It's really, really an amazing week we've had. Hashtag non-profit power. Wow, you know, I'm gonna say this really quickly. I've said it pretty much every day, Tony, and I don't know if you've been able to join us every day, you know, watching the archives, but we really had this idea a long time ago. And we thought it was just so exciting in Fundraising Academy. I mentioned it to you and you're like, I love it, let's do it. And you helped us launch this concept. And Jared, do you think it's been really fun and really successful to drill down? Yes, so much fun. So much fun, so really, really amazing. Today we have Tony Bell coming back to us from Fundraising Academy, the guru of cause selling. And we're gonna talk something a little different today and we're gonna be talking about overcoming objections, the power of mindset. And it's gonna be a really robust conversation because we've spoken so much about the tools, the cause selling piece of it, the process, but I really think we need to have the conversation about what happens emotionally and the mindset that we need to go through. Again, thank you so much to all of our presenting sponsors. If you were not with us, we would not be here. Now, Jared, how many shows in? I should look, but I think it's like 360 plus. Amazing. I know. I'm laughing because we've collaborated so much together that it was almost on cue, Jared and I both were like, you know. That's right, that's right, I love it. And having never met in person, like the three of us have never, you know, I've never met. So anyway, it's. I know. You're gonna be shocked at how tall Jared is and how short I am. Well, I'm more on your scale, Julia. It's sad, it's sad. I can never meet anybody in person now going forward for the rest of my life.  But anyway, again, thank you to Fundraising Academy for joining us on this really exciting adventure. It's been wonderful and it will continue to be wonderful. Again, I'm Julia Patrick, CEO of the American Nonprofit Academy, been joined today by the non-profit nurse herself, Jared Ransom, CEO of the Raven Group. And I judge you for math. This is our 361st episode. So what a wonderful time to bring in the discussion, overcoming objections with that mindset of success. And Tony, I'm sure I speak for Julia as well. There's no one better to talk about this than you. So thank you for joining us again. Well, thank you for that. Thank you for that. I'm looking forward to our conversation as always. So, Jared, you asked this question. Yeah, let's get started. You know, let's talk about truly how critical is this mind selling? So cause selling, or sorry, the mindset within cause selling. So we've talked all week about cause selling and the wonderful efforts from Fund Raising Academy. But now we wanna drill into the mindset of this. And how critical is that, Tony? Well, I mean, I think mindset is critical in everything that we do and in all aspects of life. Very much like when we talk about cause selling and when we are being a little bit disruptive by saying that fundraisers actually sell. It just, it is, it's that mindset. We sell every day, right? I think I use this kind of ridiculous example all the time, even if you're trying to sell, you know, your children on eating broccoli, right? It's like you're selling every day in different ways in all aspects of our lives. And I feel very much the same way about mindset.  And as fundraisers, I think it's our responsibility as best as we can. And we're gonna talk a little deeper about this as we move through the conversation to bring the fun into fundraising and to, you know, to create a joyful space and to make the experience as joyful as possible for our investors and for our volunteers and for our board members. So I think mindset is very critical. And I believe strongly, especially as a fundraiser, that it's important that you come from a place of abundance when you are meeting with potential.  You still need to come to that, come to the table with this mindset of abundance because folks want to invest in success. So if you come to the table feeling anything less than success, folks can feel that and a investor is going to be hesitant to invest. So that's just one way. So good at bringing that fun in fundraising, right?  And you're right. And I think, Julia, you say this quite a bit is that people want to be on a winning team. Absolutely. Absolutely. And a lot of time the causes that we support are not fun. It's not pretty, right? The scenario is not pretty. It's not fun. In many cases, it's sad. Yes. You know, so that's why I really lean into, you know, the cause that you're selling might again just be something very, very depressing. But that's why, you know, coming from this joyful place, doing whatever little thing we can do to add the fun and fundraising is important for us. You know, years ago I had the opportunity to interview a Fortune 100 CEO about his philanthropy. And he told me that he dealt with such sorrow and such tragedy that he in his mind always had a percentage of his total philanthropic budget specifically for the arts and culture. Because he felt that that was the link to teaching and connecting our people to positive outcomes and what could happen. And if you could see beauty, then you could understand beauty and you could understand, you know, good outcomes and striving forward. And I always thought that was so interesting. So interesting, you know, because you are right. We don't talk about that enough about how really to use Jared's word tragic the environment we can be in. Well, it's a huge advocate of the arts. I certainly lean into that philosophy, what the arts can do for our wellness, what the arts can do for the economic viability of our communities. I mean, there are many cities across the country that were in a depressed state and a leader gets in, invests in the arts and you just see from there how the economy grew. So yeah. It has a huge impact. You know, it's an interesting thing. I wanna talk to you about this a little bit deeper because we have a lot of negativity by virtue of what we're dealing with within the nonprofit sector. But then how do we navigate those negative attitudes? You know, I think there's a lot of negativity around the word selling. And we hear the word selling or we hear the word fund development and we kind of put those people in a different part of our building or our campus.   Yes. And I think I might have shared with you, Tony, before on a previous episode because we're so grateful to have you on multiple times. Even working with an organization or an organization that their volunteer coordinator brought in a Starbucks team. And I said, I would really love to address them before they start their meeting. And she was like, no way are you getting access to them? That's negative, right? Yeah, that is. So, yes, we've kind of been pushed aside. Like, I don't know, these people are scary, hold on to your purses really tight. It's true. Well, how do we get around that? How do we get around that stereotypical negative feeling that people have towards development folks? Well, that's really, that is a big question. I haven't, in my experience as a fundraiser, I haven't dealt with a lot of negativity around the work that I do. I think a lot of times that comes from a lack of understanding around the value that the role brings in certain organizations. So, in some organizations, a lot of focus is on the programs and who implements the programs. Cause if we don't have a program, right? Then there's nothing to sell. And so a lot of focus is on, these are the more important members of our team because they're the ones delivering the programming on the front line. They're really the ones affecting change in the community, right? But our fundraisers are the engine or provide the fuel, maybe that's the better way of saying it, right? Provide the fuel for the engine so that that great work can occur. Right. Okay, so we've already made a quote from you about, I said the crass thing, no money, no mission. You said something much more articulate. I'm gonna use that and create another quote slide because you're right. We are in the fundraising development team or capacity, whether it's one person or 100, we are that fuel for the engine. Gotta be looking at that. Absolutely, and in some organizations, because you have your program lane, you have your fundraising lane, you might have a separate lane for volunteers. So to be mindful of how you break down those silos, how often do you bring everyone together? So everyone gets a sense of the value that they're bringing to the organization so that everyone understands how important each other is in their role and towards the success of the positive change that you're having in the communities that you're serving. So there's lots of opportunities in terms of our organizational structures and the way that we communicate and interact that can help minimize, if not completely prevent a lot of these negative attitudes that some fundraisers may come across from coworkers or from board members. Right, well, so I love that you bring up board members. So I have a question in particular and I'm sure we've all experienced this, but board members may not be eager to share their contacts or even invite some of their close friends, family members or even colleagues to a potential event for fear, I'm gonna say fear of the unknown, right? Of how will they be addressed? How will they be stewarded and prospective thereafter? So how might you recommend in particular using this cost selling method to navigate some of these negative attitudes or really that defiance of collaboration? So I think part of it is educating board members on really what is the process of raising money? How do we really do that? And the cost selling cycle allows a lot of opportunities for board engagement, whether it's through the prospecting, right? Whether it's through the ask, whether it's through the presentation, there are many ways for board members to engage in the process so that their level of trust is such that they don't question the actions. They're not concerned about inviting someone to an event and how is our development officer going to approach them? So I think engaging them in the process in various ways is one way of building that trust. Another way is role play with the board member. I hate role playing, I'll be the first to say it.  