What's up everybody, Jason here with JaspysKazeBricks.com. We just finished this hip parade big game, Superbowl 127 box football mixer, ran on teams, and I just wanted to do a quick hit recap of just the cards that helmets and jerseys were all ripped at the end. If you guys want to see the hits for there, you can watch the regular video to square all of it to the end, and those are pretty quick. Of course, the cards took the most time, right? We ripped over like 10 cases worth of stuff, so let's show you a recap of the hits, and I'm trying to go at a good pace, but I'm not going to go too slow, not too fast either. I'm just going to grab randomly from these stacks here, but of course, a lot of hits going out. I think to my knowledge, there was two teams that went hitless, so those teams, of course, will be getting a box of Donner's football hobby ship sealed, I believe Wednesday when the box comes out, but you can double check right now too as well if you want to keep track of your hits or hitless. Here we go. So again, we've got 2020 Salah, Joe Burrow, Light Blue, Prism, Dyke, PSA 10, Little D Bill Samuel, Otto for the 49ers, David Montgomery to 25 for the Bears, Josh Allen Hollow, Rader Rookie, PSA 9 for the Bills, got a Drew Brees Rookie for the Chargers, a 1965 Philadelphia Jim Brown for the Browns, Jerry Rice, PSA 7 Rookie. I did three sets. Two more, okay. I'll click it right now, don't worry. Joe Namath, SGC 6, we got a Raymond Berry, PSA 6 for the Colts, Thurman Thomas for the Bills, Terry Bradshaw, Steelers, we got Frank Targenton, Gale Sayers, Joe Burrow, Justin Herbert Blue, Tom Brady, First Year Prism, Bass, Waddle Silver, 10, Najee Harris, Marble to 4, PSA 9, Asian Producer Refractor Rookie, Red, White and Blue, Josh Allen, we got a Tre Lans, Press Proof, PSA 10, Colpids, Genesis, PSA 9, great stuff, the majority of this stuff is actually great. All right, we got a Flawless Emmett Smith, Justin Herbert Otto, which is really nice, we got a Brady Rookie, 7, Marshall Lynch Refractor Rookie, Joe Burr, Camel 9, Justin Fields Bears 10, Dykut, Blue Scope Herbie 10, Michael Parsons Blue Scope 10, Dykut Select, Neon, Jalen Hertz 10, Colpids Marvels, PSA 9, Noel Huddle, Short Print, Justin Jefferson, PSA 10, DK McAfee, PSA 10, Red Ice, Seahawks, Mike Evans, autographed the 10 Rookie for the Bucks, Mahomesies, Raider Rookie, PSA 8, Tri-Killer, Cooper Cup, PSA 10, 101, Jerry Rice, Tom Brady, White and Blue, Devonte Smith, Downtown, PSA 10 Rookie, Roger Sparkle, PSA 10, 101 Quad, NFL Shield, Nike Tag, Laundry Tag and NFL Players, one-on-one, Zeke, Stafford of Five, Patch Otto from Impeccable, that's Lyons, Steelers, Najee Harris, 9-10, Calvin Johnson Rookie to 149 for the Lyons, and then a Jamar Chase, Eye Black, Patch Otto for the Bengals. We have a Saquon Barkley to 125 autographed for the Giants, another Najee Harris Rookie autographed, Orange to 25, Lamar Jackson, PSA 10, Raider Rookie, Norman Van Brockland, PSA 5, 1951 Bowman, Payne Manning, PSA 10 to 25 from Classics, Bob Greasy, PSA 8, 1970 Topps, Prism Purple, PSA 10, Jonathan Taylor, Tom Brady, Genesis 9-5, Christian McAfee, PSA, Pristine 10, sorry, BGS, Herbie Rookie, PSA 10, Rob Gronkowski to 5, BGS 9, and we got DK, Gold, Field Level, PSA 9 out of 10, Jaylen Hurd Sapphire, Rookie Gem, PSA 10, Trevor Lawrence, White, Select, Field Level to 35, Tua Togo by Love, Prism, XR Redemption, PSA 10 Select, Select, Field Level 10, Tyler Murray, and then we did get one Patrick Malone's autographed, BGS 8-5-10 out of 25, Patrick Malone's from 2019, Immaculate, DeMonte Smith, Genesis, PSA 10, Rookie, Seguin Barkley, Rookie Ticket 9-5-10, Christian Kirk Cousins, sorry, Rookie 9-5-10, Otto. Let me put away all these graded cards really quick. Now we're going to go to the Raw Hits, which some of them are going to be rookies, autos, patch autos, 101s, a lot of good stuff there. So here we go. Amon or St. Brown, patch autographed to 10, RPA, Drew Brees for the Saints, Ankar to 20, JK Dobbins, true RPA out of NT for the Ravens. We have a patch autographed to 35 CD Lamb, we have a freshman fabrics RPA of George Pickens for the Steelers, Kenneth Walker to the third RPA to Origins for the Seahawks, Dykut Zebra, Justin Jefferson for the Vikings, D. Bill Samuel playoff ticket to 49 for the 49ers, little Jaylen Waddle to 25 for the Dolphins, CD Lamb Origins Rookie Autograph to 99, Jermart Chase, Rookie Autograph for the Bangles out of Legacy, Rookie Ticket, Zach Wilson, we have a Mac Daddy Jones, Stained Glass, Downtown Barry Sanders for the Lions, RPA Kyler Murray out of Origins, dual autographed Kirk Cousins, Justin Jefferson to 49 for the Vikings, Rogers out of 10 auto, we got a 101 Diamond from the National, Barry Sanders, we got a Cooper Cup 101, Gold Vinyl for the Cowboys, 101 Larry Fitzgerald Auto, 101 Patch Autographed Marshall Fult for the Rams, 101 Aaron Jones Autograph for the Packers, 101 Dan Marino for the Dolphins, and 101 Ben Roethlisberger for the Steelers, 101 Nike Swoosh and Autograph, Gold Standard, Phillip Rivers, 101 Amonra St. Brown Autograph for the Lions, Kaboom Tyreek Hill, Tre Lantz, Radarookie Orange Autograph, Pay Manning for the Colts Patch Auto to 25, 101 NFL Shield, Nick Chubb, Daniel Jones Downtown for the Giants, Derek Henry for the Titans Downtown, Jermart Chase Legacy this time number to 50 Autograph for the Bangles, Justin Jefferson former the one Patch Autograph for the Vikings, Justin Fields Rookie Autograph for the Bears, Kenny Pickett Rookie Premier Patch Autograph for the Steelers, and the biggest hit of them all was a 2019 National Conventions Autograph, Patrick Mahomes, one of one and he signed it out like he used to, and it graded a 9-5-10, it did come with one of these $100 codes because it was a big hit, but that was probably by far the biggest hit of the whole break, and congratulations to the Chiefs right there. So I appreciate you guys, we did get a lot of helmets for a lot of the teams in Jerseys, if you guys want to check out the previous video, they'll be right at the end, it's pretty fast for the most part, so I appreciate you guys, that was the hit recap here for the carts in the Big Hit Mixer, thank you.


