 All right, so thank you for joining us, everyone. Welcome to this webinar. What I'd like to do is give you some information about this RFA for non-human primate developmental genotype tissue expression, or NHPDG text. For those of you who don't know me, I'm Jennifer Troyer, and I work for the Division of Genomic Sciences at the National Human Genome Research Institute. And I will be presenting today, but I want to point out that this project is a collaboration from several institutes and centers across NIH. And so first, I would like to introduce all my colleagues that are on the call. So if you can show your video for a few minutes, I will introduce, so there are several of us from NHGRI. We have Simone and Joti. Simone is the program director for GTX and Joti. For GTX and Joti is the NHGRI lead on the developmental GTX in humans. Ali will be moderating, and Lori may be joining to help her in a little while. And then Gerald and William are our IT support and are making sure that all this works and goes smoothly. We also have colleagues from NIMH, Tara and Xander. So welcome and thank you for joining us. And then the Office of Research Infrastructure Programs has Ciggy and Sherry, who I'm sure you're all familiar with. And finally, we have support from NICHD, our colleagues, Tuba, John, Bettina, and Melissa. And so during the course of this presentation, you can ask questions at any time. Your chat is disabled. However, your Q&A is enabled. And so all you need to do is type a question into the Q&A, and either one of us will answer it, be it the Q&A box in the chat, or we will, Ali will be asking some of those questions later on for me to answer at the end of the presentation. And then once the presentations are over, you may also raise your hand. And somebody will turn on your mic and allow you to ask the questions verbally. If you have any questions, you can contact Ali at ali.osgood.nih.gov. And I want to point out that this session is being recorded. So today, I'm going to give you a little bit of background on why we are putting forward this program at all. And then we'll go into some of the specifics of the RFA and what we're looking for. I will try to address some of the questions that have been asked frequently so that we don't we can cover those. And then we'll move on to any questions that you have. I want to point out that there's no need for this to take the full amount of time if all your questions are answered during the presentation. That's fine. But I'm also happy to stay on as long as needed to answer as many questions as you have. And then finally, as always, you can always send an email and we'd be happy to meet with you if you have very project-specific questions. So what is the purpose of non-human primate developmental genetic expression? And this is to study gene expression patterns in multiple reference tissues across many developmental stages in non-human primates, compare them to human expression patterns. And so the overarching what is the scientific basis, the reason that we want to do this is to determine common developmental networks and pathways, to identify patterns that are primate or human-specific, to establish widely used, we hope, data sets and samples for comparative developmental genomics, and also to inform model selection for follow-up studies from some of the other GTEX projects. So this project will sit in the context of a hopefully history and future of other genotype tissue expression projects, starting with GTEX, which was a common fun program, which is now complete. The recent paper, and as Simone will tell you, more individuals and tissues were collected, but the recent paper used 838 post-mortem adults, 49 tissues, all with DNA and RNA-seq available. This has really led the way in our understanding in humans of tissue-specific gene expression profiles, and also expression QTLs, and is one of the most highly used resources that NIH provides. So following on the success of that and the utility of that, NHGRI, NICHD, NIMH, and NINDS, we recently collaborated to launch the developmental GTEX, which is really to fill this gap of knowledge about development in terms of these profiles. And so that project will be using at least 120 infants and children looking at at least 30 tissues, DNA, bulk sequencing, also adding some single cell sequencing. And this will fill gaps in our understanding of developmentally regulated tissue expression and the impact of variation on development. So in order to expand the utility of both of these resources, we're now bringing forward the non-human primate GTEX so that we have some comparable data in non-human primates. And this is a collaboration on the call, our NHGRI, NIMH, and LRIP. But as I mentioned, NICHD is supportive and is providing us with appropriate scientific advice and expertise here. And the gaps that we want to fill here is to provide the information for the experimentally tractable laboratory animals, look in prenatal as well as postnatal developmental time points, and also build on the vast amount of developmental knowledge that it really exists for model and other organisms. Thinking about that, when we think about model systems, there really is this huge amount of information for some of the simpler organisms, including self-faith from start to end of development, some very complex and beautiful regulatory body patterning mechanisms. And then as we get closer to humans in evolution, we start to look at some fairly detailed information on organ and organ system development. However, none of these really are exactly comparable to humans. And we wanted to move at least closer in terms of model organisms. And so I don't think I need to speak to the advantages of non-human primates to this audience. If you're considering applying for this, you already know all this. But I do want to encourage you all in your applications to really think about how you're capitalizing on these advantages to inform human biology and as well as NHP biology. So what are the basics? We're proposing to fund one or two centers. And