And I hope you're all welcome back to my channel or if you're new here, my name is Cepi and in today's video I asked you guys basically on Instagram to send me in some of your questions. If you don't follow me on Instagram, stop right now, go on the app, type in Persian Bunny, it's across the screen, hit that follow button, I do lives and I talk to you guys through my stories and DMs and all of that stuff plus those pictures of me. So if you guys want to keep up with me, follow me there. But yeah, I asked you guys on my Instagram story to ask me some questions and I said that they can be as general as possible, it could be asking me about advice, you could ask me questions about my life or your life, relationships, boys, anything and I've got a bunch of questions back and we're just going to work our way through it, peeps. We're going to talk it through, let's see if I can, I don't know, dish out some advice or talk about my life and yeah, I feel like we haven't done this in ages, so the question is how old does your boyfriend have to be? I don't know why I went through this phase where I really felt like I needed to be, like if I was talking to a guy that he needed to be at least four or five years older than me because I felt like the guys that were my age just weren't, I don't know, not that they weren't serious or anything like that but I just feel like the way I thought was just a completely different way to the way that they think and mentality but anyway, time a few months has shown me that actually it's got nothing to do with age and it's really to do with like where that person's at that moment in time so there's no age I think so long as you're my age or older that's fine I feel like if I date someone who's like much younger than me that would just be like not okay because I feel like I would be much more like mentally mature than them so yeah will you be filming any makeup tutorials in the future? yes I will I've gotten so many requests for makeup tutorials recently so I really want to film like one apparently like you guys are really liking my makeup in like the past couple weeks it's really weird like as in it's only been in the past couple weeks I don't know I've changed but I'll try and film it so that it doesn't change by the time I film that video so yeah that'll probably be my next video that you guys will see how old do you love you so much love you too I'm 23 a lot of people are asking do you have a boyfriend I don't have a boyfriend yeah that seems to be the thing that stays the same with these cute nays as I don't have a boyfriend and that's not to say by the way that like I do or don't want a boyfriend it just means like I haven't met someone who I feel like is someone who I want to be with like in a long time and I don't see the point in like did I have something on my lips this whole time I'm sorry I don't see the point in like just seeing someone if there's like no future so yeah I'd want if I have a boyfriend I'd want it to be someone who I can like introduce to my family maybe even introduce to my YouTube family I don't know but yeah so that's why I'm not really in like a rush to be like yeah like let me just date you and see how it goes or whatever next question how to cut off a toxic friend trust me I've dealt with my fair share of these sis what you need to do is you need to phase them out so like don't just cut them off instantly like you need to slowly slowly like stop like just don't reply to their text messages as quickly don't answer their phone calls straight away and stuff and like don't make yourself as available to them if you go to school with them I guess it's more difficult I don't know how I'll do well or if you work with with the toxic person but yeah just like change your group of friends like let's say if like the person is like who you used to spend like all of your time with find a new person or go to like the library and read some books trust me those books will get you further in life than those toxic friends any bad breakups no touch wood nope what do you eat in a day well I pretty much just stuff my face with pasta right now because I'm having a bit of a meh day and obviously that always like can happen to anyone so don't forget that you can have like bad days and good days and you shouldn't eat your feelings I took my dog for a walk which was good and I had some pasta and for breakfast I had toast and yeah I'm gonna go Jim because I feel like Jim always makes me feel good and seeing my friends always makes me feel good so yeah if any of you guys this was kind of a joint what do you eat in a day with how to get over about a listen to positive energy tapes and read positive energy books go to the gym see your friends and yeah distract yourself that is how I get over a bad day would you ever do a meet up in London I don't know like as in is there enough people in London who want to see me I would love to the other day someone said hi to me on the train hi if you're watching this and that like literally made my whole morning but she said hi to me at like 7.30 in the morning I was like you've made the whole day great now so yeah thanks so much yeah I would love to do a meet up but I don't know how many people come so let me let me know guys if you're interested were you ever bullied as a kid being Middle Eastern or bilingual I was bullied but not for being Middle Eastern or bilingual I remember when I was in year three I joined a new school and the kids all already had their friends and stuff and they said believe me say my eyes were too big and like I was actually like the only Iranian like my school was a lot of like it wasn't really as multicultural like as you would expect to like a British school to be actually to be fair I'm lying because there was a lot of kids like in my area from like Japan and like they're like some Turkish people what many like Arabi Middle Eastern but yeah so anyway I got bullied for having big eyes and but then I found my best friend and that's where I kind of became confident but yeah oh I also got re-bullied again in secondary school and and then again I found my best friend and that's where I became really confident so there's like a silver lining with everything like being bullied is actually horrible and the people who bullied you they don't end up happy trust me I'm 23 now I've seen the lives of the people who bullied me a most of them beg friend me now like beg friend me I'm not even joking with you and I'm just like if you're an evil as a child or as a teenager do I want to be your friend now but second well they have a lot of life issues themselves they have stuff going on that you don't have any idea about everyone does so you know don't be mean to people but yeah beliefs are just you know that's just so much worse off so yeah don't don't think that like just because they look like the cool kids now that they're gonna be the cool kids they really really never ever ever end up being the cool kids I promise you I like I remember my sister used to tell me this when I was like younger she was like I promise you like these people will beg to be your friend when you're older or like they'll like wish that like you know they could hang out with you and I'd be like oh my god sharp like no they weren't and I'd be crying and all this and like I swear to God it's true so yeah it's all about the energy and vibe you give off most important thing to look for in a potential man any views on long-distance relationships hmm okay so I think three most important things for me and like this is my cardinal rule is you've got to have trust respect and loyalty and you have to see that in the person too so if they respect you like they treat you like a lady or they treat like a gentleman and they are like respect like if they're trusting like you know they don't question where are you going who you're going out with like not like in a like obviously it's normal to ask why you're going out with but you know what I mean like if they don't seem like they're constantly just like distrusting and whatever and also if they show loyalty to you you know just if someone pays them attention they're not exactly gonna like just go with the attention then yeah that's the three best qualities you can find not just in a partner but also in friends as well but yeah I think for me there's a three most important things to look for in a potential man and on top of all that you have to gel like you have to have a vibe when the two of you together like you need to feel like oh my god there is no one else I would be having this much fun with or be this happy with or be this comfortable with then I am with you right now so that's what I think as for long-distance relationships I've never like had one or like never yeah it's not pretty come up for me so I don't know like if it would like I can't I don't feel like I can say like oh it's good or it's bad if I haven't haven't got any experience on it I feel like it depends on the scenario like how long were you guys together and then were you like you know separated because one of you moved or like were you separate from the start how long until you decide to like join up and like live together like have you made plans of that have you spoken about that because I feel like those are the things that's important rather than like the trusting and stuff like that because that should have been a given from the start so yeah do you know any Albanian I really like Persian people mostly guys lol they're so hot I don't know any Albanian and I'm glad you think Persian guys are hot I will pass that note on to a lot of my friends yeah sorry I don't know any any Albanian oh my god there's so many questions and I lost my space next question what A levels have you done I did biology chemistry economics maths and Persian I got four A's and B's this question is good have you ever considered going vegan Kim and I did a vegan food trying thing video challenge watch that link down below I don't think I could go vegan as in I get like loads of people do it for like ethical reasons and this and that and I feel like it's everyone's own choice personally and now I don't think I could go vegan but never say never what's the best way to stay positive I think I just answered that before when I was saying about how I'm having like a blur day and I like to distract myself so right now I'm like filming my YouTube video I took my dog for a walk