or if you are a developer, contribute your tools to the Galaxy community. So, who are we? We're at the University of Minnesota. The other instructors that you will be hearing from today include Prateek Jagdpap, Subina Mehta, Andrew Ryszewski, and An Nguyen. We are part of the Galaxy for Proteomics, or Galaxy P team at the University of Minnesota. It is a very much a global effort. A number of other contributors have made this possible, some of whom I've listed here, as well as a few of the funders that have helped make some of this work possible over several years. So the topics of the workshop, mainly we'll be focusing on data, mass spectrometry-based proteomic informatics type of data. We'll also be using this applied to COVID-19 research, so obviously very timely application, and you'll see a little bit about this idea of metaproteomic, so more of a multi-omics approach. But in the end, what you're going to see is how we use a mass spectrometer to determine the linear sequence of the proteins that are present in a sample. So we're going to determine the sequences of amino acids, ultimately from peptides that match the proteins, and that is how we're going to determine proteins are present in these samples. So as a bit of a primer, before we move on, I'm going to give just a little background in the mass spectrometry-based proteomics, how it works, and the data type that you're going to be dealing with today. So when we talk about proteomics or protein, or mass spectrometry for proteins, there are a number of ways we can measure the molecular weight or the mass of proteins. We can look at the intact protein, or we can take those proteins, which is, again, this linear chain of amino acids that are equivalently bonded together. You can take that protein or a protein mixture, can use trypsin, which finds arginines and lysines within that sequence, breaks it down into peptides. So smaller amino acid chains that come from that protein, and then we're going to actually detect these peptides, fragment, and sequence the amino acid sequences of these peptides within the mass spectrometer. So we're going to break them into peptides and then build them back up to determine proteins. So how do we do this? So the general approach is to take a mixture of proteins from any given source, and, again, digest these into peptides using trypsin, take those peptides and fractionate them using liquid chromatography, which acts sort of as a molecular turnstile. So similar to taking a crowd through a turnstile at a large vent, we take this crowd of peptides through chromatography, which simplifies the mixture, separates out the peptides, so that they get introduced through an ionization method, electrospray ionization, into a mass spectrometer. The ionization method puts a charge on the peptides as well as puts them in the gas phase. So these would all be different peptides of different sequences and different sizes that are detected at any given moment in the mass spectrometer. They're isolated. Those isolated peptides are then fragmented, and those fragments are then the masses of the fragments that came from the starting peptide are recorded. This happens very quickly in a matter of tens of microseconds, such that the mass spectrometer records this fragmentation pattern for one peptide. It remembers those that were just there a moment ago, goes and gets the next peptide, fragments that peptide, to create another MSMS or tandem mass spectrum, with these fragmentation spectra being unique to the sequence, the amino acid sequence of the peptide that was selected. So how do you then put an amino acid sequence to this fragmentation spectrum? So this unannotated MSMS or fragmentation spectrum is put through a sequence database searching program, where it takes proteins within a database that could be predicted or known proteins that have been sequenced, takes all those potential sequences, creates theoretical spectra for those fragmentation patterns in the database and then does a match, and such that those that are observed for those that come from peptides, they will be matched to a theoretical fragmentation pattern that matches the observed. So that when this works, as we're showing here, what we get now is an annotated mass spectrum, where we have amino acid sequences mapped now to all these fragment pieces, and that gives us the full sequence then of whatever the peptide was that created this tandem mass spectrum. So once we match a peptide sequence to our MSMS spectrum, we then can relate that back to the protein because we started with proteins, digested them to peptides. So we now can, if we desire, we can take the peptides we've identified and infer a protein. Usually what will happen is that a protein, if it's present, we will detect and sequence more than one peptide off of that protein, so that gives us a bit more confidence if that protein is actually present. So you'll usually see multiple peptides when identified that map back and help you infer identity and presence of proteins. So as the topic of this workshop is, this is a very much a bioinformatics-driven analysis of this data. So what you're gonna learn about today is that there's a number of tools that are necessary to do this identification of these peptides and the inference of proteins. And we'll take it a step further is that you're going to see the application of this where we're gonna need to also identify what organism does a particular peptide sequence come from? How do we validate that these are actually confident matches? And so it's very much a multi-step bioinformatics challenge. So how do you approach that? The way that we have approached that is by extending Galaxy. And without going into too much depth on Galaxy, what Galaxy gives us in this application area is a very nice environment, sort of this unified environment to take in data, to integrate and implement tools, different software tools that can be then integrated together to work together, implement this on powerful, scalable computing resources, whether it's in the cloud or locally. And this all comes together within the Galaxy