From 4a465f02265c4c86bd224aea3663480fbb2b6fe1 Mon Sep 17 00:00:00 2001 From: Soothspider Date: Wed, 24 Apr 2024 14:11:17 -0700 Subject: [PATCH] AI Captions. April 22. Stanley Prusiner. --- ...rions 2002 STUDY HALL -- (22 Apr 2024).vtt | 5531 +++++++++++++++++ .../README.md | 5 + 2 files changed, 5536 insertions(+) create mode 100755 twitch/2128456102 (2024-04-24) - Stanley Prusiner and Prions 2002 STUDY HALL -- (22 Apr 2024)/2128456102 (2024-04-24) - Stanley Prusiner and Prions 2002 STUDY HALL -- (22 Apr 2024).vtt create mode 100644 twitch/2128456102 (2024-04-24) - Stanley Prusiner and Prions 2002 STUDY HALL -- (22 Apr 2024)/README.md diff --git a/twitch/2128456102 (2024-04-24) - Stanley Prusiner and Prions 2002 STUDY HALL -- (22 Apr 2024)/2128456102 (2024-04-24) - Stanley Prusiner and Prions 2002 STUDY HALL -- (22 Apr 2024).vtt b/twitch/2128456102 (2024-04-24) - Stanley Prusiner and Prions 2002 STUDY HALL -- (22 Apr 2024)/2128456102 (2024-04-24) - Stanley Prusiner and Prions 2002 STUDY HALL -- (22 Apr 2024).vtt new file mode 100755 index 0000000..7aa3989 --- /dev/null +++ b/twitch/2128456102 (2024-04-24) - Stanley Prusiner and Prions 2002 STUDY HALL -- (22 Apr 2024)/2128456102 (2024-04-24) - Stanley Prusiner and Prions 2002 STUDY HALL -- (22 Apr 2024).vtt @@ -0,0 +1,5531 @@ +WEBVTT + +00:00.000 --> 00:02.000 +You + +00:31.000 --> 00:42.400 +But you can tell if someone's lying, you know, you can sort of feel it in people + +00:43.680 --> 00:48.640 +And I have lied. I'm sure I'll lie again. I don't want to lie. You know, I don't think I'm a liar + +00:48.640 --> 00:54.040 +I try not to be a liar. I don't want to be a liar. I think it's like really important not to be a liar + +00:54.040 --> 01:04.340 +I'm not sure exactly who he is. His name is JJ Cooey, and I believe he's a consultant for CHD + +01:04.340 --> 01:12.040 +And I believe he has a PhD in some sort of scientific discipline from what I understand + +01:24.040 --> 01:31.040 +I don't want to be a liar. I don't want to be a liar. I don't want to be a liar. I don't want to be a liar. + +01:54.040 --> 02:01.040 +I don't want to be a liar. I don't want to be a liar. I don't want to be a liar. I don't want to be a liar. + +02:24.040 --> 02:38.040 +I don't want to be a liar. I don't want to be a liar. I don't want to be a liar. I don't want to be a liar. + +02:38.040 --> 02:46.040 +I don't want to be a liar. I don't want to be a liar. I don't want to be a liar. I don't want to be a liar. + +02:46.040 --> 02:52.040 +I don't want to be a liar. I don't want to be a liar. I don't want to be a liar. + +02:52.040 --> 03:08.040 +I don't want to be a liar. I don't want to be a liar. I don't want to be a liar. I don't want to be a liar. + +03:08.040 --> 03:15.040 +My friend Denver mixed date. + +03:15.040 --> 03:38.600 +I wanted to mix it up a little bit, maybe get some other slides out. + +03:38.600 --> 03:42.520 +We haven't used in a while, I know they're a little more reading. + +03:42.520 --> 03:44.720 +It's a little more reading. + +03:44.720 --> 03:51.520 +It's like we got ourselves some readers in the house. + +03:51.520 --> 04:01.160 +Jimmy Doar made a reference to the reader joke, very, very good Bill Hicks joint, very nice + +04:01.160 --> 04:02.160 +bit. + +04:02.160 --> 04:13.520 +Jimmy, Jimmy Doar showed a lot of courage the other night, I'm giving him another shout-out. + +04:13.520 --> 04:18.120 +It is amazing how many people think they can answer an argument by attributing bad motives + +04:18.120 --> 04:23.320 +to those who disagree with them using this kind of reasoning you can believe or not believe + +04:23.320 --> 04:30.320 +anything about anything without having to bother to deal with facts or logic. + +04:53.320 --> 05:06.920 +It's amazing how many people think they can believe or not believe it or not believe it + +05:06.920 --> 05:34.280 +or not believe it or not believe it or not believe it or not believe it or not believe + +05:34.280 --> 05:36.880 +it or not believe it or not believe it or not believe it or not believe it or not believe + +05:36.880 --> 05:43.880 +it or not believe it or not believe it or not believe it or not believe it or not believe + +05:43.880 --> 05:50.880 +it or not believe it or not believe it or not believe it or not believe it or not believe + +05:50.880 --> 05:57.880 +it or not believe it or not believe it or not believe it or not believe it or not believe + +05:57.880 --> 06:04.880 +it or not believe it or not believe it or not believe it or not believe it or not believe + +06:04.880 --> 06:09.880 +it or not believe it or not believe it or not believe it or not believe it or not + +06:09.880 --> 06:16.880 +believe it or not believe it or not believe it or not believe it or not believe it or not + +06:16.880 --> 06:22.880 +believe it or not believe it or not believe it or not believe it or not believe it + +06:22.880 --> 06:28.880 +or not believe it or not believe it Or not believe it or not believe it or not believe it + +06:28.880 --> 06:33.520 +Oh, that was a bad wolf. + +06:33.520 --> 06:34.520 +That went wrong. + +06:34.520 --> 06:35.520 +Sorry about that. + +06:35.520 --> 06:37.440 +We're just going to go through that one. + +06:37.440 --> 06:41.440 +That was an old slide that didn't need to go that fast, but that's okay. + +06:41.440 --> 06:43.560 +The stupor is definitely real. + +06:43.560 --> 06:45.440 +Brick soup is for lunch. + +06:45.440 --> 06:48.920 +No soup for you there, Mr. Mark. + +06:48.920 --> 06:54.760 +You had a really good show yesterday reading some kind of document that I actually never + +06:54.760 --> 06:58.800 +had any faith would actually exist, but you can still learn that biology thanks to + +06:58.800 --> 07:05.400 +Mark digging up some of these old descriptions of how early vaccine and let's let's call + +07:05.400 --> 07:06.400 +them what they were. + +07:06.400 --> 07:08.040 +They were, they were variolations. + +07:08.040 --> 07:11.320 +They were variolation preparation. + +07:11.320 --> 07:16.840 +But this is how they addressed smallpox back then and I think we are just up against + +07:16.840 --> 07:23.720 +one of the most demonic illusions that has ever been perpetrated on humankind. + +07:23.720 --> 07:27.400 +Very nicely done here in this graphic. + +07:27.400 --> 07:32.000 +Don't forget that they danced, don't forget that they kept people who had been married + +07:32.000 --> 07:35.120 +for 50 years apart in their last days. + +07:35.120 --> 07:40.480 +Don't forget that they, what they did to us, don't forget, you got to understand what + +07:40.480 --> 07:41.480 +this is about. + +07:41.480 --> 07:51.640 +This is a battle, you know, it's a battle between good and evil and yeah, we are trying + +07:51.640 --> 07:59.800 +to put forth some argument about biology, but even this cartoon might be largely a cartoon + +07:59.800 --> 08:02.720 +imposed upon us by them. + +08:02.720 --> 08:09.640 +Most of our understanding is dependent on their, on the very science that they have funded + +08:09.640 --> 08:13.240 +and put in front of us and this illusion is just, you know, it's something that we've + +08:13.240 --> 08:15.960 +really got to break through for our children. + +08:15.960 --> 08:17.760 +There are people who are learning biology. + +08:17.760 --> 08:21.480 +There are people who knew better and we've got to isolate those people who knew better + +08:21.480 --> 08:28.320 +and make sure that they, they don't rob us at the last little hope that is available. + +08:28.320 --> 08:29.640 +And that is in our history. + +08:29.640 --> 08:35.280 +It's our recent history and our more distant history and the people that are involved in + +08:35.280 --> 08:38.600 +it, what they did, what they said, what they wrote, what they thought. + +08:38.600 --> 08:44.280 +Those ideas are still the trap within which we find ourselves and the people that wove + +08:44.280 --> 08:50.080 +that trap and are involved in maintaining it are the people we need to identify. + +08:50.080 --> 08:54.680 +I don't think if you watch television or skillfully use social media, you're going + +08:54.680 --> 08:59.040 +to identify any of the people that matter. + +08:59.040 --> 09:03.840 +Even when paintings like this exist, you're still not quite seeing, you're seeing still + +09:03.840 --> 09:06.640 +what they want you to see. + +09:06.640 --> 09:11.160 +And only in a place where your tongues are completely free are we really free and in + +09:11.160 --> 09:18.080 +this day and age when a room like this could exist, where Twitter could be an illusion + +09:18.080 --> 09:24.280 +that is entirely created by rooms like this all around the world. + +09:24.280 --> 09:26.320 +We've got to keep swinging, ladies and gentlemen. + +09:26.320 --> 09:27.320 +We've got to keep swinging. + +09:27.320 --> 09:29.640 +Oh, that definitely was Steven Pinker. + +09:29.640 --> 09:31.600 +That was definitely Steven Pinker. + +09:31.600 --> 09:32.600 +Yes, sir. + +09:32.600 --> 09:34.600 +Rebob, it was. + +09:34.600 --> 09:35.600 +Yes, sir. + +09:35.600 --> 09:36.600 +Rebob, it was. + +09:36.600 --> 09:42.480 +Do you remember this song from way back when what's going on? + +09:42.480 --> 09:43.480 +Oh, sorry. + +09:43.480 --> 09:44.480 +Hold on. + +09:44.480 --> 09:45.480 +Let me pause that for a second. + +09:45.480 --> 09:50.920 +If you remember this song, I think you must remember this song. + +09:50.920 --> 09:53.120 +It was used in a while ago. + +09:53.120 --> 09:54.720 +It's a nice one. + +09:54.720 --> 09:57.920 +It's from the late 80s, I believe, in Australia. + +09:57.920 --> 10:04.360 +And it was for the Australian Broadcasting Company back in the 80s, I believe. + +10:04.360 --> 10:07.200 +And it's a really peppy news song. + +10:07.200 --> 10:10.120 +If you've been here for a while, you're here at the top of the wave. + +10:10.120 --> 10:13.720 +And if you're joining me for the first time at Gigo and Biological, then you might be + +10:13.720 --> 10:18.560 +a skilled TV watcher or what we call a skilled social media user. + +10:18.560 --> 10:20.680 +And you might not be staying focused on the biology. + +10:20.680 --> 10:23.760 +You might be taking their bait, and you might not love your neighbor. + +10:23.760 --> 10:30.200 +And we're going to suggest that those strategies need to be reversed. + +10:30.200 --> 10:34.600 +It could be back when ABC was probably more relevant. + +10:34.600 --> 10:36.520 +Thanks for spreading the word, ladies and gentlemen. + +10:36.520 --> 10:38.160 +Thanks to the supporters. + +10:38.160 --> 10:40.280 +Thanks to the people who are spreading these streams. + +10:40.280 --> 10:42.120 +This is Gigo and Biological. + +10:42.120 --> 10:54.040 +A high-resistance, low-noise information brief brought to you by a biologist. + +10:54.040 --> 10:57.840 +And now, since I haven't used that music in a while, this timing was a little off, and + +10:57.840 --> 11:01.120 +that thing paused, and who knows why that paused. + +11:01.120 --> 11:02.760 +I guess I had to tap that one again. + +11:02.760 --> 11:05.080 +I'm just sorry about that. + +11:05.080 --> 11:08.640 +There's some old slide combinations here for the beginning. + +11:08.640 --> 11:15.120 +I just thought I'd break out something new, old, new, old, new kind of thing. + +11:15.120 --> 11:17.200 +Welcome to the show, ladies and gentlemen. + +11:17.200 --> 11:19.920 +This is Gigo and Biological. + +11:19.920 --> 11:22.720 +The paradigm is shifting right now. + +11:22.720 --> 11:25.000 +There are a lot of moving parts. + +11:25.000 --> 11:31.520 +It's not something that can be easily understood, but definitely it is responding to our work. + +11:31.520 --> 11:36.080 +What we do is having a difference on how people try to push this forward. + +11:36.080 --> 11:40.760 +So let's move the ball forward by not participating in their illusion. + +11:40.760 --> 11:44.280 +That's really the strategy that I'm advocating for. + +11:44.280 --> 11:49.000 +This is, again, Gigo and Biological, a high-resistance, low-noise information brief brought to you + +11:49.000 --> 11:50.000 +by a biologist. + +11:50.000 --> 11:53.200 +I got my shirt on today. + +11:53.200 --> 11:55.160 +Welcome to the show. + +11:55.160 --> 11:56.920 +I look, oh, I know what it is. + +11:56.920 --> 12:01.280 +These little tiny front lights aren't on, so I look a little better. + +12:01.280 --> 12:03.280 +It might be a little better. + +12:03.280 --> 12:06.800 +Somehow I look a little, I don't know, greasy or something. + +12:06.800 --> 12:08.480 +Anyway, welcome to the show. + +12:08.480 --> 12:12.480 +This is Gigo and Biological coming to you live from the back of a garage in Pittsburgh, + +12:12.480 --> 12:14.480 +Pennsylvania. + +12:14.480 --> 12:15.680 +Thanks for joining me. + +12:15.680 --> 12:20.160 +Today, we have a little bit of work to do as far as the study hall goes. + +12:20.160 --> 12:23.680 +It's quite a long video, but I'm going to put it at 1.5 speed and we're going to take + +12:23.680 --> 12:28.400 +some notes and see what we get from Stanley Prusner's presentation from 2002. + +12:28.960 --> 12:34.160 +First, I would like to at least comment on the fact that Jessica Hockett, myself, and + +12:34.160 --> 12:47.440 +a friend of Jessica named John, scanning, hold on, that's the wrong male, why did that + +12:47.440 --> 12:57.920 +open up, see, I'm starting to get angry at a certain somebody, well, I don't know what + +12:57.920 --> 13:01.440 +his name is off the top of my head and I'm not trying to be a creep about it, that's + +13:01.440 --> 13:02.440 +the way it is. + +13:02.440 --> 13:08.040 +I apologize for that, I should have had that written down, it starts with a K and a very + +13:08.040 --> 13:09.040 +clever guy. + +13:09.040 --> 13:13.200 +The three of us presented, Jessica presented the longest in the beginning, about a half + +13:13.200 --> 13:17.720 +an hour, maybe a little bit longer than that and then John presented it and I presented + +13:17.720 --> 13:21.200 +very quickly about five or eight minutes each. + +13:21.200 --> 13:24.920 +I think the message was pretty clear. + +13:24.920 --> 13:28.960 +The message was a little bit of biology, but just a lot of what the hell happened in + +13:28.960 --> 13:34.440 +New York City to explain why we didn't investigate it yet, while there isn't a memorial yet, + +13:34.440 --> 13:38.320 +while the names haven't been released yet, why we haven't explained how this all happened + +13:38.320 --> 13:42.760 +in such a neat, uniform way and didn't happen in Chicago, even though they had a case a + +13:42.760 --> 13:47.880 +year or a month before that and so I felt like it was a decent thing, I thought it went + +13:47.880 --> 13:48.880 +pretty well. + +13:49.360 --> 13:57.040 +Unfortunately, we were briefing staffers and although I could only see two staffers on + +13:57.040 --> 14:05.880 +screen, those two staffers were, my best guess is millennials and one of them told us that + +14:05.880 --> 14:13.000 +she had a master's in public health and that any primary literature I could provide to + +14:13.000 --> 14:18.200 +back up any of the things that we said would be greatly appreciated and at that stage it + +14:18.200 --> 14:22.800 +really dawned on me one of the things that has driven me nuts since the beginning of + +14:22.800 --> 14:28.360 +the pandemic but I haven't really been able to express adequately in words but if anybody + +14:28.360 --> 14:34.400 +ever says to you to send some primary literature to them which could help them understand something + +14:34.400 --> 14:44.640 +unless it's in an extremely specific case of misrepresentation of data or a very specific + +14:44.680 --> 14:50.920 +case of a representation of a very specific situation, there is no combination of primary + +14:50.920 --> 14:55.760 +literature papers under the number of 5,000 that could be sent to somebody so that they + +14:55.760 --> 15:01.720 +could get an adequate idea of how distorted biomedical sciences has become as a result + +15:01.720 --> 15:08.760 +of the application of this p-value ritual over the last 20 or 30 years and with intention + +15:08.760 --> 15:09.760 +that is. + +15:09.760 --> 15:14.440 +It's not to say that science can't be productive and can't figure things out but what I'm suggesting + +15:14.440 --> 15:24.200 +to you is that the distortion, the intentional distortion of certain parts of the general + +15:24.200 --> 15:28.640 +scientific understanding and our education surrounding it has resulted in the place that + +15:28.640 --> 15:34.240 +we are, allowed us to be led to a place where we can no longer exercise informed consent + +15:34.240 --> 15:46.320 +because we simply don't know and trying to hit a home run in that scenario is basically + +15:46.320 --> 15:52.320 +where I went wrong, I started this, I'm going to show you the slides that I used and try + +15:52.320 --> 16:00.040 +to give the talk as I intended it but since I felt as though John abbreviated his presentation + +16:00.040 --> 16:05.320 +greatly in order to facilitate my presentation and I already felt like the staffers were + +16:05.320 --> 16:09.280 +looking at their watches and deciding whether they could stay any longer or not or whether + +16:09.280 --> 16:15.440 +they needed to listen a couple times, the kind of the head staffer or the one that was speaking + +16:15.440 --> 16:21.000 +the most into the microphone said that she's trying to get at our main idea when Jessica + +16:21.000 --> 16:27.640 +was talking and it's funny because Jessica started her presentation with the main idea + +16:27.640 --> 16:31.560 +which is that we want you to figure out what the hell happened in New York City because what + +16:31.560 --> 16:38.280 +happened there was not a natural event, it was like a bomb went off, it was a man-made disaster + +16:38.280 --> 16:43.640 +and we want you to investigate it and then she explained all the stuff and so it's weird the + +16:43.640 --> 16:48.240 +staffer was still kind of circling back to that stuff so by the time I got the mic John had + +16:48.240 --> 16:54.520 +already presented and John had some really nice slides that were actually tacked right on to + +16:54.520 --> 17:01.120 +the slides that Jessica had so they could flawlessly change to that and he had one or two slides + +17:01.120 --> 17:06.320 +in particular that I want to get him on my stream to talk about which are really slides + +17:06.320 --> 17:11.440 +that I've asked actually I believe I've asked Denny Rankor a couple times if he could make + +17:11.440 --> 17:24.800 +them which would be to let me see if that's the guy yeah that would be the guy if he could + +17:24.800 --> 17:32.880 +make a slide where you did the American data but you took the New York City event out and he had + +17:32.880 --> 17:41.520 +that slide and it's actually very striking how intuitively it felt like wow they're really + +17:41.520 --> 17:46.960 +using that curve to imply something else because of 24,000 people died in that week and he spread + +17:46.960 --> 17:52.640 +them out around America there would still be kind of a signal especially if you started that signal + +17:52.640 --> 17:59.280 +from zero right and you know in America 24,000 people have died of COVID so far but the vast majority + +17:59.280 --> 18:09.680 +are in New York City asterisk so anyway I started off already with a very good background of course + +18:09.680 --> 18:16.640 +so 45 minutes 40 minutes of Jessica's presentation eight minutes of John's statistical you know look + +18:16.640 --> 18:22.080 +if you take New York out it's crazy if you look at New York by itself it's so statistically off + +18:23.040 --> 18:29.120 +off I mean we're not talking about p-values here you know we're talking about z-score and + +18:31.120 --> 18:36.880 +we are we were way way way way and these nobody making these statistics ever thought that + +18:38.480 --> 18:43.040 +excuse me ever thought that something like this would be you know a z-score of 200 or something + +18:43.040 --> 18:49.040 +like that so my presentation couldn't really go too deep into the biology so I started with the + +18:49.120 --> 18:54.480 +idea that informed consent couldn't be exercised by anyone at the start of the pandemic because + +18:54.480 --> 18:59.360 +we were misled about the biology and about the whole experience of what was happening + +18:59.360 --> 19:03.920 +and that New York City as Jessica and John have pointed out was really central + +19:05.040 --> 19:08.560 +to establishing that there was even a pandemic happening and therefore + +19:10.320 --> 19:18.160 +I started with again reminding people of this and so we were consciously manipulated into believing + +19:18.160 --> 19:22.080 +that a crisis was occurring that it was ongoing that something was spreading in the + +19:22.080 --> 19:26.960 +background this kind of thing and this manipulation was on purpose to get us to accept that this + +19:26.960 --> 19:32.800 +crisis was occurring but the only numbers we have are the numbers that Jessica just showed you + +19:33.680 --> 19:40.240 +are almost assuredly fraudulent that John also backed up with statistics that seem to indicate + +19:40.320 --> 19:50.560 +that it's almost assuredly a man made event and I would argue that that man made event + +19:50.560 --> 19:58.320 +allowed them to imply the existence of a novel virus that that you and and Senator Ron's staff + +19:58.960 --> 20:05.520 +are very much in belief existed and the reason why you're in belief of it I would argue is because + +20:05.520 --> 20:11.040 +they fooled you into thinking that the only question was whether it was a bad cave virus or + +20:11.040 --> 20:16.960 +whether it was again a function lab league and this mystery that we've been solving over the last + +20:16.960 --> 20:22.960 +four years has bamboozled all of us into accepting that what we were told in New York City was the + +20:22.960 --> 20:31.040 +start of something very very terrible was actually a mass casualty event that was misconstrued as this + +20:31.360 --> 20:40.480 +and the beginning of this the beginning of this whole mystery and I would argue that one of the + +20:40.480 --> 20:46.720 +ways that they did it and I went from this to this was that they didn't they didn't genuinely + +20:46.720 --> 20:52.960 +describe what all cause mortality was they didn't actually tell you that you know the excess deaths + +20:52.960 --> 20:58.400 +where we're experiencing can