like going out to work and everything and then listening to positive energy tapes I like listening to music sometimes it's even like good to like let's say let your feelings out let's say if someone's made you really mad and you're angry like you know let that anger out not at someone but you know me like or if you're feeling sad and you want to cry it's okay to cry but like don't hold on to the negativity once you've let it out let it go and like start like you know being positive and going back to like distracting yourself seeing your friends going to the gym and doing all the things that you like and enjoy best tips on staying organized and balancing uni knife struggles of the law student xx good on you girl keep studying keep going and achieving your goals so like balancing is all about like splitting your time well so knowing how many hours you have in a day and if you need to wake up earlier wake up earlier if you need to go to bed earlier or later go to bed earlier or later make sure you make time to like see your friends and go out because if you just stay at home and study all the time then you're gonna get depressed so you know give yourself a solid like two three hours every evening to go and whether that's what's your favorite Netflix show or go and see your friends or whatever because so long as you're not in exam season you know you deserve that free time to relax I'd say like when you're not in exam season only aim to study like two hours a day not more because you know it's about work-life balance if you've had a full day of going to work and then you're studying two hours on top I think you've done enough but that's just my way of thinking about it yeah and go with what you think is right as well I remember when I was in my third year and I was vlogging or was I my fourth year people used to message me and be like oh my god how do you study medicine and you have time to go out all the time and vlog on YouTube and upload it and like you know do all of this like how and I remember I literally sat down at one point and I was like oh my god how am I doing all of this like crap and then I kind of open my eyes and realize wait a second girl you made it this far to get into med school get all the way through to third year or fourth year whatever I was at that point some of my final year now and you do all these other things and you're gonna let other people who can't figure out how you made it here question and doubt yourself so don't ever doubt yourself you know what's best for you don't let other people tell you how to manage your time or do what's best or blah blah you'll know yourself and yeah go with your gut also I think it's gonna sound so bad but like boys can be distracting or girls can be distracting so like make sure if someone is like in your life that they're not like distracting you from what's important because I know what it's like where like you could be like in a lecture or something like that and like you're just thinking about why did he message me this or why hasn't she text me back and they went out last night and this and that and so don't let like if you have that sort of toxicity like around you where like you're just constantly thinking like someone's on your mind for the wrong reasons you got to cut them out too because that gets in the way of your studying and at the end of the day you're still like young and you still want to get to places and you if people are already getting in your way then what's gonna happen when you have even bigger goals and dreams so don't let anyone get in the way and don't let anything distract you sometimes I feel like when I answer these questions like I'm giving so much input once like I'm jumping like I have flight of ideas oh my god guys literally hit me okay ooh this one's a long one hi seppy my questions long so I need to send it through DM okay that's fine I know a boy for two years and both of us are med students but he is three years older than me during the last two years we've been chatting a lot in the beginning we used to chat every day but as time goes on we chat a bit less maybe three times a week and when we have exams we chat at least once a week he mentions me as one of his best friends as sometimes he's so flirting calls me baby on the other hand sometimes he acts like we're just friends and I told him as I'm not officially his girlfriend he must not call me baby and it bothers me last year Valentine's Day he got me a gift I love him but I really don't know how he feels about me I'm confused and can't concentrate and I'm sick and tired of this situation I don't want to lose him but at the same time I don't want to fool myself and waste my time on him if he really doesn't love me what should I do I don't want to make the first move should I leave him okay so let me give it to real sis with guys if they're into you then they will show you that into you and usually you'll notice that at first when you know that they're on the chase they text you good night they text you good morning they'll send you memes they'll want to come pick you up from wherever on earth you are like clearly I sound like I'm coming from the blaze of experience but what I'm trying to say is if they're into you like in the beginning or whenever then they'll show you and if you're questioning if they're into you or not they're probably not into enough and it's because maybe they think they already have you maybe they've got their eyes on someone else I don't know and I know that's not something that you want to hear it's not something that all of us want to hear but any of us want to hear but if you're not sure if they like you enough you've got one of two options you can either tell them this is how I feel about you and I just feel like it's kind of obvious so you should know and yeah like what we're gonna do about it or be you can just ignore him because clearly he's not giving you all the attention that you want and when he does come back to you just you know be like well you can't pick and choose so you're either in or you're out I think that's the best two scenarios that are out there and plus when you kind of leave him to it and you'll be like sad for the first few days but then you actually feel a sense of relief like now I'm not sitting here every night wondering like oh my god is he talking to another girl like is he gonna message me are we gonna meet up blah blah am I gonna be disappointed you actually get a sense of relief so yeah that's my advice by the way guys feel free to like chime in on the advice in the comments like if you hear a question that you feel like you can give advice to I love when like all of my little Persian bunnies all talk to each other I don't know if I'm gonna call my father was Persian bunnies I don't know why I came up with that but yeah where do you see yourself in five years PS love your videos exo thank you so much in five years time I would like to think that I'm quite ahead of my career so I'd like to think I'm still on YouTube I don't know I like making videos I do it as like my hobby and I really really enjoy it and I've managed to meet so many of you guys like through it and talk to so many of you guys through it and people all over the world so I feel so special so yeah I hope to still be on YouTube and I'd like to be progressed in my medical career so hopefully specializing and this isn't the one that's not gonna be new to like a lot of you was ears but I might want to move out of the UK I'm not sure it's still just like a thought that's come to me but I'm really sitting down and thinking like what's stopping me from like moving away and like what's really keeping me here kind of thing and I'm quite young I'm 23 so I have room to you know experiment and don't know explore the world so yeah we'll see if you are stuck in an elevator and was forced to listen to only one song which would it be this is the most epic question I've been sent I actually like that one something without any words so maybe like one of like Beethoven's like symphonies or something and because then you can't get bored during if you constantly have the same lyrics going over and over and over again you'd go mad so yeah I'd pick something like that like Beethoven what are your secret skills well if I told you it wouldn't be a secret would they what's your take on the meaning of life oh my god I feel like that is like so intense for me to think about on a Tuesday afternoon but I think I don't know what I think on the meaning of life I have to come back to you on that one are you hoping to end up with a Persian man or you don't mind what nationality so like generally speaking from like people that I've spoken to like I realized I'm more like attracted to guys who are from like let's say like Middle Eastern background or even like so like Arabic, Iranian, Greek, Turkish because I feel like we have a lot of similarities like in terms of like culture but then we also a bit slightly different so I can learn new things as well so yeah that's kind of the vibe that I've always been like attracted to not to say that I will never be attracted to someone from like a different background but from my past experience I feel like that's what I've liked so far but you never know never know who you end up with oh how do you and your sisters deal with arguments with one another so have you seen those memes when they're like when you argue and then you come back and just you don't apologize that you just continue like nothing happened sometimes it's like that sometimes like it will be like we're arguing over like for example like oh I don't think you should do this it's not good for you and like the other person be like it's not your choice it's not your life and then we'll sit down and we'll actually like talk about it after and like we'll have a discussion in a heart-to-heart and all of that we actually have one of those this weekend so yeah it happens frequently I think the best thing is to be like open and like talk to one another like with your family and like always just like you know be comfortable enough to chat to them about anything and like talk through your problems don't just kind of brush it under the carpet if you get mad at each other don't stay stubborn because that's not good either how do you keep yourself away or focus from guys who want to have fun and not date I think the most important thing is first you have to realize like a hundred