I'm Megan Donaldson, CEO of the Hereditary Disease Foundation, and I'm delighted to welcome you to our first research spotlight webinar. We fund innovative research to identify treatments and cures for Huntington's disease and other brain disorders. Before I begin my introduction, I'd like to acknowledge the news from Roche Genentech that the dosing on their clinical trial has been halted. We are all incredibly disappointed about this, but it points to the need for us to continue and intensify our support for research to develop treatments and cures for Huntington's disease. Our webinars will feature brilliant scientists who are unlocking the secrets of the brain and making game-changing discoveries. We are thrilled to launch the series with a presentation by Dr. Ayama Motto, Associate Professor of Neurology at Columbia University, and recipient of the 2020 HDF Leslie Gary Brenner Prize for Innovation in Science. From a creative graduate student with HDF funding to a prestigious NIH grant recipient, to a lab of her own mentoring a new batch of young researchers, and as a member of our Scientific Advisory Board, Dr. Ayama Motto has become an acclaimed expert in her field. Her path as a scientist illustrates how HDF provides not only funding, but an international family of collaborators who help nurture and lift the trajectory of each scientist's work. It is my pleasure to introduce Dr. Ayama Motto discussing What's It All About, Elfie and Aggregates. There will be a question and answer session at the end of I's talk. Please submit any questions in the Q&A box on the bottom right hand of your screen. Thank you for joining us. My name is Ayama Motto, and I'm here today to share with you what we've been doing in the lab. And of course, one of the things that we've been focusing on is whether or not we can undo the course of Huntington's disease. So just to begin, what I'd like to do is kind of set the stage. So for us, Huntington's disease is all about a protein known as Huntington, or HTT, and what happens to it in the course of the mutation that leads to this devastating disorder. So here on the slide, what you can see is a beautiful yellow cell. This is actually a picture of a brain cell taken from the Lispata lab. And in blue, what you can see is another brain cell kind of reaching out and touching this other cell.  They kind of reach out and touch and talk to each other. And so let's pretend that this cell is inside your brain. And we're going to zoom inside there and pretend that we can see what's going on inside these individual cells. And here inside the cell, you can see this big circle in the middle, and that is known as the nucleus. And the nucleus is the place where all the blueprints for the various machines that do the things inside the cells that make the cell be able to do what it does resides. And in there resides the blueprint for this very important protein, Huntington, or HTT, here in green. And Huntington is an essential protein. That means that it's necessary for life, and it helps to make the brain and make the brain work properly. Now in the brain of people with Huntington's disease, there's a little bit of an error in this blueprint that makes HTT and makes a different form here in pink that I've called toxic HTT. Now that might be a little bit of a cruel thing to do. I mean, this form of Huntington is actually doing a lot of the things that normal HTT can do, but it's just gained a new personality as it were. It does things that it wouldn't normally do. And one of the things that it does is it becomes very, very sticky. It's sort of like if you walk through a briar patch or something and you try to avoid all of the thorns and everything, but you can't. It just keeps snagging your clothes. Well, that's what this toxic HTT does. It just starts to stick to things. And one of the things it does is stick to itself. And by doing so, it starts to collect inside the cell, accumulate, and form these clumps that we call aggregates. Now, what it is about this new form of Huntington, its stickiness, this tendency to collect and accumulate and form these clumps, what of it is toxic. What of it causes Huntington's disease is not fully clear. But what we know is that something poisonous happens, ultimately leading to the demise of these cells. Now, whatever it is that this toxic form of Huntington does is unclear. But it is so robust that we can even capture it in a humble little mouse. And this is really from pioneering studies by Dr. Gillian Bates, who found that if you introduce this blueprint, the blueprint with the mistaken form of Huntington, and put it inside a mouse. The mouse will make toxic Huntington and then start showing features that mimic Huntington's disease, such as compromises in the way the animal can move, its cognitive ability, and such. And so what I show in this graph is kind of just a picture way of showing what could happen. So toxic Huntington would be expressed over the lifetime of a mouse. And as the mouse