We'll go ahead and get started on time here, and I'll have a few minute introduction while the stragglers walk in. Really, really excited to welcome Dr. Scott Edwards to NIH and NHGRI today.  And according to his intro slide, also the department chair, so juggling many hats. Also the curator of ornithology at the Museum of Comparative Zoology, which I assume is a Harvard institution. Originally, he received his BA in biology from Harvard some time ago. And so he ended up back at his original stomping grounds. But in between, a PhD at Berkeley, a Sloan Fellowship at University of Florida, a couple visiting fellowships around the world in between before eventually winding back up at Harvard for the past 20 years or so. And not just an excellent researcher, but also an excellent educator. And Diana Proctor just mentioned to me that he has some pop-gen lectures that are on YouTube that are some of the best lectures Diana has seen on the subject. So if you'd like a good YouTube lecture taught to you by an expert, check out Dr. Edwards' videos on YouTube. His CV is lengthy, I will say. He's a fellow of every society that you've probably ever heard of, including the American Academy of Arts and Sciences, AAAS, the National Academy of Sciences, and so on, a member of many board of directors, including the really wonderful Cornell Laboratory of Ornithology, which I've had a chance to visit there at Research Station, a really cool place, Massachusetts Audubon Society, and the Adventure Cycling Association, just to throw an oddball in the mix. If you had the chance to visit his homepage in anticipation of the lecture, you would have seen up on the splash page of his homepage as a picture of him while he was completing a solo bike journey across the United States in 2020. Perhaps running away from COVID that summer, I remember that summer well, but also raising awareness for the Black Lives Matter movement and Blackbirders Week, totaling 3,800 miles, 76 days, and a fair bit of media coverage on the way. So suffice to say a little bit of an adventurous spirit in Dr. Edwards. And in addition to that, looking on his CV, he's traveled all around the world through the years pursuing birds, doing fieldwork, collecting samples, really, really clear evidence of a life well-lived in his career as a researcher and educator at Harvard. And so looking through that inspired me a little bit to get out from behind my desk a little bit more often, because it seems every few years or so you have a fun adventure. We should all strive to do that. So I'm thrilled that today he is stopped by our neck of the woods to give us a talk on something that I care deeply about. And that's discovering the functional connections between genotype and phenotype using comparative genomics approach. So with that, let's all welcome Dr. Edwards to NHGRI. Thanks very much, Adam. And thanks to everyone who organized my visit today, Monica, and all the great scientists I've met this morning. It's a real honor to speak to you here today in this beautiful auditorium. Yeah, I thought I would tell you about some of the work we've been doing over the past five, 10 years, trying to bring together comparative genomics and phenotypic evolution. And a bunch of the second half of the talk was stuff that was funded through an R01 from NHGRI. So I'm very, very excited about that. Just to introduce you, this is my lab group. And I came into biology through ornithology. I've been a bird watcher since I was probably 10 years old, growing up in Riverdale in the Bronx in New York. And anyway, it's great to be able to educate students coming from often-organizable backgrounds, but sometimes computer science or other backgrounds.  Today, we'll focus mostly on the second to the last two squares up there compared to genomics and phylogenetic methods. And I'm very fortunate to work in a world-class museum. Museums are great centers for education and training. And it's a very interesting time now because we're seeing museums try to navigate how they can be most relevant to 21st century biology. And a lot of that dovetails with genomics. And so it's been very exciting to try to modernize, in my case, the ornithology collection, but there are lots of other collections. And so anyway, if you're ever around Cambridge, I'd be happy to show you our collection. And so it was maybe five or six years ago, a colleague and I were at a meeting of the Society for the Study of Evolution. And we had just gone to a symposium talking about the uses of phylogenies. Why do we need phylogenies? And most of the symposium focused on many of the sort of ecological spinoffs that phylogenies bring, learning about biogeography, ecology, adaptive radiation. And we were all struck, at least among this community, how little intersection there was between phylogenies and comparative genomics, or genome evolution. And we ended up writing this short perspective piece and thinking about sort of trying to make our community aware of some of the potential avenues. And there were, even at that time, some really compelling examples using phylogenies to understand the genomic basis of phenotypic evolution. I'll talk about the first two examples on this list, but there have been many others. And I think it's a very exciting time right now because we're seeing lots of different approaches coming to bear on this question. And the reason why it's important is because we can't do traditional genetics on most of biodiversity. We can't do traditional crosses in the lab. And we can't set up the traditional way of allowing the phenotype to segregate along with the genetic markers, which is what you see on that first column there in this slide. What we can do, however, with the other 99% of biodiversity is look for clades that have a phenotype popping in and out or turning on and off. And we can then associate changes in the genome with changes of the phenotype on the tree. And I'll show you some ways in which we're trying to build models to do just that. And then just like standard genetics, we can then use functional tests to ask whether the candidate genes or regulatory elements that we've identified are fulfilling the role underlying the phenotype that we might hypothesize. And so it's an exciting venture because we're bringing in lots of important clades, important phenotypes that otherwise would be not amenable to traditional genetic analysis. And there have been lots of different ways people have tackled this. And so one group doing a lot of this sort of work is Gil Begerano's group at Stanford. And they've had a couple examples where they're looking for a correspondence between a particular phenotype, which you see here in the green check marks. Those are trait present and trait absent. And the presence or deletion of either genes or non-coding elements. So these red areas here are deleted. Regulatory elements or genes. And that's proven very powerful. Some surprisingly specific associations between loss of a gene or regulatory element and inability or new ability to perform a particular biochemical reaction or have a particular phenotypic trait. Another major mode of connecting genotype and phenotype has been looking for changes in the rate of evolution across a tree. And so you might have a particular set of target lineages that you're interested in. In this case, and you'll see more of this, these are some birds, some of which are flightless. And we can look for accelerations of either genes or regulatory elements, specifically on lineages with the target phenotype. And this is, I think, another way of connecting these two and has certain advantages and disadvantages. But there's a myriad of ways in which people have tried to connect these two. And so our models tend to fall into this latter category. A lot of the way in which we've approached this question has been driven by one of my graduate mentors, Alan Wilson. He was a famous molecular evolutionist at Berkeley where I got my PhD. And he and Mary Claire King wrote this very important paper back in 1975 in which they cataloged the extent of protein divergence between humans and chimpanzees. This is using very primitive databases and cobbling numbers together from very diverse sources. And yet they came up with a number, namely about 1% protein divergence, 1% or 2%. That still holds and has stood the test of time throughout all of our era of genome sequencing. And Alan had this amazing ability to turn lead into gold. A result that a graduate student might just think is just the death knell to his or her dissertation, he twists around. And in this case, one can imagine that finding a lot of differences would have been the exciting result. But it was this lack of differences at the protein level, which inspired Alan and Mary Claire King to basically say, well, it must be regulatory evolution. And so in the different examples I'll give, you'll see a few of each type of case where proteins seem to be driving the phenotype versus non-coding regulatory regions. And so the first example I can mention involves a question that a graduate student in my lab, Maude Baldwin, brought to my attention. She was fascinated with the way in which organisms sense their environment, particularly in terms of taste.  There's an umami variety. These are two heterodimers, as well as a sweet taste dimer. And there have been a number of natural experiments in which lineages have essentially lost the ability to taste a certain repertoire because of gene deletion or pseudogenization. And so for example, pandas, whose diet consists almost exclusively of bamboo, have lost their umami receptor. The umami is essentially savory or amino acids. And so not being meat eaters, Darwin's adage that if you don't use it, you lose it. Pandas have essentially lost their gene for the umami receptors. Similarly, cats, having evolved from a highly predatory meat eating lineage, have lost their sweet taste receptor. Despite their pension for milk, they nonetheless don't have a sweet taste receptor. And these sorts of natural experiments are a very nice context in which to connect genotype and phenotype. Now, birds, which has been our interest for a long time, also evolved from a highly carnivorous lineage of theropod dinosaurs. And so we now know that it's not a stretch to claim t-rex as a distant ancestor of modern birds, and of course, archaeopteryx and other extinct lineages all having evolved from a highly carnivorous ancestor. And that begs the question, how is it that birds such as hummingbirds and other nectariver species, how do they sense sweet and carbohydrates? Because we know from looking at their genomes that they have lost the sweet taste receptor. They don't have it. And so the question is, how are they able to sense carbohydrates in the same way that we mammals can do with our sweet taste receptor? And so Maude embarked on these questions. I was one of those dissertation proposals that I was highly skeptical of and did everything I could to dissuade her from going in this direction. But she persisted. And basically, through a lot of trial and error as many dissertations go, she was able to find a lab in Japan that actually was one of the few on the planet that did functional testing of taste receptors. Happened to be sort of a satellite lab for the Kikkomon soy sauce company. And they knew everything about seeing what it was that bound taste receptors and identifying those compounds. And so she was able to express the umami receptor. The umami receptor. That was her first choice for a candidate gene for being able to taste sweet. She was able to express it in mammalian cells, both in hummingbirds and in their sister group, the swips, which we know can't cannot taste sweet, as well as an outgroup chicken, which also cannot distinguish plain water from sweet water. And she was able to show that