percent that the person is only after you to have fun and not want to date so you don't want to brush them off really quickly and so just suss them out there and then if you see that you know they're just kind of not serious then you have to cut them out I feel like with boys you have to be really this is not just boys actually this is just like in general whether it's like fake friends boys whatever if you don't want someone in your life anymore then you need to really like diplomatically like phase them out of your life so like don't reply to their well I guess with boys that it didn't work out you don't really owe them much so just kind of like be really dead on text and don't answer their phone calls but like yeah with fake friends just phase them out slowly and yeah that's the way but don't don't get caught up in the whole like your feelings at first because it's so easy to be like oh my god we talk all the time he wants to take me here we've been out there blah blah and then when he starts ghosting you like oh my god but I remember when we went here and he spoke till 4 a.m. about that's like get over it so he probably does it to loads of girls if he's really good at it then you know you don't want to be just one of the 10 millions of girls that he's done this to and you'll just move on and find someone who just wants to talk to you celeb crush okay so my recent obsession I last month started watching power and surprise surprise I'm on the final season season 6 and I am obsessed like obsessed with Tommy Egan and so I was telling my dad and my mom about this and they're like you're actually crazy because I was like he's so loyal like he's just so loyal that there's nothing more attractive than that like he's so loyal to that ghost that like if anyone like fucks over ghosts he's like you're dead literally I mean I think that's a bit much but like I didn't I think that that's such an attractive character is to have and like when he was like with like Holly and like she was like he's like a one-woman man I like that as well like he doesn't mess about even though he easily could so that's my celeb crush but he's kind of like a fake character isn't he so for like physical looks crush it's got to be be young like he is a beautiful be it's for beautiful and be young I really hope he doesn't watch my videos or I'm just so frickin cringe last question I would make it a fun one what is your dream date so this is so easy hands down for me and my dream day would be to go watch Harry Potter in theater I'm gonna sound like a weirdo online but I frickin love Harry Potter so if I could go see it in theater that would be my dream date and then like the next level of that like let's say that was a like a date like just standard with dating or whatever then like the next level dream date of that would be to go to New York to the top of the Empire State Building like Chuck Bass and Blair did in gossip girl so those are my two like dream dates in my life I have to have at some point well I don't have to but as in I'd love to have at some point so yeah I hope that you guys enjoyed this kind of sit-down chit chat if you enjoyed it then please give it a thumbs up and if you want to see more videos like this tell me in the comments because I read all of your comment and I try to reply to as many as I can so yeah get chatting with me I would love to hear from you don't forget to check out my socials they're always linked down below but my Instagram is pageantbunny and my snap is sepi samai I love you all so so so so so much and I'll see you next time bye


Good morning, good afternoon everyone. I'm Alessandro, we'll see you in a few minutes for two years now. And today, thank you so much for being here to this session of live and stream and where we watch the recording. I'm so excited to be here and I would really love to thank you, all of you, for the trust that you gave me today because we will see this in a moment, why? In many of you, they probably have seen my face here and there, both on LinkedIn or something GitHub or whatever. And I'm actually working as a senior, special solution architect in Reddit. And basically, I've always worked in the open source community, both as a contributor on my project and of course, also with cognitive technologies. I started to look at Kubernetes, OBShift, and whatever it's related to. And one of the important things today is that, and this is the reason of the trust, is that it's the first time for me being here at DevConf and the first time ever presenting at a conference. So don't be so tough. Well, today's session will be mostly oriented towards Edge and how to automate an Edge using Event and Event-Driven Automation with Ansible. So I just prepared a quick agenda to guide you through what will be the steps that we are going to take today. So the first thing, we are going to start with an introduction of the Edge. So what is Edge computing? We are probably very knowledgeable about this, but it's always good to have a quick refresher and how we can automate some of the use case, the most common use case at the Edge and what are the benefits of automating the Edge? And then we are going to focus a bit more on events. So discussing a bit about events and what actually does it mean to automate with events? So what is the idea behind this different kind of automation? And I prepared a very small demo that will be covering some of the use cases that we will touch on during the presentation. And of course, at the end of this session, there will be a Q&A, so feel free to also ask questions, also during the session if you want. It's all fine, perfectly fine. So let's start from the beginning. So we heard a lot about Edge, Edge computing, and it's a way different way of calling something that we were used to deal with also in the past. When we talked about IoT devices, disconnected environments, we were already talking about the Edge, but now it's starting to get a very hot topic. Also because we have the chance to completely decentralize our computing and our different use cases can be really self-consistent, self-feeding, and basically doesn't require too much presences and too much power, for instance, or connection or everything that is created to a data center can be just completely, very little pieces. And one of the biggest benefits of this is that we can really start decoupling all of our different workloads and having different sections of our workloads, application, calculation, power, and everything, in sites that were never covered before or were difficult to cover because we had restricted protocols, we had different access to these kind of places because maybe it's a disconnected site or it's a very disconnected environment. And of course, one of the importance of the biggest thing when talking about Edge and automating that the Edge is the response time. Because sometimes we need faster reaction times in respect of other potential use cases like doing normal automation like patching a system or having configuration of a web server or configuring our infrastructure, we potentially need to react faster to anything that is happening at our site. And we don't have the control of that because maybe we cannot access that at the point of time, it is completely disconnected, or we have some limitation of any kind. So why it's a good idea to start automating Edge? Of course, one of the biggest use cases and one of the most common use cases that get Edge is of course configuration management. We want to streamline the configuration of our devices, our sensors, of our small computing units that we maybe have there for collecting data or having some interactions with local components that cannot be automated from a data center, for instance. Or we just want to have a consistent configuration across multiple kinds of devices. So maybe we can have also some sensors that require different, each sensor may require different configurations. So we need to be able to control everything from one security to the other. Security, because maybe in our sites, we want to enforce some security policies, let's see, let's tap for the CID access or anything that can make our devices and our environment compliant with our regulation or just cover the regulation if we are talking about the government agency or a government entity. And of course, one of the use cases that will also be covered in the demo is monitoring. Why? Automating Edge can be good for monitoring and remediation because actually, when we are going to take control of our devices, so having our configuration in place, being able to produce metrics based on any potential activity. So if it's a sensor, it can be the data from the sensor. If it's a computing unit, it can be how, for instance, the model that we are trying to work on for that particular device, how it is performing. If we have a weak potential leak or power leak or power outage or network coverage, we need to take care of in a very fast way and we can also implement this in a sort of self-feeding fashion because actually, and we will see this in the demo, we can also host the automation at the same site without even caring about that. Because it's there, it will be running, it will be checking for everything that is working and we can especially configure and extend this kind of automation. Another use case is the idea of having rapid developments. Whether it is a containerized application, whether it is a package that needs to be released, whether it is a firmware upgrade or a software upgrade that we need to deploy. We can do that at scale and we will talk about scaling in a second. We can do that at scale and potentially with very limited downtime because actually, we can take care of that and integrate automation in our release pipeline. We can, for instance, take decisions based on how the deployment is going so we can decide to extend the way of how our deployments is happening.  