I will speak a little more to that in the frequently asked questions part of this. Similarly, the budget I will speak to a little more later. But what we currently have set aside for this is 3.5 million per year for five years. And what we want to get out at the basis of it is really this expression data for multiple developmental stages and multiple different tissues for multiple different species. And again, I'll get into this a little more later. But there is the opportunity to propose some additional assays. And we hope that some single cell data will be incorporated as well. Again, we'll talk about the balance of those things a little later. And then this is a resource. It's a U24. It is very important to make data available and usable to the community. And because of the cost but also the animals themselves, we expect that there be real thought to how we can minimize the number of animals that need to be sacrificed and that opportunistic and bank samples are used whenever possible. So overall, what we are envisioning is that the ideal center or centers will do the following things. They'll provide samples and data for a very systematically collected set of the grid of species, at least two. And these are minimum numbers, not maximum, at least two, Old World and New World or appropriate evolutionary distances. We would like at least six stages that span both prenatal and postnatal development, 12 animals at each stage, male and female, at least 30 tissues. Now, some of you have asked me in this. I'll also address and frequently ask questions. But I'm going to mention it here as well. If you don't have exactly these numbers, is the application going to be rejected? And the answer is no. At the end of the day, what we want to do is build the best resource with the best combination and systematically collected grid we can get. But you have to tell us in your application what's realistic, what you propose to do, and why, what's the justification. And after review, we'll figure out how we put together the best resource that we possibly can. We do require that there be whole genomes for each animal, whether you generate that sequence or use animals where that sequence already exists is up to you, and that for each tissue collected in this grid, there be RNA-seq data. Other assays will be value add, and we can talk about that a little more later. And then finally, that data should be managed. QC, some basic analysis, and then submitted to appropriate resources. Oops. So now moving on to the actual RFA. Again, the purpose is to create this resource of multiple reference tissues, cross-developmental stages in the non-human primates, which requires that transcriptome sequencing be done, and then other genomic analysis of single cell and bulk tissues, and gene expression patterns be examined in both the non-human primates and compared to human gene expression patterns. And I'll get into a little bit about how that will work as well. And then, of course, as much as possible, making all of this available to the broader community, again, as a resource. So back to this question of what will we reject. And the answer is anything that's coming in the spirit of accomplishing the scientific goals we're going to accept. But if you're not leveraging, for instance, evolutionary distance and thinking about the comparative genomic aspects, that will be non-responsive. And if you are not putting forward clear plans for how this is a good resource for everybody, that also will be not responsive. And finally, if you're not thinking about how it can be integrated with human data to be useful and informative in that regard, that would be considered non-responsive. Everything else, we will accept and review. What we want you to bring in and what we want you, what we want reviewers to assess are a really thorough and complete research plan. And so all of this is in the RFA, but I'm just going to go over the headlines. There are detailed paragraphs for each of these in the RFA that you can go back and read. But you should have a high-level strategy for generating useful data and really think about how it complements both human data and data from other model organisms and how this is filling a scientific gap that we have in what's accessible to us. There should be a description of the structure, including the management and reporting. This is going to be a rather complex. You're being asked to do sort of nuts to bolts from sample collection to data sharing. So how does that all fit together? How do you work together? If you're working with other people to bring in additional species, how does that work? All of those things. A justification of the overall study design. So this is where you want to make sure that when you're talking about those samples, developmental time points, species, and tissues, you have a set scientific justification for the choices you're making in each of those dimensions. And if you're adding additional assays, rationalize them. If you're adding additional species, rationalize that. Whatever you're putting in there, just have a strong scientific justification for how this is going to help the overall goals of the project. Obviously, a plan for producing very systematically collected, well-qc'd data samples and data, that's the resource that you're providing. So how is that going to be tracked? How is it going to be QC'd all of those things? Again, the genomic data to be provided systematically. If you're doing it in multiple different places, what are the controls that make sure that that data is comparable and they're not batch effects, all those things on data analysis, data management, and then how that is going to be integrated with other resources, how it's going to be usable. What are your standards, the metadata being provided, all that, how does it all fit together? And then finally, we expect that because this is going to be a community resource that you are engaging in working with the community that you're providing the resource for in order to make sure that it is useful as