through a series of amino acid substitutions, which are indicated by those red stars there, hummingbirds have essentially morphed their umami receptor into a sweet taste receptor. And it was, I think, a dramatic example of showing how sort of rejecting Alan Wilson's hypothesis that changes in the sequence of a protein could dramatically change the functionality of an organism and was likely a major driver for the radiation of hummingbirds, which now number over 300 species. And so following up on that work, she was interested in looking at additional instances of the evolution of neck delivery and sweet taste perception in birds. And so she went to the songbirds, which are basically comprised about half of all birds and are a really impressive radiation of species. And on this slide, you simply see all of the documented evidence for sweet taste perception, either through neck delivery or fruit delivery, quite all gleaned from the literature and quite a large number of species in many lineages from things like sunbirds and honey eaters in Australia, many different repeated evolutions of the ability to taste sweet. And so she wanted to ask, are they using the same mechanisms as the hummingbirds were? And so she was able, with collaboration again with folks in Japan, to basically create these heterologous dimers. So you see on the top there the hummingbird umami receptor in orange and the honey eater umami receptor in red. And again, hummingbirds in the New World, honey eaters in Australia and Southeast Asia. And she was able to show that when you, the native dimer, which you see here, oops, sorry, both orange and both red, can provide a robust response when interrogated with a carbohydrate substrate, whereas the heterologous dimer, consisting of both orange and red, one of those two taste receptor domains, could not. And so this suggested that maybe there was some co-evolution between these two parts of the heterodimer. And then sort of screening a more phylogenetically close subset, what we've got here are the honey eater dimer on the top here in gray and red. And then on this axis here, you've got a series of different songbirds and members of the essentially perching birds, which are the larger clades. So songbirds are these four down here. Perching birds are the ones, the larger clade at the top. And what she showed was, as you can see, there's with these heterologous dimers between a honey eater domain and these other species domains, not much of a response until you get to the base of the songbirds. This brown tree creeper, which is an Australian species, has a modest response with one of the two heterodimers. And all of the other songbirds show a quite a robust response. And this suggested that it was really the very deep origin of the second origination of the ability to taste sweet and to detect carbohydrates, which evolved very deep in the songbird lineage and wasn't, in fact, present among those top two species, which are, again, part of the perching birds and part of a subclade known as the subocenes, mostly in South America. And so showing an independent evolution from the hummingbirds and a really interesting example of functional co-evolution between these two parts of the taste receptor. Now, for a lot of the work that we've been doing recently, we've wanted to expand into the non-coding region of the genome. Just seen two examples where it's really the protein sequence evolution that's driving this new adaptation. But we know that genomes are mostly comprised of non-coding DNA. And so we looked to some classical theory in molecular evolution, specifically this so-called neutral theory of molecular evolution invented by Motu Komura and his students, which suggests that functionally less important genes or molecules will tend to evolve faster, develop a faster substitution rate. And extending that to the genome scale, we could imagine that functionally less important parts of genomes will evolve faster, have a higher substitution rate. And this is sort of the paradigm that we've begun to develop a series of models that hopefully will be of use to the community in linking rates of genome evolution and changes in association with phenotypic traits. And so we've called this group of models PhiloACC. And this is, it stands for phylogenetic accelerations. And in these models, we've basically posited a series of target lineages whose phenotypes we're interested in. And we have focused most of our attention on detecting and measuring the extent to which a given gene or non-coding region will accelerate specifically on those target lineages. So here you see a binary trait, plus or minus. And we might have a series of target lineages here. Let's find regions of the genome that are accelerating specifically on those lineages. Now that's for a binary trait. We've also developed some models which complicate the simple model in PhiloACC just a little bit. I'll go into this in just a second. Here we're relaxing the assumption that the gene tree, the tree of any given gene, is exactly the same as the tree of the species, since we know that we can have a lot of stochastic variation as genes travel through phylogeny. And then finally, we've developed a method called PhiloACCC for continuous traits. Here we're trying to associate rates of evolution with variation in a continuous trait, like longevity or body size or something like that. So I'll walk you through some of the basics of the PhiloACCC model. And then we'll talk about some of the other models. And then we'll apply them to some comparative genomics data sets. So the basic model is one in which we identify a set of target lineages based on some criteria. We're actually not modeling the phenotype specifically. We're simply identifying either target or non-target lineages. In our case, these turned out to be birds that could fly and birds that could not fly. And we imagine a shift in the so-called conservation state of every branch on this tree. And what that means is that for any gene or region of the genome, some branches, it'll evolve very slowly, be very conserved, other branches that will have accelerated. And so we can imagine a gene being in either the background state on a given branch, sort of a neutral state, a conserved state, or an accelerated state. And once we've applied those states, we can then go on to actually estimate the actual rates conditional on that state. And then finally, we can apply two Bayes factor tests. The goal here is to distinguish scenarios in which we have acceleration across the entire clade, including both target and non-target species, versus acceleration just in the target clades. Turns out that's a difficult problem. It's difficult to distinguish genes that accelerate only in these target lineages, especially when they're convergent, when they've accelerated in multiple unrelated lineages. Telling the difference between that and accelerating across the entire clade is very challenging. And that's what these two Bayes factors do, basically contrasting models where the element can accelerate anywhere on the tree, versus nowhere on the tree, versus on specific lineages on the tree. So those three scenarios. That's what M1, M0, and M2 are. So that's the basic PhiloACC model. However, that model basically assumes that every gene in the genome, or every gene that you want to analyze, has the same tree topology as the species tree. And we know now that we've been able to look at many genes across PhiloACC that that isn't true. Much of the reason why genes depart from the species tree is basic population genetics and an appreciation that if you have two speciation events in rapid succession, where this time between these two speciation events is very short, it's entirely expected that some genes will not have come to fixation before the next speciation event. And so now the challenge is when you have a gene tree, such as here, where you can see how these bold lines are, actually sorry, it's this one, where the bold lines are different from the species tree. You can see here we have species 1 and 2 are most closely related, whereas at the gene level, genes 2 and 3 have a more recent common ancestor. In this tree, they're the same topology, albeit different branch legs. But we know that when this happens, that can provide a false signal of acceleration in certain lineages. So it's important to try to distinguish these scenarios. So we call this other model phyloaccgt for gene tree. It's actually now published in molecular biology and evolution. And you can see here under three scenarios of a single acceleration or two accelerations or three independent accelerations, phyloacgt does a somewhat better job than when we assume that all genes follow the species tree exactly. This is simply a measure of performance and how many times it got the right answer based on simulations. So the downside is that phyloacgt is computationally quite a bit more expensive. But it's nice to have these two possibilities. There may be situations where we have no good reason to think that genes are varying a lot across the tree, in which case we could legitimately use the standard phyloacgt model. And then the last model I'll mention is one developed by postdoc Patrick Gemel, who is interested in connecting regions of the genome and their rate of evolution with variation in a continuous trait, such as longevity. Here's a data set from Nathan Price and others looking at mammalian longevity. And you can see here that some lineages, this is sort of the body size longevity regression. You can see some lineages such as humans and bats have a longer than expected longevity given their body size. And the trick is to find regions of the genome that can associate with this continuously varying trait. And what we've done is to take, so here's the same scenario. We have a tree, we have a trait, and here's an alignment. And these red lineages here are lineages on which we see a particular non-coding region on chromosome 5 of humans showing accelerations. And so the question is, do these accelerations are they associated with, in this case, increases in lifespan? In this case of the shrews up here, it actually looks like it's the opposite. You can see the long branch of the shrew and the lower than average lifespan. By contrast, I don't know if you can read the font, but we've got a killer whale down here, which is also accelerating, but is associated with a longer lifespan. So we're trying to parse out all these signals. And what you can see here on this panel is down here we have the particular branch of the tree that we're looking at for this particular element. The y-axis is the trait change, and the color of the element here, red or blue, indicates how accelerated it is. And so we're seeing some cases, such as this killer whale, that are experiencing a lot of acceleration and have quite a long lifespan given their body size. And that's the sort of signal we want to pull out. And yeah, this is currently in review, and we're excited about its applicability to a wider number of traits. Patrick has also tried to extend the relationship between the trait and the genome segment to be a bit more flexible. And in particular, now we have ways to where the rate of evolution of the gene may be positively associated with the continuous trait, may be negatively associated, or may show a nonlinear relationship to the evolution of the trait. And so this is sort of a null hypothesis where we're seeing changes. We're seeing phenotypic sort of neutral evolution in the phenotypic state. These gray triangles are meant to indicate the variance accumulating in the phenotypic state along that lineage. But sort of variation in the rate of the underlying genes. And so in our model, we have a Brownian motion model, which means that the variance is accumulating linearly with time in the