We are talking about sometimes very deep numbers. We have some customers, also, that are starting to get more into the Edge computing and starting to, we are talking about 10,000, hundreds of thousands of nodes or sensors or devices. So we need to be able to scale in a very fast and streamlined way. And the last thing, of course, which we are really mentioning about, we need to be a panel, reaction panel. Because we cannot afford having that component starting to fail or we cannot afford having a deployment failing for one of the data collection units that we need to put fragmentation in place. If a device is failing, we need to find a way to take that device, take it offline or try to re-mediate what's happening and then we can be able to make use of it again. And, of course, all these things are pulled together, but we potentially need something different to do because the normal automation, the standard automation may not be the best fit for this kind of use cases when we are talking about Edge. Because we are talking about very definitive use cases and we don't have a dynamic adaptation of the automation. So we have some commands that we need to put and we write our labels if we are talking about Vensible because we will talk about Vensible. And, but we are going to just have a configuration in place. This configuration will be deployed, but what if we need something different? So what if we need to adapt our scenario to the automated, the optimization for our scenario? And this is how events can start to get in the game. We all heard about events. Events are potentially everywhere. When we are talking about a Mastery bucket, for instance, when you put a file in a Mastery bucket, it can generate an event. You can create a message, put a message on a queue. This is potentially another event. When the sensor is starting to ingest data in our center or in our devices or whatever, it is actually generating an event. It can be analyzed, it can be processed and we can do something based on the analysis. And one of the other use cases that we always use with our customers is also the interaction of tickets. So if we are, in our ideas, we are creating a ticket for deploying a note in the remote location that is not reachable from our data center for any reason. We can put in place some kind of automation based on events, trying to make this kind of automation reactive to what's happening. So when we are creating a request or a ticket or whatever, we can still process that and have the automation react to that message and also potentially close that message based on the automation. Respond to that message, escalate that message. And another important aspect is coming back to monitoring other things. So if we have our platform thinking of OpenShift, thinking of any other potential platform that we have in place in our environment, we can meet, we can meet to react to what's happening in the platform. So if the platform emits an alert, we need to be able to identify what's the alert is about, take a remediation on that and apply it. So we need a bit of a switch of context because actually we are used to traditional automation when we have some issued comments, we have a set of defined machines or potentially dynamic machines, but we have a potential issue in synchronous. When working with events and when working with dynamic data, we need to be able to react to each and single event that is happening in our platform. So it's not just about, okay, I need to configure this, I run my playbook, this is what's happening, I can also wait for the playbook to end because I have this task today, the playbook does this. When we are talking about events, it's quite different because maybe we have like 10, 100, thousands of events and alerts that are happening in our platform and we need to be able to manage all of those as soon as possible and of course just in time. So we need a faster reaction time even in this case. And this is actually how it is changing. So it's not about just having a command issue, but this is something that is driving by events. So it's the event driving the automation. So you have potential different sources that are emitting different kind of events all day long, all night long, forever and you need to be able to react each and every single time. So basically, if you think about events, we can define three big questions that we need to do. Is the first one is we are interested in when is the event coming from? So what is the source of our event? Is it a task vacuum? Is it a web book? Is it an MQTT listener? What's, where does the event come from? Then we need to process it because actually, okay, we receive the event. We are good to go. It's probably a JSON message or whatever, example format it, we can do this in any way. But we need to do something with that. So we need to also define how we are going to treat our event and what is the outcome of this processing? It's the work. So what do we need to do based on this one? So we need a source. We need something that processes the information and then we have something that is actually activated the automation based on information. And this is why we are talking also about Ansible. Because Ansible is one of the best bits for this kind of use cases when it comes to event treatment. First of all, because you don't have, you don't need an agent. So we all need, we all know that the Ansible is agentless. The playbooks are very easy and unagreedable content that we can reuse, implement and distribute. And we have another advantage that Ansible is community-driven and partner-driven. So also the community and the partners can contribute to the code base. And of course, covering different devices, different protocols, different connection methods. And this is crucial, especially when we are talking about a very, a very heterogeneous set of devices like it can happen at the end of the day. And the most important thing is the last one. So we have dedicated bits for event treatment automation. So we have a collection that will be also implemented in the Ansible platform controller that will be actually responsible for managing the event treatment part. So it is implementing, and we will see this in a second, new, let me say, new resources that are called Chromebooks where we are going to define exactly what we said before. So we are going to define the where, the how, and of course, the what will still be an action, a command, a playbook or whatever. So let's see how this is translated. So we have the sources. So in the event of an Ansible bit, we have the sources that can be, as I was mentioning, a Kafka, or KafkaToby. I recently created one for our demo, that is the MPTP connector. We have the community working on all the kind of connector, also, and your blog created the one for Kubernetes, for interacting the Kubernetes cluster. And this is why I was talking about it's also easy to implement. It's not something that is rocket science because actually, I'm not a programmer, so I was able to create a Python script for the MPTP source from scratch based on what was already available. And this is quite crucial because having this possibility of extending the use cases, potentially in an infinite way, only the, basically, our fantasy can be a limit because as soon as we are able to create something to interact with our source of events, we are good to go. This is exactly what we need. And of course, we have some rules. So once we have identified the sources and the event from that source, of course we need to do something with that. We will see a very small snippet from the demo later, but it's basically a sort of event processing queue where you're going to put the condition placed on the event, what is multiple condition, combination of conditions, we will be able to label our input, manipulate the event itself to make it understandable to our automation itself. So we will be able to process, to further process our events even before using it in the automation. And of course, the last part are the actions. The actions are basically a map, a playbook that can be run, relabeling our input, saving the input from somewhere. And these all together combined is giving us a real way of managing events and managing our automation with it. So just in a few seconds I will be showing you a very small demo that I created. And basically the use case, it's kind of funny because the next slide is a bit weird, but this is exactly the idea. And this happened to me last year. So with what we think about industrial use cases, medical use cases, that are of course a big thing. But what about the little things, like a fridge in our house? So we all have a fridge. And a fridge, you know, but at least in Italy, when you start to get in hotter and hotter, you can feel that the fridge is not actually working at a very good condition. So, you know, the groceries are not feeling so good. You are just starting to have a meat cold, meat hot water and stuff like this.  So what is the idea? The idea was, okay, let's think about a potential use case where you have different sensors. One in the house, one more, of course, in the house. A couple of sensors on the floor and on the roof of the room. And then one sensor in the fridge. Of course the fridge should be, let me say, a sort of intelligent fridge making it, like, just having an interaction with an external device, whether it's via wireless, whether it's Bluetooth or whatever. So you just need something that is able to interact with that device. And what is the action in this case? The action in this case is why we cannot adapt the temperature of our fridge, the working temperature of our fridge based on what's the temperature of the outside and inside sensors. So what I was thinking was, okay, I cannot implement it for real because I cannot make something here. So I decided to create the version of this use case in a sort of containerized way. So I created a purpose application for all the sensors and the fridge itself. And then we will see this in a second. So the digital version is something like this. So we have everything is based on readout device edge. So we have a potential device running in our house that is hosting readout device edge. Of course, coming back to the news session, if you were here, we were talking about the image there, how to create well, and sorry, my microchip, ISOs and stuff like this. This is exactly the same scenario. So I created my readout device edge in installation. I created the Quarkus application for all the sensors and I have a small configuration here that is based on the Quarkus application. So the Quarkus application has a configuration that is our, let me say, target of the automation. Maybe we are going to change that configuration with the Quarkus application and see how this adapts to what's happening. I've deployed a very small registration of full speedo. It's called terrorized