This is Think Tech Hawaii. Community matters here. Hello and welcome to Think Tech Hawaii and it never got quiet. This is a half-hour program that explores the Hawaiian connection with the Vietnam War. I'm your host, Vic Kraft. It seems that every outfit in the military has its own radar or Riley. Call them grave robbers, gophers or scroungers. There are one or two people in the organization that circumvent normal channels to get things done. Not all of them have higher motives. Today our guest will describe for us his fictional account of one man who manages to manipulate the system for his own benefit and along the way actually does some good by accident. He has written a book titled Loveless in the Knob. Our guest is retired U.S. Army Colonel and author Jim Borsba. Jim is a graduate of Michigan State majoring in journalism. He also attended the University of Michigan, the Prestige of Sofia University in Tokyo and the U.S. Army Command and Staff College. Jim has been an integral part of Hawaii politics having worked on several successful campaigns and also as the director of communications for the governor's office and the Hawaii State Senate. He's held several command and staff positions during his career both on active duty and in the reserves. Jim is a partner in the marketing firm of Starve Segal McCombs. He has also served a number of community activities and societies among them, Pacific Historic Parks, which oversees the Arizona Memorial, Diamond Head and three other national parks. Aloha Jim and welcome to the program. Aloha. In my family attending Michigan State and going to Michigan would probably end you up in a court of appeals. It's dangerous territory. Amongst other things, we had one of our nieces marry a guy from Ohio State and during the reception the Ohio State people were on one side of the room and the Michigan State people were on the other. So I don't know how you managed to do it and survive, but that seems like an accomplishment in itself. Well, I went to Michigan State right after I graduated from high school and spent four years there getting my degree in journalism and then it was some years later, in fact over a decade later, I was assigned with the Army near the Pentagon with Soldiers Magazine and yours at Michigan had an off-campus type of school that you can enroll in and I worked there for classes about a year and a half, got my Master's in Business Administration. Okay, we'll forgive you then. So I did it because it was going to help me and my family, I think. Well, your time in Vietnam, you were attached to what? I was with the American Division underneath, which was there were three grades. I was with the 196th Infantry Brigade, which is now stationed right up here at School Field Barracks and at Fort Shafter and then I was a Bravo Company of the Second Infantry, Second of the First Infantry. So you were essentially a private hump on the boonies? I was drafted in the summer of 68, went to basic training and AIT advanced individual training and then we shipped to Vietnam and somehow ended up in the infantry. I didn't expect that to happen but because I had a college degree but I ended up being an infantryman, I ended up being the RTO, that's the radio guy that carries the thing on his back. Plus carrying a weapon and having this? Plus the weapon and you know I carried food and ammunition and blankets and all kinds of stuff. I probably carried 70 or 80 pounds of my back when you added the radio on. But actually the radio probably saved my life once. I got blown off the back of a tank, slammed into a tree with my back and it cracked. The radio was about this big. It cracked the radio but I could still go get them to walk. You sent a letter to the manufacturer, I hope? No. We laugh about those things now and at the time it happened it wasn't that funny. No, funny at all. So what year were you in country? I got to Vietnam in the spring of 1969 and left in the spring of 1970. From the, I would say, from late April, early May I was out in the infantry and I stayed there until the fall at which point I ended up getting a job back in the rear with special correspondence unit. And again, I think because I had a degree in journalism, you know, you talk and meet people and stuff. And anyway I ended up going back there and I was signed with about five other people. We answered congressional inquiry letters, which we didn't sign. Commanders did. And we also wrote the letters to next again for people who have been shot and killed. That had to have been very depressing. That went to the commanding general who had signed those, of course, yeah. Yeah. And did you ever manage to get to Foucait? No. Oh, okay. Never got to Foucait, you had it. Or Queen Yawn in that area. I spent so much time out on, you know, LZ's like LZ Baldy and Hawkeill and those things. And we were the northernmost Army unit. The Marines were just north of us. And in fact, there were several large battles that we took apart in which the Marines and the Army were together going in to do things. Because of our location. That would be a revelation. It's the Army. No, we were friends. We were friends. It wasn't a lot of friendly fire and there's no such thing as friendly fire. Well, let's get into your book as far as Loveless in the Nom and just hold it up here and let folks see it. I'll see if I, here we go. You've created this character fictional and I asked you once before. I hope this isn't an autobiographical. No, no, this is a character. The events in the book are real events. Most of them are real events took part during the Vietnam War. But the character's made up and he's, if you read the book, he'll seem real clearly.  Never wanted to go in the Army. But he ended up getting drafted and sent over there and ends up being a prisoner of war at one point which is his own fault because he tries to escape from a battle zone by faking the heat casually and they put him in an elk hop to fly him out, he gets shot down, he gets captured. And throughout the book there are things like that. But it's all based on real events and things and I'm very complimentary I think of the soldiers and the units that are involved. But the guy himself's a character and yeah I've started a second book called Lovelace and Rising Sun and I've got a third and a fourth planned for this guy's so-called career. I'm anxious to get into it to read about it but you said it's based on incidents, hopefully they weren't your experience. But the idea of... Well the biggest battle of 1969 was Hyap Duc Hill which is near July in the northern part of Vietnam. And again the Marines and Army were both into that and this character is involved with that battle. What month was that? Do you recall? That would have been August of 69. Okay. And then I faked it. I mean President Nixon made one visit to Vietnam during the war. He had an award ceremony. There's only a country like 10 hours but this guy gets a fake award. He ends up in standing in line but he gets brought in at the last minute so that in the real news cycle people wouldn't have known he was there but the last minute the army brings him in but the president pins a medal on him and the president moves on down the line to the next and the medal falls off which it should have done. Karma. Well I don't want you to give away the plot of the book because as I said I'm anxious to get into it to see how this guy fares and maybe I can even learn something from him but the the army offers an awful lot of opportunities to... He did go to Michigan State. I'll admit that. How did you come upon creating this particular character? I mean just... I read a series of books called about a British soldier during the 1800s called Flashman series and he fought in China and India and all kinds of places like because the British were all around the world in those days and I just thought it was really unique. Those books sold really well and the character's kind of unique and I just thought you know maybe let me try something and I've had a lot of good compliments and got good reviews on this. I haven't promoted it really well so it sells just so so I think you know you can get it on Amazon, you can get it online, you can get it Barnes & Noble but that's not expensive but we'll see. My sales haven't done all that well either. If you got to remember 4,000 books are published daily so you're throwing your hat in the ring with an awful lot of other hats. That's right. So you left Vietnam. You said you were a draftee. That was a draftee, yes. But you didn't say in the field all the time while you were there. No towards the end like I said that they you're once about every third week or so we would be back at July and we get a couple days break and stuff. I ended up talking to people and somebody found out I had a green journalism and then I got contacted and said there's going to be an opening in the office would you be willing to come there and do that and said sure what the heck get me out of the infantry I'll do that it couldn't be any worse right and actually at July it was a big up big military base and had a runway and you know the ocean was there the beach was there it wasn't a bad place at all once in a while I got some rockets coming in but that's all yeah and so I just kind of volunteered to go there and work there and then when I left Vietnam I still had about half a year on my draft no time and a little less than that maybe and I you know I said they asked me where would you like to go since you were infantry and you got CIB and a couple of medals I said well or can I go in journalism so I ended up going to Stars and Stripes in Tokyo and I only wore a uniform one day a week I didn't really feel like I was in the army and I just I had re-enlisted twice and just stayed and I was a journalist. So they gave you consecutive overseas tours in Tokyo all that time? Yeah basically well I was in Tokyo or and then at Camp Zaman which was a army headquarters in the south of Tokyo. No I want to thank you because you were the highlight of the week for most of us in Vietnam when Stars and Stripes came out it was fought over. Oh everybody wanted to read it. Oh sure and it was probably one of the better pieces of journalism that was ever done. I think back in those days it was you know Stars and Stripes was all over Asia wherever our military was. Yeah and I think of the history of the of Stars and Stripes in all theaters as you know Bill Maldon the great writer and cartoon political cartoonist of the 50s and 60s he wrote up front and he was also a writer for Stars and Stripes but it's yeah it was a very reputable publication not just among GIs but I think amongst everybody. So you stayed six years there or so close to eight years in Japan yeah and what did you do? Then I had a year left and they assigned me to Soldiers Magazine which was an army's monthly publication that was over Washington DC so I went there it was a writer for