The Identification of the Missing Romanov Children

[Music] we have two distinguished folks who have dedicated a lot of their time to a very important subject especially for us Russians and for world history for that matter you know that back in the 70s a group of enthusiasts geologists and history buffs and gentlemen who made the Soviet times documentaries guinea above found the remains of the last royal family near equity Dean Borg and my mother can I had the honor of meeting these people back in the 80s when this was not being publicized we we went to Gilead albums apartment we had long conversations with him and we kept up our camp our friendship and I was found worthy to be able to serve a Vespers service at the place where the relics were really found ultimately in any case it was shown by various DNA experts in the world and these are indeed the remains of the royal family missing - and they were found two years later and also identified as belonging to the children of the last Russian Tsar and among the people who studied these these relics was dr. Michael Colville the Air Force's DNA identification lab in Rockville Maryland and he was gracious enough to come and speak to our parish a few years ago and as I mentioned just recently unfortunately hasn't learn Russian since then but we will forgive him because he's done a lot of good for Russian history and by participating and identifying the DNA of the royal family and also we have with us captain Peters that Indian hockey who we've known a long time I married them he and his wife 25 years ago in Nyack New York were actually relatives and and Peter has been very active in in the Romanov remains issue and also he is searching he's begun searching for the remains of mucolytics and original amount of in Perm and he will speak about that is well which is very very interesting and hopefully they're getting closer and closer to finding the remains of the other lamagno me so please I'm going to pass the microphone down to dr. Koval and Peter Sara Janaki I think Peter will go first to say a few words about his organization's search and then we'll go back and forth between the tune thank you good afternoon and thank you for your patience and waiting for this I we're very happy to be here with you and to tell you about our work for the past 25 years my what we'll do is first of all my purpose is talk about the history of the search for the Romanov remains their identification and then I will talk about the search for the remains of Michael Romanov my story I first learned I'm going to retire a seat American sea captain in Merchant Marine and how does an American sea captain get get involved in all this well I heard all these stories about the first heard all these stories about the Romanovs from my grandmother when I was 10 years old she started telling me how my great-grandfather who was head of the venden ski regiment in piensa Russia was a very good friend of then investigator Nicholas sokoloff and together they would go hunting and before the First World War there were very good friends when the war started my event or the regiment was transferred to the front and then later on my great-grandfather went up in rank and became a lieutenant general for the Tsarist army when the he he was saved by his troops actually and not killed at first he was he fought they didn't fight but he was part of the Red became part of the Red Army but he started working with a general chap of who started an underground railway to save as many officers as they could from and send them off to the newly formed white army so what he would do is he would send my grandmother who was then 18 years old just like a cleaning girl into the Lubyanka prison and she would go and get the laundry from the officers who were about to be killed and but still they did the laundry and in that laundry were pieces of paper that with the names and addresses of officers that needed to be saved and how she wrote it and as she told me that she went countless times into the prison getting those getting the laundry and the names and she saved a lot of officers so who were not to fight against the Reds later on so my great-grandfather escaped went over to Siberia and then started was working with a general staff his left his wife and and and two daughters in cancer and then they followed escape by using that what they call the nuns around going from monastery to monastery and so forth and my grand-grandfather gave my grandmother and the gun pistol and he said use six bullets and the last ones for yourself and that's how it was actually my great-grandmother's mother Baroness von Rosen committed suicide so that they had escaped so that's how tough it was for these families to escape through out of Russia so finally they went out into the steps and Cossacks would coming at them and it turns out that were white and they ended up so in seven days after the murder of the royal family my great-grandfather was with those troops that are right into a Katarina burger next slide please and they found that that the royal the royal family the servants and I slide had been taken first from okay first one st. Petersburg's de bolsa Ekaterinburg so go to the next slide and so when the white troops arrived into Ekaterinburg they found this double offense around the pontiff house my great-grandfather ordered the troops to tear down the fence his adjutant who later became my grandfather kitty Oh McCullers not African was went down into the basement and he saw what happened my great-grandfather did not have the nerve I guess to go see so an investigation of the murder was started next and these are the killers Peter Yermak off your offski the leader their mark off the second next slide they had taken them into the bay this is what my grandfather saw the basement that photograph from the Harvard collection next life and it's the same thing he's thinking anyway so the investigation of the murder was started and it what it was and they were heard there were two investigators now North goodness you gave who were hired by the white army except that they really they did a lot of good work and they found out where all what happened to them and so forth where they were taken they said the white army wasn't satisfied and fired them and and when my great-grandfather then introduced circle off to the General Staff as the men to do the investigation of the murder and that's how a circle of got his job cycle off went to well first of all what had happened at number three the place where the murder occurred was down down here in Ekaterinburg the bodies were taken up the gun Deena Yama number one and number two and they were the bodies were dumped into the open pit if you go to the next slide this is nicholas circle of 1919 next all right so Sokoloff then took charge of the investigation and where he's standing now he's standing all over the spot where there was one of the bonfires and according to general d-trix there were two bonfires one in the in the next slide okay Sokolov lost half a month pumping this water out of here so maybe the bodies were underneath a keep going alright so here's the open pit that if you ever go to the cutting board you'll see this open mine shaft is wider now but this little crossing is still there I went across it countless times so the bodies were thrown into this pit it was about 30 foot deep and the White Army later on found underwater they found the floor and under the floor that found the girls dog who was killed so they went to they went to a lot to hide all the information to hide everything as they could but Sokolov used to come over our our family home in France after there after they got out of Russia and told them was telling them how