So, unfortunately none of the models will explicitly come and say, hey I am a delay model or something. It will be general description and you just model it, it just so happens that it will be some form of delay or the other which you can recognize. If it explicitly says delay good for you, if not still there will be delay existing especially this example. The chemical company makes chemical called no bug used in pesticides, they dump the byproducts in the say river once a week in batches such that the concentration is about 420 paths per million. So, we can assume that the river has 1 million gallons of water that means, 420 gallons of no bug are dumped per week into the river. During the course of the week, the chemicals are absorbed by the river's natural clean up process. The average absorption rate is 2 days, model the above system. Go ahead, how will you model this? The model is will be quite simple, but description will be quite elaborate. Since, I just covered first order material delay, logic suggests that it should be similar to that. Hopefully, you have got the model this one or at least you are thinking of a model like this not a drawn it. So, it is simple again when I teach I may use words like inflows and outflows and stuff, but you can use some proper rates like you know dump rate which it represents what is actually happening, absorption rate is what they have told, absorb a clean natural river's natural clean up process. So, it will clean up whatever is in the stock, it will clean up with an average absorption time. So, I will just write the equations in red. So, here the average absorption time is given as 2 days right. So, ensure your time unit in the model, in your model time unit is selected as days ensure that. So, absorption rate is nothing, but no bargain river divided by absorption time because it is a rate stock over time it has to be. So, no bargain river divided by absorption time is what you write here. Initial value you can put it as 0 in stock, its initial value is 0. So, now to kind of simulate let us take a very simple case. Let us start with a dump rate as just a pulse quantity we already seen pulse before. So, pulse has two inputs one is the start time other is the duration. So, at time 0 there is one pulse pulse one unit occurs at time 1, but what we already know it is it dumps 420 gallons right. So, I can so I just multiply this by 420. So, dump rate is 420 multiplied by pulse 0 1. So, you can make this in Vensim. So, you have all the information you need. The units of dump rate will be gallons per day, no bug units is gallons that is amount of no bug in the river gallons, absorption rate again gallons per day, day comes from here gallons comes from here, it weakly dumping is happening, but we will just take it as only one beginning of this week it happens. So, let us just observe that then we will model the next part ok. So, this is how the model looks, N80 natural absorption time I just came up with some acronym looks like it. If you look at the absorption rate is no bug by N80, N80 is given as 2 this is easy one dump rate is 420 into pulse of 0 comma 1 just a pulse input. So, if we run the model we can see that no bug increases and then as exponential decay. So, the halving time would be about 70 percent of your average delay is a 0.7 into D, D is 2 so 0.7 times 2 is about 1.4 1.5 about a week a day and a half time it will come into half the quantity ok right. Any questions on this so far?  So, dump rate what did I do there is another function called as pulse strain what pulse strain does is as a name suggests it gives a pulse input periodically. So, in this case it gives a pulse input starting at time 0 of a duration of 1 and it that repeats every 7 days that is once a week every 7 days it repeats and the end time is 50 after 50 time units pass no more dumping happens that is after ok. So, that is a very simple model that we have. So, it is called pulse strain you can look it up in the help file. So, every week 420 gallons is dumped it ok. So, let us observe what happens I am going to run the model. So, you go to no bug let me do causes strip it nicely occurs as we see here the top graph is the stock, stock increases up to 400 and the day of dumping 420 and then gradually comes down absorption rate also happens and so nicely fits our model right here. And dumping happens only once a week which is verified by the bottom graph it shows there is a dumping happening every week. Now is the actual fun part with SD where we have to do sensitivity analysis just making this is ok, but then to get insight we can find out say for example, what happens if the natural absorption time instead of 2 days become 4 days because of various reasons it has like capacity to observe has reduced and now the natural absorption time is 4 days. So, let me just make it 2 to 4 in that case what do we expect to happen can you have the same behavior? Behavior has to be same right once it dumps its truck observing correct, but then let us see what happens let us simulate no bug I just made the absorption time 4 from 2 days the dumping is the same absorption also happens, but now you see steadily the peak from 420 has moved closer to 500 right and 480 is a peak and it does not go now below say 90 there is always some no bug in the air in the river constantly. So, as absorption time increases say logically speaking now if the absorption time becomes average becomes say 7 days itself then by 7 days we can expect roughly little more than half of the no bug would have been absorbed. So, it will start accumulating. So, the accumulation effects is captured in this stock right here. So, if we did not have the stock we may assume that as soon as a dump eventually it will get absorbed, but it is not going to get absorbed until I stop the inflows again and then allow it for some time to slowly absorb and decay completely that is interesting what? Average absorption is 4 days or 12 days of course, it is going to keep increasing instead of we have tried with 4 days you can try with 12 days the next is interesting instead of dumping 420 every 7 days what if we do 60 every day we expect some behavior a different behavior. So, quantity is the same they got permission with government to dump 420 gallons a week what different does it make if I do it every day or do it once a week let us see what happens. So, I am going to go back and take the natural absorption time back to 2 in 2 case it came up and then went down very consistently nice graph I got. Now, I go to the equation of dump rate and we are doing 60 gallons a week every day time step is day. So, I just write 60 it means every day it will be 60. So, let me simulate see the absorption time is for whatever quantity whatever quantity I put takes an average of 2 days to deplete correct. So, now if I dump every day it does not give enough time for the river to absorb everything. So, eventually the total amount of say no bug in the river actually increases to what is it about 120 looks like right that is a interesting thing, but if you think about it logically what we did is when we dumped it once a week we gave it gave time for the river to recover. In this case we are dumping 60 every day. So, by the time we start processing 60 the next 60 is coming in every day it takes time the river cannot process so much, but still so there is a some amount of about 120 gallons that is end up in the river even though nothing else changed right there is a simple act of the same quantity, but different times actually has a nice kind of so it is better when there is a time given for river to recover rather than when it is very less though the quantity is the same. So, these kind of insights you can get only if you play with it simulated try with different values and this is the kind of policy which is just a simple model can use to explain different policies people want to you know as the same example do you allow them dump once a week or dump every day though the quantity is the same we can find that actually analytically there is difference in behavior and it does not kind of not a very complicated model to explain that. Second order delay you can imagine as cascading two first order delays that is it that is called a second order delay. So, the tank example looks very similar. So, the stock flow also we may able to guess based on this. So, what we are going to do is when you do a second order delay we will assume the total delay time is divided evenly among both the levels. So, if you want to model the second order delay we just take up inflow stock 1 let us call it exit 1 stock 2 this is your final outflow this is affected by d by 2 and this is affected by d by 2. If you want to model it when same directly as a function we will do it as inflow stock outflow t here we will write the equations for outflow as delay n as your in d initial value of any then we will write the order as 2 this is your order here the equations for exit is nothing but S 1 by d by 2. So, the equation for exit is S 1 divided by d by 2. So, exit is S 1 by d by 2 outflow is S 2 by d by 2.  So, when I say first order delay there is one stock, second order delay two stocks, third order delay I am looking at three stocks and some material is conserved. So, all the stocks and flows are getting interconnected and now the delay time is evenly divided among three among three of it. So, the equation for exit 1 is S 1 by d by 3 exit 2 is S 2 by d by 3 outflow is S 3 by d by 3. So, evenly divided. So, the average delay time continues to be d here as soon as outflow happens d by 3 amount starts flowing out of exit 1 as soon as inflow happens and after d by 3 1 third of the time this stock will start to peak and then of two thirds of the time this stock will start to peak. So, this so that is the peak of the outflow when this one peaks is when this peak outflow will occur. To model it in Wensum again we can use the similar delay n function with a third order delay. So, this third order delay I just showed you delays divided evenly among three levels. Suppose delay d is divided distributed evenly among n stages then instantaneous outflow of stock I will affect the subsequent stock the delay is evenly divided by n stocks. So, for each level I have d by n so, we have fifth order or sixth order delay.


Alright, well I'm going to go ahead and begin. I want to thank you all for coming and to first off wish you a happy Darwin Day. An annual international holiday celebrating the release of on the origin of species. And there are celebrations both real world and internet every year. And what I decided to do. I think back in about 2015, I thought, you know, every year I'm going to do an annual heroes of evolution talk. And what I try and do is pick a prominent researcher dead or alive that made a major contribution in particular to evolutionary theory, but also as much as possible to the general layman's understanding of evolution as well. And so this year, I was very pleased to see that Svante Pablo was awarded the Nobel Prize in physiology or medicine. And so he is someone who's been on my mind for a long time. In fact, when I was in graduate school, getting my PhD in lecture genetics and cell biology, the news of stuff he was doing was always popping up. And I was always thinking that is really cool work he would be basically extracting the DNA of ancient organisms and it would always make the popular press. And so it was one of these things of a topic in science that ultimately I just think has a lot of great appeal. And then it's actually over time had a few interesting twists and turns, and it's something that has just become as the technology again it's important here that the technology of sequencing has really advanced, and he's been at the center of that that it's become a really interesting good story for everyone. So again, compared to a lot of my science circle talks, this one will be a little bit more light on data, a little bit more emphasis on the impacts. And I'm also not going to cover all Svante Pablo's publication and research history, just basically focus more on the stuff that led to the Nobel Prize is more specifically relevant for for understanding evolution. So he's Swedish citizen. Now interestingly, his mother was a chemist, and his father was a biochemist, and he himself was actually a Nobel Prize winner so he's one of I think about seven pairs of people where a parent and a child ended up winning a Nobel Prize not necessarily the same one. As a European molecular biology postdoctoral fellow, he moved to the United States in 18 sorry 1987, going out to again another one of their prominent research institutions Berkeley. And that he joined a lab that was interested in the genomes of extinct animals so again one of the, you know for runners in that type of field. His career advanced. Yeah, we'll talk a little bit about some of those publications during that time, his early years, and he in 1997 became the founding director of the Department of genetics at the Max Planck Institute for evolutionary anthropology in Leipzig, Germany. So again, this term, you know evolutionary anthropology is really in certain ways, a relatively young field. And then what his particular subset of that is what's known as paleobiology or paleogenetics, or even molecular anthropology is really just a new field that he was one of the prominent founders