So if you have a board member, it's like, you know, if you're sensing that they're not gonna trust then say to them, well, let's role play this. You be that donor and I will show you right now how I would have that conversation and how I would introduce myself to that individual. You can sign me up for role play as a Fezbian and a mass communication theater major. I'm all over it. Yeah, well, I'm not afraid of a stage and a spotlight but role playing is just not my gig. It's a little different beast, yes. I can see that. I like that, I like that a lot. I do too. And I'm curious, your colleague Jack Alotto that has previously been on for the Power Week with Julia and I, he shared about the Madden test and for those of you that aren't familiar, this is, gosh, I don't know how you would refer to it but it's a piece, it's a resource within the qualifier essentially within the cost-selling methodology and he shared with us exactly what Madden stood for M-A-D-D-E-N. Is that a tool or qualifier that you would educate your board on? So for instance, if I'm thinking, okay, I have this breakfast table that I need to fill and I have eight open seats, might I, board member, go through the Madden test and say, okay, who might be interested in this and how might we qualify them using that same methodology? I would send off fireworks right now if I had any because that's absolutely brilliant. And yes, it's a great tool for board members. Again, it helps them understand part of the process and how you again prioritize your work, how you determine where the greatest opportunity rests within this pool of potential investors. And what I love about the Madden concept is that really you can rename any of those qualifiers. So come up with whatever name you want. We use, Madden, it makes sense for a development officer and a fundraiser for a gala or a breakfast, trying to fill a table. Some of those qualifiers might look a little different but it is definitely an enabler for the board member to help them prioritize their Rolodex in terms of who they might reach out to and invite to maximize the opportunity, not only to maximize the opportunity, but to make sure they're inviting the right people. Right. I love that. And I think that is a really an amazing mindset approach because I think what happens is in the rubber chicken circuit you get like, well, you know, I went to their events and now they're coming to my event and you're not creating a sustainable. Yeah. You're scratching everyone's back. Yeah. Oh my gosh. I'm laughing because the rubber chicken has now become the rubber salmon. So you know, it's like. It's really true. It's really true. But I mean, you know, I think. Yesterday's rubber chicken is today's rubber salmon. I love it. I think you're absolutely spot on. I go for the vegetarian compare. It's always amazing. It's always fantastic. I'm with you there. Yeah. Whatever you have the choice, choose the vegetarian because it's going to be really good. So as I think of, you know, the end of the year because believe it or not, it is right here for us. The end of the year is now. And there are so many organizations that I know are really feeling the pressure towards this end of year fundraising. I think the statistic is roughly 30% of most operating budgets are raised in this Q4 period. So really looking at these final, final four months, right? So there's going to be news. There's going to be whether it's, yay, we made the budget or did not quite make the budget or yes, this donor said, absolutely. And oh, these donors just, they're not quite on board. Talk to us about how you would recommend when it comes to delivering fundraising news, whether it's good or it's bad because we're all feeling this. And I think now at the end of the year, coming up on two years that have been very, very heavy. What kind of sage advice do you have for us, Tony? So I shared with you guys just prior to our Chitty chat. I'm a huge fan of animation and animated movies. And one of my favorites is Kung Fu Panda. And so in Kung Fu Panda, there is this scene where it's very well said, there's no such thing as good news or bad news, there's just news. So when we talk about this kind of positive mindset, really just it's news. Now you're educated, you're informed. The real test now is what do you do with that information and how do you respond to it? And like any business, right? 501c3 tax status, not business model. In any business, you have to have these candid, transparent conversations around where the business is in terms of its health and where it's going. So if we're talking about, as a fundraiser having to communicate to an executive director or CEO, that something's not going the way that we thought it was going to go or having to communicate to board members that things aren't going the way that we thought that they would go. Really approach it that it is news. It's not good or bad, it's news. And now we have that information and now we figure out what our next step is based on that information. I love that, because I think that's a real mindset tool because if you lock yourself up with fear, then you don't really give the right information or you give it when it's far too late because you try and not broach the topic or I can just see what happens is it just, it becomes something that you don't lean into until it's really far too late. And for my own personal experience, whenever I kind of entered the arena with the mentality that I've been defeated, it prevents me from thinking creatively about the solutions because I already feel defeated.  I'm defeated, I'm done. Yeah, yeah. And that's so to me to hear the way you articulated that, I think that's one of those reasons why we have such a horrific turnover in development in the nonprofit sector. AFP reports that we have an average tenure of 18 months. I suspect that the next time that gets reported, it's going to have dropped even more. We're burning and churning our development folks. I've got to believe it's because of this. I think there's a lot of stuff to unpack there. You know, it's organizational norms. It's how some boards micromanage organizations versus being thought leaders for the organization and trusting the talent that they've hired to do the work. So yeah, so there's a lot to unpack there. And kudos to AFP in particular and a lot of organizations are doing this, but I've noticed it with AFP particularly, is one of their pillars this year and over last year is really around the mental health and wellbeing of fundraisers and how can we support that and how can we continue to encourage those types of conversations to take place in the workplace and beyond. We mentioned it was global news around the Olympics and what's going on in terms of professional sports and how those athletes are now saying, wait a second, I need to dial back for a minute. If you want me to show up on the court and be the best that I can possibly be, then I need a pause. And if that means I can't be on the court every time, then that's what it means. And I think you said it so beautifully and it perhaps was in the Chitty Chat Chat, but yes, this has become this being more of the mental health overall burnout has really become a global issue and a global conversation finally and thankfully, right? It has really become something that more and more of us are beginning to talk about and open up about. I myself am doing a self-care and how to avoid burnout workshop, actually on August 31st, but there's just been and here's my theory over the last two years, those of us that are natural helpers, right? We continue to run towards the fire, who can we help? What can we save? What can I do and how can I be of service? I'm feeling it that now two years into it, there's just so many layers, layers of weight, complexity, expectation. So going back to your comment, Tony, about those boundaries, like how can we set healthy boundaries and boundaries I think also tie into our mindset and they continue to perpetuate that positive mindset, regardless of the news, because as you said, it's news, it's not good or bad, it's news. So everyone, if you haven't seen Kung Fu Panda, that is a huge endorsement by Mr. Tony Goh. One of my favorites. And that's what kind of circling back to something we said at the very beginning of our conversation, because again, time flies, we're almost done. You know, is what can we do to always put fun and fundraising and making sure that we, because we talked about the gift, the transformational gift, but how are we celebrating the steps along the way that get us to the big wins? That's just as important as the big win. You're right. I love that. In many cases, we have the big win because we might have modified or shifted something in our organization, we might have tweaked a message, we might have created a new marketing video. So those are wins to celebrate regardless of maybe what the larger anticipated win might have done. So I really think celebrating those steps along the way helps us get there. Well, and I would say to your point, sharing that with your entire team, because I love what you said is that a lot of times these negative opinions or, you know, not embracing the fund development team is exactly what you said, it's education. They don't really understand what you're having to do. So if you can share, you know, wow, we did this, we did that, we're not just hinging everything on a yes or no.  Calendars Bill. It's true, Tony, I think I appreciate that. You know, Tony Bell, what an amazing national, actually global thought leader when it comes to this, but incredible, incredible wisdom and energy. I have to say when I'm with you, I'm in the show and I'm like, I'm ready to go ask. You know, you give me a lot of empowerment, but realistic, you know, it's a really an amazing thing. And then when we look at what Fundraising Academy has done with this cause selling cycle, it's super powerful. And to get you, as Jared and I say, you're the big get to come onto the show and get the top dog to talk about this. We wanna remind everybody that over the course of this last year, we have been working with Fundraising Academy and we have some amazing content that's in our archives. And so if you want to learn more about what this cause selling education really means, we have an amazing archive full of the different phases cycle by, I should say step, step by step going through. So check us out with our archives, because it's amazing. So tomorrow we, hard to believe, Jared, that we've like blown through a week. I know it is Friday Eve. Friday. Tomorrow we will finish up with our ask and answer. We have some really interesting questions that have come in, but we've talked about determining capacity. We've talked about looking inside at what you already have, because there's some amazing information and how do we do sorts? Getting the gatekeepers key. Who is a gatekeeper? What do they do? What do they want? How can you navigate them? That was riveting. So lots of exciting opportunities. If you're coming to us for the first time, maybe on the archive, look for these other episodes because they're so much fun and they all fit together. And I think that's part of the magic. We wanna make sure we thank all of our sponsors again. Without you, we would not be here having this amazing conversation. Thank you to my gallery for supporting. I know, it's like, join me, Tony. I love it. And thank you. I'm here for applause all the time, so. And thank you, of course, to Fundraise Academy Cost Selling Education. We are so grateful for your partnership and truly in presenting a sponsor for this nonprofit power week, nonprofit power. So make sure that you do help us get that hashtag viral. We're so appreciative for your continued support and looking forward to having you back here with us tomorrow. Thanks to Julia Patrick, CEO of the American Nonprofit Academy. I'm Jarrett Ransom, also known as your nonprofit nerd. And again, it's been such a powerful week. We are so extremely grateful. Look forward to tomorrow. Priyay's ask and answered. And as we end every episode, as the previous 360 episodes, please stay well so you can do well. Thanks everyone, and we'll see you back here tomorrow.