come from all kinds of things especially if we told doctors with fear + +20:58.400 --> 21:04.080 +and uncertainty and doubt that there was a new thing that you had to be afraid of yourself + +21:05.440 --> 21:09.360 +and that if this new thing was there you should treat these people entirely different than you + +21:09.360 --> 21:12.480 +would have ever treated them had you not been told that this new thing was there + +21:14.640 --> 21:21.600 +and all of those new things that we were told to do started killing people and nobody acted because + +21:21.600 --> 21:26.240 +there was this illusion of consensus that it was either a novel virus or a lab leak that we were + +21:26.240 --> 21:33.920 +responding to and that these people were dying from and the tools that they use to identify it + +21:33.920 --> 21:39.600 +the financial incentives that they they use to amplify it all of these things are completely + +21:39.600 --> 21:47.200 +ignored by the people that were just in front of you in that senate committee almost none of this + +21:47.200 --> 21:52.640 +part of the narrative where hundreds of thousands of Americans were murdered at the beginning of the + +21:52.640 --> 22:00.560 +pandemic in 2020 and 2021 that's wholly ignored and completely attributed only to the novel virus + +22:00.560 --> 22:05.680 +and all of these people's narrative and this is very dangerous because of course as Jessica just + +22:05.680 --> 22:12.560 +showed you New York City is this bump right there that bump right there is New York City and I was + +22:12.560 --> 22:20.480 +actually already over here down here and so the mind this bump right here is New York City + +22:21.360 --> 22:28.800 +and this could just be people acting really really badly this little rise here there's a + +22:28.800 --> 22:34.000 +lot there's a rise there right you can see that right there's not a dip that dip is missing this + +22:34.000 --> 22:38.960 +little pattern just completely gets disrupted and now all cause mortality is permanently elevated + +22:38.960 --> 22:49.280 +by a combination of confusion and stupidity bad ideas and transfection and the initial casualty + +22:49.360 --> 22:55.760 +event is something that Jessica showed you is not a natural phenomenon does not belie the spread of + +22:55.760 --> 23:05.440 +a virus but instead suggests a staged event now the reason why I think it's important to understand + +23:05.440 --> 23:12.080 +this is because these people who were in front of you in the senate not even a month ago and this is + +23:12.080 --> 23:18.080 +exactly what I said in front of your boss excuse me who are also at an event in Romania in November + +23:18.080 --> 23:25.280 +of 2023 where they heard Denny Rancor the guy in the square speak about these effects who he + +23:25.280 --> 23:31.200 +has actually been speaking since 2020 about the fact that the only deaths that are excess are + +23:31.200 --> 23:37.040 +correlative with poverty level household income serious mental illness obesity and the excess + +23:37.040 --> 23:43.600 +mortality is a direct correlation with all these things across states irrespective of whether they're + +23:43.600 --> 23:51.760 +a blue state or a red state and so he suggests that there's no evidence of spread of a particular + +23:51.760 --> 23:56.560 +novel pathogen there's evidence of bad ideas in the form of protocols and how they treat or don't + +23:56.560 --> 24:03.360 +treat people and until the vaccines are released the all cause mortality doesn't go up significantly + +24:03.360 --> 24:08.800 +with any pattern that would indicate a respiratory virus but all of those people when they were in + +24:08.800 --> 24:15.760 +front of you last month none of them said that they all spoke very very generously about the 17 + +24:15.760 --> 24:22.480 +million people that Denny Rancor said might be or estimated might be having been killed by the + +24:22.480 --> 24:28.320 +transfections but they conveniently left out that his data also shows that there's no evidence of + +24:28.320 --> 24:37.120 +real novel spreading pathogen at risk additives spreading pathogen in 2020 and 2021 and this is + +24:37.120 --> 24:43.120 +very crucial because those same people want us to interpret that what happened in the last + +24:43.680 --> 24:50.240 +four years well there was an RNA that caused a pandemic but transfection worked pretty well + +24:50.240 --> 24:56.800 +although we rushed it and so they want justice for the RNA that was released and should never have + +24:56.800 --> 25:02.080 +been made and they want justice for the people that were injured because we rushed transfection + +25:02.080 --> 25:13.520 +before it was ready and this illusion is this this this solution is a lie the reality is is + +25:13.520 --> 25:20.000 +that transfection in healthy humans was criminally negligent all along and that every biologist with + +25:20.000 --> 25:26.000 +an academic bench in any accredited university in America should have known that transfection is a + +25:26.000 --> 25:32.800 +long-standing technology even using a product like lipofectamine is a long-standing technology + +25:32.800 --> 25:38.080 +that's been used on the academic bench for acute expression of proteins in order to manipulate + +25:38.080 --> 25:46.160 +systems and understand and test hypotheses about different metabolic pathways in any + +25:46.720 --> 25:52.000 +any biological system however transfection is never shown proven + +25:52.960 --> 25:58.000 +successful in the treatment of anything that's not a deadly disease like a very specialized + +25:58.000 --> 26:04.400 +cancer and the fact that they either didn't speak up because they didn't know or didn't speak up + +26:04.400 --> 26:11.040 +because they were unwilling they did not have the principles to dictate that they should for the + +26:11.040 --> 26:17.760 +betterment of mankind for the protection of our grandkids from this illusion they're all culpable + +26:17.840 --> 26:23.200 +they're criminally negligent I lost my job at the University of Pittsburgh School of Medicine + +26:23.200 --> 26:28.640 +because I spoke out about that specific thing and since then in the four years that I've been trying + +26:28.640 --> 26:37.840 +to understand what is and what isn't well understood RNA virology I've come to the conclusion that RNA + +26:37.840 --> 26:44.080 +cannot pandemic and they've always known it in fact it's a it's a central sort of + +26:44.400 --> 26:52.400 +it's a foundational part of the understanding we have or don't have about RNA viruses that they are + +26:52.400 --> 26:59.040 +almost intractable in culture because of the nature of how RNA viruses and especially RNA viruses like + +26:59.040 --> 27:07.040 +coronaviruses are are purported to replicate and those shortcomings are overcome by a technology + +27:07.680 --> 27:14.480 +called infectious clones and that's why with regard to ignoring all of these things these + +27:14.480 --> 27:20.400 +same people that were in front of you will never really enunciate a hypothesis which has anything + +27:20.400 --> 27:27.840 +to do with that that core technology of RNA virology which is infectious clones using DNA to make the + +27:27.840 --> 27:35.360 +RNA that is infectious and so our hypothesis my hypothesis is that the who declared a particular + +27:35.360 --> 27:40.400 +a dangerous pandemic of a particular sign and that's not obviously I said it better than that + +27:40.400 --> 27:47.120 +the the who declared a pandemic of a dangerous novel virus and it was a background signal that + +27:47.120 --> 27:55.040 +they that they misconstrued as spread they used an event in New York City to to create the illusion + +27:55.040 --> 28:01.520 +that that an impending crisis was coming and at the same time they already had a plan where they were + +28:01.520 --> 28:08.800 +ready to invert to change pneumonia and influenza what would have normally been classified as + +28:08.800 --> 28:15.440 +pneumonia and influenza to a new and novel respiratory pathogen that was a was a national security + +28:15.440 --> 28:22.320 +priority because this plan was in existence because not any kind of special technology or + +28:22.320 --> 28:29.760 +decree would be necessary for this to occur and because the vast majority if not the entirety of + +28:29.760 --> 28:38.080 +the deaths that occurred in 2020 can be explained by the confusion and fear and and + +28:38.800 --> 28:45.440 +incorrect behavior that resulted from the financial incentives from the testing and from the the + +28:45.440 --> 28:53.360 +messaging on on television there is no way that an RNA molecule can sustain a pandemic however there + +28:53.360 --> 28:59.040 +is a way for that sequence to be found in many places around the world and that again is an + +28:59.040 --> 29:04.240 +infectious clone and that could have been used if that was part of the national security + +29:04.240 --> 29:12.160 +operative to make operation to make absolutely sure that there was a seamless body of evidence + +29:14.720 --> 29:20.720 +that would support the idea of a circulating novel pathogen and would support the idea of + +29:20.720 --> 29:26.880 +rolling out this national security operation en masse would support the idea of co-opting + +29:26.880 --> 29:32.080 +anybody that ran to New York City and tried to save the day and found out that it wasn't what + +29:32.080 --> 29:40.160 +they said it was on TV and it would justify the use of several different groups of optoratives + +29:40.880 --> 29:46.960 +behind the scenes to make sure that this narrative never really got off the rails that nobody ever + +29:46.960 --> 29:52.880 +questioned a novel virus so that even when we got four years into the pandemic and people were in + +29:52.880 --> 29:58.640 +front of the senate and speaking about it that the novel virus would never be questioned that + +29:58.640 --> 30:03.680 +how many people died from it would never be questioned that we would already have moved on to whether + +30:03.680 --> 30:08.160 +or not the transfection worked or didn't work and how many people were hurt by it + +30:10.400 --> 30:15.760 +and this theater is actually an orchestrated attack on the united states from the from within + +30:16.320 --> 30:21.040 +orchestrated by people outside of the united states in cooperation with people inside of the + +30:21.120 --> 30:27.200 +u.s. government and that's what i think we need to understand most is that this is not an this + +30:27.200 --> 30:32.880 +is a national security issue for americans and what's happening in canada is separate from us + +30:32.880 --> 30:38.720 +and their emergency what happens in australia is their emergency and even the people that were in + +30:38.720 --> 30:46.880 +front of ron johnson in the senate are very very particular in how they express the emergency it's + +30:46.960 --> 30:50.880 +all of us it's everyone it's the who the globe has threatened no + +30:52.960 --> 30:59.040 +the only handles of power that we have the only way that we can act is through our government within + +30:59.040 --> 31:06.960 +our borders within our legislative system that's all we've got once we start trying to organize + +31:06.960 --> 31:13.520 +with australians or organize with canadians or or or then we're actually doing their work + +31:13.520 --> 31:16.240 +for them we're we're globalizing in a different way + +31:18.160 --> 31:23.680 +we need to decentralize that means that we don't need the help of people in other countries they + +31:23.680 --> 31:32.400 +have their own country to save and i think that's part of the reason why you should be very suspicious + +31:32.400 --> 31:36.560 +of having a senate hearing where half of the people are from different countries + +31:37.360 --> 31:41.280 +where half of the people have been on stage with the other half of the people in other countries + +31:41.280 --> 31:49.600 +multiple times why were they invited them none of this stuff was said to the staff i stopped a + +31:49.600 --> 31:54.240 +long time ago already but these were the things that i used i had this slide in there but i didn't + +31:54.240 --> 32:01.440 +really read it because i didn't i just went back after i what after this slide popped up i said i + +32:01.440 --> 32:06.720 +don't think i should use that slide should leave it here let you read this and then i'll stop for + +32:06.720 --> 32:12.640 +any questions and there weren't any questions of course but that was already an hour into the + +32:12.640 --> 32:19.280 +meeting and i think the out the meeting was supposed to be a half an hour and so they were ready to + +32:19.280 --> 32:26.880 +leave as soon as i quit so some things went really well other things weren't ideal i don't think my + +32:26.880 --> 32:34.080 +presentation was ideal i was a bit riled up i was going too fast i had already become frustrated + +32:34.080 --> 32:38.720 +with one of the guys one of the people on the staff because of an earlier comment + +32:41.680 --> 32:45.920 +i'll just i'll just comment on it now at one point in time when jeff was talking + +32:47.040 --> 32:54.880 +uh he was saying that um that there is evidence from previous winters and flu seasons that you know + +32:54.880 --> 33:01.280 +we've had nastier flu seasons in the past that we've not really made any fuss about and so what if + +33:01.280 --> 33:06.480 +these are actually coronaviruses in the background this background signal is always there and now + +33:06.480 --> 33:12.560 +they just misconstrued it as something else um of course i'm parsing his words incorrectly but + +33:12.560 --> 33:17.840 +she said um oh that's really interesting because of course there's lots of evidence or some people + +33:17.840 --> 33:23.840 +are saying that it was circulating in the background before 2020 and uh we didn't have any PCR testing + +33:23.840 --> 33:30.320 +and so that's what explains all this and they kind of went on but didn't go on because i think jeff + +33:30.400 --> 33:35.760 +was just trying to hurry up and he kind of acknowledged it um tried to sort of say + +33:35.760 --> 33:42.880 +something but it was the way that she trailed off her her comment that made it kind of just kind + +33:42.880 --> 33:48.320 +of stumbled through and ended up the presentation moving on and then i unmuted really quick and i + +33:48.320 --> 33:54.320 +was like whoa whoa whoa whoa i just want to go back one step because what um she just said we really + +33:54.320 --> 33:59.920 +need to point out a little better how ridiculously contradictory that statement would be how holding + +33:59.920 --> 34:05.120 +those two thoughts in your head shouldn't be possible if you listen to Jessica's presentation + +34:05.120 --> 34:13.040 +and you heard that New York City is this you know highly isolated event with you know orders of magnitude + +34:13.040 --> 34:19.440 +more dead bodies than in any other weeks that have ever come before after that and it's a very + +34:19.440 --> 34:26.240 +strange anomaly that occurs with at-home deaths and also nursing home deaths and all these things + +34:26.240 --> 34:38.800 +are at the same time um and so it's very strange um to believe that that represents a the spread + +34:38.800 --> 34:48.640 +of a particular dangerous pathogen those deaths but it was spreading before that and we didn't have + +34:48.640 --> 34:56.160 +tests and 20 000 people for a week weren't dying because we weren't locked down we weren't aware + +34:56.160 --> 35:02.400 +we were all just going to restaurants and drinking and and nobody was wearing masks and + +35:03.600 --> 35:08.720 +and so at some point you know like there was this oh yeah but it's a variant and all it's + +35:08.720 --> 35:14.320 +this and what about the lockdowns and you know so there was a lot of discussion but the end result + +35:14.320 --> 35:22.400 +always with Jessica's data is that but you can't explain this with that hand waving because it + +35:22.400 --> 35:28.640 +essentially death starts out very normal at 4 000 people per week and then goes + +35:31.120 --> 35:39.280 +or 4 000 people per April and then it goes and there's no biological explanation for that + +35:42.240 --> 35:47.120 +there would be EMTs that are traumatized from the sheer number of bodies they had to handle + +35:48.080 --> 35:53.280 +there would be nurses that are traumatized from the sheer number of people + +35:56.080 --> 36:00.240 +there would be memorials because it's like eight nine eleven events + +36:01.760 --> 36:08.320 +and so if if it was circulating before then but we didn't know how to track it and we were not + +36:08.320 --> 36:14.080 +doing anything to close down the borders then how in the hell didn't more bombs go off in other + +36:14.080 --> 36:22.960 +cities before this and she got very defensive and was like I don't think that's what I said I + +36:22.960 --> 36:26.880 +don't I don't I don't remember exactly what I said but I don't want to repeat it because I'm + +36:26.880 --> 36:35.280 +afraid you'll get it I'll get it wrong or I'll you'll take it out of con no you can't say that a + +36:36.480 --> 36:41.360 +deadly novel pathogen was circulating but it didn't kill anybody before we started testing for it + +36:41.360 --> 36:46.960 +or we didn't kill anybody before Tony Fauci made the 15 days to stop the spread announcement + +36:53.920 --> 36:59.760 +so that's where we were um you know that we're still right here right intramuscular injection of + +36:59.760 --> 37:04.000 +any combination of substances with the intent of augmenting the immune system is dumb + +37:04.000 --> 37:09.280 +transfection in healthy humans is criminally negligent in RNA cannot pandemic we're also + +37:09.280 --> 37:15.440 +beyond our own hypothesis here we're also trying um yeah I forgot I wanted to say I'm + +37:15.440 --> 37:19.600 +wearing Jimmy Dorr's shirt and I saw Jimmy Dorr the other day and I think although he doesn't have + +37:19.600 --> 37:23.280 +everything right he still thinks there was a novel virus it's probably gained a function and they + +37:23.280 --> 37:31.440 +lied about it I do think that his courage comes from a place without expertise right so all he + +37:31.440 --> 37:35.600 +knows for sure is that he was injured by the vaccine and people didn't take him seriously and that + +37:35.680 --> 37:40.000 +pissed him off and he realized that people were lying to him and now he's people he's realizing + +37:40.000 --> 37:45.360 +that a lot of people are liars and a lot of people can hold contradictory positions in their head + +37:45.360 --> 37:51.120 +and so he's as frustrated as I am that at some point in time it seemed like Robert F Kennedy + +37:51.120 --> 37:58.800 +Jr. was a really legit possibility as a normal everyday well-read smart guy with a lot of + +37:58.800 --> 38:04.320 +you know forward-looking ideas and tell you to hear him go on about climate change or Israel + +38:06.000 --> 38:10.560 +and so you know we're all we're all a bit frustrated with the world right now + +38:11.760 --> 38:18.800 +and so I think it's always good to in those times to to take a chance just kind of listen + +38:18.800 --> 38:26.640 +and take notes and and learn or maybe not learn from people who like it or not are having a very + +38:26.720 --> 38:35.760 +big impact on today going forward I have argued that we are probably should be expecting to see + +38:36.480 --> 38:44.560 +more crowds spelled yachob's disease more more protein misfolding disease is simply because we + +38:44.560 --> 38:54.080 +are starting to roll out mRNA transfection in old people all around the world and especially in + +38:54.080 --> 39:00.480 +the United States and in Europe and that rollout as it continues with the flu and RSV and the + +39:00.480 --> 39:07.120 +pneumonia and whatever else they decide to roll out on people maybe even the shingles shot + +39:08.160 --> 39:15.040 +you're going to see more and more of these what have already been well-seeded in in the mainstream + +39:15.040 --> 39:23.200 +narrative as as ramifications of a gain of function virus and potentially a designer spike protein + +39:24.800 --> 39:29.440 +but actually it's from transfection and they knew this already for a long time they knew + +39:30.080 --> 39:35.600 +as they moved forward that the vaccine schedule as it stood was probably going to eventually fall + +39:35.600 --> 39:41.680 +apart that it wasn't going to stand this the the scrutiny of of the citizenry forever as they + +39:41.680 --> 39:47.840 +ramped it up that's part of the reason why I still find it so amazing that the average young parent + +39:47.840 --> 39:52.400 +isn't even isn't even curious about how many shots and when they give them in other countries but + +39:52.400 --> 39:56.480 +if you do that math if you do that research you're going to find out that the American + +39:56.480 --> 40:04.960 +vaccine schedule is just an exclusively early and dense and so even if some of these purported + +40:04.960 --> 40:10.400 +intramuscular augmentations of the immune system were somewhat effective the way that they use + +40:10.400 --> 40:16.880 +them and deploy them in such early lifetime is absolutely absurd and also contradictory to what + +40:16.960 --> 40:22.560 +we understand about everything to do with neuronal development and and immunological development + +40:25.280 --> 40:32.240 +in fact it's funny because you can get a you can see a nice talk by Michael Warby from before + +40:32.240 --> 40:37.440 +the pandemic when he's talking about vaccinating people for flu and given the fact that they have + +40:37.440 --> 40:42.480 +lifelong immunity to the flu that they're exposed to as a kid we probably have to start vaccinating + +40:42.480 --> 40:48.880 +people before birth if we really wanted to get ahead of this but of course that's actually the + +40:48.880 --> 40:54.000 +plan right that's why they want you to think that vaccinating pregnant women's no big deal + +40:54.000 --> 41:00.480 +yes you shouldn't eat sushi or uh unpasteurized cheese but vaccination is fine intramuscular + +41:00.480 --> 41:05.440 +injection of any combination of substances is fine for a for a pregnant woman + +41:05.760 --> 41:14.000 +let's look at Stanley Prusiner and Prions because again I think that this narrative has been + +41:14.000 --> 41:18.800 +seeded was given a Nobel Prize for goodness sakes I mean let's just talk intellectually a + +41:18.800 --> 41:25.840 +little bit about a Nobel Prize that