mosquito. And on the other side, we have two other components, one that are aggregators and one actuator. The other actuator just takes care of getting all the information from the sensor aggregated and send it to you first thing. And the actuator, we will see the connection in a second, it will be the actual generator of our event. So it will take care of monitoring the data here. So if something changes, it will send a little event that will be done in process by Ansible and Ansible will take care of adjusting the configuration. And the good thing is that the actuator, now it's just a very basic Quarkus application. As I told, I'm not a developer, so it's nothing really complex. But the idea is that this actuator can be potentially anything. It can be an application, programming, implementing a model, an AI model, or a machine learning model to just react in a better way or, of course, adapting to every single change. So the cool thing about this is that it's not just having a very simple use case. Because it's a very simple, as I told you, it's not rocket science. But this really covers most of the topics that we touched on in the last minutes. So basically, the Ansible, the event in Ansible is containerized as well. So we have an MPTT connector that is taking the information from a dedicated topic on Mosquito. And it will actually take some decisions based on what is the event that is generated. But, of course, now we've seen the picture. Now, let's just jump for a second on what's the implementation of this. I will not go into details about the actual Quarkus implementation. The repository will be shared at the end of the slide. So there will be a QR code if you are curious and you want to go through that. It's quite, it was a bit, it's a bit more, I tried to make it as small as possible, so I hope we'll be able to just recreate as needed. But I just wanted to show you, so here we are, our whole book. So getting back to the discussion that we made before, so the whole book is something that is very familiar because it looks like a playbook, but it's called in a different way and also has different, from the implementation perspective, it has also different keys that we can see here. So we have the sources and the rules. So the sources are actually the actual implementation of the plugin that is connecting to a remote source or a local source or whatever and just taking the information. In my case, it's a very simple one. It's just taking a couple of parameters here. So what is the host of the MQTT broker? What is the port and what is the topic? Of course, this can be also extended at will. What is important here are the rules. So the rules are, okay, now we are in the, we are actually in the, we have the event. Now we have the event. Every single event has this prefix, event of course, and based on the implementation, it can have additional keys that are needed to just go through the tree of the event itself. Usually, if we are talking about JSON messages or something that is browsable, it will be just about accessing it as if we are just going through a tree. And the important part, so the what, is the actual action that we are going to do. So it's, as you can see here, it's just about running a playbook. So it's just a very simple playbook that is just taking care of analyzing the event. So it will further analyze the event and based on the outcome of the event, it will just adapt our temperature. So just to make, so we have our basics. So our sensors are basically just, so we have here our sensor data. So our sensor data, as I was mentioning, we have three sensors in the room and outside and one sensor in the fridge that is, and we have, I added some humidity, temperature and stuff like this just to make it a bit more real. And from the fridge perspective, we have also the power consumption, the eco mode, if we have the eco mode on or off. And of course, what is the speed of the fund that is actually working in the fridge itself. And we have a fixed temperature here. So this is the working, the operating temperature of the fridge. And this is actually, what is the sensor getting from the fridge itself? Now, let's see what we can do here. So it's very hot outside. So we are going to just raise the temperature a bit. So it's a very simple script that is just changing the configuration of the different sensors. So to simulate a sort of change in temperature. And let's see what's going on here. So if we have any change, sorry. So let's wait here. We will have a slight change. So basically the temperature just changed. So I increased like five or five degrees. So it's a very typical summer situation where you're just jumping from a temperature to another. And here it's not happening so much at this point in time because actually the data is starting to be transferred to the aggregator. So the aggregator is writing the data here. And we have our actuator that is actually gathering this data. And it is actually taking care of trying to find a sort of anomaly detection, let me say. It's a very rudimental anomaly detection based on some thresholds and on previous iterations. But I just added myself in this side. We have what's going on in the Ansible in the Event Drive and Ansible container. So what we hear, so something happened. So basically the actuator react, notice that there was something that was changed in the temperature. So we had a very slight temperature change. So coming back here, looking at the log, we just received this event that has this structure. It's basically a JSON message. And it is basically, okay, there is something that is not working. So it is going through the processing part. So it finds that there is an anomaly. It just brings some information about what is the delta of the temperature. Again, it's a very rudimental detection. So it's nothing precise. But it's still working. And it takes some information from the action deployments, changes the configuration. So it basically changed the configuration of the fridge. So the fridge was our target. And this is basically what's going on here. And as you can see, we just see how, and it's really nothing complex because it takes a very small amount of time to just have a good automation in place. Of course, all the part behind is a bit of course, preparation and everything. But again, it's even looking at the code. It doesn't require so much effort to start working with events. Of course, it can be complicated as much as we want. But as you can see here, again, I'm not a programmer. So I kept it very basically. And I really hope this was a good use case, to be honest. And, yes. So, this was the last part of the demo, actually. So I really hope you enjoyed that. So if you have any question, discussion, or you want to raise something, feel free to do that. I'll be more than happy to go a bit deeper on that. Yeah, let's go. So you mentioned that we could write rules and the events. Sorry. And then we showed that we could write rules. Now, let's assume we have a huge scenario where we really want to automate a lot of things and write it, let's say, thousands of rules to manage it. But then at the end of the day, my code would become busy, right?  How do you manage all these rules together in one place and make it easier for the developer to bring in and change it? Okay, so that's okay, just to calculate it. So if you have... Yeah, so if we have multiple rules, how can we handle the granularity of managing all these different kind of rules when, especially when multiple developers are working on the same kind of automation? Well, this is actually how you would treat a very complex problem.  You are not going to work on the big bunch of rules, but you can, for instance, split the different rules based on what is needed by what. So in the case of having, you know, particular kind of events like, I don't know, in the case of the temperature, let me say. So you want to manage the temperature, the humidity and a lot of additional data. You can also try to split those into some smaller sub-problems, like having a dedicated rulebook that is just taking care of that particular use case. So you can also have a rulebook for the temperature and having a different rulebook for the humidity. So if you are working on the temperature and I'm working on the humidity, we can still not interact with each other, but we can still work in a, let me say, sustainable way. This is a very good question because, of course, the number of rules, but if you think of also the sources, the sources is not, we have worked with a single source, but we can have multiple of them. And each one of them can produce different events. And even the same source can produce many type of events. So, you know, it's getting, what I would suggest is try to split it into, you know, better manageable problems. And I think it will be a good idea to start with. Please. So, okay, the question, if I got it correctly, is how I actually gather the events. So do we have a listener or it's just, you know, pushed from a client deployed? Okay. So basically the sources are just listeners. Can be listeners. So they are always up and running waiting for someone to push. So instead, think of a web book. So you have a sort of, see the web server that is running continuously and you have someone calling him. So you have a client. In case of a Kafka event, for instance, you have the client also on the event-driven side because you have someone that is connected to the Kafka topic and it's just waiting for something to happen. So you also have another client on the other side that is actually producing the event. And also, sorry, okay. Okay, thank you. And so when you're talking with, you know, asynchronous messaging, you still have two different clients but it's not, you know, waiting for something to happen. So, and of course you can also extend this kind of thing because it's, as I mentioned, it's just about a Python plugin that you can just use, create and reuse for everything else. Thank you. Do we have any other question? I think you're fine. Good. Thank you. Thank you so much. This is the, this is your thing for the repo if you are interested in. And I have just one ask for you. It's the first time for me here so please make a good use of the feedback. If there is something that you didn't like, especially you didn't like, please let me know. Thank you so much. Thank you. Thank you. Thank you.