of. In 2007, he was considered one of the 100 most influential people in the world. Now a part of that of course is because of the research he was doing in paleobiology. But I think it's also important to recognize that from his early six research successes he joined many of these high throughput sequencing companies. And these high throughput sequencing companies were really coming to prominence in lots of areas lots of fields of medicine, genetics, genomes. And so that was a part of I think the influence there not just a paleobiology but the fact that he was helping enable this important. And again, useful in many ways, technology. And in last year, awarded the Nobel Prize in Physiology and Medicine, quote, for his discoveries concerning the genomes of extinct hominins and human evolution. And why these discoveries are important. And I'll come to this at the end, but I'm telling you the end of the last page of the book up front here is that by having comparative genomics. We've learned from comparing human to chimpanzee human to ape, even human to, you know, macaque, bush baby genomes that there's important things we can learn about how genes influence our biology. And this sometimes can come down to very important medical conditions, or even just baseline physiology. And so the fact that Zontepabo was so involved and the premier person at finding our closest extinct relative again we're talking about in the old days comparing genomes where we know there's a common ancestor and both exist at the same time. But very rarely, well, I would say it's unprecedented at this point to have a close cousin, where the, the cousin species was extinct and so the, but the fact that we can bring so close that common ancestor to that comparative analysis, and that means actually really useful in a lot of cases and we'll talk about some of that but I really think this is really the beginning of a lot of those comparisons. And so we'll see a little bit more of it as people even beyond Zontepabo work on it. Alright, so let's get into it. You know, the main concerning thing is ancient DNA. And, you know, the old school ways of looking at archaeology anthropology was digging up bones, comparing bones, and that was a lot of times what you're left with. Now, there's a lot that's come along the way where we can do, you know, electron microscope scanning to learn more about certain tissues and other stuff, but those can only lead to so much insight. Now, of course, in human comparisons, you can compare other archaeological finds are not biological, but in terms of really understanding genomics, this, this is the main point. And there's a challenge here, right? Because what you see here is a person in old timey dyes, their body is preserved enough that we can actually understand that there's a body there in modern day times. And what's left behind is their tissue, including DNA. Now, I want to make a quick note here. A lot of the early paleogenetics attempts were to look at mitochondrial DNA, because you have many more copies of that available in your archaeological sample. You have that tissue, and for every one, sorry, for every like two copies of the genome DNA, you might have hundreds of copies of the mitochondrial DNA. So that's always a first part. Now, there's a limitation to that, which is mitochondria are only inherited from mother to child, never from the father.  And so that chemical modification, those chemical modifications and all this contamination of other biological organisms was always one of the big challenges. And I think that this is, while there was this early attempt to do some relatively basic straightforward applying molecular biology to see if you could sample and capture some of this DNA. What we've learned from Slantepabo's career is that over long periods of time, sorry, over the development of the technology that you're actually developing specialized technology for this purpose. These are things that you, while they might be related to modern applications of developing sequencing and DNA capture technology, there are lots of examples where it's specifically developed for trying to capture this type of DNA. I think that's something where we should recognize those contributions, although I am not going to get heavily into those details because that's not typically how I want to do a Darwin Day type of talking. Okay, so what is Slantepabo's early interest? And again, there's a lot of biographical information, so I can't necessarily say much about what really spurred us on was actually being very interested in Egyptian mummies. And so one of his early publications was actually the ability to capture from a preserved mummy a stretch of DNA. Now, what was interesting is that the thought being given certain preservation practices and also desertification. Again, one thing that's nice is if you have a very dry climate, then the amount of microbes that can contaminate your sample become less because there's just they don't have enough water to live and metabolize. On the other hand, you could imagine that preservation practices would also completely ruin the DNA. And I think that's probably ultimately what happened to the most well preserved ones. And so among 23 mummies, they were able to capture one stretch of DNA. They were able to clone it and get some degree of sequencing off of it and able to just show that it was possible. So this was, again, when he steps forward, it's very interesting 1985, but in many ways using kind of classic DNA technologies known at the time. But it's the type of information that again the review and I have this in the notes, the work cited note card and that's all available on the web. People are very excited about being able to look at ancient DNA to understand the migration, the alleles, the patterns of disease that you might see in ancient DNA and also do that comparison to modern day DNA. For example, sickle cell anemia understanding is actually has helped people understand migration patterns in ancient Africa. Okay, so, but the exciting stuff the stuff that was making the news during my graduate day times of my postdoc times was this looking at ancient mammoths and sloths again nothing tends to capture the imagination of the general press as mammoths and sloths for whatever reason. Right. They're planning on reintroducing mammoths based on genomic DNA from this company called colossal biosciences just announced this year. And so the challenge though was this basic idea like I mentioned that that chemistry can mess up DNA that even if you have something that's not contaminated. Does the basic chemistry still work. And so the thought was an anything older than 10,000 years old, you basically was could not amplify because it was too chemically damaged. But the thinking of this paper was, well, if we can find some well preserved mammals that were in the permafrost that maybe those extremely cold stable temperatures would allow amplification, given the techniques at the time. And in fact, that's what they showed they were able to amplify nuclear DNA sequences so they didn't have to rely only a mitochondria they were able to amplify nuclear DNA sequences. And basically sequence and compare them. Well, no, so just just a touch on the colossal biosciences ideas that they do want to reintroduce mammoths to the wild, primarily in I think Siberia, because they think it can actually help with reestablishing the ecology and basically the carbon cycle in a much better way that actually their arguments is that they can help with climate change by reintroducing the woolly mammoths. We can I can show you my thoughts on that later about whether that will be successful. Well, let's talk about it at the end, but I it's a good conversation, we can talk about it then. And so I just want to give an example of from this paper, the types of information they're able to get out from just amplifying this genomic DNA. And so here's one of the genes I mentioned, the a to a b gene. And what you're looking at in this picture is you have the Asian elephant and consensus sequence. But notice it's only about 68 base pairs long right it's only 68 base pairs not that long. But when you look down the dots in every row below the top represent being the same sequence, the same nucleotide, but when there's a new letter introduced as saying oh it's differing from that consensus sequence at the top. And so what's really interesting here is that they were able this time to capture enough nuclear DNA. And if they were lucky about where they were finding that they could capture information that was useful for developing phylogeny, resolving ancestral relationships between different animals. And that's what was was interesting and useful about it. And then ultimately, but also other groups did more so than the font a pop up group was that once you had good primers to an amplifiable amount of DNA for something, you could then also try to understand the archaic distribution of those animals within the habitats. And so you could understand maybe migration patterns or, you know, even population levels in some cases. Now, moving on to 2003. Also looking again at trying to resolve relationships of the sloth. And what they were able to show in this case is again the ability to find the advanced here was with new techniques, able to recover single copy DNA gen genomic DNA. And that also was from a warm air climate. And so this idea that they that you have to be limited to permafrost type of samples with good preservation was apparently no longer true. And so basically this opened up new avenues new ways in which people could broadly apply this across various climates and can various ancient climates as well. And what you're seeing here in the figure is just this example of where, in terms of looking at the ancient Shasta ground sloth, they were able to phylogenetically place it as being more closely related to the three totes loss as compared to the two totes loss, which of course was an argument that had led to many bar fights among paleobiologists little known fact. Okay, so let me talk a little bit more about putting this all together to really understand ancient organisms. Oh, so actually, yeah, so I didn't even mention that one part that what a coprolite is just a reminder that is preserved fecal matter specifically. And so, again, it's one of these things that people thought for a long time might be very hard to capture DNA from that because that's an extract that has a lot of biological microorganism activity going on in it. But if it's preserved well enough, then it actually does seem to work. And actually, the other thing that's really interesting of course about it is that if you were to think about the availability of sampling ancient DNA, then corpora lights end up representing what 100 times the body mass of an actual fossilized animal that you could capture that information from corpora lights, the availability of samples are is astronomically larger. And then you also can get insight from the diet that you had not just what the animal did, what the animal was phylogenetically but also what it ate. And so those are again all things that became very exciting as people. Well, you can actually recover plant DNA sumo, you can recover plant DNA from from corpora lights, other animal DNA, but then a lot of times it's the thing about it that the challenge is it's much more contaminated by microbial DNA. But in some cases you can actually learn about the microbiome that an organism had so lots of interesting stuff. So what I've mentioned so far are generally small amounts of DNA that they were starting to be able to capture whole mitochondrial DNA sequences so you actually had the entire genome of the mitochondria for many animals. But when it comes to trying to understand again that nuclear DNA that tells you the most information about what's going on within a species that was problematic. And the reason why and what I'll show here is how these were standardly put together with the technology.  