I was very very very very very very very very very very + +41:25.840 --> 41:34.720 +cursorily connected to the Nobel Prize in 2014 was awarded for place cells and grid cells and + +41:34.720 --> 41:42.480 +spatial coding in in the brain of rodents to John O'Keefe and Edward and Maybert Moser + +41:43.680 --> 41:50.720 +I was lucky enough to have worked with and studied with and trained with Edward and Maybert Moser and + +41:50.880 --> 42:01.040 +uh probably more importantly for me Edward uh Menowitter in in Norway um for almost five years + +42:01.040 --> 42:07.040 +it's where we had our boys um it was some of the most fun that I've ever had um as a young adult + +42:07.040 --> 42:13.120 +in my life um Norway was a great place to live um and I met a lot of great people there um I + +42:13.200 --> 42:21.120 +actually got I actually got an email from the universe sorry from the Society for Neuroscience + +42:21.120 --> 42:28.400 +inviting me to register for the meeting and uh at the bottom was the you know the SFN + +42:29.360 --> 42:36.000 +meeting committee chair signature is Laura Colgan somebody who I worked with and met in the Moser + +42:36.000 --> 42:42.880 +lab um in the first couple years I was there a really nice um smart girl I shouldn't say girl + +42:42.880 --> 42:50.240 +smart woman um who's now I still believe working in New Orleans but I'm I apologize Laura for not + +42:50.240 --> 42:59.360 +knowing where you are now but being the chairperson for uh for the chairman for the Society for Neuroscience + +42:59.360 --> 43:06.080 +meeting a meeting of over 35 000 um neuroscientists every year um it's a pretty big step um I guess + +43:06.080 --> 43:12.560 +that's on the way to being president of Society for Neuroscience um she was uh always gonna be + +43:13.200 --> 43:17.280 +you know head of her department at some point so I'm really happy I was really happy to get that + +43:17.280 --> 43:24.560 +letter and just um so anyway what was I saying Norway is a place where Edward and Maybers + +43:24.560 --> 43:30.560 +Maybers Moser uh work and they in troll time and they actually won the Nobel Prize in 2014 + +43:31.120 --> 43:38.080 +um it was a year so after I had left I moved to Norway the Netherlands in Rotterdam to try and + +43:38.080 --> 43:43.120 +get tenure there and try to find a permanent job there we had intended to raise our kids um + +43:43.840 --> 43:52.080 +in in Holland but that didn't work out um and so in 2016 we moved to Pittsburgh um but in 2014 + +43:52.080 --> 43:56.080 +they won that Nobel Prize and so you would have thought that at least that should help a little + +43:56.160 --> 44:01.520 +bit with my grant applications and with my work but I was doing something in Rotterdam that was so + +44:02.160 --> 44:08.400 +methodologically close to impossible that I was so obsessed with um that I basically dropped the + +44:08.400 --> 44:15.680 +ball on getting funding and you know getting any big publications myself and so I found myself + +44:15.680 --> 44:20.320 +where I was at that time because of you know choices that I had made in mountains that I decided + +44:20.320 --> 44:26.480 +to climb so I'm not blaming anybody for those years um they were very taxing on our what on + +44:26.480 --> 44:34.080 +our marriage because I was so obsessed with with making things work at work um the reason why I'm + +44:34.080 --> 44:40.080 +talking about this is because um the year that they won the Nobel Prize they also won the Nobel Prize + +44:40.080 --> 44:46.320 +with John O'Keefe who whose first discovery of play cells was actually in the early 70s you might + +44:46.400 --> 44:54.160 +say 1971 or 1972 is when this happened and then his students actually um two students from Norway + +44:54.160 --> 45:00.640 +who came to his lab to learn the physical technique of making the electrodes that they recorded these + +45:00.640 --> 45:07.440 +play cells in the hippocampus from as the animal freely navigated in a small box or maze and uh + +45:07.440 --> 45:11.520 +they were they were keen on learning those and doing experiments with him and then they recorded + +45:11.520 --> 45:18.480 +from uh different uh upstream brain region anatomically um giving a lot of input to the + +45:18.480 --> 45:24.960 +hippocampus as suggested by meno litter and there is where they found their discovery the grid cells + +45:24.960 --> 45:31.840 +the point is is that that Nobel Prize was awarded to a total of three people essentially a teacher + +45:31.840 --> 45:40.160 +and his students who had changed the field both several different methodological ways in the + +45:40.160 --> 45:45.840 +sense of enabling the recording of these these neurons and and and show and developing the + +45:45.840 --> 45:52.400 +anatomical techniques to follow where you recorded from and and use them for leisure all kinds of + +45:52.400 --> 45:57.840 +things that essentially change the way that this kind of neuroscience was done these experiments + +45:57.840 --> 46:02.080 +were done these these methodology spread all around the world and people were putting these + +46:02.080 --> 46:08.320 +electrodes in all different heads and recording you know uh play cells and then trying to study + +46:08.400 --> 46:14.880 +how play cells encoded memory and and and how they express themselves in different behavioral + +46:14.880 --> 46:21.920 +paradigms though I mean they actually changed or even created a field that before them didn't + +46:21.920 --> 46:28.400 +really exist the hippocampus was originally really exciting because it was this one place where + +46:28.400 --> 46:33.600 +bliss and lomo were able to create this long-term potentiation that everybody thought was memory + +46:33.600 --> 46:39.120 +if you could change the electrical signal from a weak signal to a strong signal that was sort of + +46:39.120 --> 46:44.720 +like this is like the first sort of in vitro preparation of memory but the hippocampus itself + +46:44.720 --> 46:51.360 +and its functional sort of a model of its functional understanding really comes from this + +46:52.560 --> 47:01.280 +decades of work that started with the play cells in in in London in John O'Keefe's lab in the 70s + +47:02.240 --> 47:10.000 +and a and a and a Nobel Prize in medicine wasn't awarded until 2014 Stanley Prouzner got a Nobel + +47:10.000 --> 47:17.840 +Prize for for prions and the idea of prions before everybody in the field even agreed that prions + +47:17.840 --> 47:24.640 +were a thing I believe seven years after he coined the term + +47:24.640 --> 47:38.880 +it should as as Peter Thiel says it should make you suspicious because the potential for people to + +47:38.880 --> 47:48.480 +lie and exaggerate has been greatly increased and so your default should be yeah your default + +47:48.480 --> 47:54.080 +assumption should be that there is lie and exaggeration here so let's take a look at this and see what + +47:54.160 --> 47:58.320 +we can get out of it shall we + +47:58.320 --> 48:00.320 +you + +48:21.680 --> 48:23.360 +people who have dementia are demented + +48:24.000 --> 48:30.400 +and so I think it's okay to talk about demented people what I'm going to tell you about is a + +48:30.400 --> 48:35.440 +very special journey that happened here at UCSF that begins in 1972 and really is continuing up + +48:35.440 --> 48:42.240 +to the present so we'll start with this slide and I'm assured the lights will go down so I'm going + +48:42.240 --> 48:46.800 +to tell you about a saga that represents the triumph of scientific investigation over prejudice + +48:46.800 --> 48:51.760 +it's really a journey from heresy to orthodoxy it's about prions really a new principle of infection + +48:51.760 --> 48:58.320 +and disease now the diseases we're going to talk about our crew of New Guinea Natives + +48:58.320 --> 49:02.720 +Croitesville the Ocob disease called CJD. Curseman stories are shanker disease and fatal insomnia + +49:02.720 --> 49:07.520 +then scraping of sheep mad cow disease or BSE of cattle and chronic wasting disease of mule deer + +49:07.520 --> 49:13.120 +and elf now it was 1972 as I mentioned to you a moment ago that I became interested in these + +49:13.120 --> 49:17.040 +diseases and I had a patient from Marin County a 60 year old woman who was dying of Croitesville + +49:17.040 --> 49:21.440 +the Ocob disease I was a resident in neurology here and as I began to learn about the animal + +49:21.440 --> 49:25.600 +disorders scrappy and about the foray people in New Guinea who suffered of crew I thought + +49:25.600 --> 49:30.560 +this could be really a fascinating area to pursue and really the fascination came from the chemical + +49:30.560 --> 49:34.400 +point of view because there were a few tidbits of chemistry that suggested that the particles + +49:34.400 --> 49:38.480 +causing these diseases must be much different than anything that was known this is a slide + +49:38.480 --> 49:43.040 +by my colleague Steve D. Arman showing you what the brain looks like of a patient who typically dies + +49:43.040 --> 49:47.040 +of Croitesville the Ocob disease it's a rare disease about one person in a million or one + +49:47.120 --> 49:51.360 +every 10,000 deaths this is a much less common form with these huge vacuoles that you see + +49:53.360 --> 49:58.080 +you have to excuse my voice uh that's this is a virus causing it not prions + +50:00.240 --> 50:02.160 +now for many years scientists thought that scrappy + +50:06.480 --> 50:13.040 +sorry I got brought brought lunch so I'm eating in the background so first of all let's go back + +50:13.040 --> 50:19.440 +a little bit here to see that um he doesn't really tell you what this is is his brain or what + +50:20.480 --> 50:26.560 +um I presume it's brain but it's interesting how quickly you went through it not to tell + +50:26.560 --> 50:34.800 +you what to look for what to see here there's no not normal form I mean it's again are we + +50:34.800 --> 50:41.440 +are we here to objectively inform people or are you implanting an idea that you they must accept + +50:42.320 --> 50:48.720 +and and I see already a problem here and yes it is on 1.5 speed if you want me to slow it down I will + +50:48.720 --> 50:53.280 +but I think you'll get used to it really quick yes this is a much less common form with these + +50:53.280 --> 51:00.800 +huge vacuoles that you see you have to excuse my voice uh that's this is a virus causing it not + +51:00.800 --> 51:07.920 +prions now for many years scientists thought that scrappy was caused by a virus because the + +51:08.000 --> 51:12.080 +disease is transmissible the agent is small there's a rise in scrappy agent tighter that + +51:12.080 --> 51:15.360 +precedes disease so that's the concentration or amount of scrappy agent and there are different + +51:15.360 --> 51:19.120 +strains of scrappy agent that produce different patterns of disease so there was a lot known + +51:19.120 --> 51:25.360 +when we got into this and what I thought in 1972 was that we really needed to do was to develop + +51:25.360 --> 51:29.280 +a method to isolate the infectious particles so the infectious particles in this cartoon are + +51:29.280 --> 51:33.760 +represented by these little red squares and everything else is junk and we needed to separate + +51:33.760 --> 51:39.040 +these so we could under this is gonna be hard to do while I'm uh I'm gonna try and eat lunch + +51:39.040 --> 51:47.440 +here and get done so this list of assumptions was gone through way too fast for this to be so + +51:47.440 --> 51:55.280 +scrappy agent is small and filterable that sounds like you know 1920s virology a rise in scrappy + +51:55.280 --> 52:01.680 +agent tighter precedes the disease yeah I wonder what that's really based on I wonder what + +52:01.840 --> 52:07.360 +does that mean that the more you inject in the brain of an animal the quicker it gets sick + +52:08.080 --> 52:14.640 +you see these assumptions are already sketchy to me the strains of scrappy agent produce + +52:14.640 --> 52:20.560 +different patterns of disease I'm willing to bet also is based on a very small number of observations + +52:21.280 --> 52:26.080 +meant to be used to generalize to this extent and to go through this slide this quickly + +52:26.320 --> 52:35.120 +um well anyway you'll see and we got into this now what I thought in 1972 was that we really + +52:35.120 --> 52:39.360 +needed to do was to develop a method to isolate the infectious particles so the infectious + +52:39.360 --> 52:43.120 +particles in this cartoon are represented by these little red squares and everything else is junk + +52:43.760 --> 52:48.240 +and we needed to separate these so we could understand what they were made of well the problem + +52:48.240 --> 52:54.800 +became that the particles did not behave very well so instead of having a very steep curve here + +52:54.800 --> 52:59.600 +the curve was very uh very gradual I won't go into the details of some of the more difficult + +52:59.600 --> 53:04.320 +pieces of science but this shows it a little more easily so the ideal behavior of a particle + +53:04.320 --> 53:07.680 +whether it's a virus or a protein or a piece of the classic acid whatever you might want to isolate + +53:08.880 --> 53:13.520 +it would when we fractionate it had a bit one piece of whatever you might want to get a bite + +53:13.520 --> 53:17.120 +instead of having a very steep curve that you could understand what they were made of + +53:18.080 --> 53:20.000 +and so now what we're looking here + +53:25.040 --> 53:31.440 +log scraping infectivity and so this is a cartoon that's sort of + +53:36.160 --> 53:42.400 +supposed to show you that somehow or another they've tried different ways to purify + +53:43.200 --> 53:50.640 +the scraping agent and it doesn't behave the way an agent would be expected to behave + +53:52.400 --> 53:58.000 +and that for them should be a problem it should indicate that maybe there isn't one + +53:58.000 --> 54:05.600 +agent but instead they try to stick very hard in fact he will stick to the assumption till the + +54:05.680 --> 54:12.160 +end of the talk that there is a single causative agent that they just have to develop better + +54:12.160 --> 54:18.240 +techniques in order to study well the problem became that the particles did not behave very + +54:18.240 --> 54:23.680 +well so instead of having a very steep curve here the curve was very uh very gradual I won't go + +54:23.680 --> 54:27.920 +into the details of some of the more difficult pieces of science but this shows it a little more + +54:27.920 --> 54:32.240 +easily so the ideal behavior of a particle whether it's a virus or a protein or a piece of the + +54:32.240 --> 54:38.080 +plaque acid whatever you might want to isolate it would when we fractionate it have a bit one peak + +54:38.080 --> 54:42.000 +it looks like that but instead what we found was that the scrapey agent was spread throughout + +54:42.000 --> 54:43.280 +the entire uh gradient + +54:49.360 --> 54:52.560 +so again that's a problem right because it's not one thing + +54:53.760 --> 54:58.160 +now the argument I believe they're going to use for a little while at least is that the + +54:58.160 --> 55:01.760 +aggregation of a repetitive protein might produce this kind of smear + +55:03.840 --> 55:07.760 +however I think that that's you're going to very quickly see that in the end they're going to get + +55:07.760 --> 55:15.440 +back to it being a single thing and uh so just keep this in mind he's just telling us a story + +55:15.440 --> 55:18.960 +right now and so we're just listening to his story what we found was that the scrapey agent + +55:18.960 --> 55:21.280 +was spread throughout the entire uh gradient + +55:21.760 --> 55:27.840 +now the problem was compounded by the fact that when I started we needed to use 60 mice + +55:28.560 --> 55:31.520 +and we needed one year to carry out what was called an endpoint titration + +55:32.240 --> 55:36.960 +and all the red ones are positives and all the white ones are negative so what we would use + +55:36.960 --> 55:42.640 +are 60 mice as I said uh for a single sample so at each tenfold illusion we would have six mice + +55:42.640 --> 55:48.640 +and this assay took a year now we were able in the late 1970s to develop a new assay and this was + +55:48.720 --> 55:52.400 +based on the work of Richard Martian Wisconsin and Richard Kimberlin who was working with Richard + +55:52.400 --> 55:56.080 +Marsh but he was from England and we turned to the hamsters we followed their lead and we were + +55:56.080 --> 56:01.200 +able to reduce the number of animals from 60 down to 4 and the time from about 130 days for high + +56:01.200 --> 56:06.240 +titered samples to 70 days and we did this by constructing a standardized curve and then just + +56:06.240 --> 56:10.160 +simply reading the dose off of the curve and since we were interested in making more and more + +56:10.160 --> 56:13.920 +enriched preparations this worked to our advantage because we didn't have to wait to get this endpoint + +56:13.920 --> 56:17.520 +where we had the negatives and the positives after a whole year we could just simply read off the + +56:17.520 --> 56:22.960 +titer or the concentration from the standard curve this was a huge step it allowed us to do + +56:22.960 --> 56:26.240 +more experiments in a period of three years and have been done in the entire field over previous + +56:26.240 --> 56:31.280 +century now as I said a few moments ago isolating the scraping agent was a nightmare because it + +56:31.280 --> 56:39.040 +behaved I will admit that I don't really get that I'm gonna play that one more time and see if I + +56:39.040 --> 56:47.040 +can catch it better a more rapid and economical assay for the scraping agent it seems like to me + +56:47.040 --> 56:51.360 +what he's saying is if you want to find the scraping agent you have to inoculate 60 animals + +56:51.360 --> 56:55.600 +and then some of the animals will develop it and then from those animals you can isolate the scraping + +56:55.600 --> 57:08.800 +agent and with hamsters it's faster or something I don't let's listen the number of animals from + +57:08.800 --> 57:14.560 +but it was from new assay assay took a year now we were able in the late 1970s to develop a new + +57:14.800 --> 57:18.240 +assay and this was based on the work of Richard Martian Wisconsin and Richard Kimberlin who was + +57:18.240 --> 57:21.840 +working with Richard Marsh but who was from England and we turned to the hamsters we followed their + +57:21.840 --> 57:26.080 +lead and we were able to reduce the number of animals from 60 down to four and the time from + +57:26.080 --> 57:32.000 +about 130 days for high-tired samples to 70 days and we did this by constructing a standardized + +57:32.000 --> 57:36.160 +curve and then just simply reading the dose off of the curve and since we were interested in making + +57:36.160 --> 57:39.520 +more and more enriched preparations this worked to our advantage because we didn't have to wait + +57:39.520 --> 57:43.120 +to get this endpoint where we had the negatives and the positives after a whole year we could + +57:43.120 --> 57:48.320 +simply read off the piter or the concentration from the standard curve this was a huge step it + +57:48.320 --> 57:52.160 +allowed us to do more experiments in a period of three years and have been done in the entire field + +57:52.160 --> 57:56.560 +over a previous century now as I said to you a moment ago isolating the scraping agent was a + +57:56.560 --> 58:01.200 +nightmare because it behaved very oddly and also because the method of measurement when we entered + +58:01.200 --> 58:09.360 +the field was extremely difficult one year and 60 animals so having now moved away from mice + +58:09.440 --> 58:13.200 +into hamsters able to accelerate our studies by nearly 100 fold so what we could do in one year + +58:13.200 --> 58:16.480 +would have taken us a century to do and there are few investigators that live a century much + +58:16.480 --> 58:22.400 +less have a productive lifetime of the century so now what we could do is develop what's called + +58:22.400 --> 58:26.720 +a purification scheme so we could use detergents and enzymes and back to the centrifuge and these + +58:26.720 --> 58:30.880 +gradients and now we could follow up the lead that was present in the literature from a woman + +58:30.880 --> 58:35.440 +named Tig Valper who was a radiobiologist working in England and she had argued that because the + +58:35.440 --> 58:39.760 +scraping agent was so resistant to UV and ionizing radiation that it did not contain RNA or DNA + +58:39.760 --> 58:44.720 +nucleic acid and what we found was that all the procedures we use that alter nucleic acids did + +58:44.720 --> 58:48.880 +not diminish the infectivity and the procedures that modified proteins led to an activation of + +58:48.880 --> 58:53.040 +the scraping agent but we could only see this after we removed more than 99 percent of the junk + +58:53.040 --> 58:58.000 +the unwanted molecules and having done that and that's me by the way with brown hair at the time + +58:58.640 --> 59:06.160 +I thought that by 1982 actually the spring of 81 that we had enough information to say that + +59:06.160 --> 59:10.400 +this particle was not a virus or a tiny nucleic acid called a viroid discovered by Ted Deener in + +59:10.400 --> 59:14.880 +the late 1960s and that we needed to give it a new name and so I had the audacity to call these + +59:14.880 --> 59:19.120 +particles prions that's the long arm of the scientific community pushing on me and people + +59:19.120 --> 59:23.840 +say well what do those people look like and here they are at the table that's my best slide you can + +59:23.920 --> 59:28.880 +go home now and it became very very clear that prions eventually became very clear + +59:28.880 --> 59:32.640 +our infectious proteins and I'm going to take you through a lot more evidence that builds the case + +59:32.640 --> 59:39.520 +that prions our infectious proteins and then so basically I guess he won the Nobel prize for that + +59:39.520 --> 59:48.960 +hamster work where the incubation time bioassay right is where they take scraping agent + +59:48.960 --> 59:57.920 +maybe it's prepared no better than what Marx describes on his stream for the other day + +01:00:00.000 --> 01:00:04.640 +is then injected directly into the brains of mice and it used to take a year + +01:00:06.480 --> 01:00:12.800 +but eventually they were able to accelerate that in