I'd like to thank John and the organizers also for the opportunity to be here today and to speak to you about some of the work that we've been doing. We've been interested in gene expression and the role of enhancers in that process for some time now and this interest of course was boosted a few years ago as for many people as a result of the development of chip-chip analysis and all the variations on chip seek analysis that have come out in part due to the great work from the ENCODE consortium and we of course have taken advantage of that technique as well. So I just want to acknowledge people that have been involved in some of the studies I'm going to show you first here and I want to primarily acknowledge Mark Meyer who's done much of the work that I'm going to talk about along with various other individuals in this in this group. I'll talk a little bit about some of the work that we have done with Charles O'Brien and acknowledge some of the analytical work that we've done through Homer with Chris Benner who at that time was actually working with Chris Glass and we're appreciative of that effort. Now what I would like to do first off however is pose this sort of question that is why we study enhancers and of course I don't really need to say an awful lot about that to this group but I just want to point out several things and that is of course gene expression and cell phenotype are primarily governed by enhancers and so a detailed knowledge of mechanisms of regulation are necessary literally for every gene whose misexpression is known to result in human disease and this comes about from the obvious idea that if there's a human root to a gene knockout in mice that results in a phenotype that's an actually an important gene and knowledge of that regulatory capability is very important. Also disease regulatory knowledge regarding genes that play a role in disease progression and there are a number of those as well and finally we've talked a number we talked about this is regulatory knowledge about genes that are linked to RFLP SNPs or indels that increase the risk for disease. All of these are misexpression examples that are very important and therefore we actually think that it's important to actually understand the direct relationships between this regulation and these genes because these details could provide some selective opportunities for therapeutics. So I want to just first of all give you a couple of slides on our entry point into gene regulation and that has to do with the vitamin D hormone and just to remind you that while vitamin D is known to regulate mineral homeostasis and it's involved in certainly in skeletal homeostasis it has a lot of other effects some related to the immune system cardiovascular system and so forth and so it's actually vitamin D is certainly plays a role in in biology that's that's far beyond that that is related to calcium and phosphorus homeostasis. All of these activities of the vitamin D of course are regulated by or mediated by the vitamin D receptor and in fact while we had some understanding very early on that vitamin D might be part of an endocrine system it took the cloning of the vitamin D receptor in the late 80s to to really identify it as as a true member of the steroid nuclear receptor family of genes and in so doing provide a bona fide membership in this family and the vitamin D receptor through domain analysis that was done at that time suggested the amino terminus said the DNA binding domain the business end of the molecule was in fact the C terminal and then the effects of the ligand were to re alter the configuration of this C terminal and to open an act or conform an activation domain we called AF2 that led to activation of gene expression. This is simply a crystallography study that we did with the ligand binding domain to understand how the vitamin D receptor actually worked and other people have done this as well and this is the loudest to understand where the contacts for 125 D3 in this pocket this the activation domain surface with this peptide was identified and so forth so this is allowed us to begin to think about designing analogs although that hasn't actually gone all that well now I want to come back just for a minute and remind you that the primary role of vitamin D has been in calcium and phosphorus homeostasis and what you can see here is that vitamin D plays a really a very complex role in the in the intestine kidney and bone to regulate this and maintain extra cell you have calcium and phosphorus levels and it also does it by regulating PTH levels through the parathyroid gland it also regulates through bone it regulates FGF PTH is primarily involved in in the calcium regulation whereas FGF 23 is involved in phosphate and we've been particularly interested in the bone areas simply because vitamin D actually can mediate not only bone remodeling but of course it also the skeleton is actually the source of calcium and phosphorus and in situations where there's a dietary deficiency of either of these minerals so this is actually a very important area that that needs to be understood and we've focused to some extent on that now when we in the in the 90s we begin to understand three basic principles of the vitamin D receptor and quickly go through these the first one is that the vitamin D receptor interacts on DNA with a partner called RXR RXR plays a role in many other receptors as well but it's a heterodimer that functions it functions on DNA and we were able to identify response elements that were comprised of two hexanucleotide half sites separated by three base pairs so these two principles were important this is a cryo EM structure done by you know morass and his colleagues so this is sort of a contemporary view of it but the third principle was that the vitamin D receptor as many nuclear receptors and other transcription factors simply function to recruit chromatin active co-regulatory proteins at groups of proteins and this is three of them here I won't go through them there are many many of these and we yet don't understand all of them and their individual functions but I think we're we're getting there to understand at least some of them now the problem with this is that this was a very receptor-centric view of life and that is because we were studying the receptor on a couple of genes that we had begun to explore and the problem with that was was we it was very limited to these particular genes and so during the when when the unbiased of methods such as chip chip and chip seek analysis came about we tried to take advantage of that to begin to understand the properties of vitamin D action in on a genome-wide basis sort of an overarching principle again we focused on osteoblasts as bone-forming cells again because they play a large role in regulating through remodeling a bone but also because we could actually look in vitro we could actually look at the differentiation process and compare different stages of osteoblast cells and much like Evan Rosen has has told you about with respect to adipocytes in any event so we did an experiment where we took osteoblasts and just treated them in the absence and presence of 125 and did a chip seek analysis looking at the cystrom for the VDR and here what you can see is that there's about a thousand genes binding sites for the vitamin D receptor in the absence of ligand about 7000 or so in the presence of ligand clearly suggesting that in the case of the vitamin D receptor is that the hormone is very active in promoting DNA binding if we look at RXR we can see the same thing it's a much bigger cystrom probably because it has greater activities and with other receptors but in case when you add 125 D3 you do see that activity and the important aspect of this is if you actually cross these over you actually look at the peak intersex you will see that about 4,000 of these sites have both VDR and RXR certainly these this would support them the idea that RXR is a partner if you do a de novo analysis motif finding analysis in pre osteoblasts as well as osteoblasts these are early cells these are fully differentiated mineralizing cells you can see you see the vitamin D receptor RXR motif which is AGG TCA to AGG TCA motif separated by the three base pairs and in fact you can see that in in both cases there are some additional motifs that are found in these elements in these enhancers as well now this was the most interesting observation and we've talked about this already and that is the discovery in fact that by that the binding sites for the vitamin D receptor as well as many other transcription factors are in fact located distal to the promoter rather than near promoters in fact the bulk of them are either intronic or intergenic and this has had profound effects on our ability to understand the process finally when we look at pre osteoblasts and osteoblasts here's the VDR cystrom what you see is a tremendous contraction of the number of binding sites the cystrom following differentiation and this actually has some important ramifications as well now we were asking then we asked and whether the change in this transcriptome had or this cystrom had an important role to play in gene regulation we did a microarray analysis of pre osteoblasts and then the mature osteoblasts we could see some rather dramatic effects this is a microarray analysis of this as of the gene expression we've done extensive RNA seek since that time but in fact what you can see is both up and down regulated genes are changed dramatically as a result so it's pretty clear that in fact here's the up up regulated genes and here the down regulated genes you can see the contraction in both cases so it's pretty clear that there is a change in gene expression in response to 125 as a function of differentiation now one of the genes that actually goes down the suppressed by 125 because 125 D3 is trophic for the vitamin D receptor and auto regulates it is in fact the vitamin D receptor itself so it suppressed in its expression levels and that has a clear impact on the expression of the gene but the real question here is why all this occurs and in fact one of the things that we noticed was that there's still a number of genes that are targets in the mature osteoblasts and in fact if you look at this is actually quite interesting you see a tremendous change in in the regulatory capability of 125 D3 between early and late cells so for example here in call 2a1 a tremendous down regulation by vitamin D but almost no but a tremendous suppression in the case of the mature osteoblasts and almost no regulation probably a result of the loss of vitamin D receptor binding here and in fact if you look at this chip-seq analysis of this you can see that there's a complete loss of the vitamin D receptor binding activity in this gene however if you look at a couple of other genes this is an interesting one in pp1 and 3 which are right together there is a primary enhancer that we believe regulates both of the expression of both of these genes again you can see there's an incredible increase in responsibly to 125 modest changes in basal level as well and finally if you look at this last gene here you can see almost no expression and then tremendous expression and these don't really correlate with the levels of the vitamin D receptor that are bound here in in each of these particular cases so the real question here is why are these in mature cells much more sensitive to 125 D3 and in a couple of these cases and the that they're able to regulate but even though the the peak height for example is not near as high as it is and the scale over here you can see is actually rather dramatically different so there are many reasons for this and we