when he first arrived to the open pit area if you go to the next leg right here there was a fresh bed of clay and so the first circle of this pile of gravel and so for that you see was taken from the bottom of the mine shaft to see if there was anything and it was left like that actually for us because circle I've never had the time to go through it nicely they the bodies were were taken in three day there were about 50 people there trying to destroy these bodies that try to burn four of the bodies now what we found out later that you know they try to burn Alexis Maryam Bazaar and someone else and Michael determined that because of the DNA also with the bones of the heel bones were very light and thus they're supposed to be heavy and dense well the the part of the the bodies were taken that were burnt and then they were thrown back well they try to burn the phone they were thrown into the mine shaft 50 people were in this mind in this area working on this and everybody knew about it they were going back and forth into the town but he soon the whole town knew where the bodies were and so they got a shock in the head of the whole investigation came to this place and told them get rid of the bodies let's take him to a the mind on the way to Moscow so they backed them up again on carts and carriage and finally on the truck like this and took them down this road the cab tiki Road and it's still this looks like this because it's in July it's very rainy and I can understand why the truck finally got stuck next slide got stuck underneath where this wooden bridge is this is the photograph taken by circle off circle of hired hundreds of people to comb the forest to go across it and to work maybe they were going five yards apart looking for a grave many times he went through this area and never once did he think perhaps that that's where the grave is and that's exactly where the nine sets of remains were found underneath this little bridge next slide this is the photograph that we found at the archives it's just Peter Yermak off the killer and in the back of the photograph written in pencil has said I'm standing on in Russian I'm standing on the grave of the source and right away with of course this was it we knew and perhaps we thought you know when I've done Ian you know it eventually opened up the grave they found the skulls and took them for examination in this slide but they they weren't able to determine anything so and also they had to keep the secret because Russia was still communist and for another photograph it just for grand Sokolov talk right here is the Karthik Ebro but here's this spot right here so they actually found the remains of Alexis at Marea this is the southern part of the pig's meadow when we search this area our third search and I went back to this area here too because we searched the whole northern area I went back to the spot and I started thinking photographs so we had touchy key workers and so forth and I was actually standing on the grave of the children I didn't even know it so this is how difficult this work is next like Sokolov and never found the remains he concluded wrongly and I repeat wrongly that all the remains were burnt because he really had no proof to determine that what we do know is that from the archives and from the ridiculous road when the killer's took the bodies to the pigs to the four brothers mine area there was an expert that was supposed to come to to burn the bodies except that that guy never made it because he fell off his horse and Brooke is like so then Peter Yermak off was given the task of trying to burn the body says this hole and after being taken out of the water the bodies were totally frozen from the cold water they weren't even burning so so they they just gave up on that Sacco I finally made it my great-grandfather was transferred to Vladivostok where he became the military governor of the whole of more region and Sokoloff came there in 1920 in January there was a coup against my great-grandfather and the Japanese saved my family circle off left Russia with a proof of the murder whatever he had and he went with general Jana the French general a to Harbin and eventually met my the Japanese saved my family and met my grandparents and my great-grandparents in in Japan they my grandparents that Africans and the soca loves traveled together on a ship kalander the bomb and this box the proof of a murder well there were many boxes but especially this one it was about this size and and it was taken by circle off and held by cycle off and it was kept on when they traveled around the world it was kept under my grandmother's bunk so so one day while guarding the box no but they always guarded the box even on the ship and I come to here from circle of grandson I took that photograph psychosurgery has that box that massive tongue was traveling on the same going to France as well so very very interesting so next slide please in the box we had the dr. Balkans dentures in the box we had melted fat bullets the pieces of wood where the bayonet stabs weren't with blood and so forth a lot of those items now if you ever care to go are up in the museum in and the in Jordan Ville at our monastery there's a very nice Museum that you can go visit and those are what the items of the box there was also the Empress's finger next slide please and the but we have we don't know where that is yet maybe someone had kept it or something like that in 1978 Alexander of dhania that's what guardian entity above his wife and another friend decided to search for the remains of nicholas ii and and the royal family they were all antes yes and they were really but that by the Ministry of the Interior over there we have VD next and so they opened up the grave that they found the bridge they climbed up the bridge of course was covered with dirt but they climbed up on the tree and they they were able to see the you know the silhouette of the bridge and they opened it up and I found three skulls tore skulls were taken to Moscow for for them to check out and then and then and then and they were brought back and put back in the grave and until they decided to keep the secret until Russia changed and in 1991 dr. gaya above told the world what they found next and you can see the previous picture the sculpt in number four was of the Tsar in in that grave that they opened up there were nine sets of remains and they found that two were missing the one of the daughters and also say every should he see everybody else was found I can tell you right now that and father Victor knows that in 1995 when they tried to the did DNA I was given the shavings from the DNA samples that when they cut the bones and they said them to to was brought to my house gave father Victor half and I kept the other half and I put him by our wedding icons it was around midnight I went back into the kitchen and my wife Marcia also the witness and then an hour later I went back into the bedroom and the whole bedroom smell like roses and when I went to that piece of is a piece of paper with shavings or like dusts of from the bones and and but from from from the little shavings from nicholas ii and from dr. balkan the the aroma of roses was so strong that it smelled up the whole room that that was just incredible and I gave it to a priest so I felt that I didn't feel that I was where a day to keep something like that so next this is how the remains by the way that was how the remains late for seven years that morgue now my friend dr. sakeena Keeton studied doing their wicked spatial reconstruction over the skulls doctor pocket they made over next slide please organical I gone Tatiana Anastasia and next slide rope Alexandra thanks all right so we have it and I asked