hamsters I don't know if they altered the + +01:00:12.800 --> 01:00:20.480 +scraping or if they had to change the PRP protein in a certain way to get that to happen we will + +01:00:20.480 --> 01:00:27.680 +look at that in the coming journal clubs but in this summary and I apologize for eating lunch + +01:00:27.680 --> 01:00:36.080 +I'm so hungry the problem with this is though is that I think that we went so quickly to assuming + +01:00:36.080 --> 01:00:40.080 +that this is the real deal and that more importantly that this recapitulates it + +01:00:40.880 --> 01:00:52.080 +right that's the problem here is the the animal model of a gain of function virus is one where + +01:00:52.080 --> 01:00:59.680 +we go into a bat and we swab it we PCR test it we amplify all of the RNA that we see there + +01:01:00.800 --> 01:01:05.760 +whatever genes are missing we replace by something that we find in a catalog and we rebuild + +01:01:06.480 --> 01:01:10.320 +the coronavirus that was in that bat based on the fact that well if you assume the spike + +01:01:10.320 --> 01:01:15.840 +protein is a new spike protein then I guess that's a new virus we can build it we can't grow it + +01:01:16.800 --> 01:01:20.480 +we can't grow it and make a lot of it we can't culture it and make a lot of it + +01:01:22.480 --> 01:01:29.440 +but we can use computer algorithms and and nonspecific PCR primers to amplify signals in the + +01:01:29.440 --> 01:01:36.000 +background that may have already been there that may have nothing to do with the health of the + +01:01:36.000 --> 01:01:42.640 +animal that may be a a bacteriophage signal for all we know because we do nothing to differentiate + +01:01:42.640 --> 01:01:47.040 +between them this is very similar to that if we're going to have an animal model + +01:01:49.120 --> 01:01:55.040 +that has nothing to do with it if we start with humanized mice and infectious clones + +01:01:55.680 --> 01:02:01.600 +that are purported to be the same as the stuff they found in the bat and now we have this model + +01:02:01.600 --> 01:02:09.680 +of coronavirus disease where if we squirt this RNA or or the the cell culture products that are + +01:02:09.680 --> 01:02:16.080 +produced after we transfect a cell culture with this RNA if we put that material into a mouse + +01:02:16.080 --> 01:02:24.480 +that we call that a model of ARDS or SARS then then then we have a whole industry that can evolve + +01:02:24.480 --> 01:02:29.200 +from that that has nothing to do with whatever the original natural phenomenon was + +01:02:31.040 --> 01:02:37.840 +and so it's it's at least worth extreme inspection to find out whether this animal model that's just + +01:02:37.840 --> 01:02:43.920 +been sort of just presented to us as accepted and and proven and well understood + +01:02:45.200 --> 01:02:52.320 +is actually an adequate bridge model to come from what happens in scrapie or what happens in + +01:02:52.400 --> 01:03:00.080 +crowds filled yachab or what happens in kuru and notice that this entire lecture and all the + +01:03:00.080 --> 01:03:08.720 +lectures about protein misfolding have absolutely never ever mentioned the immune system responding + +01:03:08.720 --> 01:03:17.040 +to misfolded proteins do you see how significant it is that if you study protein misfolding you + +01:03:17.040 --> 01:03:22.160 +don't worry about the immune system at all the immune system is not involved this is a completely + +01:03:22.160 --> 01:03:26.880 +different thing that has to do with aggregation and cell death and you know whatever + +01:03:34.880 --> 01:03:40.480 +so now he's going to purify the protein and use using sucrose gradient which he showed earlier + +01:03:40.480 --> 01:03:46.160 +didn't work and so now we're at the development of the prion concept procedures altering nucleic + +01:03:46.160 --> 01:03:52.640 +acids don't diminish the infectivity so chemicals that are that are thought or assumed to get rid + +01:03:52.640 --> 01:04:00.720 +of all nucleic act nucleic acid still the infectivity of these preparations is apparently still preserved + +01:04:00.720 --> 01:04:06.560 +when you do what when you inject them into the brain of an animal or when you use some other + +01:04:06.560 --> 01:04:17.280 +probably not very adequate experimental correlate for what you what you would think happens in + +01:04:17.280 --> 01:04:21.760 +these natural forms i'm not suggesting that the natural form of kuru doesn't exist + +01:04:22.640 --> 01:04:28.240 +crowds felt yachab as a result of using an adenavirus encoding a viral spike protein fusion protein + +01:04:28.880 --> 01:04:34.000 +could potentially have caused crowds felt crowds felt yachab disease in 28 people like + +01:04:34.080 --> 01:04:41.440 +luke montanier's paper said i'm suggesting to you that genetic technologies like transformation + +01:04:41.440 --> 01:04:49.280 +and transfection we're already known a long time ago to interfere with and maybe even induce + +01:04:49.280 --> 01:04:58.640 +maladaptive states in healthy animals and so what this means is is that we need to be very careful + +01:04:58.640 --> 01:05:05.440 +about how many of these very recently seeded biological phenomenon biological ideas were + +01:05:05.440 --> 01:05:13.440 +actually seeded by people who have intention of confusing us and about this sounds very paranoid + +01:05:13.440 --> 01:05:21.200 +but this is the same year that sars came on the world 2002 when he's given this talk is the year + +01:05:21.280 --> 01:05:27.680 +that sars is going to be released or the year after i guess it's 2003 really when it happened + +01:05:31.120 --> 01:05:36.320 +so you can almost imagine in fact a scenario over the last couple decades where this + +01:05:37.040 --> 01:05:45.280 +this potential bio hazards from mother nature these mythologies were already these were + +01:05:45.280 --> 01:05:54.560 +mythologies were seeded and while we're being told that these phenomenon are natural we we + +01:05:54.560 --> 01:06:01.680 +have no way of knowing we have no way of knowing we do have pretty good reason to believe that + +01:06:01.680 --> 01:06:06.320 +they've been messing around with all sorts of biotechnologies for the last 30 years in attempt to + +01:06:06.960 --> 01:06:13.760 +figure stuff out and so there's no question that one of the things that could have resulted from + +01:06:13.920 --> 01:06:21.120 +are these kinds of diseases there's also no question in my mind that that kuru and eating + +01:06:21.120 --> 01:06:25.600 +your dead relatives might be a bad idea that would involve your immune system going a little + +01:06:25.600 --> 01:06:29.120 +going a little haywire can imagine that i can + +01:06:33.600 --> 01:06:38.720 +but i do think that an infectious protein is much more interesting from a bioweapon perspective + +01:06:38.720 --> 01:06:43.920 +than it is from a biological perspective not diminishing the infectivity and the procedures + +01:06:43.920 --> 01:06:47.520 +that modified proteins led to an activation of the scrapey agent but we could only see this + +01:06:47.520 --> 01:06:52.320 +after we removed more than 99 percent of the junk the unwanted molecules and having done that + +01:06:53.200 --> 01:07:00.480 +and that's me by the way with brown hair at the time i thought that by 1982 actually the spring + +01:07:00.480 --> 01:07:05.280 +of 81 that we had enough information to say that this particle was not a virus or a tiny nucleic + +01:07:05.360 --> 01:07:09.760 +acid called a viroid discovered by Ted Deener in the late 1960s and that we needed to give it a + +01:07:09.760 --> 01:07:13.920 +new name and so i had the audacity to call these particles prions that's the long arm of the + +01:07:13.920 --> 01:07:17.600 +scientific community pushing on me and people say well what do those people look like and here + +01:07:17.600 --> 01:07:23.760 +they are at the table that's my best slide you can go home now and it became very very clear + +01:07:23.760 --> 01:07:27.760 +that eventually became very clear our infectious proteins and i'm going to take you through a + +01:07:27.760 --> 01:07:32.800 +lot more evidence that builds the case that prions are infectious proteins and then as we move through + +01:07:32.880 --> 01:07:37.440 +this we'll get to some very new data and we'll talk about bse and we'll talk about other prions + +01:07:37.440 --> 01:07:40.960 +diseases of humans and then at the end we'll talk about therapies and there'll be a lot of chemistry + +01:07:40.960 --> 01:07:45.600 +at the end but it won't be complicated because i put the chemistry in the beginning you'll all + +01:07:45.600 --> 01:07:51.600 +leave so here we have there is there is something to be said about too many jokes + +01:07:52.800 --> 01:07:57.120 +have this purification scheme where we remove 99 percent of the unwanted particles we then knew + +01:07:57.120 --> 01:08:01.120 +that a protein was important and we went on to identify and find this protein and then the next + +01:08:01.200 --> 01:08:05.040 +thing we did was to try to determine whether we could separate this protein from the infectivity + +01:08:05.040 --> 01:08:08.800 +and so we used a lot of approaches so we're now measuring the biological activity enhancers + +01:08:08.800 --> 01:08:12.080 +and we're looking at the protein by physical methods and we tried to separate it and what we + +01:08:12.080 --> 01:08:15.600 +found is that every time we destroyed the protein we destroyed the infectivity or every time we + +01:08:15.600 --> 01:08:22.800 +destroyed the infectivity the protein was removed or destroyed and when we eventually figured out + +01:08:22.800 --> 01:08:26.560 +what was going on we found that there was a normal form of the protein in all of us now this is a + +01:08:26.560 --> 01:08:30.240 +gel and the proteins migrate and the smaller they are the faster they migrate through this gel this + +01:08:30.320 --> 01:08:35.360 +porous material sort of like jello and then we can stain them with antibodies and so what we found + +01:08:35.360 --> 01:08:39.920 +was that the protein was a protein in all of us which is a normal form of the prion protein + +01:08:39.920 --> 01:08:43.920 +and when we treat with enzymes that destroy proteins the protein was completely destroyed + +01:08:43.920 --> 01:08:48.400 +but in the brains of these hamsters that became ill with scrapi what we found was that there was + +01:08:48.400 --> 01:08:51.760 +both a normal form of the protein and another form of the protein that when we treated with + +01:08:51.760 --> 01:08:55.760 +enzymes that destroy proteins there was a residual piece and here they're higher-ordered multiverse + +01:08:56.720 --> 01:09:02.400 +they're larger forms that are aggregated of the scrapi causing protein and eventually + +01:09:02.400 --> 01:09:06.320 +the protein that I just showed you in the gel after we treated with proteases what we did was to use + +01:09:06.320 --> 01:09:11.600 +a lot of very complicated equipment in collaboration with a man named Leroy Hood who was at Caltech at + +01:09:11.600 --> 01:09:18.480 +the time and Darlene Groff who's been working with me for 25 years went to Caltech over a period + +01:09:18.480 --> 01:09:23.040 +of a year about seven different times and we finally were able to work out what's called the + +01:09:23.120 --> 01:09:27.360 +amino acid sequence the N terminal 15 amino acids so right in this region now this was more + +01:09:27.360 --> 01:09:30.960 +complicated because it was ragged because we'd thrown in these proteases but what we had found + +01:09:30.960 --> 01:09:35.840 +of course is that if we threw the proteases into the normal preparations there was nothing well + +01:09:35.840 --> 01:09:39.680 +now we had the possibility of doing what's called reverse genetic engineering and we went backwards + +01:09:39.680 --> 01:09:43.680 +and with Charles Weisman and Zurich we found the gene that codes for the prion protein and we found + +01:09:43.680 --> 01:09:47.280 +that all of us have this gene and it became very clear that all of us have the normal form of the + +01:09:47.280 --> 01:09:51.360 +prion protein and then by a process that we still don't understand in great detail it could be + +01:09:51.440 --> 01:09:55.760 +converted into a protein which is infectious and which can eventually kill us as human beings or + +01:09:55.760 --> 01:10:02.000 +kill animals like the cattle in Europe so now we knew that there was a gene for the protein of the + +01:10:02.000 --> 01:10:09.280 +prion and the protein of the prion we called PRP scrapey and it's and that this gene encodes + +01:10:09.280 --> 01:10:12.960 +the precursor protein PRPC there's a mistake here it's not found it's only found in six mammals + +01:10:12.960 --> 01:10:18.640 +but the gene is found in all mammals so now we had two proteins the precursor protein PRPC that + +01:10:18.720 --> 01:10:23.440 +we all have a normal protein and the protein found in okay so just notice the wording that + +01:10:23.440 --> 01:10:28.320 +he's using here a precursor protein we have to assume that it does something normally + +01:10:29.280 --> 01:10:35.440 +and that's the normal protein and that the the prion scrapey protein has to be the abnormal + +01:10:35.440 --> 01:10:42.480 +protein so it's really weird that he doesn't say it in the way that his disease model implies it + +01:10:42.480 --> 01:10:50.320 +should be said um as if the protein only exists to cause the disease in its precursor form it's + +01:10:50.320 --> 01:10:56.640 +fine in its functional form it causes disease that's really not the way we should be thinking of this + +01:10:57.520 --> 01:10:58.480 +especially if + +01:11:02.240 --> 01:11:07.760 +we have to work from the assumption that the that the that the prion protein does something + +01:11:08.640 --> 01:11:14.560 +otherwise what is it there for on the membrane a protein that's on the memory that doesn't do + +01:11:14.560 --> 01:11:20.880 +anything sounds like a very bad idea to me had two proteins the precursor protein PRPC that we + +01:11:20.880 --> 01:11:25.680 +all have a normal protein and the protein found in the animals that were ill with prion disease + +01:11:26.400 --> 01:11:29.760 +we did another set of experiments and we asked what about the messenger RNA so the messenger + +01:11:29.760 --> 01:11:34.080 +RNA is the intermediate between the gene and the protein and we said well the messenger RNA ought + +01:11:34.080 --> 01:11:39.120 +to rise as the as these animals go out toward the these are now mice that get sick at 130 days + +01:11:39.120 --> 01:11:44.320 +as incubation time gets longer and longer and so the n-terminal amino acid sequence of of + +01:11:45.520 --> 01:11:54.960 +prion protein 27 to 30 is determined isocoding DNAs used to retrieve PRPC DNA clones + +01:11:55.520 --> 01:12:01.600 +so my guess is is that they used + +01:12:04.320 --> 01:12:13.520 +DNAs that would encode for the amino acid sequence of the n-terminal then they use those DNA probes + +01:12:13.520 --> 01:12:23.600 +and they hybridized against the DNA of a human or a mouse or something like my human I guess + +01:12:23.600 --> 01:12:34.320 +and found a genomic DNA to which that n-terminal DNA hybridized to + +01:12:40.240 --> 01:12:46.400 +expected the prion protein mRNA levels were similar in uninfected controls with animals with + +01:12:46.400 --> 01:12:54.880 +scraping so when they look for the mRNA for the for the protein they find it and two forms are + +01:12:54.880 --> 01:13:05.600 +identified cellular and pathogenic and is derived from pr by n-terminal truncation let's see how + +01:13:05.600 --> 01:13:10.400 +he explains this hider the prions goes up the messenger RNA ought to rise but in fact the messenger + +01:13:10.400 --> 01:13:14.000 +RNA state exactly the same it never changed and this worried us a little bit where we're looking + +01:13:14.000 --> 01:13:18.480 +at the right protein but we got lucky because using that same technology genetic engineering + +01:13:18.480 --> 01:13:23.360 +we now looked at black mice or brown mice and we looked at these white mice the white mice had + +01:13:23.360 --> 01:13:29.280 +long incubation times about 225 days the short ones about 130 days and what we found was that this + +01:13:29.280 --> 01:13:33.360 +gene the prion protein gene that i've been talking about determined whether the mice had a short + +01:13:33.360 --> 01:13:38.320 +incubation time or a long incubation time and the mice with long incubation times had two amino + +01:13:38.320 --> 01:13:43.680 +acids out of 250 that were different and so this began to cement the idea that this protein PRP + +01:13:43.680 --> 01:13:49.120 +that we discovered was key in the disease process wow so that's a really key thing to realize because + +01:13:49.120 --> 01:13:53.920 +what is this established this is we need to find this paper so i'm going to write this one down + +01:13:53.920 --> 01:14:02.640 +because this is a really important one it's 86 through 87 and this is brown and white mice huh + +01:14:09.360 --> 01:14:13.120 +the reason why this is important is because this whole thing would have been established + +01:14:13.120 --> 01:14:19.520 +by a p-value test right if the p-value between the incubation time and the brown mice and the + +01:14:19.520 --> 01:14:26.240 +incubation time and the white mice in the experiment was significant as determined by whatever probability + +01:14:26.240 --> 01:14:33.200 +test they use that ritual would be enough to establish this as fact and then he would go on to + +01:14:33.200 --> 01:14:40.080 +say that these two amino acid differences between those mice and those mice are the reason why the + +01:14:40.080 --> 01:14:46.800 +incubation time is different and then that whole field moves on based on the idea that well i mean + +01:14:46.800 --> 01:14:56.640 +obviously think about how that that could potentially set up years and years of being wrong + +01:14:59.040 --> 01:15:02.560 +that's not definitive proof that this protein's involved + +01:15:04.320 --> 01:15:09.680 +how can that be even if there is a p-value difference there how can that be assumed + +01:15:09.760 --> 01:15:13.360 +is that the only difference between the brown mice and the white mice + +01:15:15.440 --> 01:15:17.280 +wow isn't that special + +01:15:19.360 --> 01:15:24.000 +isn't that special the lady from mit told us that all those heat shock proteins are really + +01:15:24.000 --> 01:15:28.960 +responsible for protein folding and misfolding and that they're like chaperone proteins and + +01:15:28.960 --> 01:15:34.240 +it's so cool because yeasts are eukaryotes and were eukaryotes and we have the same proteins + +01:15:34.240 --> 01:15:37.920 +that chaperone are protein folding and prevent them from misfolding + +01:15:39.520 --> 01:15:45.120 +and so it doesn't matter that the brown mice might have different heat shock protein variants + +01:15:45.120 --> 01:15:51.520 +than the white mice do it doesn't matter at all i think the lady from mit who's dead now would + +01:15:51.520 --> 01:15:59.040 +probably disagree but i don't know what's going on here that's how p-values are used to create + +01:15:59.040 --> 01:16:03.760 +the distortion of information and knowledge from noise + +01:16:06.640 --> 01:16:08.240 +it's a single experiment + +01:16:10.480 --> 01:16:17.280 +where the difference between incubation times to injection of material in their brain + +01:16:17.280 --> 01:16:22.800 +has nothing to do with looking at what was injected or where it was injected or how much + +01:16:22.800 --> 01:16:29.360 +exactly was injected it has to do with those two amino acids in that one protein that has to + +01:16:29.360 --> 01:16:33.440 +be i mean look i mean it has to be look at the other experiments we did there was like a gel + +01:16:33.440 --> 01:16:33.840 +and stuff + +01:16:38.160 --> 01:16:42.880 +out of 250 that were different and so this began to cement the idea that this protein + +01:16:42.880 --> 01:16:45.840 +PRP that we discovered was key in the disease process + +01:16:47.360 --> 01:16:50.480 +so what i've told you is that not only did the pre-on protein track with scraping + +01:16:50.480 --> 01:16:53.680 +infectivity so every time we tried to alter the protein we altered the infectivity and vice versa + +01:16:53.680 --> 01:16:58.880 +but also amino acid substitutions changed the incubation times so these findings link the phenotype + +01:16:58.880 --> 01:17:02.320 +which is called the disease characteristics in this case to the genotype that changes + +01:17:02.320 --> 01:17:07.520 +their mutations in the DNA okay so to a certain extent this is interesting because what it now + +01:17:07.520 --> 01:17:14.320 +implies of course is that function is related to structure which is one of the real overarching + +01:17:14.320 --> 01:17:19.520 +principles of reductionist biology one of the things that we all agree on as we move through + +01:17:19.520 --> 01:17:23.680 +the world trying to understand what's going on if you want to understand how a tree works + +01:17:25.280 --> 01:17:29.840 +it's probably related to its structure it's its structure in the macro sense and also structure + +01:17:29.840 --> 01:17:37.600 +at the cellular level and the molecular level but structure begets function is something that holds + +01:17:37.600 --> 01:17:45.280 +at the protein level from everyone's perspective the the amino acid sequence determines its structure + +01:17:46.240 --> 01:17:58.160 +the the RNA sequence that encoded the amino acid is to a large extent synonymous + +01:17:59.600 --> 01:18:05.600 +but we know that synonymous mutations that don't change the amino acid for which they code + +01:18:06.160 --> 01:18:10.960 +can still be highly correlated with genetic disease + +01:18:11.760 --> 01:18:20.320 +and so clearly the cartoon about yes we have DNA we translated it to RNA the RNA translates + +01:18:20.320 --> 01:18:25.600 +to amino acids and the amino acids arrange in a certain way because there are three-dimensional + +01:18:25.600 --> 01:18:32.240 +electrostatic shape inside of a three-dimensional electrostatic solvent called water and so they + +01:18:32.240 --> 01:18:40.720 +form a particular shape to try and best optimize their energy distribution in a stable way + +01:18:44.240 --> 01:18:48.080 +but that's a variable process a process that isn't understood a process that + +01:18:48.080 --> 01:18:54.560 +occurred to the dead lady from MIT