And then what you see on the right hand side you see an example in the old days at this time of basically a radiograph where you put in what are called chain terminators, so that you get a radiometric reading at every length at which you're substituting in a chain terminating C for a regular C, and then a chain terminating T for a regular T, and you get this population of modules but the key thing here. And this is what's important to recognize is you need a pretty large population of DNA to work with to have that work very well. And a lot of times ancient DNA, you just had very short segments because the chemical modifications and damage already in your template your original template already basically created chain terminators. And then assembling a genome. What I have shown here is like your standard old genome assembly technique is that as you get lots and lots of single reads. Just little short horizontal bars. In order to assemble that to a genome you need overlapping sequences, and then you understand what the overlaps are between those chunks. And then you create bigger chunks and hopefully you can keep going until you create chunks as big as the chromosomes themselves. And one of the challenges for ancient DNA is very hard to get enough of these overlapping chunks, because they were so chemically degraded or you know, not representing the best regions very well that you just could not assemble them to make larger scaffolds and contigs and all these things. So, one of the big technology revolutions was what are known as high throughput sequencing. This in many ways relied upon a relatively different strategy, which was, instead of trying to assemble, I'll come to that question in a second. And instead of trying to create multiple long reads using your standard technology, that what these machines do is use some very specialized types of chemistry to just only make short reads. On the machine on the left, it's reads size, which is only 500 base pairs. On the machine on the right, only 150 base pair reads. However, the copy number you'll notice is 500 million or 95 billion, which ends up being enough overlapping short DNA sequences that you can assemble genomes relatively easily. So, the barragon does bring up a good point, and I want to touch upon this, that this is really great for mitochondrial genomes, because you have short reads. The DNA itself is not very complicated. It's very easy to find these overlaps, even among short reads. Now, the problem that comes up is something barragon mentions as tandem repeats, but there are also other types of repeats known as repetitive elements. For me, you know, talks I've given, I've talked about repetitive elements in genomes before, those really do complicate a lot of these analyses. So, again, important to recognize that it's not an end-all solution to all this, but it can be a very powerful way to understand all of the unique coding parts of genomes by taking this very separate strategy of sequences as much as you can, and then do basically computational analyses later to clean it up. And that's what's shown here in the next slide. Again, these last couple slides I've basically stolen directly off of, I should say, fair use borrowed off of the Nobel Prize lecture website, which again is also a citation. Flante Pabbo's talk is very accessible if you want to watch it independently. But here's this very fresh strategy that now this new technology could enable, which is, even if you have a relatively small amount to DNA, maybe not very many copies of it, maybe it's highly degraded. What you can do is, if you're trying to assemble those blue segments, is you just sequence everything, right? While you try and do as much as you can to reduce contamination, what you can do is just sequence billions upon billions of copies of DNA, and then use computers to recognize where the blue, again, your target genome sequences compared to all this other stuff you're getting. And what the red might represent, for example, would actually be human DNA, right? You're extracting these from sites, and then human DNA can be one of your contaminants. But because we know the human genome sequence, even though it might be similar to the antithal sequence, we can positively identify it in many cases as being human. And then actually there are other chemical tricks you can use to sort out old versus young DNA as well. So this is kind of a good pausing point, because I'm going to get into all the genome stuff in just a minute. But anybody have any questions about paleobiology or how the techniques on this work? While you're collecting your thoughts, I'll mention that. I think one of the challenges for reviving the woolly mammoth based on sequence DNA will be they're going to try and recover and use CRISPR to basically modify elephant genomes to be more like the woolly mammoth. The problem is, unless you understand all the repetitive elements that I think have very important transcriptional regulation controls, they've been modified to do that. What's going to happen is you might have all the genes in place, but you're not going to have all the regulatory control of all those in place. So I don't think it's as easy as they say it's going to be. Okay, Phil's amazed. If Phil's amazed, then I think we can move on. Alright, so let's talk about publication history with hominin genomes. That 2008 was the publication of the mitochondrial genome of Neanderthal. And this came from an individual. This is important to recognize that the geography of this individual was from Croatia. So again, Eastern Europe. They were able to, again, put together 8,000 base pairs of sequence, but they used it from five gigabases of sequencing DNA. And so that's the important thing to recognize that, again, that technology enables this type of thing. Sorry, those are 8,000 different sequence reads. The actual nucleotide sequence is 16,000. And so, you know, they put it together, they showed the conclusion that was really interesting, and this was big news at the time, was that the mitochondrial DNA did not match anything in modern humans. And I remember there being, I think there was even a whole NOVA special about this type of stuff. But if you look at this phylogenetics down here in the lower left hand side, that little red dot is, they're saying the split of Neanderthal from modern humans. And so there's no cross-contamination, no representation of Neanderthals. And so the conclusion, and then the right hand picture is just showing this in a more graphical sense, that where is this overlap? And because the red does not overlap the black, then humans and Neanderthals, the conclusion was they were ancestry related, but never interbred. At least based on the mitochondrial inheritance pattern, right? So they're saying they did not interbreed. Which of course then made 2010 an interesting year when they sequenced up the human draft, the genome of the Neanderthal. Well, yes, Sumo, you're pointing something very interesting out, is that when they published the whole draft sequence, and again, I only have access to the abstract for this paper, so I can only recapitulate what's there, is that when they compared that Neanderthal genome to present day humans, they identified a number of regions that may have been affected by positive selection. Genes involved in the types of things you might expect, things that might be different in metabolism or cognition. And the conclusion was that we share a lot of genetic variants, which means we must have interbred at some degree of overlapping time. And one thing that they could do by doing this comparison, and you'll see this come up in the maps, I'll show you multiple times, is that the Neanderthal interbreeding with ancient Homo sapiens only occurred really outside of Africa. That however the human migration patterns worked, they left the interbreed with Neanderthals in a relatively wide amount of space, but that Neanderthals never went to Africa. There's lots of arguments to say that Neanderthals were particularly cold adapted, but that really the gene flow back in Africa was zero, non-existent for Neanderthals. And I'll talk a little bit more about this positive selection. Positive selection is a term meaning that the amount of that gene, that particular allele of it, increased in the population probably because of some useful selective purpose that was existing at the time. Okay, so the other fun surprise in this whole story was that when they were randomly sequencing a sample, again a small little bit of a finger bone, and this was in Dinosava Cave, which is in the Altai Mountains in southern Siberia, they looked at the DNA and said, well, this is not human, but you know what turns out this is also not Neanderthal. And so from that they said, hey, there's another branch of human evolution of hominids, exactly Phil. They just came along, weren't expecting it. And so what this is showing here, and there's a little bit of a graphic representation of this, is that their estimates for what happened in the branching was that about a million years ago Neanderthal and humans had a common ancestor and then branched. And then within that branch, that is what led to Dinosavans and Neanderthals. And so that's the relationship between all those. And so, and then the branch point between Neanderthals and Dinosavans was more like, I think, 600,000 years ago based on that. But the other thing too is that really made the representation, given the dating and the time of these sample remains, that it probably meant all three branches existed at the same time between 30,000 to 50,000 years ago. So that became this question too of, well now that we have this Dinosavans mitochondria, can we get the Dinosavans genomic DNA, and just like what's in the Neanderthal draft sequence, see a signature of that in modern day humans? And bearing on again, that brings up an interesting question which has not been answered, right? So the absence of human DNA in Neanderthal mitochondria, or I think what you're actually meaning is that the lack of Neanderthal DNA signatures in human mitochondria, maybe the only way this worked was Neanderthal men interbreeding with human women. And so, I've not really seen anybody give a good answer to that and we can speculate a little bit at the end, but let me just push that conversation off to the end. So what they showed in this paper, in terms of looking for the existence of Dinosavans DNA signatures in human genomes, was that, and then also having the genomic DNA, they gave them a better resolution of these divergence times, is that human diverge from Neanderthals and Dinosavans more like 800,000 years ago, and then Dinosavan Neanderthals, they had a common ancestor that diverged about 600,000 years ago. Now what's interesting is that the signature of Dinosavans relatively limited in human populations, but there's a relatively high percentage of it in Melanesians. And so this map here is showing an extraction from their paper that if you think about Micronesia, Polynesia, the islands, the larger islands right above Australia, but not Australia itself, you see a pretty good signature of the Dinosavan DNAs. But remember also, the likely crossbreeding of this probably actually occurred back in Siberia, and so something happened there that then later human migrations led to this. All the way in, we don't know all of the geographical location of all the Dinosavans either. Right, so Aurora points out something too, which is, and this is a number I wanted to just gloss over, but I think it's important to recognize, is that within human populations, if you take probably all the humans, sequence all their DNA, and asked how much of the Neanderthal genome is covered, is represented, they think we can recover about 40% of the Neanderthal DNA within the genome within the existing human population. However, the fixation of Neanderthal DNA into humans is mostly concentrated in ancient European populations, and it's typically anywhere between 1.6 to 3.5%, or something like those numbers. So almost all of this, again if you go to 23andMe, and we'll come back to 23andMe again later, they will tell you how much Neanderthal DNA you individually personally have, and again, which part of the Neanderthal genome might be very different. But we'll talk about, from an evolutionary point of view, which regions that might be likely to be. Okay, so once, so in the all-time mountains, they also were able to capture a much better purified genome from the Neanderthal. So this is known as a more high-quality genome sequence, and they wanted to make some other inferences about it. Berrigan, just remind me about that question a little bit later. And so, when they took this high-quality sequence, they make the estimate that, again, the particular proportion of amount of DNA that you have, again, now that you could resolve a lot of the smaller differences in nucleotide alleles, and really map this a little bit better, is that in general, the amount of Neanderthal drive DNA in people outside of Africa is anywhere between 1.5 to 2.1%, although that would be much depth a little bit higher within the ancestral European populations. And then they also could do a better comparison of Neanderthal to denosevins, and what they found was something a little bit better. They found certain signatures in human leukocyte antigen region, and the CRISP gene cluster chromosome 6, both of which are involved in immunity and sperm function. And so, this interesting little inference about immunity is interesting, because we will see in repeated fashions that perhaps one of the best values that you can get by having interbreeding with a more distant population are things that are affecting, and you can see these signatures because you have very strong selection from pathogens. Again, pathogens are the things that sweep across populations and can lead to very strong selective pressures. And I think the best example that people always talk about with this is the Black Death, I also know it as bubonic plague in Europe, where in its first pass it killed two thirds of the population, and then every subsequent epidemic, a small number of people passed away from it because there were genes that enhanced people's immunity to it, those were the survivors. And that's evolution, that's how evolution works. Okay, so here's a little bit of a map that they put together, which is again showing the branch points, not necessarily in time, but giving some degree of the branch points of these different hominins, and showing you in these orange arrows where there seem to be specific gene flows from introgression crossbreeding. And so you'll see some of the Neanderthal genome pass into De Noseva, but that more was only limited