What's going on guys, it's your boy DPJ here today with another Destiny video and in today's Destiny video we have Titan, Sunbreaker, Hammer or Soul, Spree's up for the week. Now if you guys have anything badass to send in, send it into the email, I've just linked in the top of the video description. Subscribe for daily Destiny and I'll catch you guys under next one. Peace out until next time. Peace! This is amazing. I can't believe what I'm seeing. Your runner has the spark. Zone B neutralized. Zone B secure. It's remaining. Triple down. You're in the lead. Zone C neutralized.


So, as a motivation, let us recall the situation to two dimensions, 2,2 supersymmetry. The situation in four-dimension will be exactly the same, conceptually is the same story without any difference, and even the proof, the strategy of the proof is the same, actually even the proof is conceptually the same for all results. But of course, in four-dimension, you get more complicated structure, richer structure, more information, more nice things, so it will be interesting to go to the four-dimensional theory. So, in 2D, start with a 2,2 super conformal field theory, and you write a super conformal index, which is elliptic genus, which has this form, and is independent of q bar, is the salomorphic function of q. J naught is the left, r-charge, and so on, and notation, I think, it is explained here, it is clear. So, this is an index, it's a super conformal index, which means that it is a quantity which is invariant, and the continuous deformations of the theory provided this deformation preserves super conformal values. You can ask if you can do something better than that. You can see this as a family of quantities parametrized by these two complex numbers, and you can ask the question if, for some value of these parameters, the index is actually more stronger than just a super conformal index, that this is invariant under a more general class of deformations of the theory. In particular, deformation which breaks conformal index. And this leads to the idea of specialized super conformal indices, that is, you specialize the index to some value of the fugacity such that that particular index is invariant under the formation of the theories which break conformal symmetry, although they should preserve supersymmetry, otherwise you have no index theory at all. So, n is an integer, and you consider this where you put this fugacity z equal to e to pi n, which is one, but in fact this is not an entire function of z, so this depends on n in a trivial way in general. This is independent of q now, you can prove just by standard techniques of written index theory, and so it's a set of numbers labelled by n, and it is, in fact, an integer just by modularity of the starting theory. You just write the index as a trace, exchanging space and time. Now, these indices are more robust than the general index here, because they survive massive deformation of the super conformal theory. I perturb the theory also by adding a relevant deformation, I go to a massive theory, I get a non-trivial organization group flow, but these indices will remain constant along the randomization group flow. At the end, we get a massive theory in the infrared, and the massive theory in the infrared is characterized by a BPS spectrum. We have many sectors, the BPS spectrum splits in many sectors labelled by i and j, where i and j go from one to m, where m is the number of duty vacuum, which has a central charge, which is anti-symmetric under the exchange of i and j, it is a complex number. And for each sector, we form a BPS operator, which is just an n by n matrix in this case, which has this property under the exchange of i and j, and which is written like this, where n i j is simply the number, the absolute value of n i j is just the number of BPS states, I mean really BPS multiplets in that sector. The sign here depends on several things, it is the corresponding phenomenon two-dimension, that in four-dimension lead to the Gaiotto-Mürnowski quadratic refinement. You have the same story also in two-dimension. Then, when you have this operator, which are defined sector by sector, you get something which combines all sectors by taking the product of these matrices in an order which is anticlockwise ordered with respect to the phase of the central charge, and the symbol here means just anticlockwise ordered. This matrix M is what we call the 2-2 quantum monodrome, it is just an n by n matrix in two-dimension. Now, in two-dimension we have also a chiral algebra associated with this problem because the 2-2 conformal field theories contains a topological sector, I just twist topologically the theory, and I get a chiral ring, and I can add the gradient operator in the conformal limit, and I can consider just the character of this chiral algebra over a module, which is the chiral algebra itself. So, I have a character here. Again, the character depends on the complex parameter, but I can specialize it to this, and just as before, while these characters are defined only at the super conformal point, the specialization f and meaning can be computed or well-defined even if the theories are massive and are independent of the massive deformation. So, in two-dimension we have a relation between the super conformal indices, specialized, the BPS Petra, and the chiral character. We have three objects, which are the three objects in the title, and they are related essentially equivalent information, and indeed the specialized indices are equal to the trace of this BPS matrix m to the n, n is an integer, n is equal to the specialization of the character of the chiral algebra, which is here in two-dimension a stupid algebra, but it will be interesting for them. So, in particular, the specialized super conformal indices can be computed in the infrared just from the BPS Petra or from the way the ring behaves, the spiral algebra. But for a given super conformal theory in the ultraviolet, we get many different BPS Petra, which depends on the particular deformation we do. We can deform in many ways. We get different infrared theories with different BPS Petra, and these different BPS Petra are usually called BPS chambers, but all the BPS Petra we can get are restricted by the condition that this trace should give me the super conformal invariant, which are independent of the deformation. In particular, trace n to the n should be continuous across a wall of marginal stability because on the two sides we have the same super conformal theory, and indeed the wall crossing formula is just a statement that the specialized super conformal index exists. They are well defined. This gives a condition for m, which is just the full information you have from the wall crossing formula. The problem of this talk is to find exactly the same relation, super conformal indices BPS Petra and chiral characters, but in four dimensional and equal to two quantum field theory instead then in two dimension. But everything will be essentially the same except that the structure will be much more complicated because everything will be infinite dimensional extension. So we start from the first ingredient, the super conformal index for a 2D for dimensional theory. I guess we are... OK, this is it, which is an expression which contains three fugacity, P, Q and T. These are the generators of the symmetry. Here we have written the dictionary. It will not depend on beta as always. This is just for converging regions, but everything is independent of beta. And what is interesting for us is that this index here can be given a geometrical interpretation as the partition function on an off-surface. The off-surface is a complex manifold which is topological S1 times S3. So compute the index with some fugacity, the partition function on S3 with some fugacity, and P and Q are just the complex structures, parameters of the surface. The partition function for a supersymmetric theory will depend only on the complex structure of the surface, not on the details. And for off-surfaces of this class, the complex structures are parameterized by two numbers, P and Q, which are in the unit disk. Well, the puncture unit disk. And this is the definition of the... of surface. It is essentially this relation with Q and P being these two parameters. But we have a further fugacity, so we have to insert a fugacity, the explicit in this game. So we have the party integral on this geometry with an explicit insertion of the operator to this fugacity here. As in two-dimension, what we want to do is to specialize this index to get something which is robust against breaking superconformal invariance. Let me start with an intermediate specialization, which is for technical reason. I set T equal QP n plus 1, where n is still an integer. So I get this. We still depend on two, a complex fugacity and an integral variable, n. And here we have various possibilities. If n is zero, this is just the partition function on the off-surface of the theory with the original... with the anthropological twist, the one which gives me the Jones polynomials. And so this is purely topological, depends on the topology, but not on the complex structure, so it is independent of Q and P. If I take n to be minus 1, I get the Schur index, which was defined by God, Rastelli, Razzamatt and Jan, it reduced to a character of a two-dimensional chiral algebra which was defined by the same people, Rastelli et alp, and since it is a character of this algebra, a two-dimensional algebra, it is independent of P. It is just a fun... an homomorphic function of Q. For general n, you should think of i and PQ as analogous to the function we have in 2D with P and Q, but variables give you general dependence, and in order to get something which is invariant, we have to specialize even further this thing. This is again... well, this is an analogy between D equal 2 and 4, you can do this argument with D, anything. So let specialize further, so I specialize just the same way I did in two-dimension, where I had the first variable set equal e to pi i n by here, for convenience, I set already n here, so this is just the same, but written in the same form, so this is the intersection, which is now a function of Q and n, and as before, this synthesis has an interpretation of this function, and it's just the partition function of the n equal to 2 in four-dimension computed in a degenerate limit of the off-surface. Just take the off-surface before and take this limit of that geometry. You get a singular geometry, in fact, a non-compact one was compact, but in this limit, they're compactified