actually goes wrong an awful lot and we waste a lot of energy + +01:18:54.560 --> 01:19:03.360 +misfolding proteins all the time and that prion protein now is a very special protein that has + +01:19:03.360 --> 01:19:10.000 +the propensity apparently as we move forward we're going to hear how it can can cause other + +01:19:10.000 --> 01:19:16.080 +proteins with a similar sequence to misfold as well so prion protein misfolds other prion protein + +01:19:16.080 --> 01:19:20.400 +even though we don't know what prion protein really does in the in the healthy animal it's + +01:19:20.480 --> 01:19:28.320 +there it's mRNA is there all the time and in in some animals like hamsters it develops faster + +01:19:28.320 --> 01:19:34.400 +than other animals like mice and in some brown mice it develops longer faster than in white mice + +01:19:38.800 --> 01:19:42.080 +now we come to a whole different approach and we spent six years here at UCSF + +01:19:43.600 --> 01:19:47.760 +trying to identify the difference between PRPC the normal form of the protein that all of us have + +01:19:47.760 --> 01:19:52.000 +and the scrapey form of the protein that's found only in the disease and we looked and looked so + +01:19:52.000 --> 01:19:58.560 +far the only evidence that they have of a different protein is that gel keep that in mind right do + +01:19:58.560 --> 01:20:02.400 +we need to go back to that a little bit if I just go back to the gel + +01:20:09.120 --> 01:20:14.880 +this is the gel right this this is the gel that he explained is the evidence that he has for + +01:20:14.960 --> 01:20:21.600 +prion protein being different this is the normal protein this is the lane with the normal protein + +01:20:21.600 --> 01:20:30.480 +after they digested it with proteases this is the protein apparently from the the scrapey infected + +01:20:30.480 --> 01:20:36.160 +animal i don't know what all this other crap is that didn't show up in this line which is also + +01:20:36.160 --> 01:20:43.360 +very frustrating how am i supposed to compare this experiment if this lane is supposed to be + +01:20:43.360 --> 01:20:47.920 +equivalent to that lane except this lane has four more bars in it than this one does + +01:20:47.920 --> 01:20:54.240 +what kind of shit experiment is this and how is he able to just get through the all + +01:20:54.240 --> 01:21:00.480 +gel it moves uh the smaller fragments move faster than the the heavy fragments and uh so uh yeah + +01:21:01.360 --> 01:21:06.560 +so what are these fragments well anyway his argument is is that this is the protease + +01:21:06.560 --> 01:21:13.840 +digested side and here they still have a protein whereas here it's all gone and so in that in the + +01:21:13.840 --> 01:21:19.040 +in the normal functioning prion protein format it can be completely digested by protease and here + +01:21:19.040 --> 01:21:25.920 +it can't that is the only evidence that they have for an alternate form of a protein that's it + +01:21:27.360 --> 01:21:29.200 +because the sequence is the same + +01:21:31.520 --> 01:21:34.800 +as best they can tell it's just one gene + +01:21:37.520 --> 01:21:45.120 +these are not as related complementary experiments as he's implying and without careful + +01:21:46.240 --> 01:21:54.000 +and maybe even several months of hard studious work you can't claim to be an expert on prions + +01:21:54.000 --> 01:22:00.320 +because nobody explains it well enough to be this is not a good enough explanation of prions + +01:22:00.400 --> 01:22:04.080 +for me to say oh yeah i guess i believe it this is hand waving + +01:22:07.280 --> 01:22:13.200 +this is how they explain to you how pandemics work and why masks work and why lockdowns if done + +01:22:13.200 --> 01:22:18.880 +correctly work and why zero virus after the end of a pandemic is a useful goal + +01:22:20.080 --> 01:22:24.480 +it's that kind of it's that kind of explanation on protein track with scraping + +01:22:24.480 --> 01:22:27.760 +infectivity so every time we try to alter the protein we alter the infectivity and vice versa + +01:22:27.760 --> 01:22:32.960 +but also amino acid substitutions change the incubation times so these findings link the phenotype + +01:22:32.960 --> 01:22:36.480 +which is called the disease characteristics in this case to the genotype the changes are + +01:22:36.480 --> 01:22:42.400 +mutations in the DNA thank you solar flare i'm clicking a whole different approach and we spent + +01:22:42.400 --> 01:22:47.840 +six years here at UCSF trying to identify the difference i'll take it between prpc the normal + +01:22:47.840 --> 01:22:51.360 +form of the protein that all of us have and the scraping form of the protein that's found only + +01:22:51.360 --> 01:22:54.800 +the disease and we looked and looked for a chemical difference because if we could have found a + +01:22:54.800 --> 01:22:58.960 +chemical difference then it would be very easy to make that chemical change on prpc + +01:22:58.960 --> 01:23:03.680 +and convert it into prpc but we never found it this was a work of a young postdoc Neil Stahl + +01:23:03.680 --> 01:23:07.440 +who was working with Mike Baldwin who's still here and who's a professor in the school of pharmacy + +01:23:07.440 --> 01:23:13.840 +and mass spectrometry so then Fred Cohn became involved and we began to think about models + +01:23:13.840 --> 01:23:17.360 +and the first thing that Fred did he said i wonder if this isn't really the case that + +01:23:17.360 --> 01:23:21.360 +the normal form of the prion protein is full of these helical structures called alpha helices + +01:23:21.440 --> 01:23:26.080 +and that some of these are reduced and that there's a lot of beta sheet these beta strands + +01:23:26.080 --> 01:23:31.520 +these blue ribbons and at that point what we did was we knew that there was a lot of beta structure + +01:23:31.520 --> 01:23:35.760 +from other studies from our won't work and from that of other people we went it off and purified + +01:23:35.760 --> 01:23:39.520 +out of brains of animals the normal form of the prion protein and we found that Fred in fact was + +01:23:39.520 --> 01:23:43.600 +correct that there was very little beta structure and then more recently Tom James and his colleagues + +01:23:43.600 --> 01:23:47.600 +here with the help of peter right now i thought it was really hard to purify the prion proteins + +01:23:47.680 --> 01:23:53.520 +from animals with the disease and now he just says we're going to go do it and we found out + +01:23:53.520 --> 01:24:01.440 +that there's more beta sheets i don't know pain dyson scripts uh from protein that was made by + +01:24:01.440 --> 01:24:05.520 +genetic engineering and bacteria and determined its structure so here we're sort of halfway + +01:24:05.520 --> 01:24:09.760 +through the protein around residue 90 and then you see this long unstructured region then one the + +01:24:09.760 --> 01:24:13.520 +first alpha helix then more of this unstructured region with a small amount of beta structure + +01:24:13.520 --> 01:24:18.320 +than the second helix and the third helix and what's called the c-terminus now what we + +01:24:18.320 --> 01:24:22.000 +hypothesized was that the first helix and in fact we thought the first helix was made up of two + +01:24:22.000 --> 01:24:25.840 +this whole region was made up of two helices and we were wrong and Kurt Rudrich and Zurich showed us + +01:24:25.840 --> 01:24:32.080 +that we were wrong much to our dislike and unhappiness and so this was the model that we had for a number + +01:24:32.080 --> 01:24:37.440 +of years so what i've just told you is that when prpc is converted into prp scraping the protein + +01:24:37.440 --> 01:24:42.560 +undergoes a profound change in shape or as scientists call it confirmation prpc is rich in alpha helix + +01:24:42.560 --> 01:24:50.480 +whereas prp scraping is rich in beta structure now if you believe alpha fold and you believe + +01:24:52.080 --> 01:24:58.480 +people who study proteins alpha helix is something that you can predict from the sequence + +01:24:59.040 --> 01:25:04.320 +and a beta pleated sheet is something you can predict from the sequence and it's not very clear + +01:25:04.320 --> 01:25:12.640 +to me that those two are are not mutually exclusive i i was i was taught that those are kind of + +01:25:12.640 --> 01:25:17.120 +mutually exclusive in the sense of we don't really understand how proteins fold but we do + +01:25:17.120 --> 01:25:23.280 +understand that they have two general motifs that of an alpha helix and that of a beta pleated sheet + +01:25:23.280 --> 01:25:29.520 +and these two general motifs of course have very specific expression but these general motifs are + +01:25:29.600 --> 01:25:34.320 +used to construct or or or or build out some of these protein machines + +01:25:36.160 --> 01:25:42.960 +so the idea that one part of a protein could be both an alpha helix and a beta pleated sheet + +01:25:42.960 --> 01:25:48.800 +is already a very novel idea in my humble opinion and that i believe is what he's implying here + +01:25:50.240 --> 01:25:53.680 +and we went on to do some studies in collaboration with Dennis Burton at Scripps + +01:25:54.240 --> 01:25:58.720 +and this is the work of David Parrots who's still here at UCSF and what we did was to make + +01:25:58.720 --> 01:26:02.320 +antibodies that reacted to different regions of the prion protein so this is the three helix + +01:26:02.320 --> 01:26:07.440 +protein now what's really incredible about this is that antibodies are reacting to a three-dimensional + +01:26:07.440 --> 01:26:12.880 +static shape they don't necessarily react to only one and antibodies are very + +01:26:15.520 --> 01:26:20.480 +non-specific so it's very very would be very interesting to go back + +01:26:21.360 --> 01:26:27.520 +and see how these antibodies were raised they're raised to different epitopes as you can see here + +01:26:27.520 --> 01:26:36.160 +they are raised to epitopes that are about 22 amino acids this is 23 right this is no that's 30 + +01:26:36.880 --> 01:26:40.880 +30 28 i don't know i can't do that math it's not very many + +01:26:42.320 --> 01:26:47.120 +so they're small epitopes presumably then what injected into a rabbit or something and then + +01:26:47.120 --> 01:26:51.680 +they take the plasma out and they get those antibodies and those antibodies although they + +01:26:51.680 --> 01:26:56.880 +were raised to that epitope aren't necessarily all going to be the same antibodies with the same + +01:26:56.880 --> 01:27:02.720 +specificity so unless they used a monoclonal antibody as they were made in the 90s that would be one + +01:27:02.720 --> 01:27:10.240 +thing to look at here and what they're doing is trying to identify using antibody binding whether + +01:27:10.320 --> 01:27:14.000 +or not particular structures are present or not present + +01:27:21.200 --> 01:27:27.360 +so i i just wanted to be sure you knew what they were doing with these antibodies + +01:27:28.480 --> 01:27:32.640 +let's see what he says hey this is helix a helix b and helix c that i showed you before + +01:27:32.640 --> 01:27:41.360 +and this is residue 90 and in this first region the the prion protein binds antibodies and the + +01:27:41.360 --> 01:27:45.600 +native form of the prion protein binds them quite well but the binding region or epitope disappears + +01:27:45.600 --> 01:27:50.000 +when prp scrape is formed so that was an important discovery and we use that information and we + +01:27:50.000 --> 01:27:57.920 +contrasted that with the green where but remember that when prion protein scrape is formed they can't + +01:27:57.920 --> 01:28:05.200 +separate it from the native prion so again these pluses and minuses are binding strength and whenever + +01:28:05.200 --> 01:28:14.720 +you have one key tip i would say that whenever you have a paper that wants to basically + +01:28:16.160 --> 01:28:21.040 +say take your word for it that okay we rate this as a three and that one is a two and this one is a + +01:28:21.040 --> 01:28:27.360 +one and that's a you know you should take it with a grain of salt here both the native form of PRPC + +01:28:27.360 --> 01:28:31.680 +and the native form of PRPC mine antibodies that's down here + +01:28:32.160 --> 01:28:39.440 +so these epitopes are regions so that's the second basic prince proof that the scrappy protein in + +01:28:39.440 --> 01:28:46.240 +its in its infectious form is a different shape than this one but in this preparation we're not + +01:28:46.240 --> 01:28:53.120 +sequencing a protein we're not starting with a pure protein in my i i don't think i don't know + +01:28:53.120 --> 01:28:56.800 +what we're doing here we got to we got to find that out we got to know we hope we're doing a + +01:28:56.800 --> 01:29:04.720 +pure protein here but is it a synthetically pure made protein is it a so that's again we have we + +01:29:04.720 --> 01:29:10.560 +have a lot of work to do here to find out where p values are essentially holding up what is a what + +01:29:10.560 --> 01:29:17.360 +is a what is a house that shouldn't be holding up we already have seen a couple examples of this + +01:29:17.360 --> 01:29:22.880 +like the brown and white mice thing with two amino acids difference and this protein means that + +01:29:22.880 --> 01:29:27.440 +the difference that we saw in the experiment had to do with those two amino acids that's the + +01:29:27.440 --> 01:29:35.200 +kind of assumptions that we're always talking about here these numbers plus two plus three is that + +01:29:35.200 --> 01:29:42.720 +really is he really saying that the native and the denatured pre-on scrappy protein are different + +01:29:42.720 --> 01:29:47.520 +by a factor of one out of three that's their scale i think that's down here + +01:29:50.000 --> 01:29:54.640 +so these epitopes are regions of the alpha helix rich precursor protein p rpc of + +01:29:54.640 --> 01:29:59.360 +prions are exposed but become buried when the protein is refolded into an infectious form + +01:29:59.360 --> 01:30:08.000 +p rpc scraping so why didn't they make um anybody see this is the problem i have with that + +01:30:08.080 --> 01:30:16.720 +they have the the protein here native pre-on protein scrappy so why don't they make antibodies + +01:30:16.720 --> 01:30:25.600 +to that why don't they make antibodies to the pre-on protein scrappy part and then show me + +01:30:25.600 --> 01:30:31.360 +that those antibodies bind pre-on scrappy but they don't bind this one why don't they do the + +01:30:31.360 --> 01:30:37.680 +reverse experiment apparently they have a pure form of this right otherwise you couldn't do this + +01:30:38.640 --> 01:30:45.920 +this assay so they raised antibodies to these epitopes but they don't raise antibodies to the + +01:30:45.920 --> 01:30:50.560 +epitopes of the pre-on scrappy because it's so hard to isolate or something i don't know + +01:30:51.440 --> 01:30:55.920 +they can't get it to form badly in a in a dish or using the right ribosome or what + +01:30:57.280 --> 01:31:00.560 +you see the problem here here + +01:31:01.040 --> 01:31:08.480 +so these epitopes are regions of the alpha helix rich precursor protein p rpc of prions are + +01:31:08.480 --> 01:31:13.120 +exposed but become buried when the protein is refolded into an infectious form p rpc scraping + +01:31:15.680 --> 01:31:21.120 +now when we treat with proteases and we cleave off the n terminal region and we form the ragged + +01:31:21.120 --> 01:31:25.920 +n terminus that i showed you that we sequenced and we call this prp 2730 this protein and this + +01:31:25.920 --> 01:31:29.040 +is the one that i showed you before where we've treated with protease so it's a little bit shorter + +01:31:29.840 --> 01:31:33.440 +that protein polymerizes into these rod shape structures that you see here + +01:31:33.440 --> 01:31:40.400 +so after they treat it with an enzyme it forms rod shape structures there's nothing to do with + +01:31:40.400 --> 01:31:45.280 +the physiology of it at all right this is an in vitro preparation with an in vitro result + +01:31:46.400 --> 01:31:53.040 +and yet somehow or another we got here by him saying something about the refolding + +01:31:53.520 --> 01:32:00.480 +now when we treat with proteases and we cleave off the n terminal region and we form the ragged + +01:32:00.480 --> 01:32:05.280 +n terminus that i showed you that we sequenced and we call this prp 2730 this protein and this + +01:32:05.280 --> 01:32:08.400 +is the one that i showed you before where we've treated with protease so it's a little bit shorter + +01:32:09.680 --> 01:32:13.440 +that protein polymerizes into these rod shape structures that you see here and there was a very + +01:32:13.440 --> 01:32:17.360 +famous man at Berkeley who died about 15 years ago roblie Williams was really the father of the + +01:32:17.360 --> 01:32:20.960 +electron microscopy of viruses and he was working with us at the time because i really thought we + +01:32:21.040 --> 01:32:24.560 +were going to find an interesting virus not a protein and we would see these rod shape structures + +01:32:24.560 --> 01:32:29.280 +and he called them rods some people would call them fibers and one day i guess we'd been working + +01:32:29.280 --> 01:32:32.800 +together for almost a year and a half i was looking at a book on the electron microscopy of proteins + +01:32:32.800 --> 01:32:37.360 +and all this looking like amyloid proteins now amyloid is a mammalian protein these are usually + +01:32:37.360 --> 01:32:42.240 +cathologically laid down in big fibers and there are many many diseases like alzheimer's disease + +01:32:42.240 --> 01:32:47.200 +where there are amyloid plaques of many of these proteins but so the first connection here so far + +01:32:47.200 --> 01:32:52.400 +seems to be that one microscopist called it a fibro and now they the people that study alzheimer's + +01:32:52.400 --> 01:32:57.680 +disease have also called the proteins involved in alzheimer's disease fibers and so now that's + +01:32:57.680 --> 01:33:06.400 +okay now they must be the same thing it turned out that this broke that what we were seeing was that + +01:33:06.400 --> 01:33:10.720 +the protein of the prion was forming amyloid and that's what these rods represent and then it formed + +01:33:10.720 --> 01:33:14.640 +plaques in the brain and i'll show you some of those plaques a little later now we also found + +01:33:14.640 --> 01:33:18.480 +and this is the work of holder willy who's here at UCSF along with the help of david agar and + +01:33:18.480 --> 01:33:24.080 +many others uh hana serven uh darlene groth that the protein forms these + +01:33:25.680 --> 01:33:30.320 +rods that the plaques a little later amyloid and that's what these were this broke that + +01:33:30.320 --> 01:33:34.080 +what we were seeing was that the protein of the prion was forming amyloid and that's what + +01:33:34.080 --> 01:33:39.680 +these rods represent the protein of the prion was forming amyloid so what i hear here is that prion + +01:33:40.640 --> 01:33:47.440 +protein is the sub component of amyloid is that what i'm hearing is that right + +01:33:53.680 --> 01:34:00.320 +hmm that's interesting that's that i didn't think that they were the same i didn't think + +01:34:00.320 --> 01:34:05.280 +they were the same protein i thought amyloid was a different protein um i didn't know that it + +01:34:05.280 --> 01:34:09.360 +was prion protein that was folded differently without an n-terminus because that's what he just + +01:34:09.360 --> 01:34:15.760 +said these are prion rods that are formed by the prion protein after you cleave its n-terminal + +01:34:15.760 --> 01:34:22.960 +off with proteases and these form under the microscope and now he seems to be indicating + +01:34:22.960 --> 01:34:31.280 +in 2002 that the amyloid plaques are actually prion protein did i not hear that did i not hear that + +01:34:33.840 --> 01:34:38.560 +on the electron microscopy of proteins and all this look like amyloid proteins now amyloid + +01:34:38.560 --> 01:34:42.080 +was is a mammalian protein these are usually depend logically uh laid down + +01:34:42.080 --> 01:34:46.240 +in big fibrils and there are many many diseases like Alzheimer's disease where there are amyloid + +01:34:46.240 --> 01:34:57.520 +many many diseases like Alzheimer's disease where amyloid plaques are laid down i don't + +01:34:57.520 --> 01:35:02.000 +don't know many many many diseases where amyloid plaques all this look like amyloid proteins + +01:35:02.000 --> 01:35:06.480 +now amyloid was is a mammalian protein these are usually Happyl logically uh laid down in + +01:35:06.480 --> 01:35:08.640 +is like Alzheimer's disease, where there are amyloid plaques, + +01:35:08.640 --> 01:35:09.560 +so many of these proteins. + +01:35:09.560 --> 01:35:12.480 +But it turned out that this broke that what we were seeing + +01:35:12.480 --> 01:35:15.120 +was that the protein of the prion was forming amyloid, + +01:35:15.120 --> 01:35:16.720 +and that's what these rods represent. + +01:35:16.720 --> 01:35:18.120 +And then it formed plaques in the brain, + +01:35:18.120 --> 01:35:20.640 +and I'll show you some of those plaques a little later. + +01:35:20.640 --> 01:35:22.760 +Now, we also found, and this is the work of Holger-Willy, + +01:35:22.760 --> 01:35:25.200 +who's here at UCSF, along with the help of David Agard + +01:35:25.200 --> 01:35:28.080 +and many others, Hana Sermon, Darlene Groth, + +01:35:28.080 --> 01:35:33.040 +that the protein forms these rods, + +01:35:33.040 --> 01:35:34.240 +and then at one end, we sometimes + +01:35:34.240 --> 01:35:36.000 +see these two-dimensional crystals. + +01:35:36.000 --> 01:35:37.920 +And we don't know whether the crystals form first, + +01:35:37.920 --> 01:35:39.760 +and from that, the rods form, or the rods + +01:35:39.760 --> 01:35:43.840 +begin to dissolve, and we form the crystals on the surface. + +01:35:43.840 --> 01:35:46.160 +But we were able to use those images of the crystals + +01:35:46.160 --> 01:35:48.600 +and then do what's called correlation averaging. + +01:35:48.600 --> 01:35:50.800 +Now, keep in mind, this is what Robert Malone + +01:35:50.800 --> 01:35:53.680 +purports to have simulatedly done at the beginning + +01:35:53.680 --> 01:35:58.240 +of the pandemic for the three CL protease of the coronavirus. + +01:35:58.240 --> 01:36:04.480 +He made a virtual X-ray crystallography model of that enzyme + +01:36:04.480 --> 01:36:07.920 +and then used that three-dimensional X-ray crystallography + +01:36:07.920 --> 01:36:12.400 +model to scan all known pharmaceuticals and nutraceuticals + +01:36:12.400 --> 01:36:15.640 +in the FDA catalog in less than three weeks + +01:36:15.640 --> 01:36:21.240 +to identify ivermectin, fluvoxamine, silicoxib, + +01:36:21.240 --> 01:36:28.280 +and remdesivir has four potential repurposed drugs + +01:36:28.280 --> 01:36:30.760 +to respond to the pandemic. + +01:36:30.760 --> 01:36:39.320 +He spun up his volunteer team and developed an X-ray crystallography + +01:36:39.320 --> 01:36:42.160 +model, computer-generated, similar to what + +01:36:42.160 --> 01:36:46.880 +is being described here, but just a computer-generated version + +01:36:46.880 --> 01:36:50.480 +of it, of the three CL protease and then + +01:36:50.480 --> 01:36:55.320 +scanned all known drugs and pharmaceuticals + +01:36:55.320 --> 01:36:58.920 +using the domain program, which was a DITRA program run + +01:36:58.920 --> 01:37:04.680 +by his friend, David Hone, and came up + +01:37:04.680 --> 01:37:07.720 +with four candidate drugs, of which at least one of them + +01:37:07.720 --> 01:37:14.400 +was tested by Steve Kirsch, through his early treatment + +01:37:14.400 --> 01:37:18.720 +fund, something something before he started his VRSF fund. + +01:37:23.000 --> 01:37:25.960 +I mean, that's neither here nor there with regard + +01:37:25.960 --> 01:37:27.320 +to what's happening here. + +01:37:27.320 --> 01:37:30.160 +This is just to say that when you can crystallize a protein, + +01:37:30.160 --> 01:37:33.160 +then you can take high-resolution pictures of it + +01:37:33.160 --> 01:37:37.440 +and get some idea of the structure of the protein + +01:37:37.440 --> 01:37:43.240 +because the crystal is a ever-repeating lattice of a pure protein. + +01:37:43.240 --> 01:37:47.480 +So this is a ever-repeating lattice of the prion protein, + +01:37:47.480 --> 01:37:50.720 +not the prionscrapi protein, which they still + +01:37:50.720 --> 01:37:54.240 +can't produce separately from the prion protein, which + +01:37:54.240 --> 01:37:57.040 +is interesting because the prion protein is not + +01:37:57.040 --> 01:37:59.200 +an infectious protein. + +01:37:59.200 --> 01:38:00.600 +And then we can average some more. + +01:38:00.600 --> 01:38:02.880 +So these are all imaging techniques. + +01:38:02.880 --> 01:38:04.240 +And we can develop these power spectra + +01:38:04.240 --> 01:38:06.200 +and get very, very detailed images. + +01:38:06.200 --> 01:38:08.160 +And so we went through a lot of this. + +01:38:08.160 --> 01:38:09.880 +And eventually, we're able to show + +01:38:09.880 --> 01:38:12.480 +that a piece of the prion protein in the middle of the protein + +01:38:12.480 --> 01:38:14.680 +is found in the center in these black regions + +01:38:14.680 --> 01:38:18.160 +and that the sugar chains are found around the edges + +01:38:18.160 --> 01:38:19.160 +of the black regions. + +01:38:19.160 --> 01:38:20.960 +And that gave rise to two different models. + +01:38:20.960 --> 01:38:23.200 +What we call a trimer of dimers, so there + +01:38:23.200 --> 01:38:26.560 +are six here, or simply a trimer, as shown here. + +01:38:26.560 --> 01:38:28.560 +And this is the right-handed model. + +01:38:28.560 --> 01:38:31.520 +This is the left-handed model. + +01:38:31.520 --> 01:38:33.080 +And here, you simply see one of these now. + +01:38:33.080 --> 01:38:34.440 +It's a much different-looking structure + +01:38:34.440 --> 01:38:35.840 +for the model of prp-scrapi. + +01:38:35.840 --> 01:38:37.080 +It has what are called beta helices. + +01:38:37.080 --> 01:38:39.080 +So this beta structure is now in a helix. + +01:38:39.080 --> 01:38:40.080 +This is an alpha helix. + +01:38:40.080 --> 01:38:41.200 +This is a beta helix. + +01:38:41.200 --> 01:38:43.740 +This is PRPC, as I talked about earlier, with helix A, + +01:38:43.740 --> 01:38:46.400 +helix B, and helix C. Here, you see part of helix B + +01:38:46.400 --> 01:38:48.240 +preserved, and all of helix C preserved. + +01:38:48.240 --> 01:38:50.840 +And the rest of this now is changed into beta helix. + +01:38:50.840 --> 01:38:51.800 +This is a current model. + +01:38:51.800 --> 01:38:52.920 +This is what I was thinking about this. + +01:38:52.920 --> 01:38:56.360 +It may or may not be right. + +01:38:56.360 --> 01:38:58.280 +In the way we obtained these images + +01:38:58.280 --> 01:38:59.040 +was using what we call it. + +01:38:59.040 --> 01:39:03.120 +Now remember, he doesn't have the crystal of this. + +01:39:03.120 --> 01:39:07.880 +So I don't really know what their molecular basis for assuming + +01:39:07.880 --> 01:39:08.840 +there's beta. + +01:39:08.840 --> 01:39:10.160 +That's a beta-pleated sheet. + +01:39:10.160 --> 01:39:11.520 +He called it a beta helix. + +01:39:11.520 --> 01:39:13.000 +I don't know. + +01:39:13.000 --> 01:39:14.920 +I thought you called that a beta-pleated sheet. + +01:39:14.920 --> 01:39:15.640 +I could be wrong. + +01:39:15.640 --> 01:39:17.880 +There could be also a beta helix. + +01:39:17.880 --> 01:39:19.720 +I mean, I'm not a protein guy. + +01:39:19.720 --> 01:39:20.800 +This is the current model. + +01:39:20.800 --> 01:39:22.000 +This is what I was thinking about this. + +01:39:22.000 --> 01:39:25.360 +It may or may not be right. + +01:39:25.360 --> 01:39:27.320 +In the way we obtained these images + +01:39:27.320 --> 01:39:29.440 +was using what we call the old electron microscope, which + +01:39:29.440 --> 01:39:30.600 +means it's no longer in service. + +01:39:30.600 --> 01:39:32.000 +But I'd simply show you this. + +01:39:32.000 --> 01:39:33.720 +And now what we're doing is working + +01:39:33.720 --> 01:39:36.640 +with a new microscope, which was purchased + +01:39:36.640 --> 01:39:38.480 +with the help of the Fairchild Foundation. + +01:39:38.480 --> 01:39:40.240 +This is a $2 million instrument. + +01:39:40.240 --> 01:39:42.000 +And you can maybe understand all this new detail that + +01:39:42.000 --> 01:39:44.280 +wasn't there in the last slide. + +01:39:44.280 --> 01:39:46.120 +So you can see that it's really essentially blank. + +01:39:46.120 --> 01:39:48.200 +And now we begin to see all kinds of information. + +01:39:48.200 --> 01:39:51.240 +So we're very optimistic that this will yield much more + +01:39:51.240 --> 01:39:53.120 +structure in the near future. + +01:39:53.120 --> 01:39:54.680 +So prions are composed of prion proteins + +01:39:54.680 --> 01:39:55.920 +that are rich in beta structure. + +01:39:55.920 --> 01:39:58.040 +And we don't know whether this is beta helix or beta sheet. + +01:39:58.040 --> 01:39:59.320 +But all the evidence so far argues + +01:39:59.320 --> 01:40:02.320 +for this new model of beta helix. + +01:40:02.320 --> 01:40:04.960 +So there are beta helix and beta sheet, according to that, + +01:40:04.960 --> 01:40:07.560 +which is good to know. + +01:40:07.560 --> 01:40:10.000 +I still don't think he's provided a lot of evidence + +01:40:10.000 --> 01:40:15.040 +that prion in an alternate form can cause other proteins + +01:40:15.040 --> 01:40:16.840 +to form that way yet, right? + +01:40:16.840 --> 01:40:18.640 +We haven't even been able to establish + +01:40:18.640 --> 01:40:21.160 +that prion scrappy exists as a separate protein. + +01:40:21.160 --> 01:40:24.520 +He's only shown as a crystal of prion protein + +01:40:24.520 --> 01:40:25.920 +in its natural form. + +01:40:25.920 --> 01:40:27.680 +Keep that in mind. + +01:40:27.680 --> 01:40:29.480 +When the prion protein PRPC is synthesized, + +01:40:29.480 --> 01:40:30.640 +it's made deep in the cell in what + +01:40:30.640 --> 01:40:31.800 +is called the endoplasmic reticulum. + +01:40:31.800 --> 01:40:32.320 +Here we go. + +01:40:32.320 --> 01:40:34.120 +It travels through what is called the Golgi apparatus + +01:40:34.120 --> 01:40:35.160 +out to the surface. + +01:40:35.160 --> 01:40:36.920 +And on the surface in these what are called + +01:40:36.920 --> 01:40:39.360 +rafts or cholesterol rich microdomains, + +01:40:39.360 --> 01:40:41.040 +where there's a lot of cholesterol on the surface + +01:40:41.040 --> 01:40:43.960 +in these little regions, the normal form of the prion + +01:40:43.960 --> 01:40:46.400 +protein PRPC is converted into PRP scrappy. + +01:40:46.400 --> 01:40:48.120 +And then it travels deep into the cell again + +01:40:48.120 --> 01:40:49.600 +and all the way down into what are called bicosomes. + +01:40:49.600 --> 01:40:52.840 +So does this look like anything that you've seen before, + +01:40:52.840 --> 01:40:56.640 +another cartoon of some intracellular biological process + +01:40:56.640 --> 01:41:00.960 +is just to be assumed to work exactly like it's drawn + +01:41:00.960 --> 01:41:03.840 +because there's a guy with a laser pointer shining on it? + +01:41:06.480 --> 01:41:08.880 +Because nothing that we've talked about so far + +01:41:08.880 --> 01:41:13.560 +in this discussion has led us to believe + +01:41:13.560 --> 01:41:16.680 +that this part of this thing is like real. + +01:41:16.680 --> 01:41:18.280 +It's an assumption. + +01:41:18.280 --> 01:41:20.760 +The cartoon is an assumption. + +01:41:20.760 --> 01:41:22.920 +And they're not going to do very many experiments + +01:41:22.920 --> 01:41:26.320 +to test any predictions that this thing might make. + +01:41:26.320 --> 01:41:29.320 +They're going to test to see if they can find data + +01:41:29.320 --> 01:41:31.160 +which fits this cartoon. + +01:41:31.160 --> 01:41:32.960 +See, it's converted into PRP scrappy. + +01:41:32.960 --> 01:41:34.680 +And then it travels deep into the cell again + +01:41:34.680 --> 01:41:36.400 +and all the way down into what are called bicosomes. + +01:41:36.400 --> 01:41:40.000 +And of course, this is a part of the biology + +01:41:40.000 --> 01:41:42.000 +that they want you to ignore that there + +01:41:42.000 --> 01:41:45.400 +are these lipid rafts that are in our membrane + +01:41:45.400 --> 01:41:47.440 +which are cholesterol-rich regions + +01:41:47.440 --> 01:41:52.360 +where most of the intracellular extracellular communication + +01:41:52.360 --> 01:41:54.560 +and trafficking occurs. + +01:41:54.560 --> 01:41:56.320 +And so it's one of the reasons why, + +01:41:56.320 --> 01:41:58.840 +besides the fact that myelin in your brain + +01:41:58.840 --> 01:42:00.720 +is made up of a lot of cholesterol, + +01:42:00.720 --> 01:42:04.280 +it's one of the wonderful reasons why statins are a terrible + +01:42:04.280 --> 01:42:08.280 +and that reducing cholesterol is just a terrible health motive + +01:42:09.800 --> 01:42:11.520 +that could have only been put out there + +01:42:11.520 --> 01:42:14.400 +by people who are idiots or people who are malevolent. + +01:42:16.400 --> 01:42:19.400 +And this is really feeling like covering up + +01:42:19.400 --> 01:42:22.360 +for people that are malevolent more than anything else. + +01:42:22.360 --> 01:42:26.480 +This doesn't feel like a legitimate biological story + +01:42:26.480 --> 01:42:28.840 +but it feels like mythology to me. + +01:42:28.840 --> 01:42:30.720 +Where proteins are normally degraded + +01:42:30.720 --> 01:42:32.040 +but PRP scrappy persists. + +01:42:32.040 --> 01:42:33.400 +But it doesn't persist forever as I'll show you + +01:42:33.400 --> 01:42:35.000 +a little bit later and it's changed the way + +01:42:35.000 --> 01:42:36.000 +we think about this. + +01:42:39.000 --> 01:42:40.280 +So the precursor of the prion + +01:42:40.280 --> 01:42:42.000 +is an alpha helix-rich protein PRPC + +01:42:42.000 --> 01:42:43.520 +that's synthesized in what's called the ER + +01:42:43.520 --> 01:42:45.240 +and a plasma-criticulum of the cell. + +01:42:45.240 --> 01:42:47.200 +The infectious form of the prion protein PRP scrappy + +01:42:47.200 --> 01:42:48.880 +of the prion is rich in beta structure + +01:42:48.880 --> 01:42:51.680 +and it is formed from PRPC on the surface of the cell + +01:42:51.680 --> 01:42:53.360 +in these cholesterol-rich micro domains. + +01:42:53.360 --> 01:42:55.120 +So this, all of this right here, + +01:42:55.120 --> 01:42:57.840 +besides the cholesterol-rich micro domains, + +01:42:57.840 --> 01:43:00.360 +all of this here is based on very, very little evidence, + +01:43:00.360 --> 01:43:02.840 +sometimes only one paper, just keep that in mind. + +01:43:05.520 --> 01:43:07.760 +Now recently we discovered that the prion protein + +01:43:07.760 --> 01:43:09.680 +was around a long, long time ago + +01:43:09.680 --> 01:43:12.880 +and that it has a cousin, actually a sister or a brother. + +01:43:12.880 --> 01:43:14.200 +It was an ancient gene duplication + +01:43:14.200 --> 01:43:16.640 +before men and mice diverged. + +01:43:17.720 --> 01:43:19.960 +And with the help of Lee Hood and David Westaway + +01:43:19.960 --> 01:43:20.760 +who was here at the time + +01:43:20.760 --> 01:43:22.320 +and later Richard Moore who came from Edinburgh + +01:43:22.320 --> 01:43:25.000 +and Patrick Tremblay who came from Montreal, + +01:43:25.000 --> 01:43:26.560 +we were able to unravel this mystery. + +01:43:26.560 --> 01:43:28.240 +So here's the prion protein gene + +01:43:28.240 --> 01:43:30.320 +and 16,000 bases downstream, + +01:43:30.320 --> 01:43:31.680 +we predicted there was a new gene + +01:43:31.680 --> 01:43:33.520 +which would be for the doppel protein. + +01:43:34.600 --> 01:43:36.480 +And when we began to look at this gene, it became... + +01:43:36.480 --> 01:43:37.440 +And so what are they doing? + +01:43:37.440 --> 01:43:39.240 +They're looking for gene homology, right? + +01:43:39.240 --> 01:43:40.800 +And so this could be a duplication, + +01:43:40.800 --> 01:43:42.080 +this could be any number of things + +01:43:42.080 --> 01:43:43.800 +that happens in our genome. + +01:43:43.800 --> 01:43:46.040 +But they're starting to make an argument + +01:43:46.040 --> 01:43:50.360 +that some of these motifs may or may not be other proteins + +01:43:50.360 --> 01:43:55.040 +in the genome with potential to be infectious. + +01:43:55.040 --> 01:43:57.120 +And clear that there were many common features + +01:43:57.120 --> 01:43:58.800 +and that helix A existed in both, + +01:43:58.800 --> 01:43:59.920 +helix B existed in both, + +01:43:59.920 --> 01:44:01.560 +and helix C existed in both. + +01:44:01.560 --> 01:44:02.760 +But there were differences. + +01:44:02.760 --> 01:44:05.000 +When we looked at individual amino acids one by one, + +01:44:05.000 --> 01:44:08.240 +there were only 25% that shared sequence homology + +01:44:08.240 --> 01:44:09.600 +that were identical. + +01:44:09.600 --> 01:44:10.920 +But when we looked at the structure, + +01:44:10.920 --> 01:44:13.160 +and this is with the work of Jane Dyson and Peter Wright + +01:44:13.320 --> 01:44:14.240 +and they're post-cycled. + +01:44:14.240 --> 01:44:16.840 +It's one existed in both and helix C existed in both. + +01:44:16.840 --> 01:44:17.640 +But there were differences. + +01:44:17.640 --> 01:44:22.000 +So helix C and helix B existed in both. + +01:44:22.000 --> 01:44:23.480 +Listen to him say it. + +01:44:23.480 --> 01:44:25.440 +And when we began to look at this gene + +01:44:25.440 --> 01:44:27.800 +it became clear that there were many common features + +01:44:27.800 --> 01:44:29.480 +and that helix A existed in both, + +01:44:29.480 --> 01:44:30.560 +helix B existed in both, + +01:44:30.560 --> 01:44:32.040 +and helix C existed in both. + +01:44:32.040 --> 01:44:33.360 +Okay, they exist in both. + +01:44:33.360 --> 01:44:36.600 +So both of these proteins apparently have the same sequence + +01:44:36.600 --> 01:44:38.560 +because otherwise that alpha helix, + +01:44:38.560 --> 01:44:40.800 +you wouldn't call it alpha helix A + +01:44:40.800 --> 01:44:43.040 +if it's not the same sequence of amino acids + +01:44:43.080 --> 01:44:45.680 +because it would be a different alpha helix, right? + +01:44:45.680 --> 01:44:46.840 +But there were differences. + +01:44:46.840 --> 01:44:49.080 +When we looked at individual amino acids one by one, + +01:44:49.080 --> 01:44:52.480 +there were only 25% that shared sequence homology. + +01:44:52.480 --> 01:44:54.800 +So if there are only 25 amino acids + +01:44:54.800 --> 01:44:56.320 +that share sequence homology, + +01:44:56.320 --> 01:45:00.200 +then how can you call it sequence the same sequence? + +01:45:00.200 --> 01:45:02.840 +It's not alpha helix A. + +01:45:02.840 --> 01:45:04.160 +It's not alpha helix B. + +01:45:04.160 --> 01:45:05.780 +It's not alpha helix C. + +01:45:07.520 --> 01:45:10.560 +Because we know that proteins use these motifs, + +01:45:10.560 --> 01:45:14.880 +alpha helix, beta pleated sheet, beta helix, + +01:45:14.880 --> 01:45:17.040 +as standard motifs. + +01:45:17.040 --> 01:45:19.680 +They're not so different these 20 amino acids + +01:45:19.680 --> 01:45:22.200 +that they don't fold similarly + +01:45:22.200 --> 01:45:25.000 +depending on their main attribute, + +01:45:25.000 --> 01:45:29.440 +which is hydrophobicity or hydrophilic. + +01:45:29.440 --> 01:45:32.440 +They either like to be near the polar solvents of water + +01:45:32.440 --> 01:45:34.160 +or they wanna fold away from it. + +01:45:37.320 --> 01:45:39.520 +And so again, those amino acids + +01:45:39.520 --> 01:45:41.520 +make a huge difference to the propensity + +01:45:41.520 --> 01:45:45.120 +by which that alpha helix curls up tightly away from water + +01:45:45.120 --> 01:45:47.200 +and bends to the left or the right. + +01:45:47.200 --> 01:45:49.520 +These differences are huge. + +01:45:49.520 --> 01:45:53.240 +And he says there's only 25% homology between these, + +01:45:53.240 --> 01:45:57.200 +but I mean, we'll still call them alpha helix A, B and C. + +01:45:57.200 --> 01:46:00.880 +Holy cow, this goes against the actual principle, + +01:46:00.880 --> 01:46:03.920 +the very foundational understanding that we have + +01:46:03.920 --> 01:46:08.000 +that the amino acid sequence has something directly to do + +01:46:08.000 --> 01:46:12.760 +with the potential structure and function of the protein. + +01:46:12.760 --> 01:46:14.080 +We're identical. + +01:46:14.080 --> 01:46:15.400 +But when we looked at the structure, + +01:46:15.400 --> 01:46:17.680 +and this is with the work of Jane Dyson and Peter Wright + +01:46:17.680 --> 01:46:19.920 +and their postdoc, Wapping Mo, at Scripps, + +01:46:19.920 --> 01:46:21.480 +comparing this to the structure done in Zurich + +01:46:21.480 --> 01:46:23.000 +by Kurt Vootrick that I mentioned earlier. + +01:46:23.000 --> 01:46:25.440 +So this is mouse doppel and mouse PRP. + +01:46:25.440 --> 01:46:27.440 +So what you see is that over time, + +01:46:27.440 --> 01:46:30.760 +the structures were preserved, here's helix A, helix A, + +01:46:30.760 --> 01:46:34.040 +helix B, helix C, helix B, and helix C. + +01:46:34.040 --> 01:46:36.200 +And yet, the sequence is diverged enormously. + +01:46:36.240 --> 01:46:38.040 +75% of the residues are different, + +01:46:38.040 --> 01:46:40.000 +but the overall structures are extremely similar. + +01:46:40.000 --> 01:46:42.320 +They're not arguing that those overall structures + +01:46:42.320 --> 01:46:46.120 +would suggest that their function is identical, but they are. + +01:46:46.120 --> 01:46:47.640 +Don't you see they are? + +01:46:50.400 --> 01:46:52.480 +Even though they know so little, + +01:46:52.480 --> 01:46:55.360 +they are already making that argument. + +01:46:55.360 --> 01:46:56.840 +We don't know what either of these proteins does. + +01:46:56.840 --> 01:46:58.400 +We don't know the function of either protein. + +01:46:58.400 --> 01:46:59.960 +Ha, ha, we don't know the function + +01:46:59.960 --> 01:47:02.880 +of either of these proteins in 2002. + +01:47:02.880 --> 01:47:05.440 +Now let's turn to the human preon diseases for a moment. + +01:47:07.200 --> 01:47:08.720 +The sporadic form of these diseases + +01:47:08.720 --> 01:47:09.960 +is called Croites Valley Aqua disease. + +01:47:09.960 --> 01:47:12.760 +And this accounts for 85% of all preon disease. + +01:47:12.760 --> 01:47:15.480 +The inherited forms account for 10% to 15% + +01:47:15.480 --> 01:47:17.120 +and are called Gerson stories are shanker disease, + +01:47:17.120 --> 01:47:19.200 +familial CJD, and fatal familial insomnia. + +01:47:19.200 --> 01:47:20.520 +These are autosomal dominant diseases, + +01:47:20.520 --> 01:47:22.000 +so half of the family members are afflicted + +01:47:22.000 --> 01:47:23.080 +if they live long enough. + +01:47:23.080 --> 01:47:25.880 +And the infectious forms of these diseases are very rare. + +01:47:25.880 --> 01:47:28.120 +And they include kuru among new-getting natives. + +01:47:28.120 --> 01:47:31.080 +It was transmitted by ritualistic cannibalism. + +01:47:31.080 --> 01:47:33.240 +Iatrogenic Croites Valley Aqua disease caused by growth hormone + +01:47:33.240 --> 01:47:34.680 +derived from human to doitaries. + +01:47:34.680 --> 01:47:37.520 +And I'll talk much more about new variant CJD in Britain. + +01:47:37.520 --> 01:47:39.760 +See, so they gave growth hormone to people + +01:47:39.760 --> 01:47:42.440 +and then they got CJD. + +01:47:42.440 --> 01:47:45.120 +So is that an immune reaction or is that a protein + +01:47:45.120 --> 01:47:46.720 +that you can only have one of? + +01:47:53.800 --> 01:47:57.360 +I don't know, I'm still skeptical + +01:47:57.360 --> 01:47:59.600 +of these things all being one phenomenon + +01:47:59.600 --> 01:48:02.800 +that is protein misfolding out of control. + +01:48:02.800 --> 01:48:04.520 +Hormone derived from human to doitaries. + +01:48:04.520 --> 01:48:08.680 +And I'll talk much more about new variant CJD in Britain. + +01:48:08.680 --> 01:48:10.760 +Sporadic CJD is a disease of older people. + +01:48:10.760 --> 01:48:12.320 +So the peak is around 70 years of age. + +01:48:12.320 --> 01:48:14.080 +This is Larry Schoenberger's data from the CDC. + +01:48:14.080 --> 01:48:15.960 +It's a 20-year experience in the United States. + +01:48:15.960 --> 01:48:19.400 +So very few young people develop sporadic CJD. + +01:48:19.400 --> 01:48:20.880 +Now, the genetic forms of this disease + +01:48:20.880 --> 01:48:23.000 +have a fascinating history because the first reports + +01:48:23.000 --> 01:48:25.360 +of Croites Valley Aqua disease were in 1921. + +01:48:25.360 --> 01:48:26.760 +And 10 years later, the first pedigree + +01:48:26.760 --> 01:48:28.560 +was brought of familial CJD. + +01:48:28.560 --> 01:48:30.840 +And in 1973, Ray Rus was currently + +01:48:30.840 --> 01:48:32.720 +chair in neurology at the University of Chicago, + +01:48:32.720 --> 