to Siberia that weren't the European Neanderthal populations, whereas the European Neanderthal populations clearly branched into the Homo sapiens, although at a point after the migration out of Africa. And then you can also see some other De Nosevan integration into very specific subpopulations of human populations, that as I mentioned seem to be representative of Pacific islands. And then what's also interesting here is that there's enough signature from the DNA that we look at the genomes and compare them, there seem to be these islands and haplotypes of yet another unknown hominin that has interbred with De Nosevans. And so there are no archaeological samples that represent that actual genome, but by comparing having enough DNA information you can say, hey, there's something extra in here that's not a part of these species. Okay, so while I think it's really important to recognize the power of these understand paleobiology, and to understand human migrations, understand this, that's not going to win you a Nobel Prize in medicine and physiology. And what's really I think the last part of the story is that the power of having these genomes and looking at these signatures has led to advances in understanding the medical pathological conditions of humans as compared to Neanderthals. So here's the first example. This is just an interesting one about a nerve receptor protein. And that nerve receptor protein is different between the Neanderthals and humans. And they did a couple things to characterize this is that one, if you look at a sample of British people, that where you have both their genomics, their allele and also some sort of other sampling data, you can ask them, you know, things about their lifestyle, and they say they experience more pain than others. And then also you can do physiology, you can basically take this back to the lab and compare the expression of physiology these with molecular chemical assays and compare the difference. And that's what they found is that the this nerve receptor shows reduced inactivation meaning it's sending more and more of a signal back to the brain. And so because the suspicion they have is that this is a pain receptor. And so their argument is, perhaps Neanderthals are more sensitive to pain than humans. Again, we don't know. But that's, but this is the type of analysis that then when you have this comparative genomics, and you have sample size data, and you can do some molecular biology on it, you can make these inferences. Another great example is the progesterone receptor. And so progesterone is important for preparing the urine lining for egg implantation and maintaining the early stages of pregnancies, as well as, again, it's involved in the menstrual cycle as well. Again, if you, does anybody know of any particular drug that might relate to progesterone offhand? What they showed is that these are different between Neanderthals and humans. And that given what we know about modern day progesterone is that this can have a consequence for carrying pregnancies to turn. And so I want to just focus a little bit on the map here for a second, what these pie pieces are showing that the orange slice represents more representation of it within that population. Again, you'll see in Africa, very small, in Europe, a lot higher, but then you'll also see it in the western world populations and within Asia as well. And Aurora points out, I think a useful thing to recognize is that genes that actually help you with directly with fertility and the amount of children you can have ends up being a very powerful thing in terms of human evolution in terms of a what's called a positive selective factor. And so in following up on this, you know, they look at that particular polymorphism, and they again use a comparison to the UK Biobank when they can identify different patients and given different descriptions of what their pathologies or why they went to the hospital, is they found a so-called negative association between the Neanderthal allele and hemorrhage in early pregnancy. And so what, and then also reporting less or fewer miscarriages within that population, if you're a carrier of the allele. And so, and then also looking at just overall census data, you can see that those individuals carrying the Neanderthal allele had significantly more sisters, although interestingly not an increased number of brothers. And so these, again, somewhat indirect logic and reasoning, and again inferring from excellent data is that these may reduce the frequency of miscarriage. Although this is in a sense competing against the fact that the lower progesterone levels and receptor levels might mean that fewer pregnancies implant well in the first place. So you might have a somewhat lower fertility in terms of implantation, but it seems like the net effect is that the trier is done, is that you do have a net impact that's positive for fertility. And also, and I think this is where the sister information comes in, is that remember one of the causes, one of the major causes of death for women is, you know, death during childbirth. And so the fact that you have adult sisters that are more prevalent from the statistical data may indicate that, in fact, less miscarriages, less issues with pregnancy. So here's something that you may have heard the news and I have to apologize yet again for bringing COVID back into the conversation. I know lots of people are sick. I'm tired of hearing about COVID. But this was an interesting set of data that came from an analysis by 23andMe. So 23andMe is a company that has a huge amount of biological genetic information about people. And they basically sent out a survey to their subscribers, people who set samples again after the fact and said how bad was your COVID. So what the graph on the left represents are signatures of correlations between severe COVID, not necessarily the prevalence of COVID or having COVID, but actually having severe COVID specifically, and a potential allele signature. And so you notice there's a very big spike on chromosome three. And the right hand image shows the set of genes that are in within that signature area. And what's being represented there in the red are the genetic variants that are correlated to the risk variant, and those risk alleles match this one particular Neanderthal genome. And that black bar at the top, that black bar, that's basically a haplotype, again, a whole stretch of DNA that is from Neanderthal. So what did we learn about that? So again, here's the signature seems to be correlated to Neanderthal of having the Neanderthal DNA makes you more susceptible to severe COVID. And then here's the geographical distribution of this, and you'll see, again, low in Africa, a decent signature in Europe, but interestingly, a pretty high prevalence of this gene in that, you know, Middle Asia region around India. And so the interesting follow up to this, and I think this is really important, is that that DNA segment may also relate to the expression of something known as CCR five. And when a follow up paper, again, slantepabo didn't publish this, but the person who published the COVID-19 paper with him published this by himself, showed that these individuals with this particular gene segment that is susceptible to severe COVID is actually protective against HIV infection. And so this is one reason why it may be a relatively prevalent gene in certain populations is that it was protective and a survival advantage in places where HIV was also prevalent. And so this is the, as slantepabo put it in his talk, the double edged sword sometimes with evolution. Now there's another variant that came out that seemed to connect with Neanderthal genome to less severe COVID. And again, I don't know a lot about the biology this one in particular. But what they showed is that a set of genes that are involved that encode work known as oligo, so oligo adenylate synthetases. And these are things that are involved in immune responses, as well as being involved in double stranded RNA. So again, double stranded RNA is something that comes from your right invading viruses. So the fact that there does seem to be this prevalence and you can see a worldwide distribution of this, there may be some positive selection. In particular, a places that have gone through other types of maybe even specifically coronavirus infections, where this was a survival advantage for them in the past, and then you've retained some of the survival advantage with a new infection to new COVID-19. Well, yeah, just to point out that again, there are two things that go on when you think about these pies is that in both cases in Africa, if you don't have the Neanderthal genes there, you can't have a positive or negative selection for them. But then when the Neanderthal genes are there and available to go through evolutionary forces, you can see different amounts of them coming back and forth. But again, that actual selection, those actual increases or decreases in percentages will reflect the living history of those individuals and the pathogens they face in this case over time. So that's kind of why you see some similarities, but then also there are going to be some differences between two. So for example, if you look back at the previous one, you know, the amount of stuff happening in the North American population seems to be a little bit less, whereas you have a lot more of it here in this particular one. So anyway, last couple. Another example looking at metabolism differences is that one thing we don't we have genes to protect us from oxidative stress, because oxidative stress is something very damaging to the molecules in the cells. And one thing that we that you could look at is the fixed the fact that the fact that there's something that was fixation, which means that at some point in time all humans had the fixed version of the gene 100%. In other words, fixed means 100%. And it wasn't until later that in Brady interbreeding with the air falls they got introgress back into humans. And again, when they look at the prevalence of it. They see a relatively larger signature in Indian Sri Lankan Puerto Ricans bangledeshes, but a lot of other world genomes don't show this illegal. Again, when you actually look at what these genes are involved with are involved in immune mechanisms, or other types of chronic diseases like inflammatory bowel diseases. So what the images show here in this case is again, while some chemical assays, you did not see very much of a difference between those particular enzymes between humans versus nanofall. But that in other cases, when this case looking at NADPH oxidation, a super oxide production, you can see the difference between the gray, the modern human version of the protein, the enzyme versus the brownish nanofall version. So again, coming back to doing molecular assays to actually compare the biochemistry of these enzymes and how they're different. Okay, so yeah, or you're asking a question about, you know, what do we consider modern human. I mean, in this case, you have a very specific consensus sequence where you can make the argument that the enzyme you're looking at is a modern human version. Now, when you talk about current North American populations, I believe and again, I would have to go through the materials and methods. Typically, that just means you're geographically locating these people and saying the North American. That's not trying to represent the old, you know, either cross ocean or cross Alaska Ice Age bridge to say North Americans are specifically derived from those right now. Sorry, that's what I was just answering that it's typically that's just going to be based on the geography of who is here is going to be based on geography, not any sort of racial inferences. Although again, there are ways that you can refine the data if you want to, but typically with these types of large global maps, they're not doing that that would take extra time and money is just geographic. This person was here. Another example gene is again coming back to metabolism is cytochrome P 450. And it's getting encoded by the gene notice C Y P to C nine. It's actually pretty highly polymorphic gene and present a humans, but this is an important time we know about a lot because if you certain people have this variance can actually get overdosed on things like warfarin, which is a, you know, blood thinning reagents and phenatoid, which ended up being direct to the end time.  