possible. So plans for community outreach and engagement. I'm going to move on now. So we have all the standard NIH review criteria. There are specific questions under each of them that we are asking reviewers to consider. And I think this is always useful when you're writing your application to have these in the forefront of your mind, is what are these, what am I writing? How is that being evaluated? And so in addition to the normal significance criteria, we want to know how the scientific knowledge gained is going to address the research gaps for understanding of development. Again, this is going to come up again and again. Is this a community resource adequate? Do you have a plan for providing this useful and usable community resource? And will it be enabling? For the investigators, the additional criteria, this is going to be a fairly large project. It is going to be part of a consortium. Again, I'll touch on that a little bit later. And so is the PI prepared to work in that mode, essentially? And then will they be dedicating sufficient time to meet the needs? For innovation, we really think, because this is going to be a unique data set in that it can be integrated in a lot of different resources that already exist out there and should be interoperable with a lot of data from lots of different species, that there is going to need to be some innovative approaches to sharing this data and also doing that cross-species analysis. And so to meet those goals, is that piece there? Is there a plan to adjust as you go along, as new technologies become available? And then are there effective and innovative plans for that outreach to the community? And then here's the nitty-gritty as the approach. So are we talking about effectively using comparative genomics approaches to understand the scientific problems? Is there a good approach for that integration with human DGTEX, understanding that some of that will come as the two projects move along in parallel and work together? You won't necessarily have a full plan for how it works, but that will be something that needs to be developed in concert. Is there an overall structure so that this project is operating really as a cohesive whole and not little parts that aren't talking to each other? And again, this comes back to the species, animal, stages, tissue types, and assays well-justified. And will there be as much effort as possible to maximize these really valuable samples, maximize the utility of them, whether that's making them available to other people doing additional assays on them? There are lots of ways in which the utility here can be maximized. Your plans for DNA and RNA sequencing, good. And if you're proposing other assays, are they the appropriate ones and the appropriate tissues? Are the tissues you've chosen well-justified for that particular part of the study? Is there an adequate plan for QC and having these data available and useful to the community? Again, I come back to that a lot. That's going to be asked a lot. Does the data management plan good? And will it facilitate interoperability with other resources? And is there opportunity for collaboration and interaction? And again, are you thinking adequately about engaging the user community? For the environment, the request for the environment is that this be done at a place where it's ready to go from day one and does not need a lot of ramp up time. So now I'm going to talk a little bit about the interoperability with the Human DG Text Project. This is a project that will be launching soon. But the non-human primate will be joining that consortium. There are two components there. There are tissue procurement centers that are managed by NICHD. And then there is a laboratory data and analysis coordinating center that is managed by NHGRI. So really the non-human primate centers will be doing analogous functions to these two different components in the Human DG Text Consortium. So they're working on collecting tissues, creating a tissue resource, generating the genomic data, doing QC analysis and management, and making it all available to the community. And so we expect that the non-human primate centers will work with the human centers as much as possible to make sure that these processes are analogous, that there's good ability to compare the data at the end of the day. And so all the data elements and standards and everything have to be working together. And then the other thing that I wanted to point out is NIMH is a partner on this call. And they are interested and may consider funding applications that have special consideration to some of the work in the brain. They're already funding a lot in non-human primate brain research. But if there are things that these projects are proposing that are very complementary and integrative with what they already fund, they may contribute some additional funding for that. So there's, of course, special considerations when you're working with the brain with the procurement, making sure that there's an expert on hand who really understands the different parts and can think about the cells properly and preserve the materials such that the single cell analysis can be done. This requires fast harvesting, for instance, and very fresh tissues. And then also in the sequencing analysis, really knowing what's already going on in both human and other primate brain resources and being able to work with them and integrate that data will be useful as well. And I know there are multiple different brain regions and brain cell types that will be of interest. So if you want to bring that expertise to the project and elaborate on that, that is also something that you can do. And we have folks from NIMH on the call to answer your questions about what they might be looking for in that regard. And then finally, I want to come back to these considerations of balance that we all struggle with. So with limited resources, when you start to get multi-dimensional, projects can get very large, very fast. And we want to keep things tractable and affordable. So thinking about