dr. nicotine how he determined that the head anastasiya because the american scientist says that you have might have the wrong daughter you Anastasia is missing and you have Maria well it's because he did also photo superimposition this is Nicky his reconstruction of an estate of Anastasia this is her photo this is her photograph this is Anastasia's reconstruction is the photograph of Maria over here and so as you can see with certain points that he measures and the points match exactly and the shape of the heads matches exactly and also we compared to Maria the points do not match up only one matches but the shape of the head does not match up there were other things involved too that I you know that the forensics that I had things that were told but basically the Russians were convinced that they had Anastasia Maria was missing and that he became a bone of contention at one point when I took American scientists to Russia in night in February 1998 before the burial of the royal family took dr. Anthony full Sethi from the University of Florida Gainesville and doctor and friends from Colorado State University both director of human ID lives both forensic anthropologists and they determined they also saw by like they also agreed with dr. William maples of Florida also that that the Russians had the wrong daughter but so then I went I went to back to the Russians and I said who I asked him who determined I didn't know that Nikitin had done this who determined that you had Anastasia and the whole point at nikitin I said well the Americans will present their papers from the Academy of Forensic Science in the year 2000 saying that you're wrong so Niketan came to America presented his paper and from the Academy of American Academy of Forensic Science proving his case that that indeed that Maria was missing and my whole thing was that will determine really that when we found if and when we find the next set of remains so so this is Nikki tooth work he did a reconstruction of Ashley this is Diane France she sent by a photo a scan of her own skull and to see how close it is how close this work is amazing so I have no doubt indicated its work really and then there was a burial in 1998 next slide so we've decided to start searching for the other two children thanks so when we came this is what we found that had cleared the forest area and and as you can see the on the right side behind the church put up a cross there and it said that all the remains were burnt I said they're wrong you know we'll find the other but and behind already the open pit circle already had opened it up more next slide please so remember from the mineshaft right next and this is us already working on that pile next this dr. Bob Dornan September 98 nice that's myself and next and I found the first bullet from the gun pistol got a shot of vodka for that and that was in that pile of stones next because they what they did is they took off all their clothing and everything and there were the diamonds and all this so we found a lot of we found on the third day I found a white topaz stone circle of found 14 I found the 15th one we found we continued we didn't find that many items but we did find continued finding was circle of phone next I found a button I found you see the snap on top the back was still in there the meaning the clothes were ripped off the rim it wasn't they weren't gentle next and they found bones by the circle of found bones as well there were animal bones next okay next so in 1998 we're still searching we also found interesting red wax in the mineshaft I think say carried red wax in this pocket and that was proof to us and nails he carried it too but he carried and wax he played with it and that was proof to us that he was there next we did a seismic profiling of the whole area and electro begged a nice electromagnetic study says well finding anomalies we checked every single one of them out and nothing nothing panned out next it started to snow so then we hit the next we moved the whole pile of stones and the pile of stones also had had charcoal and all that in there but next shape such a beautiful place so many horrible things happen next and the second search was in June 1999 next so everything's the flowers came back everything came back and what we're looking for is also the bonfires because they were the killer's world Yurovsky was that after they the two children have to be burned the bodies of us the children they buried them next to a bonfire and then they they they covered that up and then built a bonfire on top of that and so we were looking for signs of a bonfire and the only in that's one of the reasons we started working at the four brothers mine area because they were the general details map of the whole area two x's signified bonfires and that's what that's what we found and we actually I did it by compass and steps like they said you know and I found we found those two bonfires nice so we're looking next so we started searching for these these bonfires next and we found traces of charcoal everywhere that was spread out Thanks and here was one of the fires this would suck a loaf you saw in the first picture cycle of standing this is the exact spot where he was standing there was a bonfire underneath we found nothing in the because he already had gone through it all so right now there's a slide walk over it so you know they don't know they didn't know that they just put a sidewalk right over it was so this was destroyed instead of being kept go ahead one thing is remember I told you about the fresh metal of clay we dug down about 20 centimeters we found a site of a huge bonfire about the size of this table they the clay was already brick and we found this melted neck of a bottle the fire was so hot and that's where they burned the bodies and and then we saw that they had swept all the coal into the mineshaft there was coal never disappears and so this is what we found the earth doesn't lie don't keep going and this is what the four brothers mine area looks now actually these fans here but this is the mineshaft and people go it's a holy ground this holy place okay next time the third search was in July 2004 going and we went to the pigs meadow this is the actual grave where the bridge was that you saw that photograph and people commonplace a very quiet place but the church the local the church they used to go there religious processions acoustic ones through this area here but because they don't believe that these are the remains they built a separate road going to the four brothers mine area so all right next so I can wear it comparing then this is the whole northern area of so the truck came down this way and that's where it got stuck okay next so we hired the GG's tiny workers to search out the whole area plus we had next where we had a radar that's all we had at the time so we're searching the area for radar looking for signs of any taking and something like that to go we hired equipment to level out the ground so the waiter could work better but it does you know nothing you know keep going and whenever we found something we like a signal like this that means somebody had done here we would have to open it up again next so we searched and nothing was found here nice and the final search was in July 2007 that by wasn't done by me it was done by a local group of Russian army who a club who go to battlefields to try to find remains and so this is the northern area over here so we searched them and this was left over here to do this is what I wanted to do in 2007 but it didn't get a chance they did in this spot right here go ahead they found the grave of IDC and Maria nice that's all that was found from two bodies that is all and you notice this curve