And so related to C Y P to C nine is also C Y P to C eight, which encodes and codes another cytochrome gene and important also for metabolism as some other pharmacological agents. And what's actually interesting is that there are certain alleles again the asterisk to an asterisk three represent alleles versions of these genes, and they tend to co segregate. And what they actually found by with the high resolution nanosol genome is that these alleles are both co inherited from nanosol. So again this is like one of these examples of a haplotype where that whole chromosome segment of two genes that are near each other ended up being inherited, and then has maintained that haplotype through multiple generations of humans. So in this case, I just mean drugs that are known and actually prescribed to people in modern day. So warfarin is something that's commonly prescribed for people who had a heart attack, for example. Again, I don't have on the slide but like the C Y P to C nine asterisk to allele is about 14% of the European population, at least in one copy. That's his prevalence not necessarily where it's almost like us. And again, the reason sumo this is important. And it's not like xenobiotics in general what it is is the biochemistry of what these enzymes do is specifically recognizing these and breaking them down so again drugs work in different ways sometimes you actually have a drug where the active version of what you imbibe. But there are actually other examples of drugs where the active version of it is what the liver metabolizes in a way, but in all cases whatever pharmacological dose you have that always is degrading over time as you clear the drug from your body so many cases, you're just pissing it out. This appears because it dissolves it's a solute and then you you get rid of it that way, but there are lots of drugs where the dosing information is very relevant to how quickly the body specifically degrades it as well.  It's based on its removals rate. And so if that removal rate is different because the enzyme metabolizing it is defective, then over time you're do you're overdosing somebody based on those, what's called pharma kinetics. So let me give a quick summary of these genes that, you know, the nerve receptor is one that shows differences between the intervals and humans that may be relevant. I'm not sure that and people reporting symptoms related to it, progesterone receptor against something related to pregnancy that very clear signature this. I give a CCR five which I put in here indirectly because it's an expression of the allele that they looked at. Again, enhances COVID susceptibility. So it's maybe prevalent because it helps with HIV resistance. And then OAS one through three, again, involved in viral defenses and the fact that Neanderthal one maybe seems to be given a signature that makes it better and enhanced against COVID 19. They actually showed in that paper to that it helped against MERS and maybe even COVID one. And so a little bit of, again, more in depth study was interesting there. Again, and this was the power of looking at these various biobanks, as well as, you know, registries of people's symptoms and diseases and things they went to the hospital for is you again see a signature for things related to diseases related to that. But it's not as clean cut as say sickle cell anemia. And then these last two, maybe things related to drug metabolism. Now, if you go through some of the pop those publication history beyond what he talked about in his Nobel Prize talk. There are a couple other interesting genes that that they've done comparative genomics with including Fox P two, which involved is maybe in language and speech development. There's 18 a and killing one, which are involved in chromosome separation metaphase growth, which along with novel one may reflect differences in how brains develop. And so that's actually one of these big questions that you have is what really distinguishes Neanderthals versus humans could be their use of language, their use of tools. Could there be things having to do with their brain development, where Neanderthals specifically cold adapted that's why you don't see them in the north and lesser climbs all these types of things are the types of questions that are very interesting. So here's a slide that I put it in here and maybe you can review it later. I just want to make one quick point is that really. This is from a review of looking at ancient DNA analysis and the various different landmark things that occurred during that period of time. And you'll see that really he starts it at 1984 when the first arcade DNA sequences were captured. And then things that progress both in the wet lab, as well as what's known as the dry lab, another way for saying computational analysis and whatnot. And I do sometimes wonder that maybe there's still another Nobel Prize for chemistry in the works first Fonte Pablo, given how much he was involved in both ends of the companies involved in sequencing. The genomic sequencing and other aspects of these types of techniques. But this is kind of history. And that paper is a nice review of the history of modern techniques if you want to get more in depth into the technology. Now, like I mentioned, Fonte Pablo relatively quiet person, he's never been as far as I know he's never been parodied on the Simpsons. I couldn't find anything about South Park with him being in that. And so, you know, I'm a little disappointed because I wanted to show something cute funny, but he did write a biography and autobiography. The NFL man of search loss genomes were talked to me about his life. And he has been, for example, in prominent documentaries been so much interviewed. Although the, the, if you look on YouTube and look for interviews with Fonte Pablo, he's all over the place and he's represented in a lot of modern media. Anyway, so for these reasons, for the power and the investments and the knowledge and discoveries he's made in the history of analyzing archaic DNA, but then also in the ability to do comparative genetics, really get involved in the biochemistry and understanding those differences between humans and other genes and those ended up being many cases relevant. And in the future, we expect them to be increasingly relevant to understanding human medicine and physiology. Fonte Pablo is my 2023 hero of evolution. So with that, I will be happy to take any questions or clarify things and any comments that you guys might have so thank you for your attendance and coming today. Yeah, not. I'm glad I could talk about again, he was someone that I just always whenever I saw publications come out when I was going through my professional development. But yeah, just a quick mention, Berrigan is hosting naked scientists in two minutes. So if you want to go listen to that podcast from nature from, from I think nature UK again, Ergon's not getting naked. So that's not what that's about. Yeah, so you bring up a really interesting point. And that is, you know, early on, there's a lot of bias, in terms of how people thought nanophalls and humans interacted and of course that bias came down to saying, oh, humans are just superior. So we out competed them. And we just basically beat them off and we didn't interact with them. We were just better. And one of the rationales behind that was some early physiological data where people said the NFL's I think had shorter throats. And so maybe they couldn't enunciate as many things as well. So their language capacity was just not good. But I think, you know, with more time or research and even the genetics, we don't see those. We don't see signatures of obviously one being inferior to the other in the vast majority of us, although I will say that you know, when they did this organoid, if anybody saw this that came out, I think about two, maybe just last year, where they're trying to compare the NFL genes and the development of brain organoids again organoids are like a tissue like organ assembly little tissues you have in a petri dish. See a shot and tell thanks for hosting that because the human ones were a little bit more complex than the Neanderthal ones. And so there's some argument that maybe again when you think about all cognitive brain intellectual capacity. Maybe they're still as a signature that's different between those two. Yes, you're welcome Keisha. Thanks for coming. And I think it's I think it's unfair to say Neanderthal behavior, we think they were actually much more sophisticated than we were. Okay, so the term species, we have to be a little clear on what we mean by it is that we would still characterize them as separate species, because for the vast majority of their interactions that they had enough time again 800,000 years is enough time to be your own considered your own species. However, the barriers to interbreeding were not 100%. And so it's like the idea of a horse versus a donkey versus a mule. You still consider the horse and the donkey to be separate species because there's enough genetic separation to say that they are their own species. And I can't interbreed and make, you know, a still hybrid offspring that answer your question, I hope. But I think one thing that I'm. Yeah, so I kind of bypassed this because wasn't spontaneous work, but as far as we can tell, Neanderthals were red heads. And so we believe, although this is not this is not 100% the case that some of the redheadedness you have in humans probably came from Neanderthals. But I haven't looked up a lot of the paleogenetics of that particular gene, but that's one example I don't know about the blue green eyes being a Neanderthal thing that's prevalence in humans because of Neanderthal interbreeding. But again, it's not one of these ones where you when when you compare genes of Neanderthals versus humans. The ones that I gave examples of were in many cases clear examples where you could distinguish Neanderthal versus human but there are lots of examples of leels where because they're highly polymorphic and really subject to a lot of mutation pressures or selective pressures. Maybe we also don't have enough multiple read information from the genome it's really hard to exactly say where they came from. So I think that's important point to see them. Well, I think I have a second here folks. I want to go back to that business about sensitivity to work from being a different being associated with the Neanderthal gene. I mean, We're from didn't exist back then but Neanderthal, but the boyfriend is a current. And I wonder just, and, and based on what we're saying now with this in mind, does that indicate something about the Neanderthals environment or diet I mean c'morans are in like clover and stuff like that. And then c'morans. I know, or c'morans are also light sensitizers. You know, if you're too many c'morans can cause you to blister and all that sort of stuff in the sun more than anybody else. And I wonder if that's all interrelated with this part of the discussion or Neanderthals are redheaded and paleide, which also indicates a lack of regular moment which is protected against the sun. I mean I'm piling up a lot of stuff here but what I'm getting at is is what does this sensitivity to work from being related to the Neanderthal gene say about the Neanderthals. So soon you ask, yeah, this is that's really the big question that these always occur right and how much resolution can you get and really understanding it and you really need to know a lot about the specific light life history of an organism to come to good conclusions. And so we don't my answer is we don't know genetics genetics as a whole can only tell you so much about these signatures. And so, you know, but let me give you an example of like a type of analysis which is for the longest time, people did not understand why, why sickle cell anemia was so prevalent. Right. What was why do you why so many African Americans have sickle cell anemia. And then it wasn't until you could correlate that with resistance to malaria, which again, one reason you could do that is you could basically do a geographic overlay of sickle cell anemia to malaria prevalence and you could see the signature.  Like you're saying maybe there was some pathogen, or maybe there was some drug in their foods in their potential food supply like a potential poison in their food supply, where having resist having a decreased ability to metabolize it was a survival advantage but we just don't know. But that's the type of thing where once you have the genetic signature something then this can get people to ask those questions and then maybe explore it further and come to an answer. Hope that answers your question. Yeah, so brings up the northern climate adapted aspects of of Neanderthals. You know, we don't, I have yet to see papers that explicitly describe the genes that do seem to correlate with that other than of course the fact that the vertical skin that does come back to the vitamin D conversation that some people wonder, and I don't know that I've looked through all the little haplotypes for this but you know being fair skinned is a big advantage in northern climates, where in the old days you needed UV light to make vitamin D and vitamin D is really important for like bone density and strength.  In addition to maybe just being cold adapted but also to being low light adapted. And so the fact that maybe some of that got got reintroduced into humans has what allowed humans to really spread outside of Africa some people made this argument that without Neanderthals. Maybe humans couldn't spread as far as I did. Right, but I think the majority of the interactions of humans that 30,000 50,000 years ago, that that I think that's looking so late police is right is we typically consider that to be a pretty cold time that was really a little bit pre. The major ice age was 10,000 years ago but even preceding that 10,000 years ago, that was pretty cool. No, no, no, I know I know that cold adaptation of Northern climate are there. You used to be talking about cold but I wanted to bring it back to. Well something I knew more about and that was vitamin D. Great. Yeah, thanks. Thanks for all coming. Hope you all really enjoyed it. If there's anybody you think would be a great candidate for a hero of evolution. Let me know.