don't really understand it all but just let me point out a couple of possibilities that first of all is that the we had some clues as to the idea that there were other transcription factors that were present in these in at these enhancers and so we actually did a chip seek of of cebp beta and rungs to now rungs to is a master regulator of the osteoblasts is involved in chromatin opening and cbp betas is involved in chromatin remodeling and what you see here is about 17,000 sites 1700 sites where vdr and rxr actually bound or actually co-occupied by rungs to and cebp beta and all this motif finding analysis reveals pretty much the same thing and what this says when you do a further biome biome informatic analysis is in fact that there's an organization to these motifs in which if you look at on a broad spectrum you can see that there's an organization where the vitamin D receptor is bound between rungs and cbp beta and the the approximation here is quite close there's just a few base pairs in between so we've called this the osteoblasts enhancer complex it's not consistent across the entire genome here but it does suggest that these other players rungs to and cbp beta could have an enormous impact on the regulation mediated by 125 b3 now the second area that I don't have time to really go into is the the genetic and the epigenetic changes that occur during differentiation and just to to summarize these the differentiation and transgift differentiation as we and and the encode group has characterized fully is is that there are significant changes in histone modifications as selected gene loci and those are the ones of course that are changing during the differentiation process the enhancers are highlighted by signature enhancer histone modifications that change as a result of the change of these genes during differentiation these changes in histone marks and the regulatory factors can contribute to the responsibility to 125 b3 and in fact we've also noticed that 125 and other hormones provoke changes in these histone modifications particularly at the level of histone acetylation that again are very histone gene selective for 125 d3 so we think that these are clearly changing the the the epigenome surrounding the genes that are actually changed and influence vitamin D response so this is a list of overarching principles of vitamin D mediated gene regulation that we have identified I just want to reiterate three of these one the distal binding sites the locations of these because they create the complexity that we heard in the last talk and in most of the talks distal regulation where a promoter proximal binding site can be inferred to regulate a gene that's sitting right next to it but the distal regulation makes it very very difficult if 40% of the binding sites for some of these molecules is to greater than 250 kilobases away makes it very difficult to assay epigenetically their epigenetic signature marks largely identified by the end code group and these are really very important and they they mediate a dynamic nature to the genes themselves and finally the vitamin D your systems are highly dynamic based upon the differentiation process and we also believe and I think this is certainly true that disease can alter these effects as well and so these are very important now I want to go through I want to go back to one of the genes that we notice that was regulated and tell you how we look at enhancers and their activities and show that in fact these enhancers are directly linked to the regulation of the genes are many bioinformatic ways but to our way of thinking at least at this point they really are not definitive so we have looked at MMP 13 as another gene we've looked at many genes but this is one I want to give you an example of regulated by vitamin D and differentiation you can see the process over here in the early cells not much regulation here a bit but when you when you differentiate these cells tremendous increase in baseline and then a tremendous increase in vitamin D response now this is a gene called collage an ace MMP 13 a collage an ace 3 the grades extra cellular collagen it is regulated by a whole host of different factors it's certainly aberrantly regulated nearly every cancer or disease with fibrotic complications and importantly it's also regulated and important in atherosclerosis finally it's interesting in people many people here will probably agree previous work on the regulation is focused almost exclusively within the three or four hundred bases near the promoter region of the gene so when we looked at the chip seek analysis that we had had derived from some of our earlier work in early and late osteoblasts here's the gene itself we looked at the vitamin D receptor rungs 2 and c ebp beta in a series of histone modifications and I think it's pretty clear here is the promoter region right here which there is some activity but I think you can see there are three other regions 10 20 and 30 kilobases the 10k region binds the vitamin D receptor 20k region and 30k region bind the c ebp beta and then rungs is bound to the 30k distal region as well and all these are highlighted by marks that contribute to the idea of a histone signature so we were pretty convinced in fact that that it that these were enhancers a question was whether they were mediating the activity of MMP 13 and so what we have done is actually to use the CRISPR Cas9 system to generate a series of daughter cell lines from the from from the from the host cell line and actually look at those and look at the consequence by of deleting the enhancers themselves and here are the ones that we've we've actually created we created a promoter deletion with these that this approximately 400 base pairs we deleted about 200 base pairs around the 10k region that removes the vitamin D responsive elements there's two of them we've identified and then we remove the 30k region again that contains the rungs binding site we also knocked out the vitamin D receptor by that same means and we also knocked out rungs as well to see whether there was a correlation between these measurements and these measurements so this is the data it's a little bit complicated but let me go through it quickly and that is so here's the wild type cells and there's several different clones because there there tends to be some variability and we're trying to be very cautious about this this is basal activity this is the activity in the presence of 125 B3 the first thing you can see is when you knock out the 10k hormonal regulated enhancer you completely lose vitamin D induction but what you gain actually surprisingly is the ability to 125 D3 to actually suppress this gene and I'll come back to this in a little bit if you look at the promoter region you lose a little bit of basal activity but it's still inducible by 125 D3 in the fold is about the same but we're measuring RNA levels MMP 13 RNA levels in this cell and of course slight aside from the genomic modifications we're not adding or subtracting anything to these cells okay so if you look for example at the deletion of the minus 30k region now you see a complete loss of basal activity from that 30k region so and in fact if you then look at but there's some still there's some regulation of the vitamin D itself so that still seems to be a bit operable when you look at a knock rungs knockout or the VDR knockout of these cells those activities generally reflect the activities that's seen with the knockout of the two enhancers so this is the expansion of that just to show you here's the basal level here's the ability 125 to suppress and you can see that this is an expanded version of just these sets you can see a tremendous change so we actually think that they're that these are important observations with respect to MMP 13 and they establish a direct linkage between the enhancers that we've identified here and this gene so I said that vitamin D turns into a suppressor we don't understand it entirely but here's the model that we're looking at here's MMP 13 the direct effects of vitamin D to induce that to induce that gene but vitamin D has very potent down regulatory effects on runks to on the runks to gene itself and that and that also in turn has a direct effect in suppressing Ostrichs which is another key osteoblast transcription factor as well so what we think is happening is when we knock out the vitamin D response itself then the impact of the vitamin D system on down regulating a key basal regulator which is runks to leads to that kind of a suppression and we're following up on this but it has we believe it actually has significant implications for other runks to and vitamin D target gene because you can imagine all sorts of things in genes where there's a different sort of arrangement of regulation so this is a summary of what we of what we think is going on in this chromatin interaction model where this is the the promoter region itself here and we've we've centered the 30k region here because it's a primary player we have the 20 and the 10 and the promoter region interacting and the reason we have this like this is the fact that when you delete the minus 30k region it gets even more complex because what you can do if you follow up with chip analysis of these other sites delete the 30k region if you look now at the ability of the vitamin D receptor interact with the 10k region it's strikingly reduced there's a striking reduction at the 20k for cebp beta and some and and also some striking decrease and even runks activity at the promoter so this seems to be a central organizing mediator of this particular gene and I don't think these conclusions are terribly surprising but at least they do show in fact direct linkage now so where are we going with this we have basically are introducing all these mutations into the mouse and we're trying to look now to see what the consequence of that actually is and I just want to show you one one set we've deleted the minus 10k region in the mouse and when you look at the cells that are derived for in the skeleton that are derived you can see in fact a very similar activity and that is 125d3 in the wild type induces a strong up regulation of mmp13 but surprisingly or perhaps not surprisingly when you delete the minus 10k region you lose the inducibility and you gain this suppressability so we think that this sort of confirms what we've seen in the cell lines and we're hoping to see a similar and perhaps more interesting things in the in the mouse itself with the other deletions now I want to just so in the last few slides I want to go to another gene that we've studied I'm not going to go through this in in detail but this is a gene called rank ligand which we've been studying for a number of years rank ligand is extremely important because it's it's up regulation mediates osteoporosis and almost every osteoporotic disease stayed in humans as well in mice its primary role is in bone but there are many many other activities that have been seen for this particular molecule but the primary consequence of rank ligand knockout is a striking phenotype in the mouse with respect to the skeleton now this just shows you that rank ligand comes from the osteoblast so it's part of the communication that goes on in remodeling it comes and it's regulated from the osteoblasts but it promotes the osteoclast differentiation from hematopoietic cells now we originally did this work with chip chip a number of years ago fact 2004 this is actually a chip a chip seek analysis in which we can see these similar vitamin e responsive regions this is a gene that's regulated by 125 but we can identify a number of binding sites and the important aspect here is in fact that there are