environment and is also driven by a user interface that makes this usable by non-expert programmers or through the API where it can be operated as well for those with a little bit more expertise as programmers. So it brings all of this together as this really nice platform to carry out these complex bioinformatics and multi-omic data analysis workflows. So one thing that you're going to hear a fair amount about today is this idea of defining workflows and also history. So as I've kind of alluded to, one of the key pieces here is that as these analyses become more and more complex, they go from a single software tool to needing multiple software tools. And those software tools then need to be sort of chained together or integrated in a way such that the output from one software tool acts as the input for a subsequent tool. And you would like ideally to be able to develop an integrated workflow where one can put in input data and in an automated way have this data go through this workflow. And each subsequent tool takes in an input, brings an output that may serve as an input to the next tool and ultimately get to an end result that that's the desired results file that one can utilize and export off of Galaxy or even visualize on Galaxy. So that's the idea of workflows that you're going to see here. It's really these different tools that have been plugged in together and work in an integrated way. And in Galaxy, what we can do is actually save those with all the various parameters of each tool in an optimized state so that one can share and access these and make this meet more reproducible. We also will see histories. And a history is basically a workflow that is acted upon data and it's an archived record of that analysis by using that workflow, one has analyzed data and it saves all of the input data or all the intermediate data as well as the final output. So you can see sort of what happened and again, share that with others helped for these analyses to be much more reproducible as well as transparent. So those are a couple of keys that you're going to learn about today. And with that, I'm going to end the introduction section. And the next thing you'll hear about is a bit more about the specifics of the application of these workflows and tools to COVID-19 data. Now that we have learned about the mass spectrometry methods that are used to detect peptides and proteins. Let us start looking into galaxy workflows that we have developed for the analysis of COVID-19 mass spectrometry data sets. On the website here, you'll find galaxy workflows that we have developed for COVID-19 mass spectrometry data set analysis. These workflows were developed to detect peptides both from clinical as well as cell culture samples. And using these workflows, we have been able to detect peptides that have spanned the entire SARS-CoV-2 proteome. We've also developed metaproteomics analysis workflows that can be used to detect potential co-infecting microorganisms in COVID-19 patient samples. In order to understand a little bit more about the COVID-19 detection methods, let us go back to the times when methods were used in the early pandemic to detect COVID-19 from patients. Swap samples were collected and then were used to extract RNA from these samples. This RNA was converted to DNA. And then this DNA was amplified using SARS-CoV-2 specific primers using polymerase chain reaction. And then the PCR results were used to detect the severity of infection. This was primarily used for active infections. For prior infections, tests that were used were basically antibody tests, wherein blood samples were collected from individuals and the presence of antibodies against a prior infection was used as a way to interpret whether the individual was exposed to SARS-CoV-2. During these early times of the pandemic, mass spectrometry researchers also looked into the possibility of using mass spectrometry as a method to detect peptides in clinical samples. As you can see here, multiple labs from around the world collected samples from multiple body locations, including gargling solutions from the upper respiratory tract, nasopharyngeal samples, as well as, in some cases, deep lung tissue samples from deceased patients. Using these samples as well as cell culture samples, we were able to construct a panel of peptides that could be used for data analysis. In cell culture samples, for example, one could look into the deeper proteome of SARS-CoV-2 virus since you could extract cell lines that were infected by a virus and one could collect samples at various time points. So in order to generate this peptide panel, we started with some clinical samples, as has been shown here, as well as some samples which used in silico approaches to predict the presence of the proteins from the genome that was available in the early days. And these methods were basically used to detect any potential candidates for diagnosis or potential candidates for vaccination. As I mentioned earlier, we also looked at some cell culture data sets and these helped us to get a deeper understanding of the proteome of SARS-CoV-2. So using these multiple data sets, we were able to generate a peptide panel, wherein in the first step, we use these data sets, either cell culture or clinical data sets, to generate a 639 peptide panel using database search flow workflow methods. In the second step, we use the 639 peptides to interrogate whether these peptides were present in clinical samples. As a result of this, we were able to look at the entire proteome shown here in this outer circle, each of these sections showing a protein that is potentially expressed in the SARS-CoV-2 proteome. And you can see in the inner circle are peptides that we detected from our studies. So this is the 639 peptide panel that I mentioned earlier. We could also go back and look at our clinical data sets and what is shown here in the second inner most circle here is the peptides that were specific to clinical samples. Using this analysis, which are published in clinical proteomics this year, we were able to find out peptides that could be used for