over here this from a bullet hole and but you could but scientists determined that in these that have a very young male and a very young female to come okay keep going okay keep going so nine bodies were found and seventy meters away from from the set from the other side ago and so what I did then is when Nikitin called me I I decided I called up dr. Fall City University of Florida who wanted to go and study these remains anthropologists in speaking to him I said by the way do you know the head of the US Army DNA lab he says yeah I do his name is Finelli he says I said boy I would love to get up to do the DNA study of these things maybe we could organize you know I could organize it and I spoke to the Russians let you know some of you off and dr. deagle idea Valen from Academy Berg and they all you know they all agreed let's do it let's do the DMA so that's when I wrote actually false Eddie spoke to Fanelli and two Italian boys you know but very fine people believe me fine people and Cardinal Arif Finelli decided to come on board with the US Army DNA lab and hence dr. Michael Koval please talk to you about it let me know when you want to take a break take a break but talk about our search for ok thank you very much thank you Peter and again I personally would like to apologize for the late start the technology can be a bit of a problem and it reminds me that father Victor did it modest me for not learning more Russian but I'll tell you this story that a friend of mine a very good friend of mine from California he said he was in he was in South Korea and he was giving a presentation and the day before his talk he noticed that a lot of the speakers who were giving their talks in Korean they would they would constantly say you know somewhere like : social kosher so he thought maybe they're saying thank you and so that evening everyone he met he would say kosher and so finally someone said why are you telling everyone next slide so I'm afraid what Russian I may speak may not mean much so okay I think I'll go ahead and get started so that we can not again apologize for the late start so I'm going to talk about the DNA identification work that we did at the Armed Forces DNA identification laboratory this lemon story used to be in Rockville Maryland it's now been relocated to Dover Delaware so if we could go ahead with the next slide so I no longer work for the Armed Forces DNA identification lab for a fidel when the lab decided to move to dover i made a career decision to stay in the area so i now work at NIST National Institutes of Standards and Technology in Gaithersburg so the work I'm going to talk about is his work that I did when I was at a steel but again I'm not speaking on behalf of the lavatory and anything that I say opinions are strictly mine and not the opinion of the Department of Defense so when you do these types of historical investigations it's often important that you have replication of work and so we had and in this particular study we had our group a Taft ill so it's taking there we go we had our scientists at after all that we're working on this this problem and then I asked the Russian authorities if I can also invite a good collaborator from a laboratory in Austria and Innsbruck so they were in parallel doing the same work we had two Russian scientists that were staying with us in Rockville and there were two Russian scientists who were in Austria who were doing work alongside our colleagues there and then what we did is we did not talk to one another so we did our testing Austria did their testing and then we sent our work to a third party and that was dr. Peter gild dr. Peter Gill was in the UK is in the UK and he was involved with the early testing in the 1990s he was the the British lab that did the original work and so he looked at our results he looked at the Austrian results and he wrote a report that said they both came to the same conclusion independently of one another okay so I thought it would probably be best to give a very brief a very brief background on how to do forensic DNA testing and it'll be like an episode of CSI we'll do it in five minutes it really takes a lot longer to do these types of testing but I'll give you just the brief overview of how we do forensic DNA testing so if you take a look at a cell at a human cell you actually have two human genomes so you have the big blue circle there the big blue ball is the nucleus of the cell and in that nucleus you have the DNA that makes the code for proteins and the you know codes for all the various functions that you do and so forth and this is where you get a lot of very high power of discrimination when you're talking about forensic testing and you may hear then you know in a court case that there's a quintillion times you know probability that this person so that's the cut time type of testing that you're doing the nuclear DNA testing very powerful the other genome that you have is the mitochondria which are these little circular structures that one of the main functions of the mitochondria is to provide energy for the cell and interestingly they have their own DNA its DNA that's separate from the nucleus so you actually have two types of DNA in your cell you have the nuclear DNA that you inherit from your mother and father you get half of your DNA from your mom and half from your dad 3 billion bases of information and then you have the mitochondria which are actually the mitochondrial genome is quite small 16,000 base pairs but the advantage that mitochondria DNA gives to the forensic scientist is that when you have a very degraded sample we have a bone sample that's been in the ground for 5060 hundreds they're getting DNA from Neanderthals so from from thousands and hundreds of thousands of years then the mitochondrial DNA because you have so many mitochondria your cells need a lot of energy then it can provide you with some information so let's talk about the nuclear str testing that we did alright so if you take a look in the nuclear genome you will find certain regions where you have repetitive DNA and so in this case we're looking at a certain region of the chromosome again you have two chromosomes one from your mother and one from your father and in this certain region there's this repeat so you have the nucleotide bases the four bases a GC T that make up DNA and you have in this one particular region this repeat GA ta for bases that repeat side by side by side so gotta gotta gotta gotta gotta so on the bottom the maternal chromosome you have six of these repeats and the chromosome you get from your father you may have eight of these repeats so the number of repeats can be very variable in the population and so what we do is we make use of this variation in forensics and we we have to first fly this up so there's this process called PCR the polymerase chain reaction it's kind of like a Xerox machine where you take one copy of a document you put it on there and you make many thousands of documents from this so we notice that we're on this the way this machinery works is that we have these little short fragments of DNA that have this red dot attached to it these are called primers and that red dot is a fluorescent dye and so we amplify we make copies and so when you go through this process of PCR if you hit the next and hit it again you make millions and millions and millions of copies of these fragments some of them have six repeats some of them have eight repeats and they all have this little red fluorescent dye attached to it so we've been able to label these fragments with this red dye so in the next step we passed these little labeled fragments through this machine called a capillary electrophoresis so inside of the instrument you have a