took a best offer of $425. Bought two bags of stuff for $1. These were in there. Sold for $125 within 12 hours of being listed. Hey, fellow buddies, thanks for watching. Amazing, right? All right, let's get started. Hey, fellow buddies, thanks for watching. All right, one of my favorite things to sell is plush. So I decided to feature some of the Bolo Buddies Facebook group members, some of their Bolo's that were plush. So I posted this in the Facebook group. If you guys are not in the Facebook group, definitely go join it. There's a link down below in my link tree, but we are going to talk about plush Bolo's items to be on the lookout for items you can hopefully buy low and flip for a profit. This first item bought from the Goodwill for $3.99 and accepted an offer of $85 on eBay. This is what the item looks like. It is a vintage Stife Cappy dangling frog Travira Velvet and it has the little button tag on it. So yes, Stife is definitely a Bolo. Purchased at an estate sale for $1. Vintage gunned Bunky Bear tan peachy pink plush. And it looks like it's from the 1980s. And this is what it looks like. Just a teddy bear. $54. Are you kidding me? It's awesome. And the buyer paid shipping on that. Got these three gunned 101 Dalmatians from a local in person auction. I bought two bags of stuff for $1. These were in there sold for $125 within 12 hours of being listed. Wow, that is fantastic. I don't think I've seen those before. Here's a fun one. Bought at the Goodwill Bins for about a dollar. Got full asking price of $45.99 plus shipping. First time contributor, long time watcher. Thank you for all your time and effort that goes into your content. Oh, thank you so much. And thank you for coming over and sharing. I appreciate that. Oh, it's a SpongeBob. I forgot to tell you what the plush was. Vintage Patrick Starfish large 27 inch plush stuffed animal SpongeBob from 2002 Nickelodeon. This one comes from Bolo Buddies member finding fun things. And this is her eBay store. And what is a Bolo Buddies member? That means they have joined Bolo Buddies memberships on my YouTube channel for perks at level two or higher. And they get a screenshot of their eBay store or YouTube channel like this. And then I link them down in the description of the video. So definitely check the description of the video before you go and subscribe and follow these eBay stores and YouTube channels. They have fantastic Bolo's. I got this Webkins Harp seal at the Goodwill Bins for less than a dollar. I knew to buy it because I heard from this channel that Webkins with the code would sell. I sold over $29.99 plus the buyer paid shipping to Alaska. I was happy with the nice profit. Yes, Webkins. Some of them can go for crazy money. I actually have a video talking about Webkins that go for big bucks. They range from bread and butter to big money, but definitely look them up if you see them. Took a best offer of $425. It is a Folk Monus Disney Mickey Mouse Hand Puppet discontinued collector's item. And I just did a video on puppets. And this is one of the items that I talked about in the video. So if you have not seen my puppets video, definitely go check it out because some puppets go for crazy money. She did not say where she... Oh, here she goes. Paid $50 at a thrift store and took a best offer of $425. Just at a yard sale for $1 sold on Mercari for asking price of $61 plus shipping. Just one. I included the second photo so it was easier to see without the sold banner. Oh, it's so cute. It's a Disney Aristocrats Plush Barrel Berlioz. Maybe that's the name of it. I have no idea, but it's so cute. Here is a Care Bear Yellow Plush Fun Shine Sunshine Yellow Bear, another Mercari sale. And I do have a video talking about sold prices for Build-A-Bear and how a lot of times they sell for more on Mercari. So one thing I can recommend, especially if you do plush or toys, is to get your items on other platforms because a lot of times these items are flooded on eBay and you're reaching a whole different audience on different platforms. I start my items on eBay and cross posts with List Perfectly to Poshmark and Mercari. You can check out a demo video of how List Perfectly works if you're interested in a cross posting service. That's the one I use. I've been using it since 2019. If you watch that video and it's something you want to try, you can use coupon referral code BOLOBUDDIES, all one word to get 30% off your first month. If you're not on Mercari and Poshmark, I also have links down below where you can join with my referral link and you'll get money to shop. $10 on Poshmark to shop and $10 to shop when you join on Mercari and then another $20 when you sell $100 worth of stuff. So a total of $30 over on Mercari. So those referral links are down below. If you guys use those, that helps me and it helps you and it's a win-win. So I appreciate it. He said, I bought a lot of 50 Care Bears for $30. One of my sales probably profit upwards to 500 reselling them. So wow, 30 bucks for 50 of them and this one alone sold for 61. So I mean he made his money back with one and he still has 29, or no wait, 49 left to sell. Wow, wow. Bought at the Goodwill for a few bucks, sold for full asking price of $38 on eBay. This was one of my first sales as a new seller and my picks were not good. My little pony, Minty Build-A-Bear Workshop Plush from 2016. There you have it. Paid 50 cents at a church rummage sale sold for full asking price in two weeks. $179.99. It is a Prima Classic Collection Golden Retriever Dog Stuffed Plush and this is what it looks like. Yeah, those realistic looking animals, they can be big money. I got this Disney Parks Wishables Little Mermaid Ariel Plush at the Benz for less than a dollar. It's only six inches and I thought it was a TY tie at first but it had a different look. I comped it and was surprised at how much some of the wishables can go for. It sold for full asking price of $80 in one day and this sold on Mercari also.  Check it out. I have never heard of wishables and I am definitely going to be on the lookout. Found him not too long ago after Christmas so it was on clearance. Paid 50 cents I think. Sat for a little while but took a best offer of 50. It is a Zanny Brainy Zac Frost Snowman Plush from 2000 with Bell Rare Vintage Christmas Holiday and this is what it looks like. Purchased at the Goodwill for $1.99. Sold in about three months for $139.99 plus shipping. Realistic Husky Plush made by Sterling and is 23 inches. Got at the Benz for under a dollar sold for full asking price. It is a charming one of a kind handmade mohair artist teddy bear. Jointed weighted initials on foot and it sold for $100. Found at the Goodwill Benz. Paid 40 cents for it by weight and took a best offer of 63 in less than a month. Food related plush seemed to fly out of my store on all platforms. It is a Disney Pixar short happy boa dumpling plush stuffed food. I would not think that that was food. I guess it's a dumpling. Bought at the Goodwill Benz. Maybe 50 cents was asking 85. Took a best offer of 70. It is a little Snoozoms 9 inch puppy dog plush stuffed animal Nancy Cordaro 1996. $70 and that's what it looks like. Do you guys not love plush? I mean it's so easy to ship. It's easy to find. It's easy to list. Most of it is bread and butter but man sometimes they just surprise you. Cats vs Pickles by Mr. Argyle purchased at the Goodwill Benz for 25 cents and sold overnight for 40 plus shipping on Mercari. This is what it looks like. It's a beanbag plush. Here's another Mercari sale. Crazy. Pokemon Center Miao 2 plush purchased at the Goodwill Benz for 50 cents sold a couple weeks later at full asking price of $59. This is what it looks like. It is a Pokemon toy from 2013 Japan. This one comes from Nina Jules. This one comes from Nina's Jewel. She is a Bolo buddies member. This is her YouTube channel and eBay store. Jelly Cat Jaguar picked up at a church rummage sale for $2. Took a best offer of 75. Wow. Very cool. Again Jelly Cat is a fantastic brand. I have done videos on Jelly Cat. I have also done videos on realistic plush. So this hits two Bolo areas. Found at the Goodwill. Stop complaining about the Goodwill. Half off green tag $4.99. Paid $2.62 with discount. Sold on eBay for full asking price of $239.99 and took about two weeks to sell. It is kick the cheat plush homestar runner strong bad working sound 10 inches. So the sound did not even work and it sold for $239.99. So yes Goodwill is a little crazy on some of their prices but I will say that sometimes you can still get a fantastic big money Bolo. So don't quit looking. I've seen some posts on in the Facebook group of people sharing how much the Goodwill has things listed for and sometimes it's just they were one person was comparing what you can buy it for on eBay and what Goodwill had it listed for and it was way cheaper on eBay. So sometimes they're right. Sometimes they're wrong. It just depends on how much they're researching where they're researching are they looking at active listings. I don't know. I mean I don't know but what do you guys think about Goodwill prices in your area. I know in my area it really depends on which store I'm in. There's one store they're under new management and some of the prices are just whoa. I'm like okay that's what it sells for on eBay. So it's not really a great store for resellers but with that being said every person pricing items cannot know everything and you can find good deals. Sometimes you just have to look a little bit harder sold for full asking price in only a couple of weeks bought for two dollars at Sabres. It is a rare vintage Russ bunny rabbit bumpkins pink and white plush stuffed animal and this is what it looks like. Paid a dollar for this at a local thrift store sold for full asking price with no brand or tush tag listed for approximately six months. It is a vintage classic teddy bear plush stuffed animal 13 inch brown tan plaid bow tie no brand no brand no tush tag simple classic teddy bear did it take a long time to sell yes but she sold it for 48 dollars and 99 cents plus shipping. Wow I'm pretty I'm pretty amazed by that that's awesome. Bought at the Goodwill for two dollars and 99 cents and sold best I'm sorry buy it now for 125 Disney parks Splash Mountain Brara Brair BRER Fox I don't know who that is plush fox 18 inches never heard of that one there you go Disney parks neopets meow clops with ear tag purchased at a yard sale for 25 cents listed for 75 sold within a week of listing on Mercari for 60 dollars I actually bought two neopets for 50 cents the other sold for 36 yes neopets can definitely be a bolo look them up if you see them bought at the Goodwill for three dollars and sold for 40 on ebay within days of posting it is a webcam signature golden retriever by gans if you see signature on the tag those typically go for more money so do your research on webkins some of them can be a big money bolo and I do have a video on webkins I think I mentioned that earlier bought at a local thrift store for one dollar sold for full asking price on ebay in less than a week it is a 1978 Paul's Garfield plush Chinese New Year costume Daniel and company and this is what it looks like 45 dollars and 50 cents plus shipping I bought this 1992 Fern golly batty plush at a local thrift store for one dollar listed it for 75 sold in less than a month for an offer of 54 one of my best bolos ever and again sold on Mercari vintage Fern golly batty bat plush from 1992 applause purchase from a thrift store for 25 cents and sold the same day for 74 95 full asking price it is a