detection in clinical samples. The other part of workflow development that we were interested in was to find out if we could detect any co-infections in COVID-19 patients. And the reason why we got interested in this was because in most cases, especially in cases wherein patients have recovered from COVID-19 infection, there are secondary infections that sometime linger because of the weakened immune system or weakened lung function. For example, the patient could be infected due to a prior infection or also could be infected during hospitalization. These infections called as nosocomial infections can also affect antibiotic treatment plans since some of these organisms can be antibiotic resistant. In most cases, culture-based methods are used to detect the presence of any secondary infection. However, that might take time and might lead into progression of the disease. There have been quite a few cases of fungal infections after COVID treatment in India in the recent pandemic wave that it experienced. So in order to study and find out if there are any other microorganisms present, we looked at the gargling solution sample, oral pharyngeal sample, and respiratory tract sample that were previously published and were initially used to detect COVID-19 peptides. And we used a workflow that was developed within the Galaxy framework that could be used to detect any microorganisms that are present. And this was published in the Journal of Protein Research this year. So in our analysis, we found streptococcus pneumonia, which is known to cause pneumonia and respiratory tract infection. We also found some other organisms such as lactobacillus, pseudomonas species and acetylbacter species. So as a result of this work, we have now Galaxy workflows that are available, which can be used not only for data that is published, but also for any new data sets that are available from various parts of the world. We have also published this workflows in the two manuscripts that I mentioned earlier. And now it is time for Subhina, Andrew and An to take you through the hands-on session for detecting peptides from SARS-CoV-2 positive samples, as well as detecting microbial infections or secondary infections that could be caused due to co-infections during the pandemic. Thank you. Hello, now is the time for the practical portion of this workshop, where we will take you through the business of detection of COVID-19 peptides in patient data. This section will be presented by a trio of researchers in the Galaxy P team at the University of Minnesota, Myself Subhina Mehta, Andrew Rychewski and Ann Nguyen. To begin, you'll need to create an account at usegalaxy.eu using a valid email address. Once you have registered, you can use the usegalaxy.eu to access materials and you will need for the workshop. Within the usegalaxy.eu webpage, bring your cursor to the toolbar at the top of the screen and click on share data and then histories and COVID-19 inputs, which will bring you to a history containing all the necessary data to bring in your analysis. Click on the plus sign to import the history into your account. Now this will import the history into your account. Now go ahead and click on share data again. This time click on workflows and then COVID workflow. This is the automated workflow we'll be discussing. As before, import the workflow and start using the workflow. You should be now looking at the workflow in your account. To get the ball rolling, go ahead, click on play. Now you'll see the workflow in your center pane. Please select the appropriate inputs that we need for running this workflow. Select sample one data set as a raw file in the drop-down menu and look at all the tools that have been queued up. Now, once you have established that, run the workflow. Now your tools will queue up on the right-hand side pane. So I hope you were able to run the workflow on the data set provided. If not, we can go over that again. Let me basically give you a background information regarding what we just did. The data we used today was from a group which performed quantitative proteomic analysis on oral nasopharyngeal swabs used for COVID-19 diagnosis. They had patient data from both positive and negative samples. The question we are trying to answer is to determine the presence of COVID-19 as well as any other co-infecting organism. Here is the galaxy view of the workflow you just ran. It might look a bit complicated, but we have just used few text manipulation tools to make the tools compatible with each other. Here is a somewhat easier version. Let me go step by step. Firstly, the main inputs of the workflow are the protein-fastly database comprising of human proteins, contaminants and the COVID-19 proteins and the MS-MS data. The first step is to perform database searching for which we use the search query peptide shaker combo. We will be going into each of these tools later on. First, we use the peptide report from the peptide shaker to extract confident peptides, of which we perform unipept analysis to get taxonomy. We look at the organisms that have at least two peptides for selecting these peptides and perform PEP query analysis on. PEP query is the validation tool we use. We eliminate the presence of human and contaminant proteins by providing that database. Once we get the output, we perform a filtering step in which we extract peptides based on P-value and confidence. These peptides are then subjected to another step of validation, which is spectral validation, for which we use the mcidentml file, which is a peptide shaker output to create an mz2sqlite database. This sqlite database can be then viewed with the help of the multi-omics visualization platform within the galaxy. The platform has inbuilt lorikeet visualization to visualize the spectra. We then manually examine the spectra and extract those peptides and then perform unipept analysis to determine the presence of COVID-19 as well as other organisms. So having done all that, we dare say you have noticed that the workflow is not finished. This is typical. As considerable time is generally needed for these tools to