trait. And but the null hypothesis here is that the trait is changing at a constant rate, but the underlying genetic elements are changing at variable rates. This is an example of no association between the rates of evolution of the gene and the constant phenotypic rates in the phenotype. However, we can imagine a situation where we're seeing both faster evolution of a given gene or a non-coding element and faster change at the phenotypic level. And basically, the model tries to estimate parameters of three variables, A, H, and B, which describe the form of the relationship between the genomic element rates and the phenotypic rates. This is still very much in development, but we'd love for folks to try it out and see whether it yields some insight into their phenotypes. So let's look at some examples. This is an example by another postdoc in the lab, Subir Shakhya. And he is like me an ornithologist, and so he's looked at birds for the phenotypic data. There's amazing data sets now for birds for a variety of phenotypic traits, basically for all 10,000 species. And so the challenge that we're finding actually is intersecting the genomic data and the phenotypic data, because we basically need both in order to run these models. So in this case, we looked at a trait Tarsus length. Tarsus is a very easy thing to measure, both in the field and in a museum specimen. And we basically have Tarsus length for all 10,000 species of birds, and so it seemed like a fun trait to look at. And so Subir ran a few models to understand where the big shifts in Tarsus length had taken place on the avian tree. We were actually able to put together about 5,000 species and ask just from the phenotypic level, where are we seeing shifts in Tarsus length on the tree? Here are four clades, the penguins, kingfishers, swallows, and a lesser known group called the bull bulls from Asia, where we see rapid shifts to short Tarsus length. This is just one of the terminal hind limb bones. So here's a quiz for all of you. What bird group out there should be on this slide, which has a very short Tarsus but isn't? Can anyone think of a bird that just basically has no legs? You've got them here in the summertime, so at least one species. Hummingbirds again, hummingbirds. Hummingbirds are highly aerial, and they can perch, but they don't do a lot perching. And same with swallows, highly aerial, very short legs. The reason why we missed hummingbirds is because we actually had to divide the data set up into smaller chunks to test these patterns. We used a Brownian motion model developed by Luke Harmon. And so as a result, we picked out these four clades. And there's also some very interesting examples of evolution of long Tarsus length, but it turns out that in our analysis, the number of transitions to long Tarsus was smaller than the number of transitions to short Tarsus. So we thought that short Tarsus would be a little more powerful. And so we then started to look at the genomic data, and we turned to a group of loci known as conserved non-exonic elements. These are widespread in the human genome and other genomes. They often tend to be enhancers, and there's many, many thousands of them around the genome. And so here's an example of a conserved element that is not in an exon or a coding region, but is nonetheless highly conserved. And principles of molecular evolution tell us that this region is doing something important, even though it's not coding for a protein. And these have proven very useful to look at across the genome. And so Subir put together a fairly large collection of these non-coding regions using previous data sets, as well as some new data that we had. And basically put a collection of over 900,000 of these elements. They range in size from just a few tens of base pairs to a few thousand base pairs. And you can also see we've been able to intersect them with various data sets, including some epigenetic data sets from attack-seq data, which I'll explain a bit more later. But you can see that some of them are found in exons. These are just conserved regions, whereas most of them are non-coding. Most are either intronic or intergenic. And so these are likely to be putative regulators. So when we run FiloACC on a subset of these data and target these short Tarsus lineages, we come up with a bunch of different subsets of interest. So on the x-axis, you basically see the loadings of each gene on the axis where we're testing no acceleration versus acceleration in specific target lineages. That's the first base factor test. And then on the y-axis, we see the second test where we're testing elements accelerating in the target lineages versus those in an unrestricted model where they can accelerate on any lineage. And we're interested specifically in subsets along this red axis. And in this case, we found about 8,000 elements that were accelerated in all four short Tarsus lineages, about half of which were in attack-seq peaks from chicken data from the hind limb. We also found 347 elements where the target lineage model, just restricting to those four lineages, was actually better than an unrestricted model. And that means that these elements might be specifically involved with short Tarsus in those four target lineages. And so anyway, this is one way in which we can sort of narrow down a fairly daunting set of large numbers of elements to interrogate. We didn't do any functional work in this study, but we basically identified a suite of candidates that we can then look at in a more experimental fashion. This is just showing some of the sharing of accelerations across the different lineages. You can see that many of the accelerations are lineage-specific. So an element only accelerating in kingfishers, only accelerating in swallows, for example. But we do see a small number of shared accelerations across lineages. And intriguingly, we find that many of these elements are accelerating and occurring near genes that are involved with known pathways for limb development. Limbs are fairly well understood in terms of their regulatory network. And here you basically see, for any given gene, you can see the lineages, BK, P, or S, where an element is accelerated in the vicinity of that gene. And I think the take-home message is that we're seeing a situation where we're not seeing conversions necessarily at the level of an individual element next to an important developmental gene. We're seeing the same networks being affected. Maybe different genes within the same network have regulators nearby that are accelerating. And this, I think, is an interesting model to think about and one that we need to do more work to actually characterize are the same networks of genes being hit by acceleration or are they exactly the same genes? That's still an outstanding question. So this was exciting. And it seemed that we were identifying non-coding regions that legitimately could be said to have a role in limb development, known from other studies. And as I mentioned, I mean, one of the big challenges now is to really link genomic databases and phenotypic databases, because we're not seeing much overlap there. And one of the things we've been doing in the museum is doing a lot of surface scanning of skeletal elements in birds, because although we have external measurements like Tarsus length for all birds, we don't have detailed information on the anatomy or a skeleton of different species. So this has been a lot of fun. OK, I'm going to end with a final example. Again, looking at rates of evolution, this time for a binary trait, namely loss of flight in birds. And if I ever met a genetically intractable group of birds, this is it. You can't breed ostriches and emus in the lab very easily. This was the so-called paleognathy are a really iconic group of birds. They are found on all the continents, mostly in the southern hemisphere. All of them, including the ostrich from Africa, the emus, and cassowaries from Australia, the reas from South America, and the kiwis from New Zealand, these are all flightless. The one group of paleognates that actually can fly is indicated by this species here. This is a tinamoo, which belongs to a group of neotropical species, about 50 species, which can fly. They're not long-distance migrants or anything, but they can't soar. But they can definitely get off the ground when to escape predators. And so now, for a long time, folks thought that this tinamoo was the out group to all of the flightless species. And this was very convenient because it suggested a single loss of flight with subsequent spreading across the southern continents by continental drift. Very convenient pattern, which turns out not to be the case, but which is actually a boon for comparative genomics. Just to remind you that loss of flight entails a lot of phenotypic changes. You often lose the keel, which is this big flange of bone that we see on flighted species, completely lost in the sternum of these flightless birds. That's why they're sometimes called ratites because their sternum is like a raft. We see big changes in body size. And we see variable loss of forelimb elements. You know that you can see the wings slightly on an ostrich, although they're much too reduced to have it fly. Some flightless birds, such as moas, the extinct group from New Zealand has completely lost their forelimb elements. There's never been a humerus, radius, or ulna found for a moa species. So we see very intriguing differential loss of forelimb elements. Now, as I mentioned, the tinnimus were originally thought to fall at the base of this tree. And that was very convenient in suggesting a single loss of flight in the flightless lineages. We now know, not so much from our work, but from on work done back in 2008, where they relaxed this assumption of forcing the tinnimus to be the so-called outgroup. We now know that that's not true. And that tinnimus, in fact, fall right in the middle of this flightless radiation. It's one of these results that, as an ornithologist, I was completely skeptical of and downloaded the data, re-analyzed it. But I've come around. And in fact, our genome-wide data, this is based on whole genomes, all short read data, but nonetheless a large number of markers, strongly put the tinnimus right in the middle of that flightless radiation. You can see here in blue the different losses of flight, putative losses of flight. We don't know exactly how many there were. The biogeography of this group, which are indicated by these continents here, is very complex and likely involves a lot of extinction. It's gonna be basically difficult to reconstruct the biogeography of this group. And that's why the number of losses of flight is uncertain. But what we are certain about is flight was likely lost more than once. It's much more difficult to lose flight at the base and then regain it, for example, in the tinnimus. And that's another scenario that would allow the tinnimus to fly while all the other species are flightless. But we don't think that's the case basically based on developmental grounds. It's very, the number of origins of flight across the animal world is very small because it requires a huge number of changes. By contrast, losing flight is relatively common, even within birds. Okay, so we'll look again at these non-coding regions. We found about 280,000 of them and we applied PhiloACC to these data. And just a couple of neat examples. We actually inherited a draft genome of an extinct moa put together by an ornithologist named Robert, I'm sorry, Alan Baker at the Royal Ontario Museum. Sadly, he passed away, but we were able to use the genome that he sequenced in these studies. And here's an example of a non-coding element showing acceleration specifically in moa and these reas. This is after 100 million years of being conserved across all other birds. And you