with the fugacity corresponding to this factor here, and this is what is called, well, it is the Melvin cigar, which is this geometry times S1. Now, these indices is the same index which plays this, is this n equal to specialized index, which plays the same role as Ia in 2D, and is an index which you see very easy by the standard manipulation, preserved under all deformations, even the ones which break conformal invariance. And it's an infinite family now of indices with the penton Q where Q is in the unit disk. And indeed, just as in two-dimension, the fact that Ia was well-defined was the well-crossing formula. In four-dimension, you just write a condition which is well-defined, or you wish the partition function on this geometry is well-defined, and it is just on the nose the same as the concept which saw the well-crossing formula in four-dimension. So everything remains conceptually the same, but this geometry here, the partition function of this geometry here was already started before with Wafa and Nasky, and so in the massive case, and so we can borrow from that. So let me take now after the deformation of the theory, I get a massive theory and go to the Coulomb branch of this theory, and here I have BPS states as before, as in two-dimension, which are characterized now by the charge, we take the place of the sector, Ij as as before, and also they can have a spin here, charge and spin means, in fact, charge and spin of the lowest component of the BPS multiplier. It has a central charge which is an additive function on the charge lattice, the lattice in which the gamma takes value, the mass, and then you have as before the BPS phase which is the argument, the phase of the central charge. The factor that is to each charge here, you associate a quantum operator which is analog to the finite matrix MhA of two-dimension which is defined by this, it's essentially a quantum dialogue or a product, a product here, I need to take the product over all particles, all BPS particles I have in that sector with the charge. This is the element of the quantum torus algebra where the quantum torus algebra is generated by operators which satisfy this algebra here where gamma, gamma prime is an anti-symmetric pairing on the charge lattice which is given by the direct pairing, electromagnetic direct pairing. So I have this and just as before I define the quantum monodromy taking the product from all sector and ordering the product in the anti-clockwise way with respect to the phase just before and I get this. And this is the 4D quantum monodromy and again, calling it in sovereign member crossing in this formula is obvious, it's just the fact that the conjugation class of MQ is in inverse. However, in 4D if we are along the Coulomb branch we have in addition to massive BPS particle also massless BPS particle which are just the BPS photon the infrared BPS photon which F charge 0 from these and F's being plus minus 1 the first component which is the 14 and so I have to add here a factor to take care of these things which are however centrally in the quantum algebra here so it is just a C number factor which depends on Q but is otherwise totally trivial. And this is standard. R is the complex dimension of Coulomb. Now this script quantum monodromy which is just the same essentially up to normalization I take the traces to a power n these are the formations and we are crossing invariant of the theory and so should be indices of the ultraviolet super conformal theories and take the same value for all massive deformation and the analogy with 2D suggests that the traces to the hem should be the same as the specialized indices which are just this limit of the standard indices. So we can compute the specialized of the ultraviolet theory just by infrared information that is by PPS spectrum and the proof of equality is essentially obvious because both sides of the identity of an integral representation as a partition function on a geometry this is defined by this geometry this is defined by the the generation of the off surface not this but these two spaces are the same the sphugacity is inserted in both cases in the same way along the same cycle so it is just taking the generation of the off surface and see that this geometry as before when we define these things indeed a special case of this identity was checkered by Kordov and Schaub for the case n equal minus 1 in that case this is the Schur index which has a stronger property in particular you don't need to put p equal e to the 2 pi i because this will be already independent of p so it is an even stronger indices and then in that case it is just the vacuum character of the two to D chiral algebra defined by the rest of the entire so this equality was shown in this paper before and what is done here is to extend this to all integral let me see so let me consider the case of the simple class of n equal to 2 t conceptually that is the Lagrangian n equal to t which have a weekly couple Lagrangian formulation which are super conformal the indices as this form produce the gauge group have the flavor group and you write just this where IV and IH are just the three super conformal indices and the integral over the R measure of the gauge group just project on the singlets distribution so for Lagrangian it is enough to specialize the integrand because this where p and q appear you specialize it for free hyper multiplets with given fugacity you get this expression here where this is the standard data function defined like this and notice that this is just the n power where n is this n here the partition function of a complex pin 1 half fermions into the dimension theta divided by eta up to a trivial factor q to the minus 1 so this is already something we suggest relation with two-dimensional conformal field theories for free vector multiplets the story is slightly more complicated because we have zero moles which are both of fermions dependent if n is positive or negative and you need to cancel them by a procedure which is essentially the same you have in super string theory to cancel beta gamma you do just the same things so they find and after that you get the finite quantity which is like this where this is the root lattice of the gauge group for free or abelian gauge theories this equality is just autological because the same expressions just on the nose are just both the specialized indices and you just write this in this form and this is just the product I had before defining the quantum monotony in the abelian gauge theory integral and so everything is tautological what is not tautological is this equality for non abelian gauge theories think of the simple non abelian gauge theory as you do with four levels the bps spectrum consists of w bosons quark states but then you have infinite towers of dions of unbounded mass unbounded electric charges so in the quantum monotony you have infinite many factors besides the obvious one which give you a very complicated operator and you need to prove that the factor of all the bunch of dions is just one the full contribution of all this infinite tower state is just the operator one and so you can neglect the same again this was checked before for SU2 super Q2D by Kord of Ainshaw for the special case n equal to minus 1 so the story is not surprising but it is technically difficult to prove the quote the third ingredient is chiral algebra we saw in one example the free hyper multiplet which for n positive is the partition function of n free spin one half chiral complex fermions which is a character of the chiral algebra of a chiral algebra with central charge equal to n if n is negative we get the absolute value n of free spin one chiral bosons changing the partition function in one of the partition functions for a free theory changing bosons with fermions so for n positive we got a good unitary two dimensional theory with this central charge for n negative we get a non unitary theory which has the opposite central charge which is negative this result here although this is an example but we will see the general case this is at a standard general result which is true even in the nola-grantzian case and we will see that is true which is due to rastrelli et al so if I start with a four dimensional supra conformal field query which has central charges c and a then the special i index is a character of it to do chiral rational conformal field theory which has a Vrazaro central charge which is 12 n c where c is the four dimensional central charge if n is less than zero then c to d is negative and the rational conformal field theory is never is never unitary for n negative for n positive is usually unitary but we have no result in general the most important case was n equal minus one which is the sure case which is of course negative and what we get here it is minus 12 c which is exactly the central charge in fact the full chiral algebra which was introduced by rastrelli and collaborate for n equal minus one it is just the story of rastrelli but we have an infinite story for all integer n positive and negative now the special i index here is in particular a q series which is some power series in q with some coefficients i n which are integers and given a q series i can define a concept which is attractive central charge by this formula essentially I suppose this to be a character of a conformal field theory I use the card formula backwards and from the behavior of the coefficient for large n it would be central charge which in fact is not really the central charge it is central charge minus 24 the smallest dimension of a primary field with full unitary theory and also for n large n 0 in this case is just the central charge and this just and so it is given by this formula with n positive and for negative is given by this formula here we are seeing a this numbers here and this is always positive if n is negative in addition if my fourth dimensional theory is a flower group I get in this two dimensional character here a cat smoothie character all level which is minus one half k is the central charge of the fourth dimensional current algebra and so the special eye indices became cat smoothie characters of one kind or the other if I have a flavor group or related to that and we can ask since this is very similar to a character of a rational conformal field theory in two dimension if it enjoys modular properties as a character in a field that two dimensional rational field theory should have and you can show at least for Lagrangian models that modularity of the integrals I had written before are equivalent to asking that all gauge