01:48:34.280 +working with Carlton Gajosecond Joe Gibbs + +01:48:34.280 --> 01:48:36.600 +at the NIH, who did all the early work on Kuru. + +01:48:36.600 --> 01:48:40.160 +And all the transmission work on Kuru and CJD into apes and monkeys. + +01:48:40.160 --> 01:48:41.720 +They three of them wrote a paper. + +01:48:41.720 --> 01:48:43.400 +And they reported the first familial cases + +01:48:43.400 --> 01:48:45.840 +transmitted into non-human primates. + +01:48:45.840 --> 01:48:46.960 +They offered three explanations. + +01:48:46.960 --> 01:48:49.520 +First, that the CJD virus, as they thought it was in 1973. + +01:48:49.520 --> 01:48:50.520 +They thought it was a virus. + +01:48:50.520 --> 01:48:53.000 +Was transmitted among family members living in close proximity. + +01:48:53.000 --> 01:48:54.560 +Second, there was a genetic predisposition + +01:48:54.560 --> 01:48:57.600 +to a ubiquitous CJD virus that was everywhere, like the ether. + +01:48:57.600 --> 01:48:59.320 +And third, like in AIDS, there was + +01:48:59.320 --> 01:49:00.840 +vertical transmission of the CJD virus + +01:49:00.840 --> 01:49:02.160 +from parent to offspring. + +01:49:02.160 --> 01:49:04.760 +So it turned out that all three explanations were wrong. + +01:49:04.760 --> 01:49:07.320 +And in 1989, Karen Chow, who had just + +01:49:07.320 --> 01:49:08.520 +finished her neurology residency here + +01:49:08.520 --> 01:49:10.560 +and came to work with me as a postdoctoral fellow, + +01:49:10.560 --> 01:49:13.160 +identified first gene mutation in the PRPG + +01:49:13.160 --> 01:49:17.080 +in a patient who had died of GSS at UCLA. + +01:49:17.080 --> 01:49:18.960 +And from that, it became very clear + +01:49:18.960 --> 01:49:24.320 +that this mutant form of PRPC refolds into PRP scraping. + +01:49:24.320 --> 01:49:26.400 +It became readily apparent that it + +01:49:26.400 --> 01:49:29.440 +misfolds into PRP scraping. + +01:49:29.440 --> 01:49:31.040 +That's a very bold statement. + +01:49:31.040 --> 01:49:34.960 +And we're going to have to find that paper. + +01:49:34.960 --> 01:49:36.520 +It's a 1989 paper. + +01:49:47.120 --> 01:49:52.160 +That this mutant form of PRPC refolds into PRP scraping. + +01:49:52.160 --> 01:49:54.320 +Now, subsequent to this, this became an industry. + +01:49:54.320 --> 01:49:56.320 +And there are more than 30 gene mutations + +01:49:56.320 --> 01:49:58.240 +that have been identified, all of them above the line here, + +01:49:58.240 --> 01:50:01.360 +that cause these inherited forms of the disease. + +01:50:01.360 --> 01:50:04.120 +We've also identified what are called polymorphisms. + +01:50:04.120 --> 01:50:07.080 +And those are shown below the line. + +01:50:07.080 --> 01:50:10.280 +This one right here, and this one right here in sheep, + +01:50:10.280 --> 01:50:13.160 +protect people from CJD or from scraping. + +01:50:13.160 --> 01:50:14.880 +And we've been doing a lot of drug development + +01:50:14.880 --> 01:50:17.120 +with Fred Cohn and one of his postdoctoral fellows, + +01:50:17.120 --> 01:50:17.880 +Marty May. + +01:50:17.880 --> 01:50:22.040 +So he's suggesting that some of these single point mutations + +01:50:22.040 --> 01:50:25.080 +here can protect people from the misfolding disease, which + +01:50:25.080 --> 01:50:27.600 +is a pretty remarkable statement that I + +01:50:27.600 --> 01:50:29.160 +can't necessarily dispute. + +01:50:29.160 --> 01:50:32.760 +And he's suggesting that these are all associated + +01:50:32.760 --> 01:50:35.880 +with familial disease. + +01:50:35.880 --> 01:50:43.880 +I'm still at least skeptical. + +01:50:43.880 --> 01:50:45.480 +And we've been doing a lot of drug development + +01:50:45.480 --> 01:50:48.920 +with Fred Cohn and one of his postdoctoral fellows, Marty May. + +01:50:48.920 --> 01:50:50.680 +Now, let me turn to Prian's and Mad Cows + +01:50:50.680 --> 01:50:52.000 +and the transmission to people, which + +01:50:52.000 --> 01:50:53.800 +has really created a public health crisis that threatens + +01:50:53.800 --> 01:50:56.920 +the food supply as well as the blood supply worldwide. + +01:50:56.920 --> 01:51:00.800 +Just yesterday, Poland joined the food and blood supplies + +01:51:00.800 --> 01:51:02.240 +an interesting statement to make. + +01:51:02.240 --> 01:51:04.880 +It's definitely relevant for public health, then, isn't it? + +01:51:04.880 --> 01:51:07.800 +Countries of the BSE club, the first case. + +01:51:07.800 --> 01:51:10.440 +Now, we believe that BSE arose through industrial cannibalism + +01:51:10.440 --> 01:51:11.840 +in the late 1970s. + +01:51:11.840 --> 01:51:13.800 +And there's an argument whether it arose spontaneously + +01:51:13.800 --> 01:51:16.920 +in a cow, perhaps a mutation, or it came from sheep + +01:51:16.920 --> 01:51:18.600 +where scrapies endemic, but nevertheless, + +01:51:18.600 --> 01:51:22.120 +it recycled through cows and then into humans. + +01:51:22.120 --> 01:51:24.680 +Here's a typical newspaper clipping. + +01:51:24.680 --> 01:51:26.320 +And it's not only a British problem. + +01:51:26.320 --> 01:51:28.720 +This is also a problem throughout the con. + +01:51:28.720 --> 01:51:30.040 +And while there are over 100 cases, + +01:51:30.040 --> 01:51:32.000 +a new variant CJD in young people in Britain, + +01:51:32.000 --> 01:51:35.360 +there are now four cases in France. + +01:51:35.360 --> 01:51:37.520 +This is the BSE epidemic that begins in 1986 + +01:51:37.520 --> 01:51:39.800 +with the discovery of the disease by Gerald Wells. + +01:51:39.800 --> 01:51:40.560 +It takes around 19 years. + +01:51:40.560 --> 01:51:43.240 +So cannibalism would imply something + +01:51:43.240 --> 01:51:45.840 +other than protein misfolding, right? + +01:51:45.840 --> 01:51:49.240 +Because if you feed healthy cattle, + +01:51:49.240 --> 01:51:51.560 +the remains of healthy cattle, that + +01:51:51.560 --> 01:51:54.200 +doesn't mean that you're giving them an infectious disease + +01:51:54.200 --> 01:51:56.080 +unless they already had it. + +01:51:56.080 --> 01:52:00.160 +So for me, this implies a autoimmune reaction. + +01:52:00.160 --> 01:52:04.200 +It implies autoimmunity caused by eating stuff + +01:52:04.200 --> 01:52:05.720 +that's too closely related to you + +01:52:05.720 --> 01:52:08.240 +and having that go through your payer's patches + +01:52:08.240 --> 01:52:10.880 +and cause a four alarm fire. + +01:52:10.880 --> 01:52:13.840 +That seems a lot more understandable + +01:52:13.840 --> 01:52:17.080 +and more likely than the idea that, + +01:52:17.080 --> 01:52:18.680 +again, we're dealing with something + +01:52:18.680 --> 01:52:21.360 +where you get the wrong amino acids in the wrong order + +01:52:21.360 --> 01:52:24.200 +and it's just one of those is all it's needed + +01:52:24.200 --> 01:52:26.680 +and you're eventually gonna be dead. + +01:52:26.680 --> 01:52:29.440 +That's an amazing mythology that I don't think + +01:52:29.440 --> 01:52:32.880 +any of this past talk has done anything to substantiate. + +01:52:32.880 --> 01:52:35.640 +92 with almost 40,000 cases + +01:52:35.640 --> 01:52:37.400 +and then the number of cases keep declining. + +01:52:37.400 --> 01:52:38.600 +But we keep finding more and more cases + +01:52:38.600 --> 01:52:40.520 +because more and more sensitive assays are being used + +01:52:40.520 --> 01:52:42.480 +or measurement techniques to detect the prions. + +01:52:42.480 --> 01:52:44.600 +All of these animals were clinically ill + +01:52:44.600 --> 01:52:46.720 +than at least 36 months of age. + +01:52:46.720 --> 01:52:50.800 +This represents, and these little balls or circles + +01:52:50.800 --> 01:52:53.120 +are represent one case, whereas each square + +01:52:53.120 --> 01:52:56.360 +represents a thousand cases on the y-axis. + +01:52:56.360 --> 01:52:59.920 +So what we're seeing is that there is this continuous + +01:52:59.920 --> 01:53:01.440 +increase in the number of cases per year, + +01:53:01.440 --> 01:53:03.600 +this is the yearly numbers of new variants, CJD, + +01:53:03.600 --> 01:53:05.600 +except in 2001, the numbers down here. + +01:53:05.600 --> 01:53:08.200 +What it'll be in 2002, we don't know. + +01:53:08.200 --> 01:53:10.000 +So we see that this disease appears about a decade + +01:53:10.000 --> 01:53:11.160 +after the beginning of BSE, + +01:53:11.160 --> 01:53:13.240 +so we think the incubation time exceeds one decade + +01:53:13.240 --> 01:53:15.720 +and in from crew studies, we know it can be up to four decades. + +01:53:15.720 --> 01:53:18.080 +Wow, so we could be waiting right now + +01:53:18.080 --> 01:53:20.240 +for it to happen to all of us is what is impre- + +01:53:20.240 --> 01:53:21.800 +what is implication is. + +01:53:21.880 --> 01:53:23.920 +That's pretty cool because if it happens + +01:53:23.920 --> 01:53:26.720 +from a transfection that's 20 years in the future, + +01:53:26.720 --> 01:53:29.320 +he will have already predicted it's gonna occur. + +01:53:33.080 --> 01:53:36.360 +Boy, that's creating a pretty wide envelope + +01:53:36.360 --> 01:53:39.760 +for naturally occurring prion disease + +01:53:39.760 --> 01:53:43.820 +to just show up and mass across the population, isn't it? + +01:53:44.760 --> 01:53:47.960 +Oh man, we shouldn't have been eating those cows 20 years ago. + +01:53:47.960 --> 01:53:51.160 +Right, we shouldn't have been eating all that beef 20 years ago. + +01:53:52.800 --> 01:53:54.760 +Wow, I can't believe it. + +01:53:54.760 --> 01:53:56.640 +We better stop eating beef all together. + +01:53:56.640 --> 01:53:58.560 +Maybe we should just eat fake beef + +01:53:59.520 --> 01:54:02.280 +because I mean, prions take 40 years to incubate. + +01:54:02.280 --> 01:54:05.440 +Obviously, the reason why everybody's dying of prions now + +01:54:05.440 --> 01:54:07.640 +is because we've been eating meat for 40 years. + +01:54:07.640 --> 01:54:09.200 +Just listen to this talk. + +01:54:13.360 --> 01:54:16.000 +This is an MRI scan and it shows, + +01:54:16.000 --> 01:54:17.120 +if we didn't have that light on over there, + +01:54:17.120 --> 01:54:18.280 +you could see this much better, + +01:54:18.280 --> 01:54:19.880 +this hyper-density is what it's called ate + +01:54:19.880 --> 01:54:21.240 +and all the way down into the pulvinar, + +01:54:22.080 --> 01:54:24.280 +which is this dorsal nucleus of the thalamus + +01:54:24.280 --> 01:54:26.680 +and this is called a hockey stick sign. + +01:54:26.680 --> 01:54:29.160 +And that's very typical of patients with new variant CJD. + +01:54:29.160 --> 01:54:32.120 +Patients with sporadic CJD have all these hyper-intensities + +01:54:32.120 --> 01:54:32.960 +that are seen bilaterally, + +01:54:32.960 --> 01:54:35.360 +but they don't have the lighting up of the pulvinar. + +01:54:37.680 --> 01:54:39.400 +I said I would show you some of these analog plaques + +01:54:39.400 --> 01:54:40.880 +and in patients with new variant CJD + +01:54:40.880 --> 01:54:42.720 +and this is the work of Steve D'Armond again + +01:54:42.720 --> 01:54:43.960 +with slides sent to, I should say, + +01:54:43.960 --> 01:54:45.840 +sessions sent to us by Bob Will and Jim Ironside + +01:54:45.840 --> 01:54:46.680 +and Edinburgh. + +01:54:46.680 --> 01:54:48.040 +So these are the plaques around by this halo + +01:54:48.040 --> 01:54:49.360 +of sponge-formed generation. + +01:54:49.360 --> 01:54:52.680 +They stain with antibodies to the prion protein very intensely. + +01:54:55.800 --> 01:54:59.480 +So which antibody, how do you raise an antibody + +01:54:59.480 --> 01:55:01.640 +to the prion protein if it's in that form? + +01:55:01.640 --> 01:55:04.480 +You reuse the one that binds to the parts + +01:55:04.480 --> 01:55:05.280 +that are still there. + +01:55:05.280 --> 01:55:08.160 +You don't have to be specific about that at all right now. + +01:55:08.160 --> 01:55:10.320 +Given the fact that just 20 minutes ago + +01:55:10.320 --> 01:55:12.400 +you were using antibodies as an indicator + +01:55:12.400 --> 01:55:15.240 +of whether it was prion protein in a natural form + +01:55:15.240 --> 01:55:16.760 +or in the infectious form, + +01:55:16.760 --> 01:55:19.200 +don't you think you could have used that same technique + +01:55:19.200 --> 01:55:22.560 +to show what fraction of that protein was in + +01:55:22.560 --> 01:55:24.720 +in a badly folded form + +01:55:24.720 --> 01:55:27.720 +or in a dangerous folded form? + +01:55:27.720 --> 01:55:29.840 +Or do you just think that he wasn't clever enough + +01:55:29.840 --> 01:55:32.840 +to ask that question with the tools that he already had? + +01:55:32.840 --> 01:55:35.840 +BUZZER. + +01:55:35.840 --> 01:55:38.520 +This is why they hate me to be in the audience of these talks, + +01:55:38.520 --> 01:55:41.160 +why they hated me in the audience of these talks. + +01:55:41.160 --> 01:55:42.360 +Oh, very intensely. + +01:55:45.840 --> 01:55:47.400 +I've already said this seven times. + +01:55:49.960 --> 01:55:52.200 +Now, one wanted to try to figure out + +01:55:52.200 --> 01:55:53.360 +where did all of this start? + +01:55:53.360 --> 01:55:54.560 +Did it really start in sheep? + +01:55:54.560 --> 01:55:55.760 +Did it start in cows? + +01:55:55.760 --> 01:55:57.240 +And did it come from the cows to the humans? + +01:55:57.240 --> 01:55:58.440 +And if we were looking for a virus, + +01:55:58.440 --> 01:55:59.920 +we could do this very easily. + +01:55:59.920 --> 01:56:01.080 +The virus would be simple to track + +01:56:01.080 --> 01:56:02.320 +because the virus would have a nucleic acid, + +01:56:02.320 --> 01:56:03.720 +a piece of DNA or RNA, + +01:56:03.720 --> 01:56:07.000 +and we could easily track it into the sheep and see. + +01:56:07.000 --> 01:56:11.000 +Viruses are easy to track according to Stanley Prusen + +01:56:11.000 --> 01:56:14.200 +or easy, I mean, come on, that's easy work. + +01:56:14.200 --> 01:56:16.040 +Easy work. + +01:56:17.040 --> 01:56:20.440 +BUZZER. + +01:56:20.440 --> 01:56:22.200 +And it had nothing to do with the sheep, + +01:56:22.200 --> 01:56:24.400 +but that same piece of DNA or RNA + +01:56:24.400 --> 01:56:26.600 +appeared in the cow and then appeared in the human. + +01:56:26.600 --> 01:56:27.480 +We can't do that. + +01:56:27.480 --> 01:56:28.920 +So what we had to do was really begin + +01:56:28.920 --> 01:56:30.640 +to look at what are called prion strains. + +01:56:30.640 --> 01:56:32.120 +And what's happening is that PRP scrape + +01:56:32.120 --> 01:56:33.280 +is signified by a square. + +01:56:33.280 --> 01:56:35.600 +It's stimulating the conversion of bovine PRPC. + +01:56:35.600 --> 01:56:38.920 +This is from the sheep into PRP scrapey of the cow. + +01:56:38.920 --> 01:56:41.200 +And the same thing, we can reiterate this into the human. + +01:56:41.200 --> 01:56:42.400 +Now, there's a lot of data, but I'm + +01:56:42.400 --> 01:56:43.840 +only going to show you very little. + +01:56:43.840 --> 01:56:45.360 +And what we did was to use transgenic mice + +01:56:45.440 --> 01:56:47.240 +to investigate the origin of BSE + +01:56:47.240 --> 01:56:48.720 +and the transmission of prions to humans. + +01:56:48.720 --> 01:56:50.680 +Oh, so we're going to use an animal model + +01:56:50.680 --> 01:56:54.280 +to understand the cartoon that he says definitely happens, + +01:56:54.280 --> 01:56:58.840 +that the sheep protein causes the refolding of the cow protein + +01:56:58.840 --> 01:57:01.800 +that causes the refolding of the human protein. + +01:57:01.800 --> 01:57:03.920 +There's three assumptions there on top + +01:57:03.920 --> 01:57:05.920 +of three different cartoons. + +01:57:05.920 --> 01:57:07.320 +And yet somehow or another, we're + +01:57:07.320 --> 01:57:09.800 +going to move right on to a transgenic animal model. + +01:57:09.800 --> 01:57:10.440 +Let's hear it. + +01:57:10.440 --> 01:57:11.440 +Let's hear it. + +01:57:11.440 --> 01:57:12.400 +Now, there's a lot of data, but I'm + +01:57:12.400 --> 01:57:13.920 +only going to show you very little. + +01:57:13.920 --> 01:57:15.400 +And what we did was to use transgenic mice + +01:57:15.400 --> 01:57:17.760 +to investigate the origin of BSE and the transmission + +01:57:17.760 --> 01:57:18.960 +of prions to humans. + +01:57:18.960 --> 01:57:20.480 +There's now compelling evidence that indicates + +01:57:20.480 --> 01:57:22.720 +that prions and beef products were ingested by humans, who + +01:57:22.720 --> 01:57:26.480 +later developed variant CJD. + +01:57:26.480 --> 01:57:29.000 +What we began with, and this is Jim Mastrioni's work, + +01:57:29.000 --> 01:57:31.920 +Glenn Telling's work, was what we call a humanized mouse. + +01:57:31.920 --> 01:57:34.800 +So now this mouse behaves toward prions like a human being. + +01:57:34.800 --> 01:57:37.320 +And when we take sporadic CJD or familial CJD + +01:57:37.320 --> 01:57:38.880 +and transmit into the mice brain extracts + +01:57:38.880 --> 01:57:40.520 +from people who have died of these diseases, + +01:57:40.520 --> 01:57:42.520 +all of the mice get sick in about 200 days. + +01:57:42.520 --> 01:57:45.000 +But when we took new variant CJD from young people in England, + +01:57:45.000 --> 01:57:46.880 +people in their early 20s or late teens, + +01:57:46.880 --> 01:57:49.920 +we found that about 25%, this is wrong, it's not 60%, + +01:57:49.920 --> 01:57:53.080 +but about 25% of the mice became ill by 500 days. + +01:57:53.080 --> 01:57:56.320 +And when we took brain extracts from cows with BSE, + +01:57:56.320 --> 01:57:59.120 +none of the mice developed disease by 600 days. + +01:57:59.120 --> 01:58:00.240 +Now, Mike Scott did an experiment + +01:58:00.240 --> 01:58:01.760 +where he created bovinized mice. + +01:58:01.760 --> 01:58:03.440 +So these are mice that behave like cows + +01:58:03.440 --> 01:58:04.760 +with respected prions. + +01:58:04.760 --> 01:58:07.800 +And when he took mad cows, brain, 240 days, + +01:58:07.800 --> 01:58:08.840 +and all the mice were sick. + +01:58:08.840 --> 01:58:10.560 +With new variant CJD, what he found + +01:58:10.560 --> 01:58:13.080 +was that 100% of the mice developed disease + +01:58:13.080 --> 01:58:15.240 +at 260 days after inoculation. + +01:58:15.240 --> 01:58:16.360 +So this was quite a shock. + +01:58:16.360 --> 01:58:18.440 +And that when we took sheep's scrapie from the United States, + +01:58:18.440 --> 01:58:23.200 +the mice were even more susceptible, 210 days, 100% were sick. + +01:58:23.200 --> 01:58:27.400 +Now, we would have to look at what this mouse model is. + +01:58:27.400 --> 01:58:32.120 +I don't know if they have overexpression of the bovine protein + +01:58:32.120 --> 01:58:34.440 +in their brain or the human protein in their brain. + +01:58:34.440 --> 01:58:35.920 +That's my guess. + +01:58:35.920 --> 01:58:40.160 +My guess is that this time, I'm sure + +01:58:40.160 --> 01:58:41.000 +I'm right. + +01:58:41.000 --> 01:58:43.800 +Okay, I would bet my, I bet everything on it. + +01:58:43.800 --> 01:58:46.160 +In 2002, and when this was done, + +01:58:46.160 --> 01:58:48.160 +it would have been before 2002, + +01:58:48.160 --> 01:58:50.440 +there would have been no way for them to knock out + +01:58:50.440 --> 01:58:53.520 +the native gene and replace it with the bovine gene. + +01:58:53.520 --> 01:58:56.040 +So what they're most likely working with here + +01:58:56.040 --> 01:58:58.840 +is a mouse that also expresses the bovine + +01:59:00.440 --> 01:59:05.320 +prion protein driven by some genetic technology. + +01:59:05.320 --> 01:59:06.880 +And then when they inject that animal + +01:59:06.880 --> 01:59:08.320 +in the brain, it gets sick. + +01:59:11.160 --> 01:59:15.960 +And so we would have to, again, look into these papers + +01:59:15.960 --> 01:59:16.800 +and figure it out. + +01:59:16.800 --> 01:59:21.000 +But my guess is that this isn't gonna be as clean a result + +01:59:21.000 --> 01:59:24.160 +as what he's suggesting it is here, + +01:59:24.160 --> 01:59:27.800 +simply because the experimental model is not as clean + +01:59:27.800 --> 01:59:29.320 +as he wants you to believe it is. + +01:59:29.320 --> 01:59:34.320 +It's a bovine, it responds like a cow with respect to prion, + +01:59:37.000 --> 01:59:39.120 +it responds like a human with respect to prion. + +01:59:39.120 --> 01:59:41.640 +You can hear him say it again if you want to. + +01:59:41.640 --> 01:59:43.040 +Into the mice, brain extracts from people + +01:59:43.040 --> 01:59:44.320 +who've died of these diseases. + +01:59:44.320 --> 01:59:46.320 +All of the mice get sick in about 200 days. + +01:59:46.320 --> 01:59:48.800 +But when we took new variants, CJD from young people in England, + +01:59:48.800 --> 01:59:50.720 +people in the early 20s, late teens, + +01:59:50.720 --> 01:59:53.760 +we found that about 25%, this is wrong, it's not 60%, + +01:59:53.760 --> 01:59:56.880 +but about 25% of the mice became ill by 500 days. + +01:59:56.880 --> 02:00:00.120 +And when we took brain extracts from cows with BSE, + +02:00:00.120 --> 02:00:02.960 +none of the mice developed disease by 600 days. + +02:00:02.960 --> 02:00:04.120 +Now Mike Scott did an experiment + +02:00:04.120 --> 02:00:05.600 +where he created bovineized mice. + +02:00:05.600 --> 02:00:07.280 +So these are mice that behave like cows + +02:00:07.280 --> 02:00:08.120 +with respect to prions. + +02:00:08.120 --> 02:00:10.080 +See bovineized mice and then again, + +02:00:10.080 --> 02:00:11.640 +they're injecting into the brain. + +02:00:11.640 --> 02:00:16.640 +Again, how does that equivalent to eating your dead relative? + +02:00:17.080 --> 02:00:20.920 +How does that equivalent to eating another cow? + +02:00:20.920 --> 02:00:24.480 +It's just all not the experimental model that you want it to be + +02:00:24.480 --> 02:00:26.400 +if you want to understand what he was showing + +02:00:26.400 --> 02:00:28.040 +in the previous slides. + +02:00:28.040 --> 02:00:31.080 +And when he took mad cows, brain, 240 days + +02:00:31.080 --> 02:00:32.120 +and all the mice were sick. + +02:00:32.120 --> 02:00:35.040 +With new variants, CJD, what he found was that 100% + +02:00:35.040 --> 02:00:37.360 +of the mice developed disease at 260 days + +02:00:37.360 --> 02:00:38.480 +after inoculation. + +02:00:38.480 --> 02:00:39.600 +So this was quite a shock. + +02:00:39.600 --> 02:00:41.680 +And that when we took sheep's scrapie from the United States, + +02:00:41.680 --> 02:00:45.080 +the mice were even more susceptible, 210 days, 100% were sick. + +02:00:46.480 --> 02:00:47.600 +And we looked at the neuropathology + +02:00:47.600 --> 02:00:49.040 +against Stevie Arman's