can see how this might, it's plausible that this element could have played a role in loss of flight after being conserved for so long. Here's some additional examples. Again, we're able to place each branch in a rate category indicated by its color. And then we can estimate the rates of the element in those categories. And about almost 1% of the elements that we investigated were accelerated in at least one flightless lineage. And so we're able to have some sample size to work with. Now, we asked what genes are in the vicinity of these so-called rat tight accelerated elements. And what you see on the X axis here is the number of rat tight accelerated elements for a given gene. And on the Y axis, the total number of elements near a gene. And these genes in red here are those which have a disproportionately large number of punitive regulators that are accelerating in flightless lineages. And what was exciting was that we saw a lot of overlap with previous work. For example, genes like TBX5, there's some Hawks genes in here, DAC1, these were all genes that had already been studied by developmental biologists in the connection with limb development. The other take home message is that when we look at the small number of proteins that are accelerating specifically on flightless lineages, we find a modest number, maybe 200 or 250, but the functional coherence of that group of proteins was not high. We don't get any strong gene ontology enrichments. Whereas for the non-coding elements, when we look at go terms for the genes that are close by, we see very strong signals for skeletal development, cell proliferation, things like that. And so we feel that in this case, the non-coding elements, the regulatory landscape, is perhaps giving a stronger signal for underpinning loss of flight than the proteins themselves. So this is perhaps a point in favor of Alan Wilson's and Mary Claire King's hypothesis. Okay, just to end, we can show that some of the genes that are in the vicinity of these rat tight accelerated elements are indeed expressed in the developing limb buds. These are chicken embryos where we've looked at expression of particular genes with accelerated elements nearby. We can also use approaches like a tax seek. I think many of you here are probably familiar with the tax seek. It's a method whereby we can ask what parts of the genome are open and unwound from nucleosomes and available for gene transcription and binding by transcription factors. And we performed a tax seek on both the foreign high limb of chicken as well as a variety of other parts of the skeleton. What this shows here is the enrichment of the non-coating accelerated elements in open chromatin of these different structures related to loss of flight. So it was very exciting to see that during development these accelerated regions are over represented in areas of open chromatin in the foreign hind limb as well as in the keel and other parts of the skeletal apparatus. And so we can imagine intersecting the rate acceleration data, the attack seek data, as well as other epigenetic data, again to narrow down a very daunting set of candidates. Now we would not claim that these 42 non-coating elements are underlying loss of flight, but they certainly provide a very tractable set that we can then look at functionally. And that's what we have done in the early experiments. We would sort of very laboriously take individual putative enhancers and put them into a construct where we can ask whether they're driving gene expression. So in this case, we have taken the chicken version of an enhancer that's accelerated in flightless birds and shown that we can inject it. That's what the red fluorescence means and that it drives gene expression. That's what the green fluorescence means. We take the tinimum version. That's the volant paleoignate show that it too can drive gene expression. By contrast, the RIA version, which has been accelerated, has been injected but does not show evidence of driving gene expression. And so what this shows is that this acceleration has facilitated or been associated with a change in function of this element, either loss of function or a shift to a new function. We're doing a lot of comparative transcriptomics and a comparative attack seek. This is just stuff that we're currently analyzing now, but I wanna end mentioning some really neat experiments we're conducting with Emma Farley, who's a developmental biologist at UC San Diego. She has developed a high throughput enhancer screening assay. So instead of laboriously taking one enhancer at a time and asking whether it can drive gene expression, we can inject hundreds at a time. And what we do is we attach a barcode to each enhancer. That's what BC is here. We can then harvest the mRNA from the developing limb bud or other structure and sequence it and find out which of the enhancers that were present in the original cocktail has driven gene expression. And this has allowed us to dramatically scale up and in our first experiment, we've looked at about 1,000 different enhancers, putative enhancers. And so the data look like this. Here we have the amount of DNA for each enhancer. Each dot here is a different enhancer. This is the amount of input in blue across these replicate structures. The red is the amount of mRNA driven by each enhancer. So you can see that there's differential activity of enhancers in these different structures and that it is fairly replicated across replicates. And what this has allowed us to do is to identify enhancers that appear to be associated only with forelimb development, such as these here, where here we have a measure of enhancer activity as read by RNA readout. These are enhancers active primarily in the forelimb, higher readout of mRNA than the high lip. Here are bifunctional enhancers and here are enhancers more active in the hind lip in orange. And so through comparative genomics, we've been able to go from very unwieldy non-model species to putative tissue-specific enhancers, which we can then interrogate further. And so hopefully these examples have shown the power of comparative genomics for unlocking secrets about how phenotypic diversity arises. These are just some of my conclusions. And the attack seek was very useful in terms of narrowing down what we should look at. And I just want to thank my collaborators and NHGRI for their funding. And I'm happy to take any questions. Thank you. So thank you Scott. This is from Larry Brody. He says, if I understand correctly, you've identified changes in a large number of unlinked elements that contribute to the fixation of a new trait. And he would like to know if all of these individual elements are acting in concert to produce the trait, how do we get to the end point via segregation? Step-wise, bunches at a time, do we know the step size? And if known, how many generations would it take to move from the ancestral phenotype to the new one and fix it? Thanks for that question. That's a great question. I mean, we don't know whether all putative enhancers are involved with the trait. There's gonna be a certain amount of background noise and false positives. Again, the attack seek data I think is extremely important because as I showed you, loss of flight is a multivariate trait. Lots of things are changing. With just the genomic data, we don't know whether a given enhancer influences the keel, the hind limb, the fore limb, or what. And so we really need that epigenetic data to narrow down our search. As to the time it would take to go from an ancestral to a derived state, that is a tough one. I think there are probably some creative ways we could imagine looking at how fast the elements are changing. But the bottom line is it changed somewhere along a branch and we don't often know where along that branch it changed. But good question, yeah. Great talk and so many very different, exciting phenotypic changes that you're looking at. I was really struck by the repeated loss of flight in the ratites. And you mentioned the skeletal changes that must also have occurred, I guess now, multiple times. When you look at the skeletal changes now knowing that they occurred multiple times, does it actually look like they occurred in different ways or does it actually still look like it's the same skeletal change happening? Yeah, thank you, that's a great question. One thing we did do was we looked at the rate of development of tinimoo foreign hind limbs because that might help us distinguish between whether tinimoo's had regained flight or whether they had retained it from the ancestor. We found that tinimoo fore limbs and hind limbs basically developed at the same rate as did chickens. And so that was one confirmation. But I mean, your question is very, very good. And in many ways I would say that genomics and phylogenetics is outpacing morphology in this way. We have lots of really novel relationships in birds and other groups where we're wondering, okay, great, so parrots are sister group to the songbirds but is there morphological evidence now to support that sort of looking retroactively? In the case of the loss of flight, there are, as I mentioned, there are lineages have lost the fore limb elements to a different degree and so that's a certain indication that perhaps there's been imperfect convergence, if you will. But I think there's a lot more to do at the developmental level to show that maybe they're slowing down cell division at different times or in different places or by different mechanisms. So I think it's still very open, yeah. Thanks. There's a lovely talk. I found it very interesting that you see those regulatory elements over and over again on the same transcription factors that are regulating those processes. But the underlying sequences within those elements as well, are you seeing conservation of the binding sites within those elements contained also to more broadly make those changes? Thank you for that question, it's a great question. And I simply haven't been able to cajole my postdocs into looking at that level yet. It's a very good question. We do have some examples where sequence acceleration is associated with loss of binding sites, particularly for this one transcription factor ETS which is important in lots of cellular processes. And we have a couple examples where very few sequence changes seem to be relevant to abolition of a few binding sites. And we're gonna experiment now to see whether that might be actually driving some of the changes that we're seeing. No, it's a great question and you think we would have done this years ago, but yeah, it's on the to do list for sure. Awesome. So my question is sort of related to the long bones and the legs. And have you looked at sort of the opposite where you have a different set of bones changing like wingspan. So you could have long leg, long wing, short wing, long leg, short leg, long wing, et cetera. Do you see that it's the same modules kicking in just regulated differentially in the arm versus leg or is it, would you see completely different sets of enhancers being changed? Yeah, thank you for that question. It's a really good one. And this is, no, I think one of the thrills of collaborating as you work with people that know a lot more about development than I do. In briefly I put up that slide of comparative transcriptomics. And one thing we're seeing there is that the transcriptomes of foreign hind limb of flightless birds, they tend to be more similar to each other than to the transcriptomes of say foreign hind limb of chicken or something like that. It's a really interesting conundrum about homology and all this stuff. So I think we are seeing convergence at the level of the transcriptome. I think that's fair to say. Whether or not that's underlined by convergent changes or convergent accelerations in the non-coding elements is