beta function vanish that the theory is super conformal in the infrared and this is of course correct because we started with assumption that the theory was super conformal and our expressions are the same only under that case let me speak about the related setup which is different but related that has a compatification of the four dimensional equal to theory on a geometry which is t2 times u2 now this procedure give me in two dimension in this t2 always a theory which is unitary if I start with a unity theory and this is a full quantum field theory in two dimension not just an index computation it is always unitary to satisfy all the good properties so the way you do you start from n equal to 2 Euclidean super conformal this makes sense only if it is conformal because we do a partial topological twist but we twist with the u1 part of the r symmetry group two r like in Witten case so Witten twisting makes sense in general but this makes sense only if this is non-anomalous which is just the same as asking the theory to be conformal so you split this group like this and you twist by this combination of operators where n is an integer and so what you get if n is zero you just get 2,2 theory in R2 which is in some sense the obvious complication if n is equal to minus 2 you get a chiral theory which has symmetry 0,4 on this t2 or 2 and in all other case it is a chiral theory with supersymmetry 0,2 really this is correct for n even because you see if n is odd I get one quarter spins which make no sense so since it is important to extend this also to odd n's we need to have an extra global u1 in order to make a twist on that to compensate this mismatch by one quarter but in the example you get so if we start in four dimension with a free theory or wiki couple Lagrangian theory I mean a theory called Lagrangian we do this you can do field by field and you get like this the 4d multiplet becomes 2d02 multiplet so for n bigger than zero the vector multiplet becomes a vector multiplet plus n plus one chiral multiplet of 0,2 in the adjoint for the hyper multiplet became n fermi multiplet in the corresponding representations if you have n negative the vector multiplet still a vector multiplet but now you get that essentially fermi and chiral multiplets get shifted one into the other the gauge grouping 2d will be the same as in 4d and the flavor group remains the same and you have just this gauge theory with the same group as in 4d which is couple with fermi and chiral multiplets of 0,2 in the suitable way and you have also the j super potential of 0,2 which comes just this is a chiral theory in general well it is always a chiral theory so it can be anomaly anomalous and you check that the condition of being anomaly free in two dimension sense is just equivalent to the condition of being conformal in four dimensional sense as before so this is the same now we can consider the left moving of the 2d theory left moving is the part which is not super symmetric so it is the central charge you see in the elliptic genus so this means just the effective central charge of the elliptic genus and it is given by this formula you have a formula for positive one formula for and h is the number of multiplets vector multiplets this is true even if the theory is known with the all suitable definition what you mean by the number of nh and v in such a way that you get c and a correct and you check that this is the same as the effective central charge of the special index of the trace of the monodromy and for all n just identically this is always positive consistent with the fact that this theory is unitary this result is not accidental and for Lagrangian models it is obvious to understand when you have because the traces is just given by the integral this partition function which is given by the integral yet before where this is the product of all three things and the elliptic genus for this theory is given by this in the partition function of this geometry which is just an integral with the same integral but a different contour which is in this case the prescription given by Geoffrey here one for a contour so the two things are certainly different for a negative this is unitary and the other is not so it cannot be the same but at the same c effective because they are but the integral representation with the same integrant and different prescriptions and the effective central charge depends only on the integrant on general grounds not on the contour just because it is the value of the leading compressor point up to using identities from Kirilov's about the hologram you just see how many multiplicity we have you just write the product you have before I brought this explicitly and this you just do the localization computation this computation give you the same integral but the prescription from the contour is different so this is computed by localization and this is computed by saying since it is an index the theory is a marginal coupling that I can send to zero so I get the free theory essentially and it just remains the trace which is the projection implementing the Gauss law that the states are neutral here instead is really a localization computation and for Lagrangian theory it just depends on how many Ip multiplies and vector multiplies you have in which representation and they are just the same rules for the same with that dictionary and you know that this is very easy to show that the effective central charges are the same for two expressions which have the same integrant even if the contour are different and in this case since this theory is unitary the effective central charge is the central charge so the central charge of this theory is equal to the effective central charge of the other for N positive in fact you have a theory into the equation which is but in two dimension from the side of the special eye indexes we have just this character we don't know anything on the right movers and things like that the compatification give you a quantum field theory which have all the ingredients so this is different but related and so this was for Lagrangian models which are not very interesting I mean everything is almost autological you have the same integrants and you just need to well you need to prove that in Lagrangian theories you can forget of the ions that is hard but once you know that everything else is trivial so let me go to the new Lagrangian examples well we understand something particular new theorems about hypergeometric functions so as an example I can consider G.G. Primes for conformal in theories which are labelled by A.D. dinking diagrams G.G. Primes are to simply lay dinking diagrams and the particular case in which the second dinking diagram is just A.1 this is just a G. Redaglas model of type G or G is in A.D.A we know the central charges the four dimensional super conformal central charges of these models which I write here and the condition that G. and G. Primes have co-prime coseter numbers for simplicity I know that the formula in general that will take more lines to write because depend on the common divisors of the two coseter numbers in a complicated way and R is the rank and H and this is C this is A and let me list of G. Redaglas the ones which are not written here have no flavour this U1 should be A1 and this is the flavour group this is the central charge of the flavour group in four dimensions the conformal central charge of the flavour group in particular we know from Rastelli's artwork that the n equal minus the index a 2D chiral algebra the 2D chiral algebra contains the current algebra of the flavour with a level which is minus one half K where K is the 4D central charge so this is the models so computing this from the BPS spectrum we know the spectrum was computed by itself it is given by the minimal chamber where we have the lesser number of states it is given that by the product of the two ranks of the two groups G and G prime times the cosector of the second divided by two and they have these charges here I can identify the charge lattice with the type of the two root lattice and this is like this where one is simple and this is positive using this BPS pattern we can describe explicitly MQ and we are in the trace of the monogram to some N as an explicit although very cumbersome multiple Q hypergeometric sum we know techniques to compute the effective central charge of any hypergeometric sum however complicated and this is the result for N positive I get C to D the theory is unitary on the two-dimensional rational conformal theory it is given by this where two R is the correction in the case H of G and HG prime is not co-prime so this expression here which is rather complicated but it is always equal to 12 NC where C is equal to the central charging for dimension for co-prime you just check with the previous one otherwise they find C like 12 this divided by 12 for N negative it was not computed in general it was computed for this class of model you can compute but this thing is boring we did some example not the most general one and you got this which is this expression as predicted in terms of the central charge of further dimension and this equal to the effective central charge of the minimal models PQ with L plus 3 for N equal 1 minus 1 and this is as predicted by Rastrali et al in particular you check from C and see effective the value of the minimal conformal weight and this just I suspect from the generality I stress that these computations here are done from the infrared theory I know the BPS spectrum and I make computation and I get quantities that I compare to the computation done in the super conformal theory and they will develop and we get always correct matching which just means that this identity is correct which it is strange not to be since it is essentially tautological let me do some an explicit character from BPS spectrum the simple case is A to N A1 and N equal minus 1 which is the shoe index and we know what should be because it was computed with different techniques Rastrali et al computed this in the ultraviolet from the super conformal field theory we compute here from the BPS spectrum we don't need to get what we got so this is the expression for trace N minus 1 in terms of BPS states in the minimal chamber it is a very complicated Q hypergeometric sum in two way integral variables very complicated but using various techniques from the theory of Q and N identities and Andrew's identities you rewrite it as a finite product which is like this it is just N the product of two N polygymers symbols here and this expression here is exactly the