about nine or ten months I remember exactly and then I got off active duty and my family and I moved back here to Hawaii and I since I already had like almost 12 years of you know I joined the National Army Reserve they're gonna get my 20 and but when I was an Army Reserve here in the 80s and 90s it wasn't any real big wars I mean we had that one in the mid-east and the late of the early 90s but you know hardly anybody from here went to that so being in the Army Reserve you really networked well in town it was a good part-time job and plus I was saving my time for retirement and I was a first lieutenant when I came back here so I just stayed in and stayed in and in 2001 I got promoted to full colonel so it was you know a good way to try to retire then December of 03 I got called back to active duty because you know we'd went into Afghanistan late 02 and then into Iraq in 03 and I brought up several hundred soldiers from the reserves of me I was we were placed in charge of the training and the equipping for the deployment for the entire pacific basin so the Pentagon would say okay this group from Guam they're gonna go to Afghanistan the state this group from Hawaii they're going to Iraq and this group's going they would tell us where they're going but then we would do all of the training get all the equipment that we could and chip them off and we just make sure their families are all okay while they're gone and then of course at the end we bring them back kind of a big special ceremony to welcome them all yeah great outstanding yeah I want to talk a little bit more about that but first let's go ahead and take a break here and listen to these messages okay Hawaii raising public awareness I just walked by and I said what's happening guys they told me they were making music I'm Ethan Allen host of a likeable science on Think Tech Hawaii every Friday afternoon at 2pm I hope you'll join me for a likeable science where we'll dig into science dig into the meat of science dig into the joy and the light of science we'll discover why science is indeed fun why science is interesting why people should care about science you care about the research that's being done out there it's all great it's all entertaining it's all educational so I hope to join me for a likeable science welcome back we're talking with Jim Borsma retired U.S. Army Colonel and author and we were just talking on the break here about about Vietnam and motives and doing things correctly as you and I were talking earlier Jim our group that meets on Monday nights we were talking about the training involved and being prepared and it seemed as though the recruit of the army back in the 60s was rushed into combat and you were talking about grabbing units from various places within the pacific region and sending them into Iraq and Iraq and Afghanistan and you had mentioned the idea of training yeah I'm wondering how much better prepared they were than well I'm gonna give you a little bit of example up at school field barracks we had a fake village like in Iraq right they were trained in that but we had one time we would show a movie about a convoy and we'd say no this is how you have to set up your convoy this is up front this is in the back this is you know and we'd show them that that so if you're ambushed this is how you need to react to stuff then a couple days later we'd have a convoy of those guys driving somewhere and we'd ambush them and see how they would react based on what they'd seen on the screen you know so we really really wanted the details about the training now some of a lot of that came from the from the Pentagon era they came over to us and said here try this do this and so we did and it's like I say it's school field the commanding general at the time had everything set up for it really well well I wonder how successful it was because I think of the lessons learned type thing and the funny funny thing is this the 196 Light Infantry Gate that I fought for in Vietnam was called active duty about three years before I got sent up to Schofield Barracks and they were they actually we our company was in charge of the soldiers equipping them sending them the classes and the 196 was actually doing the physical training with them and so if you're in the army and you you're assigned to a unit you wear their patch on your left shoulder right you're in an army and you're in a combat unit you wear it on your right shoulder right I was the only guy at Schofield with that 196 patch on the right shoulder and I lots of people said sir you got the patch on the wrong shoulder and I said no I was infantry in Vietnam in 196 yeah that was a very strange thing but we worked with them a lot and it was very thorough training yeah and went to Vietnam we had we got training how to shoot the weapons that was about it yeah and and I'm wondering if if that contributed to the casualty rate at all I I don't know you know I didn't fight in Iraq or Afghanistan in Vietnam I could tell you it was very horrendous at times very very difficult I I remember we we we got called once ago rescue a unit in northern Vietnam and we went in there and then they called unit after unit after unit and then we ended up having like several thousand American troops and they're trying to rescue this these guys and we did most of them but we fought the the NBA the national and you know not to be at call I might even be at calling or two but the NBA was there now we drove him out of this place but then of course we all pulled back and we were told two weeks later they're back on this hill again you know it's like well what did we go I'll do that huge battle for you know take land and then then give it right back to them in a way so it was a strange kind of war yeah several of our guests previously haven't mentioned that that no clear objectives the way it was prosecuted which is another thing I think we bring up today this morning in New York City there was a terrorist action or was it yeah and several people were killed and you got a wonder I recall one of my mentors mentioning that we were going to go about this all the wrong all wrong after 9 11 that we're going to gear up the military and do all these things and go after these people and perhaps maybe that's not the way we should be doing it we went after we went into vietnam with the idea that we were going to prevent the domino theory that worked real well and we'd been through Korea we said oh we can at least do that live in north and south yeah but it was a completely different war and the way we prosecute war is different and again that gets into the training and unfortunately as you said you were getting the feedback from the Pentagon and and how to do these things and how to train people for their particular environment yeah Afghanistan we did and we had really good support in fact a more recent governor Neil Abercrombie was a congressman at the time he came over and gave a speech up on the stage there was asking me questions and a couple of troops were like sir we're not getting enough enough equipment and the kind of stuff that we need and he said you know what i'm going to make sure you get that and he did and when he ran in 2010 for governor they would made a they made a tv spot for him I mean I had soldiers coming up saying hey thanks he did this he did this force and and it's true we were having a hard time getting around all the right equipment right at first but then then it came yeah and and that's critical as far as people being in the field you want to have them well trained and also equipped properly well and I was we were shipping off National Guard and Army Reserve guys that I'd known 20 years you know a lot of my friends were going overseas there yeah so yeah yeah I think of as you said your responsibility in terms of determining what unit goes where and the people that were selected for these positions and I'm sure you're looking at their training records to make sure that everything is we even set up a medical facility so people that had mental physical or even emotional problems would stay behind until we felt they were ready to go catch up to their unit yeah and it was we're very thorough on it they went as units or the unit would go but you know if they say there's 200 guys maybe two or three might stay behind because one of them might have pneumonia I don't know they could have had an illness or they could have got hurt during training so they couldn't walk or run so we keep them behind until they were physically ready to go then they'd catch up you know a couple weeks later to their unit but but they went as a unit both going in and coming out yes almost always they went as units come in so when they came back we had a big like a basketball arena we'd march them into there and you know all the families are up in the stands waiting for their soldiers to come back that's a heck of a lot better than what we got when we came back from Vietnam yeah no we got nothing coming back from Vietnam but I will say the public nowadays is a lot more accommodating and treating Vietnam veterans much better than 40 years ago that was I was going to get into that there are some people who have made the comment that civilians don't have the attachment to the military that they used to partially because of the draft we don't have a little Johnny in uniform anymore it's a professional force and what you're saying of course you may have reservists and national guard members going and doing these jobs and they're probably far better prepared than their their drafty counterparts from 50 years ago but it's still a pressure and still you know are you doing the job right I don't know it's what do you think I mean from your experience and from your position do we have a better trained army today I think that a lot of emphasis is being put on training yeah and I think in the last 10 years of it we're being much more careful about trying to get into conflicts I don't think the army is our military is looking for wars no they're there to defend this country which we want which we need you know but I don't think we're looking to get into wars you know we went into Vietnam I mean that Ken Burns movie maybe he's gonna say the Ho Chi Man actually wrote letters to President Truman and Eisenhower saying hey you know the the the British left India after World War II you guys left the Philippines why are you letting the French come back into our country you know ask them to leave so we have our own country and our own independence so we didn't support and we ended up going into a war there that we never really fought right I don't think and and there's a lot of criticism about Iraq too you know sure Afghanistan bin Laden's there they hit us a 9-11 go get him we all all understood that but I had very senior military people come to me say look we don't all agree on Iraq but we follow the orders of our commander-in-chief so yeah that's true and so as we did Jim I want to thank you very much for being here and telling us about your book journalism is not objective no matter how hard we try we all carry some amount of prejudice towards the topic however there can be alternative journalism where an issue can be discussed without rhetoric or animosity I believe that is what we have to offer here at Think Tech Hawaii this media offers an opportunity to bring more than one perspective to the community Think Tech also provides information on a host of topics that can aid in improving your life but all this costs money we speak of free speech as one of our rights in our constitution but it requires maintenance that maintenance has been measured in the lives of those who have defended it and by those who support such efforts as Think Tech Hawaii through their contributions the staff here are not volunteers and they would like to continue to pay their rent the hosts of the programs you watch our volunteers we do this out of service to the community so please contribute you can find a way to contribute at www.thanksforthinktech.causevox.com we would love to have some feedback if you have comments please send an email to 808 vietnamvets at gmail.com I would like to thank the staff here at Think Tech Hawaii for all their support and assistance special thanks go to Ray and Robert who go the extra mile truly without them this program will not be possible please come back again next week for another issue of it never got quiet Mahalo