run, in the meantime, we would like to you to be able to analyze some data for yourself. So you go back up to the toolbar at the top of the screen, click on share data and then histories, and then finally open up the COVID sample one history. Import this history by clicking on the plus sign. We will interrogate this data shortly. Thank you for- For my desk bar. Hello, my name is Andrew Rucheski, and I'll be getting into some of the main tools of the workflow with you. First, there's the paired duo of search GUI and peptide shaker. Search GUI is a collection of peptide search engines that run together with eight to nine built-in algorithms, such as X tandem, Comet, MSGF plus, et cetera. We provide it with all the identification parameters for database searching, such as the spectrometer resolution, fixed modifications, variable modifications, et cetera. Search GUI then generates a report. Peptide shaker takes search GUI outputs and ranks and filters the peptide spectral matches based on their quality, selecting those peptide spectral matches that are better than a 1% false discovery rate. Peptide shaker then creates tabular peptide report files, follows an MZIdentML file. We use the peptide report and the MZIdentML file for downstream processing. The next major tool is called UNIPEP, an open-source application that was purpose-built for metaproteomic analyses. It can perform both functional and taxonomic annotation, but for this tutorial and workflow, we are performing only taxonomic annotation. We select organisms with at least two peptides, as this is a general standard for confidently assigning peptides to a target in protein mass spectrometry. Generally, this strategy is used to assign peptides to a protein, but in our case, we assign peptides to an organism in the community. We then extract those peptides for peptide validation analysis. There are two main steps we use for validation. First is the PEP query search engine, where we eliminate the presence of host proteins and extract validated peptides for our duly identified organisms. With that accomplished, we then manually assess the spectral quality of these validated peptides. This is necessary as peptides which can pass quite PEP query to not always have the highest quality, making them ill-suited for reliable automated detection. This figure summarizes how the query search engine works. It was generated by the creators of the tool, the Zhang Lab at Baylor College of Medicine. First, our peptides of interests are compared with the MSMS data to generate PSMs with matching spectra. These are then compared to a reference database, the score based on whether there are better matches to the PSM spectra amongst the reference peptides or not. Then the PSMs are compared to a library of randomly generated peptides to see whether a match to the spectra can be found at random, increasing the robustness of this match. Finally, if engaged, the PEP query performs what is called unrestricted modification searching to see whether the reference proteome can form a better match to our PSM spectra, allowing for post-translational modifications to be counted. Peptides of interests with PSMs that have no match to the reference proteome with or without post-translational modifications and no matches to random peptides are considered to have passed. When the query is finished, it returns a number of tabular files, including the PSM rank. We can then filter the contents of this output to eliminate those peptides that show too close a match to the reference database, which in our case is the human host proteome supplemented with common mass spectrometry contaminants. Theoretically, those peptides with a P value less than 0.05 correspond to peptides which haven't had a match in the reference database. When the more stringent unrestricted modification search is performed, there is an additional column called confidence which designates the peptides as being yes or no. We will filter the PSM rank file based on those peptides that are designated as a yes. These are our validated peptides. However, there is one last step to validation and that is manual evaluation of the peptide spectra. Hello, my name is Andrew and I will take you through the final validation step of the COVID-19 data. Spectra of peptide past the validation by PEP query will be manually annotated by MVP, a multi-omic visualization platform that includes the Bolarikid visualization tools. Bolarikid utilized the SQLite conversions of MCIDEM5 generated earlier in the workflow. We first manually look at the signal-to-noise ratio of the product ions MSMS spectra and spectra-containing product ions with at least a pre-fold higher in intensity than noise peaks were retained. Next, we inspect the degree of completeness of the B and Y ion series with passing spectra determined to have at least a three consecutive B or Y ions. Peptide with spectra fulfilling these two requirements will be considered valid target for the detection of SARS-CoV-2. So here we present two examples of spectra. The first spectrum doesn't have three consecutive B or Y ion, which is why we rejected. The second spectrum, on the other hand, has more than three consecutive Y ions and product ions are noticeably more intense than the noise peaks, so we accept it. Next, let's visualize the process and see how it works. In the completed history, click on data set 10 MC to SQLI to expand it and then click on the background icon to visualize the data. From there, you can click on the MVP window, which will open the large heat visualization window. Rather than go through all of this data, you can select load from Galaxy, select the list of validated peptide and then click use for filtering. This will narrow down the list of peptide from several thousands to a handful. From there, you can select a peptide of interest, click on PSM for selected peptides and then inspect the spectra presented below. So with all of this background presented, we encourage you to run the workflow by yourself. Take a look at the PepCurry PSM rank five. Visualize the MC to SQLI output using MVP and annotate the passing peptide by yourself. Now you decided what you think is the COVID and co-infection status of this sample. With this, we conclude our presentation. We would like to thank you for attending and engaging in this workshop at the 2021 Galaxy Community Conference. Below is a collection of links that you may find useful, which include our contact information, a specialized Galaxy 4.0 mix platform host by Galaxy EU, repositories of our workflow, as well as training materials for the use of the Galaxy platform. And finally, publications detailing our COVID-19 ProTill mix research. We hope you enjoyed the workshop.