work. + +02:00:49.040 --> 02:00:51.480 +What Steve found was that if we began with sheep's scrapie, + +02:00:51.480 --> 02:00:53.360 +there was very little of the prion protein deposited + +02:00:53.360 --> 02:00:54.480 +in what's called the corpus callosum, + +02:00:54.480 --> 02:00:56.520 +the structure that connects both sides of your brain. + +02:00:56.520 --> 02:00:58.400 +And some people with intractable epilepsy, + +02:00:58.400 --> 02:00:59.480 +this structure is cut. + +02:01:01.520 --> 02:01:04.080 +When we looked, if we began with BSE in these bovineized mice, + +02:01:04.080 --> 02:01:05.680 +we saw large amounts of the prion protein + +02:01:05.680 --> 02:01:07.160 +deposited in these big plaques. + +02:01:07.160 --> 02:01:08.640 +And if we took new variant CJD, + +02:01:08.640 --> 02:01:10.040 +the image was indistinguishable. + +02:01:11.040 --> 02:01:12.360 +So when we put all this together, + +02:01:12.360 --> 02:01:14.040 +the indistinguishable neuropathology, + +02:01:14.040 --> 02:01:15.440 +the very similar incubation times, + +02:01:15.440 --> 02:01:17.200 +it became very clear that there was little doubt + +02:01:17.200 --> 02:01:19.400 +that the people had become sick from the cows. + +02:01:19.400 --> 02:01:20.960 +And that's in addition to all the human epidemiology + +02:01:20.960 --> 02:01:23.080 +showing the disease has confined largely to Great Britain + +02:01:23.080 --> 02:01:24.680 +with four cases in France, as I said, + +02:01:24.680 --> 02:01:26.280 +one in Italy and one in Ireland. + +02:01:27.680 --> 02:01:29.480 +And now there's actually one in the United States, + +02:01:29.480 --> 02:01:31.640 +but this is a woman who, 22 years old, + +02:01:31.640 --> 02:01:33.640 +22 years old, living in Florida, + +02:01:33.640 --> 02:01:36.200 +who lived in Great Britain for nearly 10 years. + +02:01:36.200 --> 02:01:37.560 +And it's a British citizen. + +02:01:39.080 --> 02:01:40.560 +So all of this put together, + +02:01:40.560 --> 02:01:41.840 +Mike Scott began to think about this + +02:01:41.840 --> 02:01:42.760 +in a slightly different way. + +02:01:42.760 --> 02:01:44.640 +And he's begun to wonder if, in fact, + +02:01:44.640 --> 02:01:46.840 +all sheep with screpy prions, the blue ones, + +02:01:46.840 --> 02:01:50.080 +also have a few BSE prions that multiply more slowly. + +02:01:50.080 --> 02:01:51.720 +And that what happened in the late 1970s + +02:01:51.720 --> 02:01:53.080 +was that when they reduced the temperature + +02:01:53.080 --> 02:01:55.560 +and also the process, they did less severe, + +02:01:55.560 --> 02:01:57.280 +what they call the rendering process, + +02:01:57.280 --> 02:02:00.040 +the screpy prions were still destroyed, + +02:02:00.040 --> 02:02:01.720 +but the BSE prions survived. + +02:02:01.720 --> 02:02:03.760 +And now they multiplied in cattle + +02:02:03.760 --> 02:02:05.360 +where they became pathogenic for humans. + +02:02:05.360 --> 02:02:06.640 +And there were no more screpy prions, + +02:02:06.640 --> 02:02:08.000 +the blue ones, the whole down the amount + +02:02:08.000 --> 02:02:09.240 +of the red ones that could be made, + +02:02:09.240 --> 02:02:11.240 +because whether that's true or not, I don't know. + +02:02:11.240 --> 02:02:12.240 +So this is a hypothesis. + +02:02:12.240 --> 02:02:14.160 +But there's no way for that to happen + +02:02:14.160 --> 02:02:17.280 +unless you're talking about prions moving separately + +02:02:17.280 --> 02:02:22.040 +as animals breed over time and building up separately + +02:02:22.040 --> 02:02:24.600 +in the population, even though they're not passed on + +02:02:24.600 --> 02:02:25.680 +through birth. + +02:02:25.680 --> 02:02:27.600 +That's incredible. + +02:02:27.600 --> 02:02:29.760 +Or maybe they are passed, I mean, wow. + +02:02:30.760 --> 02:02:32.360 +That's a pretty funny model. + +02:02:32.360 --> 02:02:34.120 +It makes a lot of predictions that I bet + +02:02:34.120 --> 02:02:35.680 +they won't bother testing. + +02:02:35.680 --> 02:02:37.680 +We're trying to pursue this. + +02:02:37.680 --> 02:02:39.360 +So variant CJD is caused by prions + +02:02:39.360 --> 02:02:41.080 +in beef products from mad cows. + +02:02:41.080 --> 02:02:43.040 +Over 85% of all human prion diseases + +02:02:43.040 --> 02:02:45.000 +are sporadic 10 to 15% are inherited. + +02:02:45.000 --> 02:02:47.200 +So I just want to point out that while there are about 5000 + +02:02:47.200 --> 02:02:49.000 +cases of CJD across the planet each year, + +02:02:49.000 --> 02:02:51.760 +we're talking about 25 cases of variant CJD so far. + +02:02:51.760 --> 02:02:54.800 +So just to give you a little perspective on the numbers. + +02:02:54.800 --> 02:02:58.880 +Let's now turn to therapeutic approaches to prion diseases. + +02:02:58.880 --> 02:03:00.240 +So there are a large number of approaches + +02:03:00.240 --> 02:03:01.600 +that we've taken here at UCSF. + +02:03:01.600 --> 02:03:03.440 +And I won't talk about rational drug design based + +02:03:03.440 --> 02:03:05.080 +upon what is called dominant negative inhibition + +02:03:05.080 --> 02:03:05.880 +or gene therapy. + +02:03:05.880 --> 02:03:07.360 +And I'm also not going to talk about what + +02:03:07.360 --> 02:03:08.280 +is called enhanced clearance. + +02:03:08.280 --> 02:03:10.640 +But I come back to talk a little bit about clearance of prions. + +02:03:10.640 --> 02:03:11.760 +I am going to talk about antibodies + +02:03:11.760 --> 02:03:13.920 +that were used to inhibit prion replication. + +02:03:13.920 --> 02:03:16.160 +And then I'm going to talk about a drug called quinacrine + +02:03:16.160 --> 02:03:18.080 +that we're giving to patients in a study with Bruce Miller + +02:03:18.080 --> 02:03:22.240 +and Michael Geshwin and many, many other people here at UCSF. + +02:03:22.240 --> 02:03:24.320 +So this is David Parris' work using these antibodies produced + +02:03:24.320 --> 02:03:27.040 +by Dennis Burton and Anthony Williamson at Scripps. + +02:03:27.040 --> 02:03:29.200 +And again, the same blue antibodies react out here, + +02:03:29.200 --> 02:03:30.520 +more toward the N-terminus, the green ones + +02:03:30.520 --> 02:03:33.000 +at the C-terminus, and the red ones in the middle region. + +02:03:33.000 --> 02:03:36.880 +That's Helix A, Helix B, Helix C. + +02:03:36.880 --> 02:03:38.600 +All this looks a little complicated. + +02:03:38.600 --> 02:03:40.360 +But let's just look over here at this graph. + +02:03:40.360 --> 02:03:42.040 +So we're adding increasing amounts of antibody + +02:03:42.040 --> 02:03:43.040 +to cultured cells. + +02:03:43.040 --> 02:03:44.800 +And we're seeing the decrease in the amount of PRP + +02:03:44.800 --> 02:03:46.280 +scrapey, the protein of the prion. + +02:03:46.280 --> 02:03:48.240 +And the red antibodies work better than the blue antibodies + +02:03:48.240 --> 02:03:49.080 +but only a little bit. + +02:03:49.080 --> 02:03:52.440 +But both red and blue work much better than the green antibodies. + +02:03:52.440 --> 02:03:54.040 +And if we look here, now instead of having + +02:03:54.040 --> 02:03:56.280 +the concentration of antibody, it's the duration of antibodies. + +02:03:56.280 --> 02:03:59.080 +Are you at all surprised that one + +02:03:59.080 --> 02:04:00.840 +of the therapies he wanted to talk about + +02:04:00.840 --> 02:04:04.200 +was using monoclonal antibodies to fight prion disease? + +02:04:04.200 --> 02:04:05.880 +Are you at all surprised? + +02:04:05.880 --> 02:04:08.240 +This is at a time when monoclonal antibodies + +02:04:08.240 --> 02:04:15.680 +are right at the hottest biologic there is to make. + +02:04:15.680 --> 02:04:19.680 +And the IP law is ripe for the writing. + +02:04:19.680 --> 02:04:20.800 +Antibody treatment. + +02:04:20.800 --> 02:04:22.960 +So we picked one concentration of antibody. + +02:04:22.960 --> 02:04:25.600 +And now what we've done, and you see that the red ones + +02:04:25.600 --> 02:04:27.720 +work quite rapidly compared to the blue ones. + +02:04:27.720 --> 02:04:29.800 +And what this allows us to do is to look at when + +02:04:29.800 --> 02:04:32.400 +we see a half maximal decrease, when we've gone down 50%. + +02:04:32.400 --> 02:04:36.000 +And that's about at this point at about 30 hours right in here. + +02:04:36.000 --> 02:04:38.840 +It was actually IMM, UNITY. + +02:04:38.840 --> 02:04:41.880 +That's what I got, bodies anti. + +02:04:41.880 --> 02:04:45.840 +He spelled immunity not by antibodies. + +02:04:45.840 --> 02:04:48.720 +And in 1990, Bruce Cheesebrow and Byron Coe + +02:04:48.720 --> 02:04:50.240 +working at the Rocky Mountain Laboratory + +02:04:50.240 --> 02:04:52.280 +did a lot of work on the synthesis of PRPC + +02:04:52.280 --> 02:04:54.040 +and defined what is called the Half-Life for formation. + +02:04:54.040 --> 02:04:55.480 +Very rapid. + +02:04:55.480 --> 02:04:58.360 +And David Borchelt working here in San Francisco confirmed + +02:04:58.360 --> 02:05:00.560 +that and then showed that the Half-Life time for degradation + +02:05:00.560 --> 02:05:01.600 +is about six hours. + +02:05:01.600 --> 02:05:03.880 +And that PRPC was made even more slowly + +02:05:03.880 --> 02:05:05.560 +with a half-time of formation of about three to 10. + +02:05:05.560 --> 02:05:08.400 +OK, so here they're transfecting these two. + +02:05:08.400 --> 02:05:12.880 +And now PRPC grapey actually must have a different sequence + +02:05:12.880 --> 02:05:16.360 +because otherwise you're expressing the same protein. + +02:05:16.360 --> 02:05:17.960 +It gets folded in two different ways. + +02:05:17.960 --> 02:05:19.600 +So either you're trans, how can you + +02:05:19.600 --> 02:05:23.120 +transfect neuroblastoma cells with this protein + +02:05:23.120 --> 02:05:24.640 +unless it has a different sequence? + +02:05:25.480 --> 02:05:27.200 +What's going on here? + +02:05:27.200 --> 02:05:30.400 +What is going on here? + +02:05:30.400 --> 02:05:33.720 +David Borchelt working here in San Francisco confirmed that + +02:05:33.720 --> 02:05:35.480 +and then showed that the Half-Life time for degradation + +02:05:35.480 --> 02:05:36.600 +is about six hours. + +02:05:36.600 --> 02:05:38.800 +And that PRPC grapey was made even more slowly + +02:05:38.800 --> 02:05:41.080 +with a half-time of formation of about three to 10 hours. + +02:05:41.080 --> 02:05:45.000 +Now, I had thought that PRPC grapey was complete granite + +02:05:45.000 --> 02:05:46.560 +and that it was never degraded. + +02:05:46.560 --> 02:05:47.960 +But in the last slide that I showed you, + +02:05:47.960 --> 02:05:48.960 +I don't get that at all. + +02:05:48.960 --> 02:05:50.880 +This is really represents the degradation of PRPC + +02:05:50.880 --> 02:05:53.800 +the loss of prions from the culture. + +02:05:53.880 --> 02:05:56.000 +They couldn't separate the two + +02:05:56.000 --> 02:05:57.920 +because they're the same weight. + +02:05:59.080 --> 02:06:00.400 +They can't separate the two + +02:06:00.400 --> 02:06:03.320 +because they exist together in their cartoons. + +02:06:06.120 --> 02:06:08.440 +But then they shouldn't be able to exist together + +02:06:08.440 --> 02:06:11.000 +because the one causes the other to become it. + +02:06:13.640 --> 02:06:16.080 +And yet they have the same amino acid sequence. + +02:06:16.080 --> 02:06:20.800 +So how would you express one or the other in a cell culture? + +02:06:20.800 --> 02:06:22.440 +You'd have to use the same RNA. + +02:06:22.440 --> 02:06:23.440 +I don't get it. + +02:06:23.440 --> 02:06:24.320 +I don't get it. + +02:06:24.320 --> 02:06:25.880 +I don't get it. + +02:06:25.880 --> 02:06:27.280 +And so we were able to calculate a number, + +02:06:27.280 --> 02:06:28.240 +as I mentioned before, + +02:06:28.240 --> 02:06:31.640 +where 50% of the PRPC grapey disappears of about 30 hours. + +02:06:32.600 --> 02:06:33.680 +This has important implications + +02:06:33.680 --> 02:06:34.680 +for thinking about all the purposes. + +02:06:34.680 --> 02:06:35.520 +I'm just an idiot then. + +02:06:35.520 --> 02:06:36.360 +I got more reading to do. + +02:06:36.360 --> 02:06:37.200 +That's all. + +02:06:37.200 --> 02:06:39.120 +See, this is the reason why I haven't started teaching it yet + +02:06:39.120 --> 02:06:40.120 +because I've got more read. + +02:06:40.120 --> 02:06:41.400 +I don't understand that. + +02:06:42.600 --> 02:06:44.600 +And that contradiction seems pretty big. + +02:06:44.600 --> 02:06:46.960 +The whole talk has been explaining + +02:06:46.960 --> 02:06:49.200 +about how difficult it is to sort this out + +02:06:49.200 --> 02:06:50.520 +because it is a pre-... + +02:06:50.520 --> 02:06:52.840 +It is a protein that has alternate forms + +02:06:52.840 --> 02:06:55.480 +that alternate forms are hard to study. + +02:06:55.480 --> 02:06:57.000 +And yet now we can express them + +02:06:57.000 --> 02:06:59.480 +in different models when we want them. + +02:07:01.920 --> 02:07:03.120 +Tells us that cells are capable + +02:07:03.120 --> 02:07:05.640 +of degrading both PRPC and PRPC grapey. + +02:07:05.640 --> 02:07:07.400 +And it raises the question whether PRPC grapey + +02:07:07.400 --> 02:07:09.720 +is normally found at very low levels in normal cells + +02:07:09.720 --> 02:07:11.840 +and has a physiological function. + +02:07:11.840 --> 02:07:13.880 +And that is a very appealing way of thinking about all this + +02:07:13.880 --> 02:07:15.120 +because it makes much more sense + +02:07:15.120 --> 02:07:18.040 +than thinking that PRPC grapey is something totally apparent. + +02:07:18.040 --> 02:07:19.360 +It raises the question of whether or not + +02:07:19.360 --> 02:07:20.200 +we really have an issue. + +02:07:20.200 --> 02:07:22.520 +It's a kinetic race between the formation of PRPC grapey + +02:07:22.520 --> 02:07:24.960 +and the cell's ability to clear PRPC grapey. + +02:07:24.960 --> 02:07:27.240 +And that when the cell can keep up with the formation + +02:07:27.240 --> 02:07:30.720 +and clear it like it does with all other proteins in fact, + +02:07:30.720 --> 02:07:33.280 +when that happens everything is functioning fine. + +02:07:33.280 --> 02:07:35.320 +But when the cell gets out of balance, + +02:07:35.320 --> 02:07:39.200 +it can no longer clear PRPC grapey at the rate that it's formed. + +02:07:39.200 --> 02:07:40.360 +I'm talking about very, very low levels + +02:07:40.360 --> 02:07:42.880 +that we can't detect even by these animal assays. + +02:07:42.880 --> 02:07:45.000 +Then something goes awry and we begin to accumulate + +02:07:45.000 --> 02:07:47.440 +more and more PRPC grapey and eventually the animal gets sick + +02:07:47.440 --> 02:07:49.320 +and goes on to die or the human being. + +02:07:50.200 --> 02:07:53.000 +Now, I promised you a little chemistry at the end. + +02:07:53.000 --> 02:07:54.480 +And if you just look down here at chloropromacy + +02:07:54.480 --> 02:07:55.920 +and this is thoracine, this is one of the first + +02:07:55.920 --> 02:07:57.600 +antipsychotic drugs and this is the structure of it + +02:07:57.600 --> 02:07:59.200 +and it has these three rings. + +02:07:59.200 --> 02:08:02.240 +And when Karsten Korth added one micromolar, this amount, + +02:08:02.240 --> 02:08:04.880 +he still saw these protease resistant bands. + +02:08:04.880 --> 02:08:07.400 +So the three bands are the ones with no sugars, + +02:08:07.400 --> 02:08:08.800 +one sugar chain and two sugar chains, + +02:08:08.800 --> 02:08:09.800 +they're shed a little better here. + +02:08:09.800 --> 02:08:11.320 +So two sugar chains, one and none. + +02:08:11.320 --> 02:08:12.160 +I'm gonna go back. + +02:08:12.160 --> 02:08:13.320 +Or even up in here. + +02:08:13.320 --> 02:08:15.000 +It's a very appealing way of thinking about all this + +02:08:15.000 --> 02:08:16.640 +because it makes much more sense than thinking + +02:08:16.640 --> 02:08:19.160 +that PRPC grapey is something totally apparent. + +02:08:19.200 --> 02:08:20.520 +It raises the question of whether or not + +02:08:20.520 --> 02:08:22.120 +we really have an issue, it's a kinetic race + +02:08:22.120 --> 02:08:23.680 +between the formation of PRPC grapey + +02:08:23.680 --> 02:08:26.120 +and the cells of the clear PRPC grapey. + +02:08:26.120 --> 02:08:28.360 +And that when the cell can keep up with the formation + +02:08:28.360 --> 02:08:31.880 +and clear it like it does with all other proteins, in fact, + +02:08:31.880 --> 02:08:34.440 +when that happens, everything is functioning fine. + +02:08:34.440 --> 02:08:36.440 +But when the cell gets out of balance, + +02:08:36.440 --> 02:08:40.360 +it can no longer clear PRPC grapey at the rate that it's formed. + +02:08:40.360 --> 02:08:41.520 +And we're talking about very, very low levels + +02:08:41.520 --> 02:08:43.840 +that we can't detect even by these animal assays. + +02:08:43.840 --> 02:08:46.480 +So very, very low levels, kind of like asymptomatic. + +02:08:46.480 --> 02:08:50.560 +You're asymptomatic for CAD, all your life. + +02:08:50.560 --> 02:08:53.320 +And if you lose the race, then you become, + +02:08:53.320 --> 02:08:57.160 +so it's actually not what a lot of these worst case scenario + +02:08:57.160 --> 02:09:00.520 +people said, which is you just need one molecule. + +02:09:00.520 --> 02:09:04.160 +You have one molecule all the time by this hypothesis. + +02:09:04.160 --> 02:09:06.400 +And it's just a question of whether you fall behind + +02:09:06.400 --> 02:09:10.280 +with degradation or whether you inherited too much + +02:09:10.280 --> 02:09:13.680 +to be ever catch up in the beginning or something like that. + +02:09:14.680 --> 02:09:17.680 +It's extraordinary what is being said here. + +02:09:17.680 --> 02:09:19.680 +Then something goes awry, and we began to accumulate + +02:09:19.680 --> 02:09:21.680 +more and more PRPC grapey, and eventually the animal gets sick. + +02:09:21.680 --> 02:09:22.680 +And it goes on to die. + +02:09:22.680 --> 02:09:25.680 +And still we're talking about a very specific protein + +02:09:25.680 --> 02:09:29.680 +with a very specific sequence in a very specific structure. + +02:09:29.680 --> 02:09:32.680 +And then sometimes we're not. + +02:09:32.680 --> 02:09:34.680 +Sometimes we're talking about a very specific protein + +02:09:34.680 --> 02:09:37.680 +with a very specific sequence in a very specific structure + +02:09:37.680 --> 02:09:39.680 +that has two structures. + +02:09:39.680 --> 02:09:42.680 +And those two structures, apparently, one of them + +02:09:42.680 --> 02:09:45.680 +is more stable than the other, so much so that it can cause + +02:09:45.680 --> 02:09:48.680 +the good one to form the bad one. + +02:09:48.680 --> 02:09:53.680 +But that part of this cartoon is almost wholly missing + +02:09:53.680 --> 02:09:56.680 +from the explanation. + +02:09:56.680 --> 02:09:58.680 +And instead we have incubation times + +02:09:58.680 --> 02:10:01.680 +and p-value differences between brown mice and white mice + +02:10:01.680 --> 02:10:04.680 +with two amino acids difference in this protein. + +02:10:04.680 --> 02:10:08.680 +And then that being a sign that the protein is involved + +02:10:08.680 --> 02:10:12.680 +in whatever happens. + +02:10:12.680 --> 02:10:14.680 +Okay, it is 335. + +02:10:14.680 --> 02:10:16.680 +I need to go to basketball. + +02:10:16.680 --> 02:10:20.680 +I'm going to play the funny opening that I started with again + +02:10:20.680 --> 02:10:24.680 +as a way out in case you didn't see it because it's just funny. + +02:10:24.680 --> 02:10:29.680 +I got a new toy, which is actually a new toy for + +02:10:29.680 --> 02:10:36.680 +it is a new toy, which I am planning to use on the wheel. + +02:10:36.680 --> 02:10:40.680 +And also maybe on my motorcycle if we get a little better weather. + +02:10:40.680 --> 02:10:45.680 +And that toy, of course, was a little camera + +02:10:45.680 --> 02:10:47.680 +that allowed me to make this video. + +02:10:47.680 --> 02:10:49.680 +But anyway, thank you very much for joining me. + +02:10:49.680 --> 02:10:52.680 +This has been Giga Ohm Biological, a high-resistance, + +02:10:52.680 --> 02:10:55.680 +low-noise, oh, that's not the right song. + +02:10:55.680 --> 02:10:57.680 +You funny man, that's not the right song. + +02:10:57.680 --> 02:10:59.680 +This is the right song. + +02:10:59.680 --> 02:11:01.680 +That's it. + +02:11:01.680 --> 02:11:07.680 +This has been a production of Giga Ohm Biological. + +02:11:07.680 --> 02:11:11.680 +And this was, of course, not meant to make you take me seriously + +02:11:11.680 --> 02:11:15.680 +as a basketball player, but at least to let you know that I do + +02:11:15.680 --> 02:11:17.680 +actually go to the gym with my kids. + +02:11:17.680 --> 02:11:23.680 +And I'm going to try and figure out a way to make neurobiology + +02:11:23.680 --> 02:11:27.680 +shorts and immunology shorts while I'm shooting around. + +02:11:27.680 --> 02:11:29.680 +And I thought this camera would kind of help me. + +02:11:29.680 --> 02:11:32.680 +It actually works surprisingly well. + +02:11:32.680 --> 02:11:36.680 +It also works surprisingly well on the one wheel. + +02:11:36.680 --> 02:11:39.680 +So I think I'm going to be able to make some shorts + +02:11:39.680 --> 02:11:43.680 +that I haven't been able to make before because I just don't like + +02:11:43.680 --> 02:11:49.680 +putting three GoPros on something and having to edit for hours afterwards. + +02:11:49.680 --> 02:11:52.680 +And that thing seems to have solved that problem a little bit. + +02:11:52.680 --> 02:11:55.680 +I can give a GoPros to my wife for her yoga. + +02:11:55.680 --> 02:11:58.680 +Anyway, thanks very much for joining me. + +02:11:58.680 --> 02:12:03.680 +And when Giga Ohm Biological, we are trying to distribute the knowledge + +02:12:03.680 --> 02:12:07.680 +that can dispel this big E enchantment. + +02:12:07.680 --> 02:12:10.680 +Thanks very much, and we'll see you again soon. + +02:12:28.680 --> 02:12:31.680 +Thank you very much. + +02:12:58.680 --> 02:13:01.680 +Thank you very much. + diff --git a/twitch/2128456102 (2024-04-24) - Stanley Prusiner and Prions 2002 STUDY HALL -- (22 Apr 2024)/README.md b/twitch/2128456102 (2024-04-24) - Stanley Prusiner and Prions 2002 STUDY HALL -- (22 Apr 2024)/README.md new file mode 100644 index 0000000..ed05c3c --- /dev/null +++ b/twitch/2128456102 (2024-04-24) - Stanley Prusiner and Prions 2002 STUDY HALL -- (22 Apr 2024)/README.md @@ -0,0 +1,5 @@ +# Stanley Prusiner and Prions 2002 STUDY HALL -- (22 Apr 2024) -- Gigaohm Biological High Resistance Low Noise Information Brief + +## Streams +- https://twitch.tv/videos/2128456102 +