Welcome to Microterrorz. Scary stories for kids. Where it's always the spooky season. Full of chills. Thrills. And spine-tingling spooks. Microterrorz are family-friendly frights for those ages 8 and up. And while our stories are for younger years, we are still talking about things that go bump in the night. And some children may not be able to handle what others can. Parental consent is recommended. Now for tonight's Microterrorz. Lovebug. Every desk had one sitting on them. A tiny red box with a sparkling ribbon tied elegantly on top. They were no bigger than a necklace box, which had the girls in the classroom hopeful. However, sixth grade girls wouldn't normally receive such a gift from sixth grade boys. Not to mention every desk had them, girls and boys alike. Suspicion immediately fell on the teacher, Mr. Mavis. But he denied it as well. Those who knew Mr. Mavis well enough knew it wasn't out of character for him to surprise the class with special gifts. He'd done it in the past with other holidays. Snickers on Halloween. Candy Canes on Christmas. Why would Valentine's Day be any different? The only thing was he always admitted it up front and it was never some kind of ultimate secret. These little red boxes, however, did appear to the entire class to have come from a mysterious origin. A secret admirer, even. Well, the entire class with the exception of one. One of the ten students sitting in Mr. Mavis' sixth grade classroom was not who they seemed. A wolf in sheep's clothing with a secret agenda. A girl in the back of the classroom, Jenna, was the first to flip the small red box over in an attempt to find out who it had come from. She noticed a tag taped to the bottom of it. Love Bug, it said. She quickly nudged Bridget, a brunette girl with blue Disney princess-like eyes who sat next to her, to let her know about the tag. Upon seeing it, Bridget wondered who Bug was. He or she had to have been the one who left the gifts. She looked around the room, her thought process instinctively picking out the boys who had names which began with the letter B. Brendan Love would have been too obvious. He barely said a word to anyone in class all year long and wasn't much for one showing affection, despite the last name. Plus, he also had a red box on his desk which he appeared to be shaking and smelling in an attempt to identify its contents. Although Bridget assumed that since Mr. Mavis wasn't responsible, then the culprit, as they were, would have also supplied themselves with a false box to throw off suspicion. This is kind of thrilling, Bridget thought, trying so hard to suppress her excitement. Bridget then focused her attention on Benji Blink. He was definitely a stronger candidate for the mysterious Cupid that went undetected in the classroom. Benji was as much a ladies man as a sixth grader could be. Well, he tried, at least. He was a little blonde flirt that wouldn't go down without a fight. Fights that he always seemed to lose because of his unknowingness of when to stop. One girl ignored him, he just moved on to the next. Rinse and repeat. In the front of the classroom, Lucas Quinn had also noticed the tag at the bottom. Love bug, he read softly to himself. He immediately turned to his right and looked down at the socks the girl next to him wore. They were black with red ladybugs speckled on them. Emma Sophia was always known as the bug girl. Of course, there was a bug girl. She considered them cute, like most kids would consider puppies and kittens cute. On most days, her attire consisted of something that incorporated bugs. Today it just happened to be the ladybugs on her knee-high socks. Emma fell Lucas' gaze and flashed him an uncomfortable smile. She didn't like being looked at with such a curious eye. She didn't like not knowing what people were thinking about her. If you're thinking something, you might as well say it out loud, she thought. I'm not a mind reader. At least when Benji Blake, the legendary failure of flirts, thought something, he said it out loud. Best to be denied immediately than to drag this out any longer than it needs to be, she thought about Lucas' fanatical stare. Lucas' stare was something that also caught the eye of Jaden Burke halfway across the classroom. He had been crushing on Emma Sophia since just before Christmas, but had yet to find the courage to make his feelings known to her. Even though he was sure she would deny him just as she had Benji, he still longed for the desire to talk with her about her favorite bugs, sit on the playground with her, or even share a lunch table. Holding hands would have been the ultimate, but Jaden wanted to play it slow and cool. In time, Jaden, he would think to himself, Do you think there's candy in here? Jackson Gruber, who sat behind Jaden, said to him, tapping him on the shoulder and holding up the little red box. Jaden shrugged. I hope it's candy, Jackson said, a glimmer of hope in his eye for a sweet treat. I don't know what else it could be. Jackson sat back in his seat and studied the box. He wanted to pull the ribbon and unveil a surprise, but no one else had yet. He didn't want to be the first one. Or did he? I could get this party started right now, he thought. Of course you wanted to be candy, the boy behind Jackson laughed. The boy kicked his foot on the back of Jackson's chair, immediately souring the mood. Rowan Davis was a bully, always picking on the others in class. He made fun of Emma Sophia for her love of bugs. He picked on Benji Blake for never landing a girlfriend despite his endless attempts. He scoffed at Lucas Quinn for being a hopeless romantic. Jackson Gruber was his favorite target, even though there was no real reason for it. That was just the raw nature of a bully. Jackson closed his eyes and wished Rowan away. Jaden could tell that once again Jackson was bothered by this bully the same way most of the class always was. He was a terror. Lastly, sitting by the window that looked out onto the playground was Mia Hart. She was one of the more quiet ones in class, although very attentive. From the moment they all entered the classroom and were greeted by the little red boxes, her inner accusations were ping-ponging off each and every student as she tried to narrow down the mysterious Cupid. And it didn't take her long to figure it out, either. To her, it was obvious within seconds who the culprit was. The only thing she now wondered, what could have possibly been in the boxes? The students were growing impatient. All but one of them had thoughts about the box's contents and who could have left them. Mr. Mavis could see how antsy everyone was becoming. Okay, Mr. Mavis said, before we jump back into the lesson, go ahead and open the gifts. I don't need these little things to be a distraction all day. Mr. Mavis had barely finished his sentence before the entire class minus one tore into their boxes. Candy, jewelry, the anticipation was killing them. Mia Hart, however, was slow to open hers. The student that she suspected of leaving the boxes was the only one not opening theirs. Theirs was nothing more than a prop. Mia was now concerned that there wasn't a lovey-dovey reason behind the gifts, but more of a malicious one. Mia stopped opening her gift and pushed it aside. That's when the screams of the children ripped through the room, demonic bugs with dozens of legs, fiery red eyes and jagged fangs burst out of the boxes, screeching and skittering like an unstoppable torrent of terror. Each box contained the endless flood of the demon bugs. They gushed out in continuous streams, covering each of the children from head to toe. Their screams eventually went silent as their bodies were encrusted with a thick husk of bugs. Mia was the only one still screaming. She had stopped opening her gift box, but the insanity that unfurled in the classroom was enough to entice her box of infernal insects to burst out as well. She was completely covered within seconds. Mr. Mavis, frozen in fear, remained in front of the classroom. He watched nearly all of his students be consumed by the wicked, creepy-crawling evil. The only students that remained sat as still as a statue, almost as if it weren't really him to begin with, but instead a copy of him, a malevolent imposter. At his home, Benji Blake, the real Benji Blake, lay in bed, quote-unquote, sick for the day and not even once regretting his decision to summon the vengeful spirit that he did. Every girl denied him. Every boy laughed at him for it.  A person could only take so much. Soon, the vengeful spirit's real-world manifestation that sat in the classroom would be back, confirm the deed is done and collect its payment. However, the spirit didn't know yet that Benji would not be able to pay up. He refused to. He couldn't bring himself to the level of sinfulness required by the spirit as payment. But none of that mattered to him. Just knowing the laughter and constant denial was gone forever? That was all Benji Blake wanted. Whether he could afford it or not. You can find us on www.microterrors.com where we will also have spooky games you can print out and play, like wicked word searches, mysterious mazes, and more. Microterrors.com is also where you can find us on your favorite social media and even send in your own scary story for us to tell. Plus, you'll learn more about our author, Microterrors for both young and old. I hope you'll join me again soon from Microterrors. Scary stories for kids.