capillary go ahead and it's kind of hard to see but this capillary is about the size of a hair and so it's very thin and inside of this capillary is a polymer it's kind of like a pancake syrup so it's very viscous so because DNA has a negative charge we put a positive charge on one end of the capillary and the DNA starts swimming to that positive end so as the DNA is passing through the capillary the little size the six size fragments the side the fragments with the six repeats they're able to move a little bit quicker than the eight repeat fragments so we're getting separation of the six and the eight and they eventually get to this little spot in the capillary where a lane sir shines through and when that laser hits that red dye it gives off a red signal and that signal gets captured and gets transformed into a red peak that you're seeing here so this is the six repeat and this would be the eight repeat so we're able to actually visualize how many repeats were amplified from this particular individual now the FBI decided back in 1997 that the US would study we would all look at the same 13 markers so these are called the co2 slow sigh this is what makes up our national DNA database so we look at 13 pieces of information and when you look at 13 different pieces of information you can get the type of power that can give you an individualization so we could tell my profile would be different from anyone else's profile in this room if we look at those 13 pieces of information in fact the only way that you would predict that it would match is if I had an identical twin in that case you do share the exact same DNA okay so this is an example of a kit that's used in forensics it's actually looking at 16 pieces of information so the first marker I want to point out is this marker it's amelogenin it's used as a sex type marker so we can tell whether the person is a male or female and in this particular sample we only see one X that's because this is a female females have two X chromosomes X X males are XY so the reason you only see one peak why don't you see two peaks is because both of those X's are sitting on top of one another so there's actually two peaks there but they're sitting there the same size so they're both sitting on top of one another if this were a male sample you would see an XP and a white peak so you would see two separate piece and then we have 15 pieces of information the 13 code is slow side that we all test in the US and then there are two markers that are unique to this kit so there are 15 total markers okay now what's the probability that I would find someone else in the world who has this particular type 1 and 129 quadrillion so that's the basically that's saying is the same probability of taking 129 quadrillion white marbles and putting in 1 red marble and then stick your hand in there and picking out the 1 red that's about the same probability for this particular profile now this is actually my daughter's profile my daughter Sophia who my wife and I adopted Sophia from Russia and her little brother Matthew so whenever Sophia asks me dad am i special I'm like yeah under 29 quadrillion I'd say that's pretty special but she doesn't quite get that she doesn't get my joke someone said actually she does mostly so now we have the nuclear DNA but there's also again a couple of things I want to point out go ahead those are the autosomes it's just a nuclear DNA that we're using for STRs that we do in this type of testing we also have the sex chromosomes we have xx again if you're female to X's males XY and there's also outside the nucleus in the cytoplasm of the cell we have mitochondria which have their own genome the mitochondrial DNA and there's a little bit of a relationship between mitochondrial DNA and the Y chromosome because of the way they are inherited so if we take a look at your DNA and we take a look at your great-grandparents as Peter mentioned his great-grandfather you are about 1/8 of your all of your great-grandparents so you have eight great-grandparents and you are about eighth of all of your great-grandparents and we can keep going back you know 1/16 your great-great grandparents and so forth we can keep going back in time you can figure out what percentage of your family you inherited from all of your relatives immediate relatives in your family tree but when it comes to the y-chromosome men only get the Y chromosome from their father who got it from their father who got it from their father and from their father so even though you have four great-grandfather's only one of those great-grandfather's passed his Y down to you so so that's a really unique feature about Y chromosome testing is that you can find relatives that share the same Y type when you're trying to do a human identification okay so this is a there STR so they short tandem repeats are also on the y chromosome and for the most part you only see one peak and each marker here so there's one peak and there's one peak there's one key and that's because you men only have one Y chromosome so you don't have to so you don't see two peaks like you do with the nuclear DNA where you have your mother and your father have given you DNA now y-str testing has been used a lot in forensics and a lot of historical cases like the the case of Eston Hemings who esting Hemmings mother with Sally Hemings who was a slave of Thomas Jefferson and so one of the descendants of Eston Hemings said you know I've been told that my father was my great-great grandfather was Thomas Jefferson and I'd like to go to this party that they have a Jefferson family has every year in Charlottesville I'd like to come and they said no we don't we don't think I don't think that's right I don't think you actually are and so they said well let's do this why testing and when they did so if you take a look at so I have to say you know Jefferson had I think he had one daughter or two daughters he didn't have any sons that we know of and so if you'd look at other relatives they had the same Y chromosome as Eston Hemings and it's a really rare Y chromosome so it's not like there were a lot of beans so now we can't say with definitive proof that Jefferson Thomas Jefferson was the father of Eston Hemings but we can't say that someone with the Jeffersonian Y chromosome was the father of esta so there's a limit to how much real identification you can do with Y's it's really powerful for just sort of saying it's definitely not him so like for example Sally Hemings have another son Thomas Woodson and he had a different Y chromosome so we can conclusively say that was not he was not fathered by a jeffersonian chromosome now on the other side of the coin is the mitochondrial DNA and it's kind of like the Y in that the mitochondria is only passed from mother to daughter and so all of the women in here got your mitochondria from your mother and you'll pass it on to any daughters that you have now men also have mitochondria the mitochondria are very important for functioning we all have mitochondria but when it comes to passing it on to the next generation only the females will pass their mitochondria on to the next generation so again it can be quite useful and these human identity if you could find relatives that belong to this family tree you could use as comparison now we look at the sequence because the mitochondrial genome is so small we simply look at the sequence of the DNA and we do a comparison what is the differences between the sequence that you're looking at and a reference sequence so we would say this particular person at position 263 the reference sequence has an A this person has so that's the way we would report mitochondrial DNA and I'll take just a second to say that this this worked really