ages, they'll start showing various symptoms, such as the ones I just described. Now, even at the level of the brain, you can start seeing how this toxic form of Huntington starts getting sticky. So I'd like to show you a picture of that. So this is an image from a patient brain. And you can see here labeled something called an INI pointing to this brown clump. This is one of these protinaceous clumps that I was talking about, about Huntington sticking to itself. There's also things known as CI's that form cytoplasmic inclusions. And these are similar clumps that are forming outside of the nucleus inside the cytosol. Now we can actually see very similar structures inside of the mouse brain. So here's the picture of that, with the black showing these INIs, intranuclear inclusions, these clumps that are forming inside of the nucleus where all the blueprints reside, and then read the cytoplasmic inclusions, these clumps that are forming throughout the other parts of the cell. So as I mentioned, we can capture all of these features in the little mouse. And because of these studies by Dr. Bates, we were able to really propel how to study the relationship between this toxic form of Huntington and these symptoms that are associated to Huntington's disease. And so way back when over 20 years ago, when I was a graduate student in the laboratory of Renee Henn, together with a postdoc named Jose Lucas, we create our own mouse model to try to understand the relationship between toxic Huntington and these symptoms.  And what we had found, of course, is that if the gene is on, this toxic Huntington is formed and the mouse gets sick, much like Dr. Bates' study. But of course, we were able to turn the switch off. And when we turned the switch off in mice that were already showing symptoms, the mouse got better. So this finding really set up the question of, how did it happen? And can we figure out what happened so it can bring this about to patients? Well, one of the things that happened was the clumps. So here we have, as I mentioned before, this form of Huntington that gets sticky, forming this protonaceous clumps throughout the cell. And then this mouse model as well, we could see these clumps. As you can see these dark kind of brown dots kind of littered throughout the image from the mouse's brain. Well, when we turned off the expression of this toxic Huntington and the mouse didn't make it anymore, we were very much surprised to see that these clumps went away. And so we were wondering, well, where in the world did they go? And does this mean that if the clumps go away and the symptoms go away, are those two things related? Does it mean that if we can get rid of these clumps, can we somehow also get rid of the disease? So to go about this, we had to first try to understand where those clumps could have gone. And that kind of took a while for us to figure out. But ultimately, it brought us to the sky. So for those of you that might remember back in the 80s, there was this creature that kind of was all over the place known as Pac-Man. So Pac-Man is this creature that lives inside of a dark box and goes around and gobbles things up and eats anything in its path. I wonder if it would surprise you to tell you that we actually have a Pac-Man like thing in every single cell of our body. Of course, we don't call it Pac-Man, we call it autophagy. So what autophagy does is actually clear out the debris throughout the cell. And it can clear out all sorts of things, including these progenacious clumps. Now, the problem about autophagy, to try to harness the power of kind of clearing out these clumps, is that autophagy can degrade all sorts of things.  So for me, I don't know which macaroons to begin with. I would want to eat the strawberry ones, the lime ones, the chocolate ones, the coffee ones, all at once. But of course, we're trying to stay focused here. We want to focus Pac-Man autophagy onto the clearance of these progenacious clumps, like the chocolate macaroons. So how can we do that? Well, studies suggested that actually Pac-Man doesn't just randomly eat things. In fact, it eats things in a very, very deliberate way. And it does so because the cell has a way of labeling those things that need to get eliminated. And so we wanted to understand how is it that autophagy knows how to eliminate these clumps, these progenacious aggregates. And that introduced us to something known as alfie. So alfie stands for autophagy link five domain containing protein, which is not as fun to say as alfie. Alfie is actually like a little flag. What it does is that it will find the aggregate, the clump, label it. And as it labels it, it sort of acts as a beacon for, in this case, Pac-Man, our autophagy, to eat the inclusion and clear it away. So we identified the flag, how to attract Pac-Man to eat the clumps. And so now we wanted to test whether or not we can actually take advantage of that. So can we take the autophagy that's in all of ourselves and now label the progenacious