still something we need to look at. But the signal from the expressed genes is, it's sort of as if the foreign hind limb are co-evolving. They're sort of clustering together. They're species, they tend to be lineage specific rather than say all the hind limb transcriptomes clustering in all the forelimb. And maybe that's what's been seen in other sort of serial structures, but it wasn't what I was expecting. So yeah. We'll squeeze one more in from Elena Ostrander who thinks a lot about dog phenotypes. So she might have this in mind and asking that the examples you presented all assume that the trait has some commonality and mechanism. How often do you expect that to be true? And for other phenotypes, there might be possible mechanisms that would permit a particular phenotype. Yeah, thanks, Elaine. That's a great question. Absolutely. I think this whole comparative framework at some level depends on there being common mechanisms. Although it's also reasonable, we can quantify now the extent to which changes in the non-coding genome are lineage specific versus common. That's sort of an empirical question. And as we saw in the TARS example, there's quite a lot of lineage specific stuff. And I think the real challenge is figuring out well, how much of that is related to the phenotype versus other kinds of noise. And so, yeah, and they're also, as I was saying, there might be sort of commonality at the level of the network rather than the individual non-coding region. And that's, I think, a big task for the futures. How do we kind of collect that data and analyze that sort of data? And last, from Shorjo Sen, he wants to know about the bike that you rode cross country. Is there anything special about it? Like gearing, tires, an old trusty bike? Oh, man. I think I'm more known for that crazy bike trip than for any science I've done. I would highly recommend it. I came back basically saying to my graduate students, look, it only took two and a half months. Just do it. I'd love to chat with folks about it. It was a life-changing experience, a good way to connect with the communities and that an urbanite like me might not connect with very often. And it gave me a lot of hope, honestly. I think if we can get past our political divisions, we actually have a lot in common, more than divides us.


Everybody forest here being 2022 and NT and 2023. I'm at collegiate football hobby two bucks. Make sure number six random teams It's randomized right now. We'll get right into the rip as well same recording Those seven times names first one two three four five six and seven Christopher s down to Derek s and teams now One two three four five six and seven commanders down to lions Christopher s commanders Ryan v Ravens Felix and with the Browns Jeremy Y Broncos Chris G Seahawks Preston E Jags Gordon P Titans DW Vikings Collin T Falcons Chris V Packers Simon H Panthers Alex S Niners Felix and Jets Gordon P Bills DW Bucks Jonathan L Chiefs Dylan C with the Raiders Chris V Eagles Mark S Bengals Shelley E Texans Ken S Cowboys Brandon C Colts Pats Eric P Rams Sean M Bears Troy D Cardinals Kyle C Chargers Patrick F Steelers Joe W Dolphins Ryan V Giants Gordon P Saints Derek S Lions it was posted Randy what's up, man? And we're gonna get started right now Here comes your link. Alrighty. I'm at collegiate first There's just one more case of NT after this and we'll do our promo random. That'll be it for the night That is to 35. It is a Dak Prescott the Cowboys Bengals Jumbo swoosh from I think a hat Chase Brown for four Bengals. Yo, yeah hats whoosh cat cat swoosh Since the next up is a Chargers auto to 99 of Mike Williams. What's your up to your Randy? Randy Mike will Chargers. Oh to 99 Parker Washington for the Jaguars Yep Jags Parker Washington after that is a Clayton tune to 65 on the rookie Jersey Auto Clayton tune Cardinals May get some work here The rays on the West Coast are you watching a replay and we got money Manziel to 99 autograph for the Browns. We got money Extra innings in LA nice. I Guess even for West Coast it is late now the pitch clock rules NT football 2022 To 99 is an Ezekiel Elliott for Dallas It's a 25 Michael Strahan Superlimitality Giants Tony it's a 20 that's a 49 is Brandy cooks Texans Brandy cooks Texans two-color patch it's a 49 rookie dual-patch bills James Cook 35 Cook Rook rookie tremendous treasures Jersey to 25 is a mirror white Raiders triple Jersey Got chiefs Bengals and bills to 99 Mahomes borough and Allen If anybody has a majority here chiefs Jonathan L bills is Gordon P and Bangles is mark s random at the end Field pass rookie auto Titans Malika Willis 20 of 49 Jets on the auto Quinn and Williams the 99 Titans again here NFL gear dual rookie relic auto to 49 Traylon Berks Tennessee and Rookie material signatures Kenneth Walker the third Seattle out of 99 Three-color jumbo patch rookie material signature Seattle Kenneth Walker the third Got one random to do we'll do it real quick. No recap short break We got one more break Sean's Sean's loony now chiefs bangles bills Four and one five times service up top gets it one two three four and five. Let's go into the bangles And we'll do it. Thanks y'all very much. We're moving on now to the final break of the night


this video. Odds are, you are always focused inward on how you feel and what you're thinking. So today on Recovery Monday, episode number 15, we're going to talk about how to recognize that and some ways you can begin to change your focus. So let's get cooking. All right, guys, there's my little 15 second intro for Instagram so that people watching on my Instagram stories get like a really slick intro. We can be go. And let's get into it. So I'm going to put the chat overlay up when you guys pop in, just say hello, let me know that everything's seems to be working just fine. No technical issues today. Thank the gods of technology.  Welcome. If you're coming from the Facebook group, just know that I'm only I'm not going to see your name. I'm only going to see Facebook user. Sorry about that. That's just the way restream does it. So we are doing another Recovery Monday. This is episode 15. We've been doing this for 15 weeks. Can you believe it? So and we will continue to do it until we are through this book. So we are essentially teaching recovery lessons out of this book every Monday here at two o'clock. If you don't have a copy of the anxious truth, then you can get that on my website. I will put that up on my on the screen right now. So if you need a copy of the anxious truth because you want to follow follow along, you can get it right there. The anxious truth comm slash books. And before we get cooking, I will remind you if you have not subscribed to the anxious morning, which is my new email newsletter and podcast that comes out every weekday morning, hop on over to the anxious morning dot email and check that out. Everybody is digging it. Anyway, welcome everybody. We're up to about 30 people here. We're not quitters. Love it. Love it. Love it. Okay, so let's see here. Welcome everybody. Welcome everybody. Welcome everybody. Hello, Long Island neighbor. How's it going? Love it when I have local people pop in. That's always a good thing. Let me close this. So let us see what do we got going on here? I'm just going to see who's here. All right, let's get cooking here. So let's talk about focus. Today we are talking about changing your focus. If you're watching me, like I said in the intro, the odds are I want to look at what I'll sell. I'll start this video just like I started this chapter in the book, which is I'll ask you over the last two hours, tell me how much time you have spent thinking about how you feel like physically your symptoms and what you're thinking. So over the last two hours, I would bet that if you're watching a considerable amount of the last two hours have been spent focused inward on how your body is feeling, what sensations you're having and what thoughts you're having. This is really super common. Like people in the grips of an anxiety disorder have an almost obsessive drive to focus inward all the time. How do I feel now? What are my symptoms feeling like? What am I thinking? Am I thinking those thoughts? Are they going to come again? How can I get rid of them? How is my heart beating now? Am I dizzy now? How are my legs working now? So it becomes really an obsession with how you're feeling all the time.  How am I feeling now? Now how am I feeling? How am I going to feel? So this is really common. If you're stuck in this, don't sweat it because everybody gets stuck in that. I used to be stuck on it too. I used to spend all my time poking and prodding and checking my pulse and like just all the time I was just always scanning to see was there any new symptom? How was I feeling now? What was going on? I was always aware of what I was thinking. Should I be thinking this now? What can I think differently? What can I do? What can I do? So it's just relentless drive to always focus inward. One of the podcast episodes I did, I don't even know when, was about, it was actually called Stop Checking In With Yourself. So I want to address that for just a second before I go on. This is in a long topic, so I'm going to go for another few minutes and then we'll start taking some questions here. But one of the podcast episodes I did was called Stop Checking In With Yourself because one of the generic mental health bits of advice that you get online, especially from people that are coming from like that healing space. I'm going to use air quotes. I'm not trying to diss the healing people. I get there's value to healing too, but especially when people who approach this as an emotional healing or spiritual healing kind of thing, often you'll hear check in with yourself.  That is awful, awful, awful advice for people like us. I'm not saying it's universally bad advice, but when you get told do the check in, take five minutes and check in with yourself to see what you need. That advice is not for us. When you get better, you can use the self-check in. I know people swear by it. You got to take care of your emotional health. You got to take care of yourself. I'm with you on all those things, but when you are stuck because you can't not check in, you're always checking in. Being told that the check in is good for you is terrible advice. If you have heard that advice because yes, check in with yourself. What do you need? What do you need to see what your anxiety is telling you? If that has thrown you for a loop, then don't sweat it because it was going to throw you for a loop. You're not broken if that's not working for you. Let's talk about you're always focused inward. One of the ways that that expresses is that you will take everything that happens in the universe, everything from a dog, the neighbor's dog is barking, something is up with your kids, there's a big project at work. Let's see, a dinner plan just got canceled, the pizza delivery guy is late. Anything that happens in the world, there's a supernova in a galaxy, 72 billion light years away, everything that happens in the universe, you will internalize and ask yourself, okay, how will this make me feel? Is this going to trigger my anxiety? Will this make my anxiety worse? Will this make my anxiety through the roof? Now, this may be things that you are personally involved with, like how am I sleeping? What did I eat? What do I have to do today? Do I have to leave the house? How will this make me feel?  If my kids are acting up, how does it make me feel? Me, me, me. So your anxiety, not because you're selfish, you're not in any way selfish.  