Welcome to Calibrating a Depth Micrometer. In this video, you'll review the steps for calibrating the length of depth micrometer measuring rods. The depth micrometer extends its range with measuring rods. A typical set measures from 0 to 6 inches using 6 measuring rods, each with a 1 inch range. To change a measuring rod, unscrew the thimble cap and switch to a new rod, removing all dirt or chips as you seat the rod into the thimble. Every time you change a rod, make it a steadfast rule to recalibrate the depth mic against a surface plate or a gauge block. Holding the base down firmly, advance the rod just until it touches the surface plate, but not so much that it moves the instrument up. If you do find that the depth micrometer doesn't zero out, change the length of the rod via its adjusting nut. Clamp the rod in soft jaws in a small vise so it won't bend when you adjust the length. With the small spanner wrench, back the nut off one half turn and then turn it forward again to its new length. Admittedly, this trial and error process can be frustrating. That's the process of recalibrating depth micrometer measuring rods. Congratulations! You have completed the video Calibrating a Depth Micrometer. Thank you for watching. If you liked this video, give us a thumbs up.


My idea for a video game? It's a rhythm game.   They're twisting and coiling all over the place, which makes you have to do hammer-ons on a guitar at your own controller. You can hear your parents fighting in the other room, even though you're listening to sleep. It's dark outside.


Ladies and gentlemen, we are back. And I feel like I've lost my hair. It's very stressful as a business owner to not have product when you have demand, especially for six months. Maybe the supplements have kept me from losing my hair. And I'm excited to announce this reopening, grand opening, because I'm bringing something no other company has. Formulas and products developed based on what the body needs, not what makes a profit. For years on YouTube, I would just recommend other companies, user referrals on Amazon. But after I realized I could provide a better product at a more affordable price, and some stuff wasn't even available, I decided to do it myself.  I create products that I deem crucial to human health. For the meat, I was tired of recommending people to eat wild, them getting ripped off by farmers. So I started Frankie's Free Range Meat. Frankie's Naturals, I was making hygiene products for myself, because I couldn't find anything in the store. Organ supplements, same thing. There's additives and chemicals in just about everything we consume. So if you make it yourself, you can remove those factors. And Wi-Fi shielding was more of a matter of making it more affordable to the average person. Previously on organ supplements, I only had six products. The beef testes, the full spectrum organs, beef liver, and then for vitamins we had D3, K2, and iodine. Now I have 21 products total, 12 freeze-dried powders, and nine vitamins and minerals. And today I'm just gonna do a brief overview mainly explaining how to use them, the nuances. And in a future video, I might go in depth on each of these if you guys want to and what specific health issue you would use them for. But a lot of my past videos do explain why you would want these. The animal glandulars are powders and the vitamins and minerals are liquid. That means the powders need to be blended into something, water, smoothie, milkshake, whatever, or put in a capsule. And some of these liquids don't taste that great. Most of them are fine though, small amounts, and can be taken orally on their own. But as with the powders, you can blend them into something. I really didn't want to put high quality freeze-dried, like adrenal powder, which is hundreds and hundreds of dollars per pound, in a feedlot gelatin capsule, mixing quality with crap. The glandulars are basically food. It's the original organ freeze-dried into a non-perishable state. Since it's freeze-dried as opposed to vacuum dried, it preserves all of the nutrients, those glandular properties that work in your own organ, and the taste is much milder. When you consume this and digest it in order for the thyroid material or the liver material to be used by your own cells, it has to be in that raw preserved state. That's why I didn't opt for the vacuum dried versions of the supplements. Thyroid, for instance, is actually prescribed as a medication called Synthroid. I'm sure a lot of you are taking that. Synthroid is lower quality. It's just like conventional pig thyroid, and that works very well. This also contains the nutrient iodine, which is crucial for the function of the thyroid, in addition to those properties of the organ you consume, being digested and going into your body to help the function of your own organ. So we still have the full spectrum organs, which is formulated for the correct ratios in our body. Any other product you see out there, they just throw even amounts of the powder in there, no rhyme or reason to what they're doing. And I was able to figure out what ratio we should be naturally consuming based on our ancestral diets. Most people don't need all of these organs on a daily basis, so I do encourage you guys to try each of these different amounts, see how you respond. When you freeze dry a food, the weight is between 20 and 25% of the initial product. So if you're trying to gauge how much of this you should consume, think of it as 1 fourth or 1 fifth of the typical organ you're consuming. So when you eat the freeze dried stuff, it's five times as much. A tablespoon of freeze dried liver powder is the equivalent of five tablespoons of liver or about four, between 20 and 25%. Moving on to the vitamins and minerals, and I made these because I'm taking them myself every day. I suggest practically all of my clients get most of these. And there's two issues with regular supplements. One, the dosage is not based on nutrition, science or what naturally occurs in food. And two, they have fillers and stuff you don't want. Magnesium, stearate, oxide, silica, overall things that you don't have to have. These are just spring water and the mineral. And the dosage on the glandulars for other companies isn't as big of a deal because they are whole foods and have complex vitamin and mineral profiles. You know, when you have thyroid, you're getting 15, 20 different vitamins and minerals. But when you have a concentrated mineral, something like selenium or magnesium, if you take them at high doses, it can really cause imbalances. And it makes you wonder why in the case of something like selenium, there isn't a supplement that is safe to take on a daily basis. Most of them are dosed way too high. And I formulated each of these supplements to be what I deem as an acceptable amount to take on a daily basis. The frequency can be higher obviously if you're nutrient deficient, but for the most part it's a maintenance dose. And I'm really, really excited the most about the trace minerals right here. We have a B complex. We have an antioxidant support. The trace minerals are dosed to be what naturally occurs in high quality plant foods. The antioxidant support is great for headaches if you're temporarily in a high Wi-Fi environment. And the B complex is unlike anything else on the market. It has all of the B vitamins naturally occurring in meat in natural ratios. Any other B complex supplement has usually reported toxicity for certain B vitamins such as B6. And to be clear, I don't mean natural as in they're derived from meat. I mean natural as in, you know, if you ate like a pound or two of steak a day, that's the ratio that these B vitamins occur in. I guess the overarching problem is that most supplements that are available online are just dosed too high and you can't really tell how much of it you're supposed to take. So definitely let me know if you have any questions down in the comments below guys. I'll kind of leave you guys at it. You know, I've done videos on copper, magnesium. I have to touch on selenium. I think I spoke about it in the iodine video. The trace minerals are boron, manganese and molybdenum. Stuff you usually don't get on carnivore that occurs in plant foods. The B complex is simply all the B vitamins. Antioxidant support is N-acetylcysteine, glutathione, coenzyme Q10, vitamin C. So organsupplements.com, check this stuff out guys. Let me know if you want me to go more in-depth on each of these. I would be pretty involved. But you know, most of this stuff can be taken at face value and assumed what it's for. So thank you guys for joining me today. If you can please drop a like on the video. Leave me a comment down below. Subscribe so that YouTube can unsubscribe you next week. And if you want to check out everything besides organ supplements, you can go to frank-tifano.com. Thanks again guys. I'll see you for tomorrow's video.