clumps and say, OK, Pac-Man, go eat it. And clear out those clumps. And let's see whether or not it can undo the course of Huntington's disease. So how do we go about doing that? Well, Catherine Croce and my laboratory is doing just that. What she did is she's taking a mouse model of Huntington's disease. And now, instead of adding a genetic switch like I did so many years ago, she's actually introducing increased levels of alfie. So now we hope that all of those aggregates will get labeled, right? And then we can see whether or not autophagy can go and clear them out. So let's see what happened. So here is a picture from a Huntington mouse brain. Again, you can see those dark brown dots and kind of like pepper flakes, kind of like sprinkled around the sample. These are, of course, those aggregates. Now we next like to look at a brain that has a lot of alfie. What happens to those progenacious clumps? And look at that. Where did the aggregates go? It was very much like what we saw when we turned the switch off. The aggregates went away. So it looked like alfie is doing its job. It's finding those clumps and helping to attract that Pac-Man to eat it all up. All right, so mission one was accomplished. We can clear out the inclusions. But of course, the reason we wanted to do this to begin with was to see whether or not we can have any impact whatsoever on those symptoms that are associated with Huntington's disease. So in order to do that, Catherine next started and is continuing to do some very painstaking studies of trying to understand how this is impacting the course of disease in the mouse. So the place to start is looking at movement. And so we do a variety of tests. So one of the tests that we use is known as the Accelerated Rotorod. And that's just a picture shown here. What you can see is a dark bar kind of going through the center there where the mice are sitting on top of it. This bar is like a log roll. And it just starts to roll, turn, turn, turn. Accelerating, accelerating through the course of 300 seconds or five minutes. And what the mouse has to do is scurry along and try to stay on top of this rod. Otherwise, kerplunk, it will fall down just like some of the ones that you see here. And so what we can do is actually quantify how long the mice are able to stay on this rod and be able to successfully do this task. So here are some examples I show here. So what you have is we're measuring how long the mice are able to stay on the rod. And this is a two month old animal. And what you can see in blue is the control mice, a normal mouse. And normal mice are able to do this task with no problem. They just stay on for 300 seconds. Well, then if you look at the Huntington's disease mouse here in red, what you can see is that the mouse has a more difficult time staying on the rod. And they have a tendency kerplunk to fall at about 220 seconds. Well, now what happens if the mouse has Huntington's disease but also expresses a lot of alfie? Remember, these mice have fewer clumps in their brains. Does this have any impact whatsoever on the movement? And so far what Catherine is showing, very excitedly, of course, is that indeed, these mice look just like the normal mice. They're able to stay on the rod for the full duration of the 300 seconds. Now, we don't only look at this at one age. Catherine's been looking at this continuously across many ages. And I hope you can appreciate that if you look at the green line, that they are able to look just like the blue line. That is to say that the Huntington's disease mice with a lot of alfie look just like normal mice and are not showing the motor symptoms that we see in the Huntington's disease mouse. Now, this is already very exciting. But as scientists, we tend to be very, very cautious with our conclusions. So we had to, of course, do another test. And luckily, it's a relatively simple test. It's known as the open field. The open field is literally a box. And inside the box, you put a mouse, just like I show here. And what we do is we just measure how much it runs around the box, explores the box, to try to get a sense of how well it can just simply walk around in its environment. Here is an image just depicting some of the data we have so far on that. Here, again, in blue are the normal mice. And they tend to walk about 8,000 centimeters over the course of two hours. That's a lot for a little old mouse. Now, in the Huntington's disease mice, they still can move around. But they move around a lot less, closer to about 6,000 centimeters. And so what happens if we have alfie, the mice that can do the accelerated rotor rod, the mice that have fewer clumps in their brains? Well, in this task, they don't do as well as the normal mice. But they sure do a lot better than the Huntington's disease mice, suggesting that alfie seems to have an overall impact on the way the animal is able to move, that somehow, in the absence of these clumps, these mice can move very much like control. So if I take it back now