Feggis-Nexchiani-Auguri way of writing the vacuum character of that BPS automotor so this is just the fact that this is equal to that proved that what we got is equal to what Rastrali got and everything is correct and everything is analytic here you can do the sum explicitly for even the situation well it is the same but now I am computing M plus 1 trace where we have no prediction one computed from the other side you get this expression here where C is the Carton matrix of A to N minus 1 you go down by 1 in rank and this sum was computed by Feggis-Stoyanovsky for any Carton matrix here of any simplest Lie algebra and what you get is that this is the vacuum character of the loop algebra of the maximum important sub-algebra of G1 at level 1 in other words you take your algebra G you write like the standard Carton, the composition then you go to the loop algebra and you make the central extension and you take only things which came from here except to say this will return in a moment and so in this case we get just the the vacuum character of this algebra where G is the affine SU2N if I go to the opting case which is more complicated to compute but you get the same story with just in terms of the rank it is just the same for instead for trace m to the minus one often even are very different let me go to this case probably I have here considering instead the characters of the Archirae Douglas of type DR which are much more here we have a phenomenon of saturation if you recall on trans we had a talk about any equal one extension of supersymmetry of 2N equal to 2 cases in which N equal to 2 and the story was like this you start in the with N equal to 2 theory you give a perturbation a very special perturbation with preserve only N1 and you get here a thing that a priori should be an equal one but instead you discover that is an answer to N equal to 2 so the infrared fixed point is magically N equal to 2 this the example to offer a certain class of N equal to 2 super conformal theories in the ultraviolet that you start here I am talking exactly on the same class same characterization and we will see what are this so in this language you start from one super conformal theory in the ultraviolet which has some character you end up in the infrared with another N equal to 2 which has also some characters and you can ask is there a simple relation between the two characters so what I call cheese factorization is just the definition of what are the good ultraviolet fixed point we start and they are good for my courses too so in 2D we have the Sugabara formula like this and for a unitary theory if this is in equality in general for unitary you have here because you can have other degrees of freedom besides the current algebra but if you have equality then the current algebra is the full rational conformal field theory and the characters of the rational conformal theory are just characters now since we have a 4D to D dictionary due to rasterality between the two dimensional central charge and the four dimensional central charge which is like this if I have a flavor group GF we call that the group is the flavor group so if this equality holds for an N equal to super conformal field theory I say that this is cheese saturated why I call cheese saturated because you have this experimental fact that if this is true you can always find a finite BPS chamber that is a chamber with finitely many hyper multiplies something else as BPS stays such that the hypers are 12 times C that is if you compute C just from the BPS spectrum thinking of them as free fields you get the current C although A will be not correct but for C the BPS spectrum saturate about and in general you have inequality so this is the equation of inequality if this is true you can see that for N positive you have in general formula like this the trace of the monodromy to the N with N positive is a theta function of some lattice with a penton N and so on these are the flavor fugacity divided by Q to 12 and C so this you have essentially eta an effective central chart which is the central chart in this case so you have just a bunch if you wish free fields if you bosonize the theory as many fields as you have central chart so for this class of theories this is which are that you have this formula here and the only thing you need to find is the precise lattice you have here because in general it is a very complicated story this is what you expect from this story of C saturation on the other hand what you have from the actual spectrum here is a very complicated Q sum of the infinite many things well many things so reshuffling this in this form is very obvious and in particular for this class of models as we will see they are C saturated so you have you expect an identity like this that the Q sum from the spectrum of this class of agiridaglases of this form and this is very similar to an old conjecture in this function theory that you should have an identity between a sum of the four similar to this with some decoration and see the carton of the A and a theta function of this form but this conjecture was very old because it was motivated by the geometric interpretation of this Q function but was obviously wrong because you just expand the two sides of the proposal conjecture and just the first term is not correct so the problem was what is the correct conjecture the precise conjecture the conjecture was essentially correct but there was a small information in the formula a very small change but no one was able to find what that variation was so what this formula says is well one side is what we want to go this is on general ground of that form essentially from Namward with appropriate definitions and they should be equal so this is the correct conjecture we replace the not precise conjecture people had and indeed when we come to this through the monotomy operator we just give to Wagner we have the conjecture so this is the precise conjecture in two weeks was pro of the general case conjecture it is easier to prove than say there is a very conjecture something should be true so now what I say is really new theorems and proof in full ok so let me start with Algiridaglas with Autrank Autrank FSU2 flavor have this central charge for the SU2 flavor in two dimensions C is just N over 2 this are classical you put here the number and this inequality so it is saturated the SU index is just the vacuum charge of SU2 at that level because if it is saturated the two dimensional is just pure current algebra and then it is just the vacuum charge of SU2 of whatever level you get you get a negative level so it is some less it is in the mathematical literature you have and you check and everything is correcting trace MQ is more interesting since you expect to be of this form and indeed after we proved in the physical sense and then Wagner proved in mathematical sense that you have this equality where this is very complicated to write explicitly in the original form I will not write and here C2N is just the carton matrix of A2N so this is just a specialization of the theta function for the SU2M-1 root lattice the most beautiful form you can have the carton algebra here is just SU2 2M-1 at level 1 plus some free scalars in order to get this A-tas in front so this in this case and this even better you see for M positive you get unitary theory for M negative you get non-unitary theory in two dimensions strangely enough people prefer negative ends but M positive are better because they are positive they are really two-dimensional rational conformal theory if we consider even even is trickier and was not correctly studying the literature because it's Flavre SU2 times Q1 with this central charge for SU2 and this central charge for the anomaly conformal anomaly and it was thought not to be saturated but it is saturated you want carton algebra is just a free scalars so if you put here the one corresponding to the one this is just saturated and you get the same formula well now this depends on two fugacity just because the rank of the Flavre group is two so you can put two Flavre fugacity you have a certain explicit lattice that I will not write that give you this and this is really a theorem because Wagner proved it in functionality to give you a flavor of this function if you take the coefficient of v to the zero here this is just the standard function of the SU2 and root lattice if you want other example in the paper we computed a very large example so let me return to this one particular case this was the R A1 model here and you get here AR-1 the carton algebra here for any plus one is just affine SU well, specializing something in the odd case for R odd is just this otherwise times you want to not consider free the couple free field also here and here is SU are restricted to the nilpotent part so the relation is just that you take instead of the full the algebra just its nilpotent part and take the character of that and you get the infrared story from the ultraviolet one I don't know how this is related to realization group and so on and so forth but the map between the starting data and the final data is easy, it is just you have the same the group and instead of having all currents you set to zero all currents but the upper triangular one and P and that is it both level one nothing else change, it is just that you take a subalgebra instead of taking the maximum nilpotent subalgebra instead of taking the full algebra that is it okay