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నిర్న్నిన్ నినిల్నంవాడిల్దరా. Today we will see the exercises regarding test cases and scenarios from a set of requirements which we had studied about the requirements in the master class and try to study that with requirements and the corresponding test cases. So we had also gone through the example SRS it is called embedded unit instrument. We will just try to recap that then we have a test review checklist this is based on high level testing so there is a sample here this typically being done as a completion step in auto typical rate of the embedded software testing high level testing that is been done on the set of requirements. So this checklist will talk about the rules and whether rules are followed and whether that embedded software testing can be taken for in next level like this it will be used so that is how the rules are being used. So once so these two exercises are done we will try to continue the exercise in terms of identifying and establish traceability matrix for the developed test scenarios from the set of embedded unit instrument requirements. So you know the traceability, traceability basically is required to see and report that from the requirements to test cases how the coverage is there how we are able to trace different requirements to test cases similarly test cases to requirements so two are different basically I mean though we call it as a traceability so both are required because the coverage aspect is different so the test cases requirements is called as upward traceability and requirements to test cases are called as downward traceability okay. Now let us try to recap the requirements so as far as one talks about the embedded unit instrument operating in the five operational modes such as the unit operational mode and then here you can see different modes of operations so how they can enter and exit so this is a parallel mode of operation is fail which can go from any of this unit operational mode at any point of time based on the different conditions so those conditions are nothing but the modes of operations typically SRS 2 onwards we have modes of operation requirements I think we had gone through this SRS 2 regarding modes of operation SRS 3 as well with any to mode SRS 4 over operation mode 5 over maintenance mode and the 6 again maintenance mode so likewise we have different modes of operation SRS 2 to SRS 5 6 other similarly we have performance requirements in terms of SRS 7 to 11 here we know that the booting type requirements and the workload performance of the processor or the CPU how much it should be and also along with the timing and the workload we have the memory related performance requirements such as flash memory RAM memory stack memory so how are we going to verify those let us try to go with some few example test cases so that there is a framework on which the entire testing can be taken up further in terms of test scenarios as well as the telequation similarly we have a communication requirements in terms of how the instrument unit can communicate with the external world it is through can messages we know that because the first requirement talks about the different messages it is receiving and based on different messages different modes of operation so there is a watchdog requirements you know that watchdog should be pulsed less than 500 milliseconds so this are some of the requirements for which test cases have developed so let us try to understand the test cases in one of the session we tried about how a test spec or the test cases should be existing so we need to have the requirement document test plan we assume that test plan is available and test spec which is nothing but the test cases scenarios corresponding to requirements then we are going to group it then we are going to press it these are some of the specific activities that we do for developing the test cases and in the test specification we are going to have a strategy in terms of how we can do it then practical steps which are nothing but the test procedure this how we do so these are some of the things that we need to do we know we have gone through a example of template how it should look like one of the theoretical class so let us try to understand the test case perspective how it can be developed yeah so we have about three levels of four levels of requirements in terms of modes of operation performance requirements communication requirements and watchdog requirements so we know the templates how we are going to use it the first section should talk about the test case identification for each of the requirements you can see as each requirements we need to first identify this case I guess we have gone through this template how it should look like like it should identify with the test ID it should be unique for each test case and what is the input for that particular test case so what is the condition that for this input which should exhibit then what is the expected result that we are going to have similarly for each of the requirement like we are going to take up SRS 2 SRS 3 SRS 4 likewise we are going to have for each of the requirement so that is how we develop test cases and the grouping is based on different functionalities like first few cases we will concentrate on the modes of operation next one more performance requirements and next grouping is based on the timing and next group could be based on the watchdog likewise you can group it but before grouping first of all we need to have is the test cases individually identified for each of the requirements is very important and basically we can adjust like what test case can go into what and what can be from the or if it duplicate or redundant any aspects that we can do first the first and foremost thing is develop the test cases for each of the requirements once we are done with all that then we are going to have a grouping once the grouping is done and we know that for each group there is a set of conditions that need to be existing before we start the practical step or the test procedures so what are those conditions they are called as pre-conditions so pre-condition for example I have put for one of the test case we can have a test group one which is nothing but relating modes of operation so what are the things that we need we need to power on the unit and set the below values below values could be something like what are the inputs or the unit should be hooked so what are those one could be can message adapter should be up similarly any interconnected interface delivery subunits should be powered okay so what will we need some discreet we could need should be up likewise we are going to have the pre-conditions so the pre-condition can be unique or different based on the type of group that we are using so for modes of operation we can message should be coming because without that we cannot test the modes of operation because similarly we need to apply power a voltage of 5.0 so like this we are going to have it and some of the requirements we may not be able to do dynamically or when the target is running so those list for static analysis where we need to do it static execution is by seeing the code so like performance requirements some of like memory map those things you don't need to execute on the target so those cases we are going to list it out here telling that that group of that specific scenarios can be tested statically or static analysis based on that similarly the template talks about the scenario functional or operational requirements next to test cases scenarios categorized and non functional requirements etc that is basically whatever the user or business based functionality means the instrument should be doing some function like we should rotate the motor that is the actual requirement so the non functional requirements such as you should power up within 5 seconds something of those test cases they are not this is related or user related that is all performance or supporting sort of a requirement so likewise we are going to have the appendices updated for the test case that is one section that we have and the first sections what we have seen is test cases identification for each requirement so this is what we are going to do so for example we will study a few requirements and few test cases per se okay so as far as to we know that what it talks about it talks about the modes of operation like we know that the instrument shall set the mode to failure mode in the following cases there is a power logic so one of these conditions one of these three conditions if it occurs it is going to enter into failure mode that is what the thing so here what is the input contents of this having the inputs so let us try to understand one contact or one condition when mode is in metastore so mode is a input and received message is an input so what is the output fail mode that means again mode set to fail mode condition is what this itself is the condition so first condition second condition third condition one of these condition exits it is going to enter into failure mode okay let us try to draw an example cases first test case let us try to define so we know that mode is an input received message is an input why because scan message is a dependency right so message is one input mode is another input if those two these two are inputs we are going to apply some conditions for that input and we are going to get the experience so what will happen if mode is in maintenance mode correct if this input is satisfied and when we get a receive message zero once we are enough do not worry about what is it CRL message just assume that because we are focusing or our emphasis is to test this portion not the actual data that may be a robustness in terms of different message other than zero once around what will happen that may be we can define later but we will start with a normal case saying that we have fed the two inputs maintenance mode and the message with zero and serial CRL mode directly as per the requirement we should go to fail mode so this we have tested here and in the next case you can see we have to test with the CRL message zero to CRL in that case also it will go to fail mode similarly the third condition when the mode is in init mode if the status is in error state that is during initialization if any error occurs it will go for failure mode so these are state forward test cases that we have seen for this requirement similarly if more conditions are there we are going to apply as per that okay the state to define now this is our logic so that means any of these conditions or multiple of these conditions occurs so we have something like a or b or c equal to b b means it is a failure mode so we are going to have a true table sort of a thing where either a can be set to true or b can be set to true c can be set to true in that condition this will go to fail and next one a b set to true a c set to true a b c set to true in this condition also it is going to go for failure so likewise we are going to have this is called that ncdc or the modified condition decision logic because based on the condition different decisions are there here when a a becomes true the condition will be fail when b is true condition will be fail c is true condition will be similarly a and b both are true condition will be fail right both are true fail both are true fail similarly a b c last case which becomes to true to be fail next we have what else left everything will have right everything we have written so in these kind of things it is good to have something like a true table we can define that way it will be easier actually maybe I will try to have one more column added so a b c and the output is what we are trying to set for different values so we know that combinations of a b c we will