what there's maybe a little bit too much well it's going to be macho but it's you don't want to talk about it all the damn time so no no in fact being i'm really getting sick and tired of alpha i don't know about you i'm getting really sick and tired of alpha bullshit well alpha alpha people can go the other way they can they can get very arrogant and braggadocious and always competing you know me i i don't compete with anybody i don't care about you for shit i don't care what other people do you know you know like i don't compete you went already if i exercise i compete with myself i compete you know i compete with the james the james bond from what i did last week yeah that the guy that worked out last week yep me is the one who i compete with do do am i stronger this week be better than yourself right am i stronger this week than i was last week never mind him fuck him never mind you fuck you how much better are you today yeah it's like it's like a big dick contest yesterday's gone yeah yeah that's wrong yesterday's done well dick swinging is is only gonna hurl you back quite a bit all you know it's a bad crappy evening you know who who likes to compete and stare at everybody else and people who go to a nationally advertised gym a health club that they're so preoccupied with what other people are doing it's a thirst trap i can't i i i actually it's a total thirst trap i i like to exercise alone i like to do it so well you don't have people saying i'm gonna beat the shit out of you well there's no judgmental people when you work out alone you know there's no people staring at yet there's no people you don't have a gym gas lighting you saying how oh don't worry nobody's fucking with you and then you hear about how people at the gym say they hella they hella say on their even on their phone recording without permission oh i don't know that but it's so this guy i'm i'm gonna kick your ass it's like whatever it sounds like be the be the bevis and butthead episode you know it's it the thing is and then they say oh you're doing it wrong here let me show you how to do it nope don't need your help let me show you how to do curls hey you're not doing it properly yeah let me show you yes i am go away you're not talking properly you're also not even working out you're just telling other people how to work out how come you're not working out you know i i used to laugh when i was a personal trainer these young chicks would come in to the gym the health club they'd be wearing all this like designer spandex and everything oh and they had big earrings on and they had all their makeup on i'm going careful don't die who puts all this makeup on to work out need you here i mean i mean a girl who is really dedicated she comes in wearing rags no makeup and rags because she she's there to work out how can anybody go ahead look at it they're looking at themselves in the mirror they're looking around at everybody then they take a sip of water and then they go to the machine and they do a few reps and then they stop and then they sip some more water and they they're looking in the mirror again sounds like a couple of jumbos going around a couple bazookas that's that's just why would anybody ask them out i don't know no it's like it's like they're not with some burpees the narcissists or they they know they're checking out i think they come in peace they want to see who's looking at them they want to see how many guys are checking them out i'm i'm looking at myself don't worry so they're really they're really not there to train yep you know and then oh then there's the people who are supposed to be they're supposed to be working yeah and they're always on then they're on their phone at the gym what the hell texting texting texting texting texting texting you know so that's the story hey at least at least you got topics for tomorrow oh yeah um i mean uh but James like what's the point of doing all this stuff in public if you know it's like every time you're in public you have to deal with bullshit i love it i got my my i call my personal training studio 54 i what i i got everything i need it's called studio yeah i made my own sandbag for doing certain exercises i got i got one of my new duffle bags small one and i worry i got a 40 pound bag of washed play sand from home depot so now i'm going to make my own weighted sandbag for doing it has the straps oh congrats yeah why pay a lot of money for exercise equipment i can make my own yeah especially if you're gonna anytime you're going to be there you're going to be all can i work in or i'm gonna i'm going to beat the shit out of you people saying that at the gym oh my so my shoulders my chest my chest people hell mean mug you at the gym what's that about i'm not trying to i'm not trying to be better than you it's like what did i do and what do what do what do they do besides hang out with your buddies and make fun of other people i mean why are they always yeah they always have this nasty scowl on their face yeah why why are they why did they pay money to join a health club or a gym to just go for go for round or something yeah yeah especially that especially to go around with this nasty scowl on your face but yeah now now speaking of alkalizing even walmart is selling in the pharmacy the organic freeze dry juice concentrates the green the green the green concentrate the green concentrate where you got like everything the wheat grass the barley grass any freezer burn on that huh any freezer or fridge burn or something