Hello everyone and welcome to today's Bitesize talk. I'm very happy that with me is Simon Hoimus from the cubic in Tübingen and he is going to talk about a NFCore pipeline called NFCorePan genome. Off to you. Thank you very much for the introduction Francesca, I am happy to be here. I will talk about cluster scalable punchline graph construction with NFCorePan genome. Thanks to advances in sequencing technologies, very high quality genome assemblies will become basically default in the near future and then this will offer us an opportunity to study genomic variation as never seen before. However, how can we represent and work with hundreds of genomes at the gigabase scale and one solution here could be so-called punchinom. It is able to model an entire set of genomic elements in the population in contrast to reference-based genomic approaches where we relate all the sequences to one linear consensus model of the genome. Punchinom creates each new sequence to all other sequences represented in this punchinom and in particular this helps us to mitigate reference bias. Now, how can such a punchinom data structure look like? And one way here is to encode it in a graphical data structure. As you already know, in the linear genome world we take the reference genome and augment it with variation. However, a punchinom graph now can compress the shared and variant sequences into one graphical representation. And in these punchinom graph models, DNA sequences are incorporated as nodes then with edges connecting the nodes as they occur as sequences representing the graph. This is basically this visualizing variation that you can see here in this figure. Also, if now sequences represent identical regions such as paralogs or orthologs then they will share actually the same nodes while the variants will be added as new branches in the graph. Like what you can see are the insertions, same letter change or inversion. We can also visualize such a punchinom graph in the tube map like way. Let's stay there for a moment to get a better idea of a specific punchinom graph model or implementation, which is called a variation graph. And here the idea is that actually all our sequences are paths that go through the nodes and these paths can be contigs, haplotypes, reads, or even whole chromosomes. In this example you can see two genomes and actually they share some sequence both actually visit this node number one because they have the same sequence but then their sequence diverges and so they both visit different nodes. This is a good visualization however if we have hundreds of genomes it can become hard to read and so the idea is that we can actually project this into a 1D visualization. What we do actually is we just concatenate all the nodes nucleotides into a so-called punchinomic sequence and then we write it from left to right, which you can see below in this figure. And then with a binary matrix we can actually encode the genomic sequences from both genomes. So if the sequence is present within the genome then we just draw the color and if not then it's actually not drawn. Now the question is how can we scale this up to potentially gigabase scale punchinographs and here the idea is actually binning. So here you can see a large scale punchinograph 1D visualization of a certain HLA DRB1 gene from the HLA region in human and what we are doing here is basically the same, we again arrange the nodes of the graph from left to right to find a punchinome sequence and if there's a coloring then the path has that sequence but if there's no color then actually the path does not have the sequence. You can place the path names on the left and basically what we are now missing are the edges but we draw them as black lines on the path to so visualize somehow the graph topology and the key idea is binning. So of course one pixel now represents hundreds or even thousands of nucleotides and so we bin them basically together and that's how we can visualize these large graphs. What is not clearly understandable here is the graph topology so there actually is also a way to visualize these things in two dimensions. This is extremely important to easily grasp large structure variations but also to take a look at certain kind of bubbles which can indicate regions where paths highly diverge or maybe could be hinting at the repetitive flow side. Now these are the key concepts to understand all the visualizations that are coming in this presentation. Now the question is how can we build such graphs that already exists is the so-called PGGP algorithm. It's called the Punchinum Graph Builder Pipeline and it basically comes with three major steps. The first one is the all versus all alignment step. So as input we have your sequences in fast A format and then we do an all versus all alignment which is quadratic so it can be quite heavy and then basically here it is visualized what you will get out of it. Once you have these alignments we can use a secret to actually use this kind of alignment graph to fold it together into a variation graph. So the idea is that when we have the aligned sequences and they share certain subparts of the pairwise aligned sequences then we can collapse these together into a node in this variation graph. The third major step is a normalization step. What we do here is we take this raw variation graph, we sort it in one dimension. So basically we sort it so that the 1D layout is guided by the nucleotide path positions given in the graph and we then normalize it by applying local MSAs across windows of this Punchinum Graph. So what we obtain in the end is this smooth graph. And usually in PGGP this is done around three times because at the edges of these windows we also need to ensure that things fit together. Then we remove some redundant nodes and we are also able to covariance against any path in the graph. So you can choose your reference of choice or any other genome you are interested in. Additionally we also provide, this tool provides graph statistics, 1D and 2D visualizations which are all aggregated in the Wild Dixie report. And the PGGP algorithm was already used to build one draft human Punchinum reference graph. And this was published in Nature this year I think. And therefore it's already really well tested. However, it comes with the caveat that currently it's a bio script. There is also a docker, one huge docker container for it, but it's only able to utilize one node at the moment. So it's hard to scale up. And that's why I then choose to implement the NFCore Punchinum pipeline. And here in the middle is its core workflow and it's basically directly disabled for that I just showed you it's the PGGP workflow. However, now I implemented some new features to make it faster or more scalable across the cluster. So one major bottleneck when you have a huge amount of input sequences are the always alignments because they are quite tragic. So what's possible now is that we just do first and almost all approximate mapping. And then we split these into chunks, let's say for example 20. And then for each of these chunks we can run this the heavy base level alignment and parallel across nodes of the cluster and this then leads to a much shorter runtime for this step. All the following steps then are as already presented. Another interesting feature is what we call community detection. So when you have like an input of an organism and you know it has comes with let's say eight chromosomes but you input all the sequences from our chromosomes and you did not know in advance which sequence belongs to which chromosome. The idea is that we can do this automatically. So here what we can do is that we do an all versus all approximate mapping. And then we translate this into a network and from this network we can apply this light and clustering algorithm to then detect all our communities so that by expectation we expect eight communities when we input eight chromosomes right and so for each of these communities we then can execute the whole workflow again. Once all of them were run through we can combine them again back into one huge graph. So they all stay in one file. And we can calculate the statistics and visualizations of the final graph. That's how the pipeline works in general. Let's take a quick look at what comes out in the magic issue report because I think it's a little bit more customized compared to what you usually observe in in your daily pipelines it was like R&B, C or so forth. So I implemented an Archie Maticus module, which basically takes the Archie stats table and then neatly visualizes certain features of the graph like the name, the length, the nucleotides, the number of nodes, etchers, paths, and number of components. So assuming now we as input we had given an organism with eight chromosomes and the humanity detection worked really well then you would expect something like eight components here because you would have eight distinct graphical components in your full graph. So first what you can also see is the number of bases and what you can also optionally do is like add here something like present and or present CG content if you're interested. The report also comes with lots and lots of visualizations so that you can better understand how your graph looks like and if it actually made sense what you did and maybe if you need some parameter tweak it or something like that. So here on top here is a so-called compressed 1D visualization and here the idea is that we collapsed all of our paths together into one row and then indicate by color coding like a heat map where a huge amount of paths actually have that sequence like here in blue or here in red. This basically means that this is a very unique sequence and this is not seen so often in the whole graph. And actually you have like thousands of paths later in your graph. This can give you a great overview of where homologous regions are or where are really unique regions on your graph. This default 1D visualization I already told you about. And here the 1D visualization is colored by path position. This means light gray is a very low nucleotide position within that path and black basically is the highest nucleotide path position. What's interesting here is that this path compared to all the others actually goes the other way around. And so it's the reverse complement. And this is because it apparently was assembled in a different way, like maybe from a different chromosome strand compared to all the other sequences. Here the information is basically the same. So all of these nodes are traversed in forward orientation, except here for this one it's traversed in reverse and that's why it was colored in red. So the 1D visualizations that are by default in the Maticus report are visualization by node depth. So Cray means that the node is visited once, but here in red actually it means that this node is visited twice so this helps us to detect like repeat regions or otherwise complex regions in the graph. As we have seen in the statistics table we have some ends in the graph and we can also highlight them in Cray and so specifically apparently only this path has some incident which are then here to get then the novelty of the topology of the graph. We also have a 2D drawing in the Maticus report. Now let's take a look at some real life examples when we actually applied the pipeline. I did that to an organism called Lodolomycesium longisporus. It's a very underestimated yeast fungi pathogen. And like when old people with a weak immune system somehow are infected with this they can die within days and so this can be quite an issue. Like Lodolomycesium longisporus short Lodolome comes with 8 chromosomes and some MTDNA. And the input of the graph that I will show you are 11 