try to set zero zero sorry one zero zero output will be fail similarly zero one zero output will be fail this becomes zero this becomes zero c next condition zero one one typical true table we are going to have it for all the combinations the more over we may not need because we know the result of this why we need why we may not need is the independence we need to prove I will try to explain that okay here you can see the multiple two table combinations right c is true output will be false and b is true output will be false similarly both are true will become false and now we are going to set a as one and we are going to vary it here this is going to be still true the last condition is zero zero zero it will become true that means true means there is a failure or not a failure here in this case there is a failure right okay one more condition is all should be one should become failure yes means basically failure so for each of this we need to develop the test cases adding the inputs accordingly okay now we have tested about is normal cases of with a true table modified condition next within each of this we can identify other test case or other group of test cases such as okay I will try to draw that only okay so input is more maintenance more and receive message is zero one CRLF right that we have already tested this will become failure okay next condition within this test case input mode is not of maintenance mode that is not we do not know what mode it is so it will become no fail correct because we are not all there in that actually the next one input is in maintenance mode but the received message is not zero one CRLF it is something else x x x whatever it is so this also will become no fail so like this the combination that we need to have that is very important once this is done then we are done with the complete test cases and we are going to apply it okay the next thing that we need to take care is the independency very important term you can see in the above case abc is set here it has occurred false now we have proven only the b has changed from zero to one whereas the remaining things are constant let also we need to prove right because remaining thing is not constant here we have seen b has changed along with c also we do not know because of what the failure would have occurred so better to have the independency such as setting it back to zero so that we are proving the independency with this because of this only the failure has occurred and not because of this or the combination of this so that is how independently we are going to prove similarly for other cases also like we have a zero zero zero no fail and since this has toggled these two bits because of this they are getting the fail so that is how the effect of the inputs we need to tackle it appropriately so that independently this is one of the important thing that they look for when we are going to write the test cases so that is how we are going to have for different sort of combinations and inputs for that particular requirement okay next one next one is SRS 3 this talks about what when the mode is unit mode and the status is no error state that means no error when the unit is in the when the mode is in unit mode the instrument shall set into operational mode so two inputs mode and status unit mode no error it will be operational mode mode received message mode and received message so this is not a right test case maintenance mode okay let us not write to this we will try to put a simpler one mode status non-init mode okay let us put it more simpler it is in unit mode status is error so what it will become it should not be not of operational mode that is for sure because the requirement says it should be there in operational mode only when so by going through the operational mode requirements this is clear for us probably by seeing if it is not of operational mode what is the mode we are going to take care let us try to go through the requirement again we will come to know requirement is yes it is going to go for the failure mode because if it is going to be no error state then it will be operational otherwise it is going to go for failure mode that is what it says right okay so nothing but it is nothing more fail mode so likewise we are going to add this case while looking at the requirement so for three we have done okay four is regarding maintenance status status alive messages and six is about the next set of messages zero four likewise we are going to add it I can see particular SRS and for that test cases we are going to add one more thing you would have noticed is interface requirements tested so what will happen is some of the signals that are having a dependency for particular requirements suppose for this function requirements for the signal dependency is there some discrete and that we need to list and by listing that we are also trying to test that particular interface requirements so that also we need to cover it as an input that may not be a full-fledged functional requirements but there is a dependent interface requirements okay the next one is performance requirements let us try to understand what performance requirement talks about the entire process of booting up till operational mode shall not take more than one second that means upon power up we should not take more than one second to become operational mode so how we are going to do there is a two input that means we need to measure right measure the time that is what it says between boot up till off mode okay this is something like a assertive test case where we do not have a negative or where what is it called up we are expecting something with some actions here action is the thing but between boot up boot up how it is going to happen upon power up that is an input input is power up and mode and condition is power up entire process of or boot up boot up how it happens with a power up button power on we can say the action is called as power on or boot up so output is time difference between operational mode entry from power on time so something some instrument called tautiloscope okay with tautiloscope we need two tapping parts or we do not need two tapping parts or depending on what sort of a process we suppose power up has a pin number 20 where you can see the glitch of the signal pulsing at a certain level by words if it is we can observe that with one pin captured there and operational mode if it is tied up with a led something say led 2 it will have a port something like a GPIO port or whatever it is so these two we can see it so that we can see the reference as one with a power up pin other one as in the led port when it is going to become operational mode so the difference we can find out with the help of this so that will become a test case in terms of measurement so input is power up so we need to give a pulse to or pulse or reset to pin 20 booting the processor measure using tautiloscope what will happen is expected result is less than one second that is what is the right so that is how we are going to measure it with the force of play and try to validate the test case okay okay next type of requirements we have communication requirements okay here you can see two requirements are there instrument shall communicate through can messages with outside interface the instrument shall communicate its alive status at one second at least one second that means two things are there we can combine this requirements to have multiple places under one group saying that proving this also can prove that it is communicating and it is also communicating at which status that one second so it is just a validation of this basically there is no verification like there is no input that we can tweak or we can modify so what sort of a test cases we can have test case one will have observe the output can message check whether you can message are received at every one second next one check whether the can message belongs to alive status these are the important steps to validate this requirements right okay next negative type or when the instrument is powered this is going to happen so switch off ui what will happen is no messages are observed no message should be observed outside so this is proving that instrument will communicate only when it is powered off outside the box and the rate at which the can messages are coming how we can verify the rate is based on the timestamp because every can message will have a timestamp a logger outside in the host with that logger we can see a difference between message to minus message one there should be one seconds of course it may not be exactly one second it could be something like 980 seconds it depends again to one zero zero here tolerance of plus or minus 20 is there you can see it can fall anything between 980 to one zero two zero so it depends on type of system tolerance also will be there so you may not get exactly so they will specify in the requirement accordingly saying that for this tolerance it should work so that way we need to verify the such requirements any timing requirements or any one log or any of the outputs that has a tolerance we can verify that so this is how communication requirements can be tested with the test cases this way the last one being the watchdog requirements so there are two requirements here the instrument shall configure the interval what about generate a reset to microcontroller with a threshold duration of five minutes what it means is it has a configuration saying that when it is up and operational it should be configured for find a millisecond such that the watchdog will be satisfied with a pulse of less than five milliseconds so it is the next requirement if it is not the watchdog is going to reset it because it is configured for find a millisecond right okay so here cases we need to verify in the sense that first one is verify the configuration of the watchdog this is theoretically or by seeing the code or by seeing the registers we can validate dynamically we can validate such a way that we can capture the pulse of the watchdog usually watchdog is a circuitry where it has a counter and that counter is configured for 500 and there is a decrement counter the 500 becomes 499, 498, 494, when it becomes 0 it is going to it is going to reset or pulse the reset pin of the processor such that the processor gets reset how are you going to emulate this somehow you make sure that you reconfigure the watchdog or you make it hang the program that is there in the this thing or capture the pulse so what is responsible for pulsing the watchdog so that should be less than five milliseconds so that is what the second requirement talks about at least once in every five milliseconds or less it should be there so one is configuration other one is whether it is pulsing properly or not we are going to verify this is how we can verify the watchdog sort of a requirement okay so likewise we are going to have the test cases defined with test ID inputs conditions expected results and appendices we have studied now having seen all these let us try to understand the review checklist we have done the testing of high level requirements whatever I have demonstrated so far is writing the test cases and the requirements and how procedures can be written on this is what we have understood now having done this we need to understand the checklist usually this will be a getting criteria for the test plan or test cases to make sure that the rules of the particular verification of the test cases are adhering to the requirement or as per the need that is what we are going to do okay okay so these are an example template generally they use it it is again specific to the organization or specific process that is being followed in the type of embedded let us try to go through this and try to fill out the checklist on what we