for every developmental stage that you add or number of individuals you add or number of tissues or assays or species, that increases things exponentially. And so thinking very carefully about what is the right balance of these different things will be important. And as I said, justifying them well. And then in the realm of providing data and samples when there aren't a lot of resources available as much as possible, leveraging existing resources and integrating with those will be encouraged. But there might be places where something just doesn't exist and you want to create something new. So again, that's something that you just have to think about in terms of the balance of this application. Again, what we and NIMH are looking for is the resource that will be of most use. And so some of that is going to be provided by the applicant. Some of that is going to be provided by the reviews. And some of that is going to be how we choose to put together the program at the end of the day. And so that brings me to some of the frequently asked questions. A letter of intent is not mandatory. We appreciate it because it helps us to start thinking about who reviewers might be and manage conflicts early. So if you have not sent a letter of intent, but do plan to apply, if you could just send me an email with the people involved and what institutions they're at, that would be helpful. It's not necessary. It's an open competition. So anyone can apply. The one restriction is that it is not open to foreign applicants, although you may have foreign components. A lot of people have asked if we're going to once the data is there, will there be RFAs for analysis? And the answer is probably so you can keep your eye out for future calls for those. And this is a resource EU24, but can there be a research component? And the answer is yes. We hope this data that you would be generating will be useful to you and your research, as well as to the rest of the community. And so having something in there that is proof of principle for the utility of this data, if it's not a huge part of the cost will be accepted as part of this application, it is not required. So that's up to you. The 3.5 million a year that we currently have committed is for total costs. That's what we have in hand. It is not direct costs. It's total. But then the follow up question is can you submit applications that request more than that? And the answer is there's nothing stopping you. It says to ask for the amount that you need to complete the project. So as you all know, our budgets are year by year. We don't know exactly what our budgets are going to look like next year. As you also we've all learned in this year, we never know when our priorities are going to shift. So it's possible that when it's time to make the funds, the funding decisions, we have more money to commit. It's possible we have less. So this is just what we're planning on. I will say, if there are seven applications that get really good review scores and reviewers like them and they come in at all different costs, we're going to select the one that's the most efficient and will provide us the most bang for the buck. So you want to keep the cost reasonable and down. And I would suggest if you're thinking of asking for more than that 3.5 million that you really be putting something for that extra money that is value add to the basic resource that we've asked for. Because that's what we have to spend on the basic resource if we are going to spend more it is going to be because there is significant value add to that particular proposal. And then how many applications will be funded? Well, again, that's the funding envelope we have targeted right now. So it is one complete resource. We might make that complete resource by putting together two different centers if they have complementary strengths. And so you could see that happening in several different ways. Maybe there is, one group has great samples, not so much experience with the analysis. Another group has great analysis, but is lacking in samples. We might put those together. You could see it running by two different species coming together. There are lots of ways in which we might put together two centers. But we're not going to double the amount of money that we're putting into it. So the fact of the matter is probably it also then would be at a reduced funding level. So just thinking about that. But we want, again, our programmatic priority is to put together the best resource that we can. And so we'd like your help with that. Just to take you to the page where you can find this funding opportunity if you have not yet read it, you certainly should. And the application due date is coming up. I realize this is a short turnaround time. It's what we've got. So if you want to have a call with us or send specific aims any time before that, you're welcome to reach out. And then finally, information will be on the developmental GTX expression page at genome.gov as we move along. If there's anything to post, we will post it there. And with that, I think we will move on to our Q&A. Allie, do we have any questions in the Q&A? We have one question. Someone asked, since human DGTX can leverage adult GTX, it would seem to analyze data across the non-human primates to human lifespan, at least one adult time point is needed. Is that correct? So I think that's a really good question. And I think that's up to you in your application to decide if you think that that's necessary. I think I would also refer you to what's already available and already might be being done through other non-human primate resources. So we are not requiring an adult time point. If you want to put in a strong scientific justification to use one, that's certainly one way to approach, again, this multi-dimensionality, which could be expanded or contracted in many different ways. I see that Siggy's on. Siggy, do you have any comment about that or thoughts about that? And it's fine if you don't. Yeah, I don't have any additional comments. I think it's fine. I agree totally