assemblies from this winter wet lab school this year and they were generated from nano point Illumina data. Two of them were fully assembled. And the other nine ones are still on the contic level basically. What I first did is that I applied the pipeline in this community detection mode. And the good news was that hey, we have 8 chromosomes plus the MTDNA and we also get 9 humanities. So this is amazing. And also most chromosomes are linear. However, some of these have these very long thin tails like here or here. And this is unmapped sequence. And this was somehow strange and not ideal. Also this chromosome B and especially chromosome H are not very linear and so I was not happy with the result. So what we tried next is something called reference guided community detection. What we did here is that we took all of the reference sequences which are each one sequence per chromosome. And then we mapped or aligned all the contics of all the other sequences to that reference and then we were able to actually place each of these into their respective community. And so for each community I then ran NFCropanogenome. This is the result and in general, most chromosomes now look much more linear. Even chromosome B looks beautiful now. Our chromosome H still looks messy. And so the next step was then that we got back to the assembly team. And they actually told us, hey, we see some interaction between chromosomes C, H and G. And so we just put all of these sequences together into one graphical component and voila, we have our beautiful graph here actually. We currently think that this is due to some RDNA region which somehow forms this interaction of these different chromosomes. I also told you something about cluster scalable. So let's take a look at an E. coli graph of over 2000 sequences. Here the quadratic or versus all alignment problem actually becomes a problem. So I ran WF mesh map it takes one hours 30 minutes that's really good. However, when I do it WF measure line, it takes 1000 times 20 minutes, and it generates over 600 gigabytes of path files. So this means that subsequently unsurprisingly sequished and run out of scratch disk space. And so I thought maybe I can actually use our network storage to build the raw graph, but it was so slow, it wasn't doable. So luckily there's an option WF mesh that allows us to retain only a subset of all the mappings. And it is configured in a way that although we get less mappings for the base level alignment step. The idea is that we will still get one huge graphical component so we can still get a really decent graph of it. And after doing this WF measure line and took 100 times five minutes so much faster than before much less resources required. And now with the default settings now see which complained about not having enough RAM. So there's another parameter there, which I decreased by two orders of magnitude, basically the idea is that it uses less RAM but it takes longer. And so after five hours, I had my raw equal and functional graph. So actually, on such a huge graph can really take its time. And so after 62 hours, I did one round of smoothing, and I couldn't do more rounds because of the seven days time limit on our cluster so here. So there's still somehow potential maybe to improve the tool smooth XG itself to make it more cluster scale. There are ideas for this but it directly involves to code in C++ and not to some optimization steps here in the NFCore function graph pipeline. So you can see the result in graph. And again, we have this compressed mode and you can see a huge amount of links so all of this black lines, each of them is an edge and because we have so many edges in this graph basically everything becomes black here. And that's because the bacteria interchange and change their genes with each other, right, huge amount of times. And I also put the genes at different positions in their respective genomes. And that's why this is such a mess. And that explains this in 2D this Harry ball down here because they have so many interactions between different regions of the different E. coli genomes that it's really hard to to see what's going on. And for sure here for example is that there's the like the majority of the sequences in this punch them are actually covered by, let's say basically all of these sequences, and then there are some unique sequences or roughly unique sequences in there. But you can also see here on the left hand side in this 2D visualization are these tiny graphic components. These are sequences that did not align to any other sequence in the sewage plot here. Yeah. I think that's all I wanted to tell you and I want to think all of these people here they all help me out a lot. And yeah, thanks for having me and thanks for listening. I'm happy to take questions. So, I am now allowing allowing people to unmute themselves and also to start a video so if there are any questions please ask them now. In the meantime, I was wondering what is happening after this pipeline let's say you have your pan genome what are you going to do with the data and yeah what is it going to be used for. This is a good question. It, and I think it also depends on the use case so for example with the landlord data you can take a look at all these SVs and how the chromosomes interact so if you're interested in chromosome interaction. This can help on last structure variation detection but also you can take the graph and it will improve your mapping for example if you the agent human is really diverse. So if you have short reads and specifically map them to a punch in a graph which already has lots of variation in there then it can improve your mapping, for example. And you can do sort of all kinds of things in downstream analysis with a tool called Archie. You can detect complex regions or, for example, what you can also do is then create a function at the tree, for example to get an idea of how close the sequences actually are that you, what you've created a punch in a graph from. Yeah. Thank you. Are there any questions from the audience. It seems everyone is happy. Then I would like to thank you again for this really nice talk. I would like to thank the audience for listening in and as usual, the John Zuckerberg initiative for funding bite says talks. So, thank you everyone. Thank you.


You're going to get to make something that's called the big production. It's going to be a production of some sort that will encourage young people to be more active or to eat healthy. Stand by and action. I'm Saida Gilley, but you can't call me Albra. And I'm here with Honey Johnson to have an interview about school lunch, nutrition and fitness. You don't find it. You don't find it. Remember when someone else is reading the line, you have to be very quiet because your mic will pick it up, okay? One, two, three. Let's go shorty, let's go. Let's go shorty, let's go. This message is about not to starve yourself or stuff yourself. What should we eat? Eat fruit. Acid, vitamin C, yellow number six, niacinamide. What's that? We're going to be going to a grocery store for a scavenger hunt. We are going to do some exploring at the market for what is good for you to eat. When my group first entered Rick's Market, we were like, wow, how are we going to do this? But finally, we found our way around.


What is going on guys welcome back to the channel and I do appreciate you being here today I bring you another destiny 2 video and today guys I want to bring you this short video showcasing a new trailer under Lumina Which is an exotic hand cannon which quest starts tomorrow also guys bring you these perks It's intrinsic perks while it does and so much more But before we go any further guys if you enjoyed the video I would like to show you support you can by hitting that like button if you are new around here and enjoy Okay guys so bungee just dropped a trailer for this new exotic the Lumina check it out on screen now Okay, so wow then how epic does this thing look now? We already know from the trailer that this thing is going to be Basically a support weapon much like the reverse version of the phone and to be honest a support weapon is kind of something Unique to what we are used to in destiny. That's for sure now I do have further details on this weapon including the type of hand cannon it will be and the perks it will drop with so Let's check them out on screen now. We can see stats. We can see it's a part of a 150 rounds per minute hand cannons same as the phone in fact when stats are compared to the form which you can see on the Screen now there isn't much difference between the two So what about each perks well, it's intrinsic is called noble rounds kills with this weapon leave behind remnants Absorbing a remnant convert your next hit by a shot into an ally seeking noble round and partially refills the magazine And its trait is blessing the sky Using noble round on an ally heals them and grants both you and them a weapon damage bonus for a short duration So yeah people this is definitely going to be a support weapon And it's basically the exact opposite of what we know the phone is which is cool We know the weapons are kindly related with lore and tomorrow I expect we learn much more about the Lumina with the quest and how to obtain it Which I'm looking forward to I'm also looking forward to doing tests do these remnant stack Can multiple be attached to the same one garden increasing their damage even further Well tomorrow people if the quest isn't time-gated, which I don't think it is we will see now I have made a video on the entire quest line Which was leaked a while back if you do want to check it out you'll find it linked within a video description I'm pretty sure Bungie did state that the quest for this Lumina will start in the EDZ and my video on this covers every other step In which I do believe the weapon starts out to be a standard white hand cannon And we kind of build it up into the Lumina, which I'm looking forward to doing tomorrow it lands and right here on my channel I'll bring you a full guide into getting it so stay tuned for that but on that note Let me know what you think about a Lumina and how it seems to be a support weapon But on that we have come to the end of the video guys I hope you enjoyed if you did leaving a like really does help out if you're new around who and enjoy daily destiny videos Be sure to subscribe and if you never want to miss a video upload a content notifications and by hitting that bell button But guys thanks as always for stopping by hopefully you enjoyed the video and hopefully I will see you on that next one