on that no no you could you actually i keep my powders in the freezer nice i keep them in the freezer and they they don't they will never go bad on you the freezer is good because they can freeze half of what you buy the week before and then you don't have to go back for a while yeah well your powders they they they're still powder in the freezer oh powders powder is nutrition i think yeah like like there's a company from california that makes powdered juice concentrate from every anything oh yeah anything anything you want you want barley grass they have barley grass juice powder you want wheat grass you want oh that's really healthy it got me i i use moringa powder that's really healthy moringa is like i'm is on top of mount olympus splat no when it comes to nutrition it's second to none by the way did you did you hear about those jumbos uh that the james web space telescope discovered these kind of think gas planets rogue gas planets out of never the james web telescope space telescope is right breaking it's finding new groundbreaking things every week oh my gosh every week i on the show i show photos of one of saturn saturn has 145 moons one of the moons looks like a ravioli it's called the ravioli you all right no yeah uh that's that's cool that's cool yeah it looks like a ravioli a ravioli moon and saturn has 145 moons and 145 moons and one of the picture looks exactly like a giant ravioli how come saturn has so many fucking moons i mean saturn how come the gas giants are always better than the terrestrial planets yeah well look at look at the the moons of jupiter europea there's fantastic there's water there's oceans on there there's paradise the same thing with saturn there's like the moons are like little little planets paradise oh yeah probably because the planet itself is so big so the nebulas are gorgeous the beauty the beauty of the nebulas the uh wonder what life is like in a nebula like if you lived in a nebula wonder what that's like i i i they took a they took the most high definition photo of the andromeda galaxy that's our nearest our nearest neighbor andromeda getting near yeah i i think i still i think i might still have it we have some time before we collide with andromeda and our planet gets impacted yeah well what i like is that they they're actually able to deflect a killer asteroid before it hits earth don't nuke it right no because then you'll have like a thousand one you know it's like a shotgun with a buck shot but no they'll they want to they they tried they actually succeeded in deflecting an asteroid wait what do they do they they sent uh a probe or a space they sent a probe i don't know what else to call it and it it landed on the asteroid and it uh it caused it to to change its course whoa that's crazy so if its course if its course is to destroy us then it has now it has a new course nice india landed on the south pole of the moon the other day yes they did they and and the funny thing is they keep saying oh there's lots of valuable metals and minerals and precious metals on how you got how you gonna ship how you how are you gonna mine a moon or mars or an asteroid how you're gonna bring all that back to earth yeah especially without it burning up in the atmosphere or something they're ridiculous they're talking about they can make a fortune after fortune by mining the moon or mining an asteroid yeah try to bring that back it's not gonna happen especially maybe non non entities might non entities may try to may attempt to get there first yeah yeah there's uh oh i read an article where this this professor of this famous professor of a certain university is yeah is he's in he's a doctor right he has a phd and he's insistent yeah there are no aliens aboard ufo is how the hell does he know does he have made you the queen of england does he have x-ray vision could he see inside of the ufo and say oh there's nobody in there well it's like who died and made you the queen of england he he doesn't know of course there's somebody operating it there's some entity operating it yep you know what is what does he think they you know i mean they they move at the speed of light because there's no way any any aircraft yeah nothing can move faster than the speed of light don't think really much of anything from earth can move at the speed of light so don't even try to engage with ufos if you're if you're a pilot in the air force you're never you're never gonna you might not have a choice you might have to engage but you want but you won't you won't succeed don't think so if it's if it's what you think it is tech so much more technologically advanced yeah anything that can move across the sky at the blink of an eye mock speed i wonder what the mock of ufo is is wondering what the mock speed of that is like mock 20 maybe even what it called warp warp speed warp speed like in star trek warp warp maybe they're not even there maybe it's just kind of they're whizzing by because they're like really kind of like kind of a version of a wormhole or something and it's just kind of a flicker i think i think they're they could be inter dimensionally traveling perhaps you're right about the wormholes uh uh because you know like einstein said a human being couldn't cannot survive the speed of light the light you sure you you think uh human would die if they went the required speed to reach certain parts of the universe i think they have to take shortcuts through wormholes i think i think that's how that's how aliens travel i think they take shortcuts you know they'll go through a wormhole end up in so many thousands of light years away that's more practical and there's dimensional what about all the the cryptid creatures that people are seeing nowadays you know they could be inter dimensional uh beings yeah and also just we're already seeing hints of life on other planets far away but still yeah there's uh i was i was watching a video about dimensions said there there really are many dimensions above the third yeah you know i mean we only know of the third dimension but they're they go they go up pretty high you know i'm sure and uh you hear