a lot of the work the the analysis work can can go quite quickly a lot of the work depends upon the type of sample that you're looking at so as Peter mentioned these bones that have been buried for a hundred years and the frozen you know the ground and in Ekaterinburg is frozen for about eight months of the year so you go through a little bit of free-stall over a hundred years the types of bacteria that are living in the soil bone is a very nice source of calcium so their minerals there so there's a lot of feeding that happens on the on the material so you take the bone and you try to first try to sand away any of the you know in 30 anthropologists in the room I apologize anthropologists love to pick up bones and you know so you have to be careful that you make sure you're getting rid of any external contamination and then you grind the bone up into a powder and then you digest this into a buffer and that will help to release the DNA there and so you're hoping that you're getting the DNA that's actually from the bone and not from some outside contaminant so as Peter mentioned because of the work that of Dovan did we have the excavation in the early 90s of the of the first grave that was discovered and as Peter mentioned there was some debate because the American anthropologists believed that Anastasia was missing they both agreed that there was a boy missing that Alexi was not there but the the other daughter that was missing the American experts believe that it was Anastasia and the Russians believe that it was Maria and so based upon the work that the Russians did the the facial reconstruction will for this talk will move forward say yes there was Maria that was missing so go ahead so I'll talk very briefly about what Peter Gill did in 1991 and so he published his paper in 1994 in Nature Genetics and he used the STR testing that we're using today at the time they were only looking at a few looking here at five STRs remember today we're using 13 and we're getting ready to move in the u.s. to 20 starting in 2017 so we're getting ready to look at more information but at the time there were only five STRs that were tested and what Peter Gill could show is he could determine there was some of these skeletons look like they're related and there's some skeletons that are absolutely not related so these were the four servants that had different profiles okay and so based upon the DNA work that was performed in the early 1990s this is what Peter Gill was able to determine using mitochondrial sequencing and STR testing is that you had remains that were consistent with Alexandre the czarina and three of her daughters and they used as a reference someone in that maternal lineage His Royal Highness Prince Philip as a reference he is a it was a cousin of the Czar Dina and so or I should say Empress and so they asked him for a sample of his blood and he donated it and they looked at the mitochondrial sequence and it matched now for the Tsar it was a little bit different because when you looked at his reference the Czar had a condition what we call heteroplasmy and that is that some of his mitochondrial DNA at this one position 161 69 had the T base which I haven't read and some of his mitochondria had the sea base and so when you put those two together you get in the sequence what looks like a mixed a little bit of tea and underneath a CP here so at this one position it looks like a mixture of two people but you didn't see this anywhere else only on this one position and so when Pavel Ivanov came to the u.s. he's the Russian scientist who was involved with this first round of testing and he met the director of F dill at the time and dr. Victor Wheaton and he said yeah there's a lot of people that are having doubts about this heteroplasmy because at the time scientists didn't really know a whole lot about heteroplasmy so he said can I come to a still and we do some more testing so so he did and what he did was they exhumed Georgi bizarres brother who died of tuberculosis and they exhumed his remains and when you looked at the mitochondrial DNA of the Czar and Jurgen you see the same heteroplasmy the same position but in different ratios so this is where the Armed Forces DNA lab first became involved and we were able to show that that this was not such a rare event that it did happen especially in this in this lineage okay now despite the overwhelming evidence there were a lot of concerns by people who were trying to discount the DNA testing they were saying well you know this heteroplasmy we're not really sure what that's about at the time the mitochondrial DNA databases were quite small just a few hundred people so the statistics were not that overwhelming but they were still substantial and STRs were in their infancy so fast forward now 2007 as Peter mentioned in the news came out that we think we found the the two missing children and that's a picture that I took of Peter with the the men who were the the man standing the sight of Peter were involved with the actual discovery and this is a local archaeologist who was who did the dig who actually did the excavation as Peter mentioned this is pretty much what we had these were the largest bones there there were 44 bones or bone fragments and I think there were of those 14 I think there were three teeth there now if you think about it you have over 200 bones in your body and there are two people here so out of over 400 bones you have less than 40 bones fragments that were still there so much of that material is gone has been burned or has disintegrated over time so there were only a few bones that were really sufficient for DNA testing and so that was the question are these the remains of the two missing children so we have the sequence the mitochondrial sequence from Peter Gill okay go ahead this is a bone from a leg that's a from a fragment of the leg one thing I will mention about this is that these are only portion of a leg bone so the anthropologists didn't have the luxury of doing these measurements to determine the height and maybe then they could definitively prove or at least suggest that this was you know Alexander Anastacia or Maria so the Anthropology was quite limited and the the remains that were recovered here's another fragment from a right femur from a right leg bone thigh bone that was tested and the sequence we got was the same that Peter Gill generated back in the 1990's and I remember when this happened we had Alexis saccharin was our train later he stayed with us the entire time because the two Russian scientists spoke no English very little English and he was there with us and when this sequence came up on the computer everyone started crying I mean even though we had all this definitive this proof I mean we had all this information that it was them that was the two missing children but once we saw this sequence everyone just broke down and was crying because it really meant a lot that we finally found the two missing children so of all the fragments that we tested that we could get DNA from again if we tested ten pieces of ten bones the bones that we could get DNA from they all had the same sequence all right but that doesn't tell you whether that was from Alexi or Maria it just tells you that you know of the bones that were recovered we had pretty good idea that one was from a female once was from a male based upon the anthropology this time we didn't have a database of two or three hundred we had a database of twenty one thousand we've never seen this sequence before so it's a unique sequence again which means that it's pretty rare okay we also did STR testing and so we took the fragment from what's believed to be a male and we see the two peaks x and y which means that's from