four or five of them and in fact that the reason we could never identify a regulatory mechanism for this gene was in fact that the that in this particular case this binding site is 23 kilobases upstream and these sites are in fact about 88 kilobases upstream so there's a whole cluster here but they're they're broadly up regular up up beyond the gene now we've done a lot of work and I'm not going to go through this in detail except to say that we know where there's a lot of binding proteins that are going on here in all of these and we've actually classified these as two types one mesenchymal or osteoblast like enhancers from D1 to D6 and then we've actually identified hematopoietics cells T and B cells that express rank lag and from an even more distal set between 123 and 157 a 53 kb even even further upstream and so these and interestingly D5 seems to be one that mediates both mesenchymal as well as immune cell activity now this is just to show you that when we use that construct or that segment that contains all this information we can fully rescue this run to rank lag and no mouse which has all sorts of skeletal defects and all sorts of in lymph organogenesis there's a whole host of these things but we can fully rescue virtually all of it if we use smaller segments of this gene they are unable to rescue this this mouse so what we've done now is then I'm not going to go through this but we've made four five genomic deletions in the mouse one near the promoter where we deleted about seven kilobases and then some of these other regions that correspond to some of these enhancer regions I've just summarized all the data here I won't go through it in detail to say that we've just finished the phenotyping of literally all of these and each one of these has a basal a regulatory and a cell type specific phenotype that is consistent with what we had identified in the cells themselves so let me summarize this by saying the vitamin D system represents a good paradigm for the regulation of genes by systemic endocrine signals differentiation alters the epigenetic state we think this is actually very very important I've showed you a little bit of this section of the control of of MMP 13 through direct linkage between enhancers and the target gene and then we've done a similar analysis with this rank lag and TNF SF 11 gene that is complicated and we can actually see right distinct regulation in mesenchymal versus a metabolic enhancers and I will stop there thank you very much it sounded from your talk like RXR might be prebound at VDR sites or at least a subset of them yes and at those sites where VDR joins in and our XR is prebound do you see histone modification changes or not so I I would say that we haven't drilled down that deeply but it's absolutely true that RXR appears to be marking these sites we don't know the status of RXR that whether it's a homodimer whether there's another nuclear receptor that's located there or or what the role is but but years ago we began to think that in fact the prebound RXR might be player play a role in perhaps even directing the vitamin D receptor to those sites and that rather than forming in solution in the in the cytoplasm


All right, thank you so much. So let's kick this off. If slides will cooperate, we got it. All right. So take a trip down memory lane with me. Do you remember the first time you ever did anything in the social change space? Go back. Maybe it was a club, maybe it was an organization, some project you got involved in. Do you remember that really clearly? I know I do. I think many of you probably do, too. Let me tell you about my first time. This was mass CPR training. I was 19 years old, and I didn't know anything about CPR. I wasn't in the health and safety. I was studying astronomy in university. Not really the best fit, but a friend of mine that I knew from where I grew up said, hey, we need someone to help out with this CPR training program. So I didn't know what it was. I want to do good. I jumped in and did it. And I think many of you might resonate with this. Six months later, 17 people, 1,500 hours, and we had our first training done. 151 people trained in adult CPR. It was a good time. We did some good. The program is still going on. It's 10 years old plus now. So I'm somewhat proud of that, but also I'm not. I learned after I started doing this three years in, turns out CPR is actually not that effective. So even with best trained people you can get, right good timing, chances of survival are very, very low. Not to say you shouldn't do it if you're trained or you know what you're doing.  So turns out CPR is good, but maybe not great. So what is great? Well, here's one that is extraordinarily good. This is one of the best things humanity has ever done. We eradicated smallpox in 1973. So this killed 300 million people, which is outrageous. But by eradicating this, this global effort, we'd saved another 60 to 120 million people after that. But then in context, if we had achieved world peace, we would not have saved as many lives.  Gives you some perspective. Definitely could not have saved as many lives with CPR as doing smallpox work. But this was taken out before I could get to it. So what about something more current? How about malaria? So maybe you're not interested in health, but if you want to save lives, there's an organization called Against Malaria Foundation. And they're really good at distributing bed nets. And what these bed nets do is they help protect you from malaria. And their lower bound costs for doing this to save one life is about $3,300, which is an extraordinarily cost effective way of doing it. Now, if for some more context, US government occasionally will have to rate or value a human life. You have any idea how much that's valued at? It varies, but oftentimes it's around $7 million or so.  So AMF, Against Malaria Foundation. That's one powerful thing that I discovered that if you think strategically about what you're doing, you can make a lot more impact. And so this is exciting to me, but it can be exciting to you, too, just to rethink things. So I'd ask you to go back to that first moment and imagine that you could start over from scratch. Imagine you have a blank slate, and you can do pretty much anything you want to do. How many of you would do the same thing back when you were younger? How many of you would do the same thing now, like this path that we all get on? It's a sometimes strange, serendipitous path. You don't necessarily know how you got there. I know I would do things very differently now, because I've learned a few things after doing this for more than a decade about how to do good better. So this brought me to something I found extremely exciting. This is the effective altruism community, so EAs. This is why we're here to learn a little about this. I discovered this a few years ago, and it changed my path dramatically. So effective altruists combine evidence and reason to do as much good as possible. It's a new philosophy and a social movement that is really picking up now. So it combines both head and heart. The heart motivates us to be empathetic and do good, and the head guides those actions to try to make them well-directed and strategic. It's really the scientific way to doing good. So it's brand new. The term was only coined in 2011. This was out of a group of philosophers in Oxford coming up with terms to describe what we're doing. There was like super-do-gooder and whatnot, and eventually they came up with this term. Now, obviously there's a lot of history behind social impact and doing it well, but this does seem like a new phenomenon is happening. And so Peter Singer, who's here, gave the first TED talk on this in 2013, and he's one of the most famous moral philosophers around. And more recently we've had people like Elon Musk and Peter Thiel and Kerry Tuna and Desmond Moskowitz come on board with their time and their funding, actually putting in billions of dollars to doing EA organizations and causes. So it is a really exciting time to be in this space. And how do you do this? This is just overall general. What do you do? Because it's such a new field, a new concept, we're still learning. We're still defining this. But there are five basic questions that seem to be helpful for thinking like an EA. One is just something we ask ourselves a lot, how many people benefit and by how much, right? We're trying to get as many people as we can and help them the most amount that we can. Pretty simple. Second, is this the most effective thing that I can do? Given my experience, my skills, and given the need, is this really the best thing I could be doing? This one is actually hard to confront sometimes. Another one is, is this area neglected? So if you're jumping in and doing great work and other people are doing great work too, your marginal impact might be smaller than you expect. So it's something that happens. Another consideration is the counterfactual. If you didn't do this, what would have already happened? If you're digging a well and you leave tomorrow, will someone else take it for you? Like, is there someone else that could do it? You have to think about what actually happens if you're not adding your effort. And then lastly, what are the chances of success and how good would success be? So thinking in terms of expected value and getting as much as you possibly can, these are exciting actually to think through this. So I thought through these questions even recently this year and pivoted dramatically from what I was doing before, which was behavior change events and interventions to really getting involved in the community and at the honor of coordinating effective altruism global this year. So from two years ago when there was 60 people crammed into a single house to last year, was 180 people crammed into a conference center in Berkeley to this year, 825 people across three cities, San Francisco, Oxford and Melbourne with Elon Musk keynoting, it's been quite the honor. And what this showed me helped me rethink my mono efforts is that there's a massive opportunity to do good with corporates and funders. So we talked to over 100 and got them introduced. Many of them were not yet thinking about effective altruism but they were interested. They came, they spoke, they partnered. So they wanted to get their feet wet which is very encouraging. So this leads me to my final point is that I think effective altruism is this extraordinary opportunity for all of us to rethink how we're doing good and try to do good better. It's humbling to think about this, rethink about this and try to get deep into the research and to the critical thinking to do it better. But the people we've talked to recently are interested and especially the funders because it's a lot easier to move around a little bit of capital if you just think more strategically but also you, everyone, how you're spending your time. So I look at this as an opportunity to find ways of doing more good within our own domains or switching out to something completely brand new. And that's just an exploration that we all get to take. This actually may be the best time in history to be alive to do social change because there's so much need and there's so much capacity now. So if you are at all interested in this sort of thing, as I am now, please check out the websites, effectivealtruism.org is the portal in to the community. There's a few thousand of us around the world and growing fast. And because I now think this is so important to do, I'm happy to help you on that path and guide you to what might be something that's powerful for you. And with that, thank you for your time and I hope to see you on the social change path. Cheers.