with you, since this is most focused on development. And adding the devider about the time points might be challenging in terms of how many time points you can cover, the developmental stage. Exactly. So it's a trade-off. It's always going to be any of those points or a trade-off. Any more, Ali? Yeah, there are a couple more coming in. Someone asked, as the genetic heterogeneity of animals in centers is less outbred than other populations, is this a concern for 2TL-based analyses in non-human primates? Yeah, that's also a great question. And I agree that that is a limitation. That being said, I don't think we're asking at this point to go out into natural populations and collect that variation. And I'm thinking as well that our numbers are not going to be probably high enough to really address the EQTL issue, although there might be some opportunity to do some preliminary work with known human EQTLs to see do we see the same trends in non-human primates? For instance, if there are some variants that overlap. But I think that we're not asking you because we are not providing sufficient resources to really get at that EQTL question. However, as much genetic heterogeneity can be managed, we'll be helpful for planning future studies, I believe. And then someone also asked, since this is a transcriptal resource, what is the expectation for the required whole genome sequencing? So I think that it's still important to have the underlying whole genome sequence. And like I said, I think for some primate centers, some of that is already available. I think that you can make the case that perhaps genotypes data is sufficient. So this is a little bit of that. It's up to you to decide and justify. But we do definitely want as much information about the underlying genome so that the transcriptome data can be interpreted correctly. And someone asked if the use of banked tissues are discouraged or would hurt any proposal because of potential quality control issues? So I'm going to throw that one to the reviewers to decide for you to think about and in an application to, again, explain that trade-off of using banks or samples and how much you know about how well they were preserved and what the quality will be and how are you going to monitor that quality to make sure that the data you're getting from those bank tissues is comparable to the data that's coming from fresh tissues, for instance. I mean, all of these are practical considerations and can be managed. So we actually are encouraging you to maximize use of tissues that you have if they are appropriate for the study. But that has to be determined by you. And then we have a few other questions. Someone asked, how would you recommend you coordinate with human DG-tex? So I don't know if Jyoti was able to join. She was somewhere where there is not great internet. So it looks like she was not. So I'm going to do my best to answer here. So you will be part of the DG-tex consortium and expected to attend meetings. There will be probably some working groups that do not involve the NHPDG-tex, but a lot of working groups will involve members from both human and non-human. And so the coordination will be expected to be pretty close. It will be a consortium. And then another question that just came in. Is there an expectation that the investigator team will include investigators of existing National Crime research centers? So again, this is a question. And by the way, I just want to encourage all of you, if you want to ask your own questions, you can raise your hand and we can unmute you. So you don't have to use the type in. You can also raise your hand and we'll go back and forth between the two or something. But so Sugi, I see you've taken off your video. So do you want to answer that one? Yeah, sure. I can answer that once. My suggestions would be, actually, I think there are some expectations of including the investigator from NPRC. And the major reason is because these NPRC people who investigate are very familiar with the non-human primate and also to have very high standard operation procedures to collect the high-quality samples. And then also some of the investigators are highly involved in a genomic sequence of a few of the non-human primate. So there's a lot of experience and expertise that the programs can leverage the non-human from AHP Research Center. Yeah, I would totally second what Sugi said. I mean, it's not required. This is, again, something for you to propose and for reviewers to evaluate. But it's hard to imagine a project that isn't leveraging those resources in some way or another. So that's my thought. I was checking to see if there are any raised hands. And I do not see any yet. Someone also commented, the RFA tends to indicate a desire for single cell analyses on all samples. But it does not seem that this is an absolute requirement. Do you want to? Yeah, so and I might ask Tara or Xander to pipe in here about the requests for brain single cell. But certainly doing single cell on every tissue for every animal would be cost prohibitive. And that's not what we're asking that you propose. I think as much single cell is reasonable and affordable and targeting places where it's where that data is most informative is what we're asking for. And then I'll turn it over to Tara for thoughts on brain single cell, which is specific. Yes, so for NIMH, what we would consider a value add where would be that a single cell now single cell proposals for the brain that coordinate on a level with what we're producing in the BICCN with NINDS. So if the cost NIMH is looking at these applications for what they're adding to the brain and considering whether to contribute funding based on what is proposed for the brain. But our commitment is not part of the money that is already committed. So if you are strongly considering adding a section of the brain for single cell and working on that as a strength, that would be a value add sort of proposition for us. So that would be one example of where you might go over the costs that we've put in there thinking that