a male and we see the one x peak here from a female and the one thing that you notice when you look across this profile I'm only showing you here three markers the sex marker and then two other STR markers when you look at the DNA profiles you notice that go ahead there's a lot of sharing going along the one peak here the brother has the sister has or the putative sister has also that peak so that suggests that these two individuals may be brother and sister so we did this statistical test we generated the profile of the of the female and the male and we did this statistical test that's done for paternity testing and and kinship testing and we simply asked two questions this is called a likelihood ratio we simply has two questions calculate the statistics as if these two individuals here are in fact brother and sister and then calculate the statistics as if these two people were two people I grabbed at random in the population that are not related so we have two different statistics that we calculate and then we take the ratio which one makes most sense which one explains the data better is it more information that they are related or that they're not related and when you did this test you found that it was 5.6 million times more likely that the remains are brother and sister than if the remains were two people we picked out of the population so very strong evidence that these are related individuals brother and sister as Peter mentioned because of the previous testing that was done the lab in Ekaterinburg kept some of that material that DNA material from the Tsar the czarina the three daughters so we're able to get STR testing on all of the family and when you do that you notice here on top is the Tsar here on the bottom is the czarina and in the middle here we have Alexei and when you look at the peaks go ahead you can see that every P and this child can be explained if the Zarn there czarina where the mother and father so as you would expect child gets half his DNA from the mother and half from the father and we'll do this again for the female or the skeletal sample we get at every peak we can explain her profile from this the czarina so we developed profiles for everyone and when we did that likelihood ratio test when we asked the question these two individuals the boy and the girl do they belong into this family tree with the Czar the czarina and the three sisters well for the girl her probability was four point three trillion and for Alexi it was 80 trillion so the the results are four point six trillion to 80 trillion times more likely if these two individuals aren't in this family tree then if there were just two people at random in the population that would fit into this pedigree so since we did have Alexi and we did have the Czar we decided to do Y chromosome testing back in the early 90s there was no y-str testing at that time so when you look at the profile from the Tsar which is on top oh I'm sorry from Alexi and from the Czar they match at all we look at 17 markers they all matched exactly at all 17 and then as a relative we used Andrew Romanov from California he's a cousin of both Nicholas and Alexi and when you look at his profile he matches it all 17 so we had a perfect match between a living relative and the remains from Nicholas and Alexi as far as the rarity how often do you see this well in 20,000 a database of 20,000 y-str types we haven't seen this this haplotype before so it's quite rare so I'll take just a second and talk about the work that the ladies in Ekaterinburg these are the the ladies that work at the DNA lab and Canberra again two of these ladies came to that on the right and left came to the US and two of the scientists went to Austria now they did DNA testing along with dr. Raghav on this shirt bloody shirt that Nikolas wore so when Nikolas was a young man his father sent him on a world tour and during a stop in Japan he was attacked by a someone who was mentally disturbed who I think was dressed as a policeman but he had a saber he came and he hit Nikolas on the side of his head and you know created this huge gash and so Nikolas was bleeding all over his shirt in fact Nicholas's cousin was able to stop the attack he had just bought a bamboo stick as a souvenir just just minutes before and when he said when he looked around and saw Nikolas being attacked he hit the guy with the stick and was able to disable him so that they stopped this guy from killing him and so they the wounds were present there the you know as far as the Anthropology goes but I won't Mitch I'm not an anthropologist so I won't talk too much about that but the shirt that Nikolas was wearing was sent back to to Moscow for after the attack and was was as proof of what had happened and so eventually the shirt made its way to the Hermitage Museum and so you know here in the 2009 2010 we're doing DNA testing and mr. solo have says hey you know I calls up the Hermitage as it says you know you guys have this shirt with blood from from Nikolas you think we could do some testing on that and they said no we can't find this shirt we have no idea where it is it wasn't there what they had not didn't have a lot of good record-keeping and so forth they knew at some point the 1930s that it had it was in some Museum of the revolution or something and then it was sent it so they no one had really any idea where it was well at about the same time there was a lot of pilfering of art of that was going on at their Herrmann shoes so they decided let's and thanks to donations from from Friends of the Hermitage Museum and so forth they were able to get them a computer system where they could document everything and so they're going through and they opened up this drawer and they see something wrapped in this brown paper they open it up and they found the shirt and the Hat the derby hat that he was wearing so so the Russians were quite excited wow you know what can we do can we do DNA ties it well what do you plan to do well we're gonna you know we'll cut out a little bit of the you know no it'll be no cutting so they said it was okay to take like a q-tip and some water and sort of rub the blood and you know there you go that's what you can do so that's what they did and just to show the beauty of this shirt that the craftsmanship craftsmanship and you know some this is the tag 1931 is the tag that was on it go ahead and so you can see they took a sample up here from the collar that had a little bit of rust on it there were the blood had sort of crusted over time and so again you can see where they took a little bit of a q-tip and a swab and sort of swab this area where the blood was dripping down onto the shirt they also swab this area of the sleeve where Nicholas was holding the side of his head so the profile that the Russian scientist got was the exact same profile that we got from the bones and they also did y-str testing and they got the same answer as we did for the Y's so interestingly you have a DNA profile from Nicolas when he was living from the blood on his shirt and we had the DNA profile from his bones after he died and they matched you have a question no he didn't die he survived the tack and he went back home but he he survived that attack okay and so he had complete concordance with so a lot of times you know DNA tends to get all of the glory all of the honor because it's used a lot in fighting crime but I'd like to point out in this case that DNA is actually just one piece of the puzzle when you take a look at all the other artifacts like the bullets that were recovered and within those 44 bone fragments that Peter has talked about there's a piece of pottery which they know was used that sort of uric acid was purchased and used to try to