about the heishan world the the ocean world that might harbor life they say there's a there's a there's an ocean deep in in the earth oh no there there's a planet there's an ocean planet there is a planet with an ocean and it's it might harbor life there there might be life well it only makes sense since uh since they look at look at the vast amount of galaxies that are out in the universe i mean uh i mean there's who's to say there's not a planet here and a planet there that are in the goldilocks zone goldilocks zone like we are not too hot not too cold ah you're a smart cookie that's right thank you not too hot not too cold yep my my my my head is is is the sun all right of course venus third and mercury are way too hot no goldilocks but then then mars and semi goldilocks and then you have earth all right that's what that's what we want to find yep that's not flirting that's a real deal right and and and that doesn't come often and and uh of course has to be a solid planet you know order rock can't guys and giants don't have a surface can't have any any toxic gas like like a lot of gas planets uh too much but some of them are beautiful like urinus urinus urinus urinus neptune is a beautiful blue oh blue planet yeah jupiter is too easy everybody's favorite planters jupiter mine is neptune how about how about you you know i like neptune i i love the i love the color blue it's like um it's like a it's like a cross between a it's like an uh like it's like a royal blue yeah to be honest i thought i thought of i always thought neptune was navy blue it's well urinus is a lighter blue uh it's lighter it's pretty shade gray it's light i thought it was green thought yeah well yeah well urinus is could be like a turquoise it could be like an ocean an ocean of blue is like a combo blue and green right turquoise wait hold on i'm gonna play the movie contact okay i'm gonna watch i'm gonna watch contact now okay no problem oh no but i'm still here so all right yeah yeah uh yeah neptune is definitely as far as looks go neptune is a beautiful planet now pluto yep they keep on demoting pluto and then they make it a planet again but pluto as small as pluto is and it's pretty damn small yeah is a beautiful planet they show the photos of pluto i'm telling you it's it's like a it's pretty pink like a purplish purple and purplish pink of some kind sure it's a beautiful planet but some say it's smaller than our world yeah this world so small dude yeah but there is activity on pluto you know mark uh there no yeah for sure uh but pluto is just it's too cold it's too far away uh it's just it was there's no sun there's no warmth um there's um yeah it's just it's um just picture how cold antarctica is and then think about pluto you know but at least antarctica has like uh has life i don't know how the life survives but you know there's life for but you know um yeah pluto forget it it's i would be shocked if there's any alien life on an ice planet like that oh idea you know what i gotta do tonight i should check netflix and see if there's any any new horror movies on or any even action movies you like horror i like i like horror in terms of like spooky stuff you know like spirits and and uh monsters i like monsters i used to watch all the jeffet monster movies when i was a kid i am legend pretty good tolo productions yeah i like i like i like monsters and then i like any kind of monster movie you'd like i it's a bit of a new movie but you would like i am legend i am legend yep i remember that the special effects in that in this movie were amazing uh aquaman oh but i watched aquaman and uh you know of course because of computer animations they're they're able to do so many things yeah mars has two moons you know there are two moons they're basically asteroids but mars has two moons demos and phobos so mars has two moons the earth has one really mars has two moons mars has two moons i don't think venus or mercury have any moons they're small right yeah mars is smaller than the earth um how are we so small yeah they're they're they're somewhat small they're they're not they're not a lot smaller but it's somewhat small in the earth yeah it's like that it's not perfectly round mars has a one side that's like like flat we're almost like an asteroid hit it one time yeah yeah and uh you ever see the polar polar ice caps on mars i think i have yeah yeah they have polar ice caps um they used to be more habitable yeah at one time maybe they were it would have huge asteroid maybe it was in the goldy lost zone got repositioned and then maybe it got knocked off and then maybe that's why i don't know very it's very possible that um i don't know um that they were um they well the top scientists that are in the spotlight say that um that asteroids have life on them some asteroids do have life you know it could be uh bacteria algae light whatever likens and life was brought to the earth so then we try to mine on asteroids and then we end up getting our asses kick yeah i think the only way they could help mars if they terraformed it they have to put yeah they have to put uh very primitive tiny forms of life that create oxygen that you know there's something that life requires or there's something that's the only if it's a building block it can only be there it's forming life that's the only yeah why it exists yeah they need to they need to rebuild mars's atmosphere see mars mars's atmosphere is very thin they have an atmosphere but it does not much of it it's very thin yeah so if they terraformed mars with life that would build up the atmosphere you would like the movie contact and you would also like ad astra those are two good space movies ad astra is i only you know it's it's 2019 it's new but it's so good space movies are really good oh yeah yeah space well especially space movies today they're so good they're automatically going to be fantastic they're not they're not going to be low budget like the star trek series oh no no they're their quality space movies are totally quality and it's it's there those movies are for hustlers that's just every time watch those get really motivated you