have done so far okay first one are templates followed during the development of test cases this we had a document this template which we have followed right yes are the valid equivalence classes and it is applied to all inputs of high level requirements yes we have seen the equivalence classes in terms of the complete truth table and all and set of inputs we have considered are limits values used for exposed remove here there is no exposed signal but can used so there is a limit in terms of the single flux here the values coming from there we have used for inputs structured in a table are bounds and one intermediate value of the table equivalence class there is no structured input table so this is not applicable we generally see the functional analysis used for the logic equations yes are particular test cases created for time related functions and state transitions yes but we have them only for the time related there is no transitions we have is one the questioner test case created for each external inputs yes we have the negative or the different values of kind of inputs that we try to put it like robustness so that is what is there is identification format of test cases we have seen that every test case has its own unique ideal is each high level requirement covered by at least one test case and we will correct one it is very important correct one is also so this based on the review we need to fill this whether correctly with the yes we are done for each of the requirements for each one test case you reach test case created covered by scenario and by the correct one this the test cases like we have our test case for each requirement and that requirement of multiple scenarios and the correct scenarios we have identified are values of inputs and outputs defined for each test case is each scenario qualified as normal is probably we are not done this needs to be done based on the validation of test cases this will be taken care since we are not completely valid if we review and all that once we have that we can define these things that is what in this justification we should be able to report it saying that we know what is my action for this whether it is correct or not is it cost test coverage or not it is commercialized itself I think we do not have that so there is a two section one is guideline other one is rules rules are mandated to be followed as yes there is no solid test case and also called as deviation of the result which the report will not be accepted by the designated authority or after a customer as we have seen guidelines is a description of these scenarios here and there somewhat it is I will say more if you can add no description are the expected results defined for each test scenario or it is bench specification defined for each test case what we have done is we have come to broken and identified the test bench or the setup that we required the preconditions defined for each test case yes we have seen that it exists so this is how we are going to fill the test case checklist it is called deviation of test cases it is objective description and there is a saying that is in the following so that is what we have to do or set the develop test case from the requirements so that is how checklist will be filled this ability this will continue in the next practical session in terms of upward test ability as I said from test cases to requirements is called as upward test ability or downward test ability which is required to test cases


So the next talk is from Mike Snyder, again, on non-coding genome in cancer. Sure. So I'm going to continue the same theme. Like many others, we're very interested in understanding non-coding information and its contribution to disease. I'll tell you about our cancer work, but we have work in other fields, particularly in autism in the space. So I think all of you appreciate until recently, most disease studies were really focused on exomes, because it was cheaper, easier to get lots of data, and I think this is from about 18 months ago when, I wouldn't say when we launched it, but towards, I guess, when we were getting ready to publish our first paper in this area. This is the relative size of the genome to the exome, but when you look at the sequences that were determined at the time, it's much, much more skewed the other way. There are many, many more exomes than cancer genome sequences. And, of course, this is shifting now that the cost of genome sequencing, human genome sequencing is dropping considerably, and, in fact, it's at the point where we pay about $1,200 for a human genome sequence. And so there are getting to be large projects out there, and, in fact, the PAN cancer project has just released 2,800 whole human genome sequences, so there'll be a lot of sequences in this space. What we've been doing is using several approaches of trying to identify recurrent mutations or highly rich mutations in regulatory non-coding regions, as well as within the genes themselves. And to do this, we've been using a variety of approaches, I'll mention two today. And we've been basically using the cancer data, but we've also been taking advantage of this database Colin told you about yesterday, regular MDB, which is primarily ENCODE data, road map data, but it's also other what we felt were high quality data sets that were in the literature that we processed with the exact same pipelines and put into this database, and it's a joint project between our lab and Mike Cherry, and now Alan Boyle, who started, who's at Michigan. And so what's nice is that, again, it has a bit more information than just ENCODE itself, and it does have the information annotated in ways that I think Colin told you about yesterday. So we've taken these whole cancer genomes, and we've used a fairly stringent mechanism for identifying the recurrent mutations. We're taking them, the called variants, and there are several methods for calling variants that are enriched in cancer samples. Mutech is a common one, the Variscan's another one. We've taken the intersection of those, which is throwing out a lot of good mutations, so to speak, but it does give us a more higher quality stringent list. At the time we started the study, there were 700 genomes that were done at the time in TCGA.  That was a new concept, and that created all kinds of problems for us and others. But then gradually they said, yes, you can do secure computing in the cloud, so they gave us a break and let us continue. So anyway, that's why it's an odd number. It won't match anything you'll see in the literature, but it is what it is. So these are the tumors. There were eight types that were there, and you could probably guess from the name what most of those are, and we'd call mutations in these various ones. But we are a bit underpowered because it's only eight tumor types and spread across. So we did do a lot of analysis on individual tumors, and there were marginal signals there. But to be honest, the best signals really came out from the aggregated data. Although as I say now with the larger data sets, we should be able to drill into individual cancers, much like the analysis you just saw. So basically what we did was we started looking for mutations that would be enriched in regions of interest using a variety of methods. And our control of most of these cases is using simulated regulatory regions that have similar characteristics. And specifically, the controls you need to run for cancer analysis are varied. There is one, the different kinds of mutations differ with the different kinds of tumors. So that, as I say, lung cancer, as you might imagine, especially for smokers, has a certain distribution. Melanoma has other kinds, and skin cancer, and so on. So we actually control for the mutational spectrum across the individual tumor types that we normalize per tumor. We also control for variation in gene expression. We control for replication timing because it turns out there are more mutations. Many of you may know this in the late replicating regions versus the earlier ones. So we control for that. And then we also control for base composition as well. So all these are taken into account for simulations. So then what we do is we do start looking for enriched regions, either regions for different kinds of analyses. In one case, like this one here, we basically look to see whether there are bound regions by transcription factor using regular MDB or ENCODE and seeing whether we can see enrichments in certain mutations for different kinds of transcription factors. And these are some of the positive results we've obtained. So basically for these different factors, in certain kinds of tumors, you will see enrichments in mutations in the binding sites for these particular factors, again, implying that they do have a role in cancer. We can also drill into the individual sequences, in this case using motif analysis, to see where those mutations may lie. And specifically, here's two examples here that were successful where you can show that the mutation counts, they don't just lie anywhere in the sequences. They can lie in very specific residues. In this case, the essential two seem to be the most mutated ones. In this case, it's actually more varied across where that, for SB1, varies across individual residues. What we found for ARAs, which differs for the other case, and if you think about it makes sense, most of our mutations are actually loss of function mutations. Not all, there are some that seem to be gain of functions, but most appear to be loss of functions. And so you can show relative to the PWM, most deviate from that. And so let's see, well, I guess here are the various factors that we've analyzed. And bottom line is they're, again, mutating away from the PWM. There are exceptions, individual cases. So then we can ask, is there recurrence happening at very specific genomic loci? And I think I won't get into the details of this so much, but basically the bottom line is you're looking for an enrichment over certain regions, and this is done, we use fixed windows for this. Varying sizes say 50 or higher, and surveyed the genome. And when we did this, we came up with about 123 sites, sorry. 