disintegrate the remains so we found pieces of pottery with Japanese writing on it so there's a lot of information there so if we take a look at the forensic DNA evidence you have you have evidence from the mitochondrial DNA which agrees you have evidence from the forensic STRs which agrees that these are in fact members of this family and you also have the y-chromosome evidence so taken together it's very strong evidence here from the DNA that we have the two missing children Peter wanted me to mention about the work that Roe gyeom did on hemophilia so we know that hemophilia ran runs through the royal families of Europe because of Queen Victoria and with with Alexandra being a granddaughter of that Queen Victoria we know that Alexi had hemophilia and so regard is actually able to find a lot of people think that that the hemophilia was a deficiency in factor 8 will show that that was not the case but before we do that one more quick review the way that you make a protein in your body so if it's a protein like insulin or something like that you start with a DNA that you and that gets transcribed into a message and we call it messenger RNA and that messenger RNA then gets made into a protein so you have DNA makes messenger RNA makes a protein does a little soundtrack going on and so so this is the way the DNA is organized it seems a little crazy but that's just the way it is when you look at your DNA and you want to make a protein like one of these proteins that are used for clotting blood the way that DNA is organized is you have these sequences called exons which are in blue that's what's actually transcribed into the message the message RNA and then you'll have these red regions which are called introns which are just junk DNA doesn't code for anything is just a bunch of gibberish so it's kind of like reading a book and you're reading and you're reading like five or six pages and then you've got like ten pages of gibberish and you're like so you have to skip up and then you pick back up and you start reading again so this is a way that our DNA is organized and they think that it's probably because if your DNA was just simply pure this is the code for how to make this protein and if a mutation happens well then you're pretty much in trouble because there's no way to correct that so if a mutation happens in an intron no big deal we can still get the information from the exon so what happens is when you go to make this message the red parts are cut out and the blue parts are put together to make this one message that then gets made into a protein so the way that happens is you have enzymes that are reading the DNA and they know exactly where to cut we're going to chop here and we're going to cut out this intron okay so if you take a look at blood clotting disorders when you cut your there's a lot of things happening there's this cascade of enzymes will cut this this protein and then that activates this and then and so you eventually with all these little factors that are getting these these factors that are being created to create the clot you requires a lot of enzymes and proteins to do so so one way that you can get haemophilia is if you are missing or if you have some issue with factor eight which is one of the proteins in this process the other way is if you have a deficiency in factor 9 so if you if you knock out either one of these then the pathway to making a clot is broken and so you have issues when you cut yourself you can't stop the bleeding or not very efficiently so what they found was that when you look at here in this red the red here represents all the exons that are necessary for making factor 9 which is again one of these protein products and what they thought rug I have found was that there was a mutation at this point here in the fourth exon for making factor 9 so basically what happened is instead of having the normal AAA G which is what the enzyme recognizes a AG cut here okay instead of it being an ahe Aleksey had an a G a so the the mutation occurred and that the consequence of that is that the enzyme could not cut that exon from the intron so the next slide what you get basically is this no cutting you produce a protein that has this both the exon and intron and then the rest is good but because you have all of this gibberish junk when it comes time to make that protein you don't make a proper protein it makes a non-functional so you can't clot so that was the problem that Alexi had so rogue I have was able to discover this and he found that not only was it president the czarina she was a carrier so usually females are protected because they have two x-chromosomes so if one copy of the gene is bad then females have a backup copy have another X but for males it's usually usually afflicted because you only have one X chromosome you don't have another X you have a Y so it's a it's it's that's why mostly males are afflicted by these blood clotting diseases so Alexi had only one X Anastasia had she lived would have been a carrier for this hemophilia she would have passed this gene if she had had sons they would have been afflicted or there's a 50/50 chance she would have passed it to any of her female children and I'll finish up with of course there's always this mythology about the missing Romanov children how they you know somehow they were actually survived that night and the most famous at least I think was Anna Anderson who who believed that she was Anastasia and convinced a lot of people that she was Anastasia when she died she died of colon cancer she wanted her body cremated and so this is before DNA testing so that wasn't such a big thing but so once the DNA testing had happened you know we were in the early 90s with the discovery of the missing family the researchers were actually able to disprove that she was in fact a Romanov because the biopsy that she had taken at the University of Virginia Hospital for colon cancer they kept that biopsy material and so they were able to go back and do DNA testing and it's kind of hard to see here but the the the scr testing here is well let me see if you'll pass my if you can pass my test alright so here's your i'll give you a certificate for friend being a forensics a junior forensic scientist if you can answer this question is it possible for a father who's an eleven thirty two and a mother who's a thirty to thirty six have a child that's a fifteen eighteen no okay end of story there's no way an Anderson could have been anastasia because she does not have the same DNA type that you would expect to be from a daughter of the Czar in azzurrina so in since 1918 there have been a lot of people who have claimed to be one of the missing children been eighty one man who claimed to be Alexi Peter's friend Nikita and once told me when we were in Russia he said yeah you know it's the funniest thing that were five children went down into that basement and two hundred came out so so I think we can with at least with the DNA testing we can now conclude that all of the children were executed that night that early morning [Music] [Applause] [Music]

Channel: The Romanov Royal Martyrs

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Keywords: romanovs, romanov, romanov royal martyrs, Nicholas II, Tsar of Russia, Royal Family, Romanov execution, Romanov dynasty, Romanov trailer, Romanov family, Romanoffs, Helen Rappaport, Four sisters, Last Tsar, Ekaterinburg, Olga Shirnina, Anastasia Romanov, Alexandra Feodrovna, Grand Duchess Elizabeth, Royalty, British Royalty, romanov family today, romanov family death, romanov coronation, romanov sisters, romanov jewels, romanov saints, romanov book, National Geographic

Id: qud87s2c1II

Length: 86min 37sec (5197 seconds)

Published: Sat Jun 27 2020