know what i mean oh yeah some movies just they get they get you going um oh yeah they you're at the edge of your seat with some of them yeah and you're like holy shit want to want to be like that that's kind of like top gun top gun's kind of like sci-fi it's kind of like action scott it's also got some sci-fi top gun it was science non fiction but top gun's good top gun yeah okay yep that's that's that's an old movie but i was uh i've been reading i've been showing a lot of robotics videos at the beginning of the show that the robotics are getting advanced at a very fast pace oh slow down they are almost to a scary in a scary way because they are they have humanoid type robots where you can have a conversation with them and they they show emotion sense of humor uh intellect they i'm telling you it's like careful it's incredible yeah well then they they i could see them getting together if they can think for themselves they'll get together and they'll say you know these humans and they're nothing but trouble they're yeah they can't manage themselves or what what they said in the terminator or something like that and it's it's just uh yeah it's kind of like uh the animatrix they said uh humans made robots in their own likeness kind of quoting the bible think and then with that they created their own demise yeah the humans are our our creators and we are superior to them they are absolute like this try light zone episode how can you make something absolutely wonder how you can make something that is better than you well they don't have a soul uh even though the soul has well i don't want to say the soul hasn't been proven but yeah their their spiritual beings have been proven but yeah it's like machines machines though they can get stuff done and stay focused a human being can get stuff done but then still have a flip out in public well machines are like mr. spark they're logical yep they they they have a mission yep no emotional bullshit they have um what do you call like the clidesdale they have blinders yep and they their function is to get the job done it's funny how it's funny when a computer says to hey i'm not quite working properly i i think you're loading too many files onto my onto your computer you're not deleting enough shit you are loading too many files i am you're not deleting enough shit i am experiencing overload you must delete delete i'm gonna give you shit until you delete some shit you know they said that within 10 years from now yep okay the humanoid android robot yep will be mass produced for that where the average person could buy his or her own robot and they will do all your chores around the house outside the house i like to do my own chores you know they will i mean the thing is they have to have very sensitive dexterity in order for your android you want to you can call mini me let's just call them mini mic okay hey could you make me fettuccine carbonara with jumbo with jumbo shrimp i feel like having fettuccine carbonara and the robot says robert robert i mean michael no my robot will be named robert okay all right okay master michael i will make your dinner now and if if the robot had the dexterity to handle kitchen tools and actually make it the right way yeah then um then it it's very possible uh now i'm afraid if i get the room but if i got the robot the vacuums the one that finds its way back to the charging unit and recharges itself well what if what if i have to make sure it's turned off late at night what if i get up to go to the bathroom and i and i trip over it it's and it's go it's zipping around the house at night and i step on it and i go flying oh no that's not good i gotta make sure it's off yep but um i can picture robert your robot robert robert the robot roby the robot that master michael we need bacon to make fettuccine carbonara where is the bacon do you want to make it yes it's all in the back robert look for it get your own go go shopping how fast could you run robert could you get to the store like like the flash could you run like the flash of course master michael i can run very fast i will go to the market no no i think i think that robots will help but we just can't treat them like we treat students or teachers well there's um there's an android named sophia and if you looked her videos up on youtube fantastic you will be amazed that people are actually talking to a robot fantastic that's that's how advanced they are now named sophia yeah yeah fantastic she has a she has all right how's your conversation she's got everything personality sense of humor sarcasm she's a nine she's yeah the only thing that she just she's not in a solid nine physical like in other words they have androids that have very humanoid physical functions trauma so fia is can only move her arms she yeah she she's mostly talking and and the and the brain and but she she has spatial expressions that's the amazing part these these humanoid robots have facial expressions whoa just like us man yeah stop it's mind blowing oh it's not that face yeah it's not that face of ages now yeah yeah it's kind of like bill murray face yeah bill bill murray you like um um um now that the writer strike is over um it's over yeah um um bill mar you like bill mar yeah i think he uh he went on strike what what do you think they all they all went on strike like bill mar like bill mar yeah yeah i like i like he does what the like uh the daily show the original guide john stewart you said you know uh mixing comedy with with real news real political news yeah so people get their news from that well actually they can yeah and it's it's it's true and it's not like it's not like exaggerated or something i don't know like uh hey james i gotta go yeah i'm gonna close up shop and i'm i'm getting a little hungry so i'm gonna go yeah it's midnight what's wrong with you peace dude okay peace talk i'll see you tomorrow you too see you tomorrow bye bye bye okay everybody thank you for stopping by everybody that's so me from japan mic van raven from chicago western mic from san francisco thanks to everyone have an enjoyable sunday and have a safe and enjoyable