So the disorder is selfish, you are not, but part of internal focus is to recognize that you may be internalizing everything to see how it's going to make you feel in relation to your anxiety. And you will also evaluate future events. This is where anticipatory anxiety comes in. If you have something coming up that you and you are going to have to do or something is going to put, place some demand on you, but how small that demand may be, you will internalize that as well. How is that going to make my anxiety react? Is that going to bring my anxiety up? Am I going to do it? Am I going to panic? So very, very common also. So one of the, I want to touch on two other things. I'm going to talk about distraction for just a minute and then we'll get into some comments and questions, right? One of the ways, and by the way, I wrote about this in a lot of details. There's much more detail in the chapter in the book than I can go through in a video. So one of the common ways that people try to get out of that constant inward focus and checking and evaluating is by distracting. And I'm going to flat out say like distraction works. Like distraction, it gets a bad rap, but it works. The reason why I give it a bum rap and the reason why people who sound like me and talk like me about these things will give it a bum rap is because it does work, but it reinforces the wrong lesson. So people who will run immediately to, I have to call my safe person. I have to snap a rubber band. I have to top pop a pill. I have to sniff a sense or an oil or whatever it happens to be. Distraction, I got to get my coloring book and start to color or I, you know, I immediately start, I know people who will do things like I count backwards by three from 100. I count prime numbers. I've heard all of these things, right? Distraction will work temporarily, right? It will work temporarily.  So I must get a coloring books for a while. I don't know if you guys are still into coloring. Who still raise your hands if you enjoy the coloring thing. For a while, adult coloring books for anxiety, like that was typically, these are adult coloring books for anxiety, were all the rage. That actually goes back a few years. Everybody was talking about their coloring books. Now, look, if you like to color, there's nothing wrong with that. Like I'm a fan of coloring. If you did coloring because it's relaxing and it's a hobby and you like to be creative, then knock yourself out, color. But I remember when coloring books were, everybody was talking about their coloring books and I know people that literally ran to the coloring books, they would travel around with their coloring books and markers or crayons or colored pencils. I know people that absolutely, I know women that bought bigger purses. I'm not kidding, bought bigger purses so that they could carry around their coloring books and their colored pencils because if panic hit, no matter what they were doing or they started anxiety rising, they would feel like, oh, I got to get out my coloring book and start to color. Now, why does that work?  So it's a good example of why getting out of your head and changing your focus outward is what we want to do because you may get some temporary relief that way except if you're coloring at 1pm because you try to stave off panic, you know, really by 1.45, it's going to happen again. You're going to need the coloring up again, right? So that's kind of the way that works. So that's why we don't want to use distraction because distraction is temporary relief that doesn't actually teach us anything. Right? So what does work? How do we want to work on changing our focus? Well, first thing we want to do is want to recognize the problem, which is what we're talking about now. So recognize what's going on. Catch yourself when you're doing these things. Like, oh yeah, I'm doing that thing again, right? And that's okay. You can do that.  I got to work on this. Okay. So I recognize and I'm doing it. Now, what can we do? Essentially, this is a, changing your focus is pretty much the process of kind of turning your back on the thing that you feel.  I can't handle this. This might never end. I'll have that never ending magic panic attack. That doesn't exist, by the way. Whatever that fear is, you have to kind of turn your back on that and say, well, I can't really scan for that. If I feel like I should be scanning for that, I should be evaluating my state to see if that's coming to get me. I should be having inner dialogue to try and stave that off and prove it to be untrue. But really and truly, you have to kind of leave that and turn, it's turn your back on it, right? You have to turn your back on that. So that's what that looks like. It's really uncomfortable and it will feel wrong and it will feel like this is not right. I'm supposed to be protecting against that but changing your focus literally means first thing I know is that I have to just turn my back and leave that fear unanswered and then I'm going to find a way to engage with the outside world the way non-anxious me would do. What would I be doing right now if I wasn't in the grips of all these intrusive thoughts? What would I be doing right now if I wasn't having a racing heart and feeling dizzy or my stomach wasn't in knots? What would I be doing? I'm going to do that. Sometimes you don't necessarily even remember what that look like, which means you're going to have to start to try. You're going to have to try to try different things. I guess I'll make lunch. I guess I'll take a walk. I guess I'll read a book. I guess I'll brush the dog. I guess I'll wash my car. I guess I'll send some emails. Whatever it is, you might not know what that looks like because maybe you've forgotten. That's okay. Just try different things. So, you're pretty much just going to you know disengage with those that fear, turn your back on that and engage outward and every time that anxiety starts to pull you back in like in the godfather every time I think I'm out they pull me back in. You're just going to have to keep repeating like nope I don't have to I don't have to scan for this. I don't have to have inner dialogue about this. I can work on making my tuna salad right now. All right? So, just a quick recap before we get into some of the comments and the questions which we can do right away. The disorder will have us continually in a state of inward focus almost obsessively. We're constantly focusing on how we feel and evaluating our state our mental our emotional and our physical state because we think that keeps us safe. We're trying to guard against that bad thing happening which would be panic or anxiety or whatever you think the bad thing is. We will take everything that happens in the known universe internalize it and then evaluate it based on how it makes you feel. How will this make me feel? Then you may try to get out of that cycle by using distraction techniques which will work but only temporarily and then you have to keep going back to them again and again and again and so what we really want to do is start to understand that the process of changing my focus from inward to outward will be to start to engage with the world the way a non-anxious person would and leave that fear in the background chattering at me. That it's going to be right in your in your ear. Got to hear it anyway. That's why we don't try and distract from it. We're going to hear it but we're going to engage elsewhere even while we hear it. So that's what we got. That is basically what this chapter is. What's chapter of the book that is this? I don't even know. This is lesson 3.3 because the book is broken into five chapters each of which has a bunch of sub lessons in it right. It's kind of organized as if it's a course like you're taking a course basically. So that is the deal on changing your focus. That's the the big overview that like I said there's more detail in the book if you read it but that's what's going on. All right. So let's let's go through anybody have any questions on on this sort of stuff. So I'm going to scroll back to the top here. Look at this. I'm 15 minutes in and you're getting into comments ready. I love it. Let's see questions anybody who is guilty of distraction who's guilty of that stuff. I saw some of the coloring books things pop up because when I mentioned coloring books yep all the time okay zero only because I've been watching stage. I don't know what stage does. So you haven't been focused on yourself. Put this up on the screen. I'll have to Google that now. Maybe that's something that I'm missing. I'm out of the loop on stage, man. But there you go. So you have not this is good. You have not been focused inward on yourself at all because you have been outwardly focused on on this this program that you like clearly. It's entertaining. It's keeping your interest and so your your interest is what's going on in the program. Not what's going on in here. That is a perfect example and I would say along those lines are the last 14. Somebody I see we have stage fans here. Colleen jumped in on that. I'm going to have to check it out. One of the things that I always try and teach people too is think about the times because it does happen even for the most anxious person in the most distressed situation right. It does happen where things come up during the day and you will get a few minutes here and there where you do accidentally start to focus on something else. It's an accident because you're trying to stay focused inward because you think keeps you safe but something comes up. You get into something. You forget that for a second and all of a sudden you discover oh wait a minute. I just had three four five ten minutes where I wasn't focused on myself. So think about how that feels because I know you have a few minutes like that here and there throughout the day. That's what you're going for. That's what you're going for. So there you go. That episode stop checking in with yourself actually because I know Allison threw that up there. I don't remember what episode it is. If you go to the anxioustruth.com there's a search function there in the menu.  Search for check-in. Stop checking in. You'll find it. That episode generated quite a lot of dialogue with some of my mental health friends that are not necessarily anxiety specialists and they were really didn't like that I said that. The check-in. The check-in you got to do the check-in. I'm like no you don't understand these people can't stop checking in. So let's see here. Okay. I'll throw this up on the screen I guess we'll address this. Somehow or other. You know what this is a little bit meta. So a little bit let's go a little meta here. So we are taught today we're talking about inward focus and always focusing inward and being obsessed with how you feel always and how will this make me feel. How will this make me feel. And invariably when I do these videos when I do any live video and whether I'm doing it by myself or I'm doing it with Josh or Cam or whoever or Lauren and it's people can ask questions. Invariably people will completely ignore what the topic is and go right to tell me about my thing. I have it I need to know about my thing forget the topic. I'm not angry about that but this is a perfect example of that. I start screaming for help and I get out of the car I lose control. This is a perfect example of I'm trying to actually address a thing that would help with that but instead of paying attention to that information and maybe taking a lesson I'm not not not angry I'm not picking anybody here because I don't know who that is. But nope I'm just going to go right back to the thing that I care about right now. Like my thing right now my thing. So it is a good example of how the disorder will literally grab your attention and drag it wherever it wants to go. It's just not it's not a good way to live that's that's terrible I don't want anybody to live that way I used to live that way. But it will literally drag you around and I know I'm going to tell you where you're going now if you let it. So completely ignoring