123 regions, let's see, I thought it was 200. Okay, I don't remember what the difference is now, between the 123 and 200, I should. Colin, are you here? Yep, I lost them at key time. All right, anyway, so we found, I believe it's 123 sites, regulatory regions. And the top two are actually telomerase regions. And these are well-known sites that are mutated. In this case, they're actually gain-a-function mutations that activate. And then we found, again, 121 others, regulatory regions. I think the difference for this, the 200 is that we can map them to more than one gene, and I think that's what the point of this slide is that even some sites will actually, many of them are not necessary. In fact, the majority are not in promoter regions or immediately adjacent to a gene, so they can map to one or more gene. And so I think we assigned about 200 genes that were either in the vicinity or using some of the interaction data. And a number of these were, in fact, known oncogenes or genes implicated in oncogenesis. And so quite a few of them were, but the vast majority of them had not either weren't assigned to known genes or maybe were not mapping them to the right locus at the time. So similar to what was presented before, we took nine of these regions and tested them in reporter assays. And two of them, which were predicted to be loss of function mutations, actually do, so if this is the original reference locus, and these luciferase assays actually lose activity, that's just two different constructs. Here's another one. And this is the telomerase control, which is a gain of function mutation. So that one goes up. So the vast majority of ours didn't validate in this simple, simplistic assay. The other thing we've done that was a little more sophisticated after this came out is actually, rather than use fixed windows, we look for increased density overall, so we let the size of, if you will, the window vary. And then we just basically looked for enrichment across any size regions for very small, even single bases up to several hundred base pairs. And then in this case, we applied it to exomes. We wanted to get more power. And so you might think, why is this in an encode talk? Well, it turns out, even in exomes, there's plenty of promoter regions that are sitting right upstream because they go just beyond the five prime end in many cases. And in fact, we actually do pick up quite a few promoter ones. And we've run all the same corrections I mentioned before about sequence composition, replication timing, et cetera. Because there are many exomes, now there's many more than this, or 4,500 a time. I think these slides didn't come out that great, sorry. But I think I can still explain what's going on here. Bottom line is you can get, we call these significantly mutated regions. And you can get really huge enrichments when you have this kind of power of 4,500 exomes of data. And basically, I won't go through all the details of the scoring pipeline, but we wind up getting a strong enrichment in many, many different oncogenes. So again, this is probably hard to see, but I think for some of the points I want to make, you'll see in a minute. So the bottom line is that all the top ones are very well-established oncogenes. It was a case, as I mentioned, that many of these enriched regions, these significant mutated regions, do lie in binding sites, either in three prime UTRs, five prime UTRs, three prime UTRs. And also in promoter regions, and this is really hard to see, sorry. Well, maybe you can squint or something here. And the bottom line is you will see a lot of them are in the promoter regions. And you can assign this either based on motif scores or using ENCO data to help validate the motifs are the ones you think are really hit, you can basically see that these are mutated in regulatory regions. Although I think you do have to be a bit careful about assigning, at least for the UTR ones, some of them that landed in UTRs, we don't think they're necessarily affecting translation per se. They actually may be affecting gene expression. And these are some of the ones we had. We had several that landed in UTRs, as they say. What I liked about the variable approach is that you can actually drill into specific locations, whether you're outside the gene or inside the gene. So the example I'm going to show you here is that you can drill in, you can see hypermutated regions. Typically this has been done for cancer across the entire gene. But what we did here was because we're finding recurrent mutations, you can drill within subsections of a gene. And it turns out that a lot of the mutations actually landed at protein-protein interfaces, which was quite interesting. And then some of these actually landed in a way that they would be mutated in some cancers but not others. And here's where having 4,500 exomes is very valuable. So as an example, this is just the PIK3CA gene, which has mutated many different cancer types. These are the different cancers, and these are the different regions that are mutated. So you'll see this particular region is mutated in virtually all cancer types that we've analyzed here. But other regions are only mutated in some cancer types. And so you're seeing very specific subregions of this protein that are obviously more important to one kind of cancer versus another. And so it's nice to be able to get this kind of resolution. And I think the way you get this is by having huge amounts of power and lots of genome sequences. So in summary, what I've shown you is we can identify mutations in regulatory regions that are enriched for mutations in cancer. They often lie near known oncogenes. They can disrupt specific motifs of very specific factors, and we can implicate certain kinds of factors in very specific cancer. As is mentioned before, the density analysis is nice to be able to identify very specific coding regions. And of course, now we want to incorporate these in the whole genome. And actually, we're trying to see if some of these regions can be put into a clinical interpretation as well, which I think is promising as you get larger numbers. And with these 2,800 genomes, that's the next set that's coming along. And of course, there'll be tens of thousands after that. So I think we should be able to get highly annotated non-coding regions. And I think, again, the recurrence information and the ENCODE data will be extremely valuable for these sorts of annotations as they are for other kinds of diseases. So these are the folks who did the work. None of it was done by me. Colin Melton, you've heard yesterday he did the first analysis I mentioned. Density work was done by Carlos. Help from John Sennick and Jason Ruder on reporter assays. Alan Boyle is the one who made a regular MDB. And the last analysis was collaboration with Will Greenleaf's lab. And of course, if you want to learn more about this whole space, I'm making a plug for my new book that's come out. So feel free to go buy it and have fun with it. All right. So anyway, that's my story. If you have questions, I'm happy to answer them. Great work. So I'm interested in the transfer factor binding side mutations that you mentioned there. I'm curious because even though there's consensus, but the transfer factor binding motif can tolerate a lot of different variations. So if you see, for my understanding, the transfer factor can tolerate a lot of the differences inside this motif. So why the poor mutation inside the motif, but a less concerned position can have a functional impact? I'm curious about that. Yeah, it may be the specific kind of mutation. I didn't see what they were switched to. Yeah, you're referring, presumably, to this one here. There it is. Yeah, the CBPD. Well, that C is clearly a highly conserved one. And presumably, I guess all I can say is, first of all, maybe the C isn't the only important thing. That is the reference one, maybe the tube. But if you think about it, it should be both these residues, because it is a symmetrical sequence. So offhand, you would think both should be important. So I'm guessing this is the more likely conserved one. And so our data would suggest those are probably the two most important residues of the bunch, at least in this context. I mean, your point is well taken otherwise. And certainly the same things here. The highest ones are in these Gs, which conserved. What's going on here isn't clear. And I think that illustrates, though, a good problem, which are understanding these motifs in the native context. So a lot of these PWMs, they come from in vitro experiments, which doesn't really match the chip experiments for those who've done this sort of thing. Sometimes they match, and sometimes they match pretty well. But there are plenty of cases where they don't match that great. And there are plenty of cases where you will find alternative motifs besides the one that's in the literature, suggesting there may be a different binding site in context with something else. And so I think the in vivo ones, I do think that would be a good lesson for this group to take away. Don't assume the PWMs you see from a select experiment are going to match what you see in the chip ones. They definitely, you know, they generally kind of overlap. Exactly. If you look at the strongest of the chipsy pigs, there's a conditional team inside the pig, actually, minority. Yeah, and we believe there's probably a biological reason for that, especially when you're turning genes on and off, you don't want to cement your transcription factor on permanently. You probably don't want the strongest binding site available if you want to turn that gene off later, if it's an activator. It's a great question, though. Were the sites that you found in the 3-prime UTR, typically microRNA targets? Yeah, I don't remember the answer to that. I know we did look at it, but so maybe. Very nice presentation. I would like, having worked with Luciferase reporter assays for many years, and it appears like that the bioinformatics people are now beginning to adopt some of that. Maybe they've been doing it for a while. But I would actually just voice a word of caution about using those. They're very quickly done, and I understand the screening aspects of this. But the false positives and false negatives of putting together these heterologous promoter regions and then doing the transfections, as I'm sure you're well aware, is very problematic. And I think that the assays and the plasmas that you derive and all those sorts of things all impact the results. So it gives you a warm and fuzzy feeling if it looks like things are up or down. And we still use them just to gain that warm and fuzzy feeling. But in reality, I think it's something that you have to be extremely cautious about, particularly when you're putting active segments or potentially just random segments next to a heterologous promoter. So just a word of caution. Yeah, no, as you might imagine, we're pretty aware of this. And the purists hate those assays. It is an assay. That's how I treat it. It's just one more piece of information. There's no perfect experiment for anything we do in life. But it is an assay. We have a lot of controls that we're on.  We always do the scrambled sequence as well. But it is what it is. You know, the problem with the knockout assay, which is a pure assay, if you go in and crisper it out, is that there's a lot of redundancy. And so if you don't see something, you don't know whether it's there either. So, you know, it's a bit of information. I don't disagree with what you said. And we're cognizant of that. But yeah, OK. Yeah, so actually, I have a very simple question. You mentioned that you choose eight cancer types for their study. Are there any particular reasons for that choice? Yeah, they had the most sequence genomes at the time we were downloading the data. So that's why we chose them. Because we were hoping to get a strong enough signal, even within a cancer type. And I think we were marginal with that number. And they're roughly equal. So it's a 436 divided by 8 is roughly how many we had of each. So, yeah. Well, by the way, I thought of one more thing on Luciferase assay. You'll never see this kind of thing reported. But we actually did one where we were so sure it was a hypermutated region. It was actually coming out of a cancer study as well. Very, very heavily mutated in cancer. And it sits right upstream of an AWG. So we were so sure it was going to affect translation. And we did a zillion reporter assays and had zero effect. So it actually turned us off thinking it was having an effect on translation, like we thought. So rarely people use information that way. They use it to show what they're looking for. In this case, it was very disturbing to us. Because it think of us the result we were looking for. So that one's not published, as you might imagine. So, yeah. To your left. There's one more. Oh, yeah. Yeah, we're here. Yeah, hi. I'm just wondering whether the mutations within regulatory elements are mutually exclusive for mutations in the putative target gene? Yeah, we did look at that. And I don't know the answer to that. I'd have to go back and look. Yeah, it's a great question, though. Yeah, you would think they could be separate. Although they don't have to be. Because there are plenty of cases where genes are mutated and amplified as well in cancer, as you probably know. So it's conceivable that they could co-occur. That's a great question, though. Thank you.