little brush of loves brush plush stuffed doll by matchbox 1988 yes that's awesome paid almost four dollars at the Goodwill and sold for 52 99 and a week or two of full asking price of 52 99 it's a rare hard to find jelly cat bashful Mary Day tiger this is what it looks like paid 15 dollars at a thrift store and sold for 239 dollars and 99 cents in two days at full asking price posheen gunned 19 I'm sorry 2019 deluxe fluff sheen fluffy plush stuffed animal hard to find rare wow 15 dollars and 239 99 found this cutie on the cart at Goodwill and I paid 299 for it plus I'm sorry paid two dollars and 99 cents sold for 31 with an offer on Poshmark so here's somebody selling plush on Poshmark it is a 19 I think 85 tonka pound puppy with carrier and sorry it's really small but this is what it looks like this find was actually a dump dumper I think it's dumpster find Goodwill throws out a lot of beanies and I save them all most I keep but a few I sold and this was the best sell of them all sold on McCurry for 20 it is a lollipop tie beanie baby so I'm not sure what this means a dumpster find at the Goodwill can you dumpster dive at the Goodwill I guess you can I don't know maybe check that would make me really nervous but dumpster diving in general would make me nervous I think different states have different laws and I you know just check your state be careful with that bought at the Goodwill bins for 43 cents and sold for full asking price of 64.95 in three hours I think I may have underpriced it but was my quickest sale to date it is a vintage 1970 1971 animal fair Henry puppy dog stuffed animal plush soft name tag this is what it looks like got this in a plush grab bag for six dollars at the Goodwill there was none listed but many solds around 80 and since I had the only one available I was able to raise the price and sold for a hundred and four ninety three yes if there are no others listed I either say start it at an auction at a high price so if I saw solds around 80 I prop and no none were listed I probably would have started an auction at 80 and just seen what happened but what they did was also smart they priced it higher and it sold so this is what it looks like it's a blues clues rainbow puppy plush stuffed animal toy bought at a local thrift store for 50 cents there were comps for a recent sale of 65 so I put it up for auction for 64.99 as start as starting price and a hundred dollars buy it now sold for buy it now in 48 hours so I probably priced it too low but very happy with the return it is a gunned Buford plush stuffed animal puppy floppy brown dog so yes I I probably would have done the 64.99 and let the auction ride I would not have had to buy it now on it but still excellent excellent return on a 50 cent purchase bought these at a thrift store for two dollars and 49 cents they were together in a hanging bag they sold in eight days for a full asking price of 50 plus shipping the buyer was all in for $60 and 53 cents and it is a road runner and Wiley coyote beanbag plush Warner Brothers Looney vintage 1999 with the original tags bought at st. Vincent's DePaul for two dollars this is a character from department store in our area in south Louisiana I have not sold it yet but it has several comps around this price point it's mr. bingo but for this video it has to be sold so all right you guys thank you so much for being here thank you for watching I hope to have more plush videos like this in the future I have tons of plush videos on this channel where I research different brands and do category videos so check out some of those they will be under the playlist bolo category videos so check those out thank you so much for being here if you are new here let me know how you found the channel if you've been here a while let me know how you found the channel and just post that down in the comments thanks so much be sure to like comment subscribe and share and thanks for watching now I forget what I was going to say oh


So welcome to week eight on autoencoders and GANs and as we had a very serious seven weeks so far Let's just have some fun there. The first one is GANs in German means goose but I will be talking about a Generative Adversial Networks, but still it's fun to know that it's the case and now once I told The TAS and specifically Richard about it. He pointed out that VAE means there is alas and Latin So well, let's start with a little recap from week seven. So what did we learn last week? We learned about Alex Knapp the beginning of the modern era of deep learning and we thought a lot about skip connections and of course about the other innovations of Alex Knapp Alex Knapp which is Which includes dropout and reload we saw a lot of great application of confnets For example, we learned about face net and we learned a lot about domain attiptation And while we use it so much when we do actual deep learning and we pundit about lifelong slash continue learning As an important area for future development in deep learning So now let us talk a little about last week Talk with your part about what you learned last week what you're still hoping to learn and what was most surprising. It's time for discussion


Hello everybody, welcome to day 29 of the 30 day shadow work challenge. You are literally there. You are at the end of the tunnel and I hope you guys are feeling really, really proud of yourself. I know I'm proud of each and every one of you. The signal group is still popping and I'm going to say this again before anybody asks, we are leaving the signal group up. It's going to stay up. It's not going anywhere because shadow work isn't actually going anywhere.  We have over 200 people all over the world in that group helping each other, cheering each other on and so we are going to be leaving that group up for anybody who wants to jump in that group and join in on the friendship and the support, absolutely go ahead and do that. It will be the same signal support group for the 60 day challenge which is coming in 20 at 23 but with that being said, let's go ahead and look at your challenge for today and for tomorrow, the final day. So once again, you are getting up, making your bed up. You're no snacking after 7 p.m. You're either doing the 45 minute kickboxing or the yoga or the bar. You've got Reiki meditation again today with Emmy. You've also got journaling to do and then let's go ahead and look at tomorrow because this is the grand finale for this challenge. Your last meal again should be between 5 and 7 p.m. And you're going to be doing the 45 minute bar because guess what? That was the exercise you did on the very, very, very first day of this challenge 30 days ago. So I really want you to experience and really be present with this video and present with your body and look at how your body has changed over these 30 days. How has your mind changed over these 30 days?  And then something happens at some point where you're practicing and you go, you know what? Maybe I can do that. Maybe it's starting to seem possible. So the magic of this is that things that seemed impossible are now starting to feel possible.  So I want you to think about that in the 45 minute bar. What has changed? Was there something the first time you did this video where you thought, I'm never going to be able to do this? Are you now doing it? Are you now enjoying the burn, the sweat? Do you feel thinner? Do you feel stronger within your own body? How is that different? Where is that difference? Alright. So you're going to have a 5 minute cold shower again, same as normal. Do your food journal. Now for tomorrow, journals to ask yourself, how do you feel completing this challenge?  What is the biggest thing you learned about you?  What will you continue to do going forward? Which activities changed you the most? Which ones will you leave behind? Your final challenge. Pull out a calendar for the month of December. For the next 31 days, create your own challenge just for you. Each day schedule an exercise, a meditation, journaling, etc. Feel free to use this challenge as a template but in your own personal December challenge add something that really scares you. Maybe it's joining a yoga shawl or taking a rakey class to get it tuned. Maybe it's going back to school or taking a course to learn something new that you've wanted to learn. Maybe it's starting a YouTube channel or as I said, maybe it's signing up for a 5K training to do a 5K or a 10K or a marathon or maybe it's picking up the phone and calling someone you need to make a menswear. It's all up to you. You have the power to create this. You've had the power all along. Let me reiterate that. It's all up to you. You have the power to create this. You've had the power all along. Closing meditation. I want you to reflect on all that you've learned over the month. Take time to just be while listening to this chant. This is a very, very powerful chant. The mantra means in this chant that I am one with God because you are. Remember on those hard days as Marni Alton says, reach down and feel the earth and remember she will always be there to catch you when you fall. The earth will always be there to catch you when you fall. Same closing ritual going to bed before 10 turning off all your electronics.  You are all so incredibly powerful and I am proud of you for sticking with it. I am proud of you for all the tears that you've shed.  I'm proud of you for all the forgiveness that you've granted yourself. I'm proud of you for all the laughter.  I am so proud of you. I hope at this point you've learned that it's not about the workout. It's not about the actual workout. Yeah, your body is going to change and get stronger but it's about what the workout is showing you.   So I thank you from the bottom of my heart for all the hard work that you've done because your hard work has affected me as well as your neighbor, as well as your spouse, as well as your children. So I thank you for getting up every day and facing this battle because you are here on this earth for a reason. You are way more powerful than you know and you are way more special than you know. You are the light of God. You are that fractal of goodness and sometimes we don't know that until we face the darkness. We can't know our own light until we face our own darkness. Alright you guys, this will be the last video or the challenge on my channel because tomorrow, as I said yesterday, over on Aquarius Rising Africa or excuse me, Solutions by Aquarius Rising Africa, we are going to be doing a live show at 11 o'clock Eastern time. So that's 11 o'clock for Atlanta, Georgia and New York City. So wherever you are in the world, just look and gauge how many hours apart you are for me at 11 o'clock. It's this live show. We're going to be dropping the leak into the chat and so if you participated in this challenge, you can click that leak and you can come on screen with us and talk about your experiences and talk about everything that you've learned about yourself, about your own spirituality. Your job now is to pay it forward. Pay it forward. Keep taking care of yourself but also pass this challenge, what you've learned from this challenge on to others as well so that we collectively can all walk each other home. I'll see you over on Solutions by Aquarius Rising Africa tomorrow around 11 o'clock Eastern Standard Time. I will put a link to that channel down in the description box below. If you're not subscribed, you can go ahead and get subscribed and again, as Marnie Alton says, feel the earth and know she will always be there to catch you when you fall.