sometimes a topic and jumping in with nope I got to answer my question my question. That's a perfect example of how the disorder will keep you focused on how you feel and what is scaring you right that minute all the time. It's a really good illustration so again I'm not picking on whoever answered that question or made that statement because I didn't even tell who it is but it's just a good illustration of how this works. If you were not anxious that way then you would probably not be thinking that way you would not you would not do that you would be maybe paying attention to like what is this crazy guy saying or you wouldn't need to be watching the video which would be great but it would be a little different animal. So let's see here um the supernovas are hard to ignore I mean that is that is true that's any points we all love a good supernova I know I want to pay attention to them. Alright so let's say this is a good one too I've seen I've done enough checking on on me to last the rest of my lifetime which is true I got to tell you so for folks like us when I was able to start to break that habit and I was doing better on a consistent basis and I would find that those times when I would go you know longer and longer without checking in with myself oh it was the greatest thing ever was like winning the lottery it feels so good when you get relief from that you know in a big way. So let's see here I see that we have a bunch of people who raised their hands on the coloring thing um it's hard to come let's throw the coloring distraction thing up here it's hard to color when you're so anxious you don't even want to sit down okay that's pretty fair so Bethany thank you for making that comment because the coloring books were super popular but I do get that sometimes finding a distraction can be really hard because you don't just don't even know what to do with yourself so even if you want to distract which we would say is maybe not the most productive response in the beginning I can understand it but you start to learn not to do it finding an effective distraction when you just don't even know what to do and you're just trying to get away from yourself can be difficult so thank you for bringing that up let's put this up on the screen I'm going to I will address this um okay I know this is not a topic but that's okay I'm going to address this so listen brother I know that you think you have transmitted your anxiety to your daughter I know I know that you are terrified that you have somehow given your anxiety problem to your daughter first thing I want to tell you is number one you didn't do that you cannot transmit it anxiety is not contagious so I know you know that on a logical level but on an emotional level I know dude that you are terrified that you have somehow accidentally transmitted your anxiety to your daughter you did not do that you are off the hook on this and I know that a lot of the concerns that you have are based on the fact that you should not feel this way this is a defect why can't you get this I'm not getting it right I know there's some issues that come up recurring like that and that's going to make you feel even worse that not only are you failing because you are anxious but now you are failing because you transmitted your anxiety to your daughter you did not do that did not transmit anything to any member of your family okay so first of all huge kudos to you and a fist bump for caring so much thank you for being a good dad so good I'm glad that she's listening the thing that you can do to help your daughter is to help her understand and if you're listening welcome hello that none of what you're feeling is dangerous right now and right now you are surrounded by 66 other people watching this video with you from all around the world that absolutely 100% understand how it feels when you have all those scary anxiety symptoms and sensations and your stomach is not and you get sick to your stomach and it seems so terrifying and you just want to be saved you're okay you're really okay we all understand how that feels so you're just going to have to really start to understand that sometimes life stresses us out and makes us feel a certain way sometimes things trigger this sort of stuff but we don't have to call the anxiety itself a disaster even though I know that it's really super scary and when it's new to you you don't know what to do with it so just your dad knows all right he knows he's been hearing all of these things and just because he may struggle now and then doesn't mean he doesn't know so he will be able to give you some good information and point you at good information take it and understand you're surrounded by people who know what that feels like and you are not in any danger I promise you right so there you go I hope that helps brother but again you're not responsible for this you're good dad thank you for caring I've been practicing my okay let's see pop through let's keep going here this is good we got extra time for comments I'm loving it I've been practicing mindfulness through the day and it's easier in the morning but as soon as it gets dark my anxiety shoots up okay let's address this this is a good question so what's the difference between mindfulness in the afternoon and mindfulness at night well the clock it's dark like it's dark half the time on the planet that's the way it happens so the first thing that I would tell you in that situation is like well my anxiety shoots up okay well it sounds like you're hoping that the mindfulness is during the day is meant to keep you at a at a low level you're welcome you're welcome my friend don't worry about it it's gonna be okay I promise so back to the mindfulness during the day thing you're trying to trying to get the mindfulness to bring you down like I didn't need to this is gonna calm me and keep me cool and then somehow or other you've decided well nighttime is a different animal the mindfulness doesn't work and my anxiety goes up but really truly sometimes that's a function of I'm hoping that my mindfulness activity during the day will keep me at this calm level trying to hold my anxiety at bay through mindfulness but really mindfulness is meant to get you through it not to keep it at bay so even though it's dark at and you've decided my anxiety goes through the roof it triggers at a certain time when it gets dark number one you're anticipating that and you don't like when it happens but when it does happen you just have to use the same routine that you're using during the day it just means you're gonna do it while you're uncomfortable and you will learn that you can actually move through that it doesn't matter what time of day it is so a lot of times it's just a you know that's an expectation thing right this is good I gotta put this up grease belly garage one of my favorite YouTube names I'm gonna call you GBG it's important practicing changing your focus when you're not anxious thank you thank you dude this is huge it is a huge thing you guys hear me talk about meditation practice mindfulness practice breathing progressive muscle relaxation I talk about it all the time it's all over this book like I have a chapter in here about learning new skills if you go to the anxioustruth.com slash skills you'll find I've linked tutorials on basic meditation and progressive muscle relaxation and diaphragmatic breathing the reason why I tell you guys to do that is not because I want you to become expert meditators or I'm trying to turn you into mindfulness gurus I don't care if you become mindfulness gurus or not we're learning how to focus our attention in this situation meditation and mindfulness are attention training tools this is how we exercise our attention muscles our focus muscles and so when we need to I can flex that attention muscle and put my attention where I want it as opposed to where the anxiety is trying to drag it so that is a really really good point if you don't practice those things or you never take them on and then you hope that when the shit hits the fan you can magically relax and breathe and be mindful and it'll go away you're going to be super disappointed because it doesn't work that way so you got to practice when you're feeling good so that you can use those tools to move through the times when you're not feeling so good really important really important but I know for a lot of people look I gotta tell you I'm not a big fan of like I didn't want to do that stuff I didn't want to do it I had to learn by trial and error that oh if I if I refuse to do it because I'm stubborn I'm a stubborn mofo so I'd be like I don't need to practice practice I'm that guy like I never practiced I never studied I still did really well so I'm like I don't have to practice anything I learned the hard way like I guess I actually do have to practice it sucked man so practice practice when you're feeling good okay so let's pop this up JJ sensory motor OCD you can't stop noticing a numbness sensation that's okay I'm not in any way an OCD expert nor what I claim to be but I'm pretty sure that my qualified OCD therapist friends would probably tell you in this situation and you should always seek those people out that it's okay that you notice it the problem is not that you notice it the problem is what you do after you notice it all right so noticing it is okay even when you are you know trying to disengage outside which I'm sure you've probably been taught if you're working with an OCD therapist to try to get you to engage elsewhere while you feel that you will still notice it it's okay you're not trying to make the noticing go away okay people to all right I'll throw this out here let's see here Colleen what's up Colleen my fear is I'm sure people with health anxiety do have heart attacks how do we know when it's appropriate to go seek help okay that's fair so in this situation if you want to talk about you know focus your heart anxiety or some people will call it are calling it cardiophobia now will continually want to bring you back to that continually want to bring you back to my heart has my heart doing now is it racing how do I feel do I have any pains that indicate a heart problem what are the odds let me read about healthy people who have heart attacks let me try and get a grip on this we'll always try and bring your attention back all right here's reality it is true that raise my hand and being stubborn off of yeah joining the club brother I don't learn some hard lessons in recovery I really did but here's the deal on that like nobody gets a hundred percent guarantee nobody if I I think even if I could be the world's greatest cardiology right now Colleen I would not be able to offer you a 100% iron clad assurance that nothing will ever happen to anybody's heart including yours we don't know that but right now nothing is happening to your heart nothing is happening and it becomes an uncertainty intolerance problem that's really what this is so the health anxiety is the intolerance of uncertainty if you cannot get a hundred percent guarantee about your heart then it's nothing 0.0001% chance that you may suffer heart attack is too much for you so we we are swimming an uncertainty all the time we live our entire lives absolutely immersed 16 feet over our head in uncertainty and most of the time we don't care but you've latched on to one area where uncertainty is intolerable so you really got to work on that it's not about being assured that your heart nothing will happen to your heart it's about that it's about the uncertainty okay let's see here this is I'm gonna scroll down a little bit let's see just changing the focus to throw it on here yes this is about rewiring your brain the changing your focus to outward rewire the brain to ignore the inward focus because that seems like what's what's happening to me yeah I mean we a