if it is sufficiently well justified, if it reviews well, and NIMH sees it as this value add, they might contribute some additional funds on top of what's already there. So just to restate what Tara just said. So thank you Tara for clarifying. And again, I think that that is if you want to make a case for pursuing particular other organs more deeply and want to contact other ICs and see if they would be interested in also supporting that work, that is another way that there might be some additional funds available for some of these additional studies. So within the funding envelope that you have, think about what is the most valuable single cell. You can also propose additional and we can see if we can find funds for it. I hope that answers your question. And it seems there's sort of a follow up to this. Someone said, we have access to excellent frozen tissue samples for adult chimpanzees. Is there any value to include single cell RNA seek from brain and just this species? So if you're asking me personally, not as a representative, but as a scientist, I think sort of almost anything that you could propose along these lines would be value add. So I think that again, this is your scientific thoughts justification, bringing that in, seeing how reviewers felt about it. It could certainly be a value add proposal. And I'm just going to answer. There's another question about micro RNA and link RNA data sets. And again, that's similar. It's not in the basic request of required elements. However, it could be considered a value add so scientific justification for why those would be important in the athlete. You can put them in the application. It's not precluded, but it's not required either. And then there's also, can we make the slides available? And actually, this whole thing has been recorded, and the recording will be made available. It looks like there's another question that came in. Someone said, we have fibroblast cells in culture for many primate species. Would characterizing these be a value add in addition to the stated goals? OK, so same answer as before. It's like, scientifically, I find all of this very interesting whether it would actually be funded as part of this remains to be seen. I certainly think that if you find that compelling, it's worth putting the argument for it in your application. Again, as value add. The other thing that I would mention just for all of us to think about is, again, we have limited resources. Whatever that envelope is, it's never going to be as big as we want it to be. So is it because you could do a million things on any given tissue? So as much as you're also thinking about storage for future use, if additional funds come available from either us or other sources, as much as you're thinking about, again, how do we make this resource as useful as possible considering we're using these animals? That will be an important component to this. And it looks like someone has a question related to review. They said, given that tissue sets will be negotiated to harmonize with the funded human DG tax, how will reviewers be instructed to score the proposed tissues in the proposal? So I think that's a really good question. And that's a question that applies, of course, not just to the tissue harmonization, but to the harmonization across the board between human DG tax and this. And I think that the reviewers will be instructed to consider the value of the resource, but also how well the applicant has described their ability and willingness to work with the human resource to make sure that those are as comparable as possible, knowing that they won't know exactly what's happening in that resource yet. And that some of that remains to be seen. So is there a good plan? And does it have sufficient flexibility to be interoperable? I'm not seeing any more questions coming in right now. And I'm also not noticing any hands being raised. But in regards to the earlier question about slides being made available, where would oh, and I see John is raising his hand as well. But in regards to where could people find the slides once they this information is available? So those will be posted on the developmental GTX page at genome.gov. And we can put the link, I think, in the chat for that. Gerald has done it. And there's a hand up, you said? Yeah, John is raising his hand. John, go ahead. Yes, I'm sorry. You know, I'm having a problem with the mute button. Since the RFA single cell seek is not required, wouldn't it be wise for the applicants to consider storing samples for single cell RNA seek for future studies to correlate with developmental GTX of human? Oh, absolutely. And I would say I would encourage everybody to consider some single cell sequencing. I just acknowledge that it's not going to be possible to do the amount that you might want to of all animals, all stages, all tissues. And so selecting judiciously will be important. But single cell certainly goes along with everything else as how can we maximize the utility of these for the future? And so if they can be processed and stored to do later single cell analyses, that is ideal. Do any of my other colleagues have anything they would like to add at this point? Tips, things that your ICs are particularly interested in seeing? Well, I thank you all for joining. I am going to stay on until everybody drops off. So if you want to ask additional questions, you may. You're welcome to stay. You're welcome to leave, whatever. And like I said, I am, of course, happy to talk to any of you on a one-on-one. Some of you I already have. I recognize some names. So hi. And if you want to send some draft-specific games or have any questions that are specific to your application, just send me an email. You can always get to me at jennifer.troyer.nih.gov. And we can put that in the chat as well. And I hope this has been useful. And it looks like an anonymous attendee said thank you with two exclamation points. All right, so it was useful for at least someone. That's good. And thanks, everybody, for joining and helping. I really appreciate it.