lot of times we use the term rewire your brain and technically like if you had a neurologist here or somebody who's involved in neuropsychology probably say yeah that your brain literally does rewire itself we know this to be true people who suffer head injuries or have strokes learn to do things again they learn to walk again they learn to speak again they learn to use their hands again it's your brain is amazing and it's plasticity it does literally rewire itself so when certain connections are are interrupted it will make new connections so it does literally rewire essentially the process that you're going through is when you keep turning your focus outward instead of inward you allow that that worst case scenario to be there without addressing it the inward focus is trying to stay safe from the the disaster right when you allow it to to just all right I'm gonna have to surrender to the disaster and I guess I'll have a heart attack while I'm eating lunch and then you don't have a heart attack and then you know it again and again and again you essentially are learning oh there really was no danger that's what that is if you want to call that rewiring yeah probably technically probably so let's see here and to follow up answer Colleen how do you know when to seek help if you've already if you have already been seen by a doctor like cardiologists you've had your heart tested everybody tells you that it's okay then then you're good then there's no more help to seek there's no more help to seek and understand that people who legitimately have physical impairments like heart attacks they know people who have heart attacks know that something is wrong the difference is I I know I feel something is wrong and different about my body versus I'm afraid that something might be afraid afraid with my body right that's the difference there oh David Tennant is on stage David Tennant was my favorite Dr. Who by the way I'm not a huge Dr. Who fan but you know getting there let's see here okay let's see here I'm going to scroll scroll scroll I walk in right through my time here in a good place thinking about with the kid getting in right okay this is good Jenny thank you for bringing this up Fitbit I'm this is a guy wearing an Apple watch so I kind of get that when my mic is holding up getting in my Fitbit really helped me with heart rate anxiety it was fixated on those numbers if you have heart rate fixation then Fitbit or an Apple watch or any of those things that's rough that is essentially just pouring gas on that fire in a big way in a big way right so let's see here I will throw this up here you're asking two different questions my friend my opinion to stay your is your opinion you're staying in the car and screaming well that's a different opinion that's a different topic than this you're talking about what to happen what to do when you panic you this is going to be unpopular but I got to tell you anyway because you need to hear this in between the moment where you feel oh my god that first flash of fear and the moment that you begin to scream then you actually make a choice so one of the emails that I had last week on the anxious morning was that the power that we find when we recognize that even though that's a tiny little space that's only a couple of seconds wide you do actually have the power to make a choice there so you choose to begin screaming that you're panicking I understand that that is real fear I'm not invalidating the fear but you have to understand that you may initially have that reaction but we get to decide if we continue that reaction or not so my you're talking about what to do when you panic in the car and the answer would be first of all recognize that you do not have to scream you don't have to do that you will have that initial reaction that's you can't stop that that's okay it's not wrong but you don't have to go into I'm going to scream and lose control and flail about you don't have to do that so you have to see what it looks like when you don't do that you're doing that because you think that's somehow going to save you or that that's that's an escape in some way some way shape or form so you don't have to do that you can't changing your focus this is a little different animal I'm talking about focus in general focus during a panic attack yeah you have to put your focus back out of that it's really hard during the panic attack that's why I was talking earlier about practicing that before it happens but little different animal doesn't necessarily apply but generally yes your your panic is telling you you got to start screaming and crying and flailing and doing all those things when and truly you don't have to do that you know let's see I'm going to scroll down a little bit more okay I'll answer this one really quickly again you know I don't want to get too much into the mechanics of OCD because that wouldn't be fair to you guys but can it turn into a void? what I'm always what I always hear from the very smart people is that when you're dealing with OCD you have to be careful that your coping tools don't become compulsions so sometimes that becomes a concern so you kind of want it to make you want it to look as organic as you can as much like life as you can being mindful of the fact that if you start to make it a compulsive thing every time I every time I have this thought I have to do this activity that that could be that could be something to look out for so you got to be careful about that it's not avoidance people think that like well if I if I focus away from the fear then I'm avoiding the fear no no that fear is still there right so what I would say is whether it's a sensation that you're fixated on whether it's a thought or some other panic sensation or an anxiety sensation it's not avoidance to focus away from it because you still fear it you're not running away from it you're still afraid right that's not avoidance at all it's not that's not avoidance at all let me scroll down a little bit more here okay I'm gonna throw this out here both of my parents were big warriors so I did get my anxiety from being raised in an anxious environment you were shown cues that say I should worry but worry is not the same as somebody who develops an anxiety disorder so it's important to understand the difference here did that contribute to the way you generally see the world and respond to it it totally did I'm not I'm not denying that but your parents do not do not pass along to you an anxiety disorder they may teach you certain ways to relate to the world which may tend you toward the interpretations that lead to an anxiety disorder but it's really important that we we draw a distinction here because many many people come from households where there are anxious parents or parents that are excessive warriors that do not develop anxiety disorders so does it have a does it play a role it totally does we can't we don't want to discount that or I'm not going to invalidate that but just be careful about just saying well my parents taught me to panic and run from it they they didn't they taught you to maybe be more fearful of the world but that's a thing that you can work on that that's you can work on that all right hope that helps Renee I'm going to do a couple more because we're at 35 minutes and I get to get cooking but what exactly is mindfulness this is this is an easy answer people sometimes think that mindfulness is a very magical spiritual esoteric new agey kind of state that you get into it is not mindfulness is nothing more than paying attention without judgment that's it that's all mindfulness is so mindfulness would be when I drink out of my Snoopy mug here I I pay attention you're welcome Carol no worries I pay attention to the mug in my hand I pay attention to how it feels it's cold because there's cold water in it right now I pay attention to the grip I pay attention to how it feels as I drink the sip how it feels going down that's all it is mindfulness is just paying attention without judging what you're paying attention to and without judging your ability to pay attention that's important too so mindfulness is purely just the act of paying attention that's all it is directing your attention somewhere and learning to keep it there and being non-judgmental about what it is and how you're doing with your mindfulness right really important because sometimes people get into that like oh I'm I'm not good at this I suck I'm not good at I'm getting it wrong if you're paying attention we all get it wrong like we're not good at that humans are not meant to pay attention that way we suck at it like we're really bad at it I meditate every damn day most of the time and I still suck at it like I try I try and you know maintain my focus on for me it's my breath when I meditate sometimes it's other things but and on my mind will wander so we're all terrible at it don't judge yourself for that all right so let's see here so I'll follow up on this one then I'm gonna kind of end it because I can't get to everything today sorry guys it is when you stay in the present okay that part is true if you're paying attention then by definition you will be in the present now the act of slowing down and when I wrote seven percent slower my most recent book seven percent slower believe it or not is a mindfulness manual it just takes the mysticism out of mindfulness because this statement instead of fearing the past and worrying about the future we do not qualify mindfulness so I would take that part away it is staying in the present how do you stay present you slow down and pay attention and that's it so we don't qualify it as we we enter into a state where we're not worried or we're not concerned or we're not thinking certain things we might think things we might worry that's okay that is a very important part of mindfulness practice is understanding that there's no specific state that you're after all right anyway guys I kind of I'm sorry I couldn't get all of them I would could what if I could but we're at 36 minutes and I got a boogie because I got stuff coming up in about 20 minutes I have to be ready for so thank you so much for coming by I love doing these next week we're gonna do I don't know what the next chapter is in the book let's look how about we look because it's right in front of me so I can do that okay Lesson 3.4 next Monday we are going to do stop avoiding going into fear we're gonna talk about why avoiding it's no bueno so that will be next Monday the same bat time same bat channel we got a bunch of these we're gonna be doing this into the spring what else will remind you again let me put up some maybe just some shameless plugs before I go because that's that's what I do now I'm getting good at the shameless plugs I've learned so if you want to read along and you don't have this book the anxious truth you can go right there to the URL that's on your screen anxioustruth.com all my books are there and if you have not subscribed to the anxious morning which is a free email newsletter that comes at every morning with a little three-minute podcast attached to it go ahead and do it theanxiousmorning.com or theanxiousmorning.email everybody is digging it and I am gonna tell you flat out I have produced a lot of work in my time doing this those email newsletters in the morning and the podcast that is the best work I have produced to this day I'm gonna go on a limb and say it that is the best work I have put out to date is the anxious morning what does this put do I put this on my Instagram I will put it on my Instagram so these videos stay in my Facebook group on my main Facebook page on my YouTube channel and I do post them to Instagram so in about an hour it will go up on my Instagram and it will stay there as part of my video feed so thanks guys for coming by I will see you next week have a good one