to this graph that I've been showing you, what we think is happening is that in the presence of alfie, even though toxic Huntington is continued to be expressed, the clumps go away thanks to alfie, thanks to that pack man that can gobble those things up. And then when it comes to the actual symptoms that we've measured so far, it's significantly delayed. And when we do start seeing it, we anticipate that we'll see a very, very blunted symptomatology. Of course, it's going to take a while before we can really complete our studies to see the lifecycle of this mouse. But so far, we're very excited about how promising it looks. Now, this in and of itself is very exciting for our Huntington's disease research. But I wanted to just simply raise the point that there's more to this than just Huntington's disease. The appearance of these abnormal proteinaceous clumps, the accumulation of protein, is something that we see across a lot of different diseases, from Parkinson's disease, ALS, and Alzheimer's disease. This is just a mosaic I like to show of various images I've taken from different sources showing how these proteinaceous clumps can be found in different diseases of the brain. And so our next goal also as we continue our studies with Huntington's disease is to really try to understand to what extent alphae and autophagy can impact proteinaceous inclusions, these clumps, inside brains of different diseases and see whether or not it can have similar reparative effects as we saw in HD. So that's all I have for you today. I just like to mention that this work was not literally done by me. I don't do it anymore. I have all these people that trust me with their futures and they really work hard for me and I want to mention them. I only have here highlighted the people that have contributed directly to the work with Alfie and Catherine's work is really what I highlighted today. Leora, Kyrgyz, Joanna, Aveline, they've also all contributed significantly to the progress that we have made over the years with Alfie. Also, there are people outside my laboratory that we work with very closely and Siemensen is the person that named Alfie so we can credit her for giving us something fun to say. William Yang, we're doing this to phenotype the mice that is to characterize the symptoms more deeply. David Hausman, Nancy Wexler, Jean-Paul von Sattel, where we're actually looking at how Alfie might be impacting age of onset in HD patients. And Bev Davidson and Frank Bennett represent a new line of work where we're trying to actually augment Alfie therapeutically in people. Thank you very much. Well, thank you very much, I, for that outstanding presentation. We have quite a number of questions here and I'd like to start with the first question that someone wrote that's not directly related to your presentation but I did want to just address it in case anyone else is thinking about this. The question is, I'm not sure if this is the right forum for this but I'm coming close to early onset age with Huntington's and my family. What is your recommendation for testing and are there any benefits to finding out early? What I would say to this is if anyone is having similar questions about this topic, please email me, Megan Donaldson, M-E-G-H-A-N, Donaldson, D-O-N-A-L-D-S-O-N, at hdfoundation.org and I'd be happy to speak to you about this. Now with that, I'll leave it to I to answer the questions about Alfie. Thank you. Great. Thank you, everybody. First, thank you all very much for coming to the webinar. I hope you enjoyed it and I'd like to thank, of course, the Hearty Territory Disease Foundation, their supporters, which is many of you out there, for your continued support because without you we could never have gotten this work off the ground. I'd also like to thank the Alexander Boyd and Jane Starboy Charitable Foundation for their support, especially of this ongoing work we've really entered. I hope you all agree, an exciting phase of this research. So there are several questions that have been imposed. A lot of anonymous attendees, I was going to list everyone's name, but of course, I'm happy to start answering them. Some of the questions are similar, so I'll put them together. So I think one of the continued themes in the questions has to do with, if this looks so promising and nice, how are we going to bring this to humans? Can we label these protein clumps in human cells? Can we ever test this in humans? Can we bring this to a therapy? How would we go about doing that? And that is, of course, a very big, typically, actually, to be perfectly honest. First of all, we would usually say we just put the ideas out there and hope someone brilliant will come up with a plan. But thanks to Nancy Wexler, we already have a plan. So I'll get into that in just a moment. So first of all, this protein is present in all of our cells as well. It's not something unique to mice. And so this was one of the reasons why we really started to focus on it is because this is something that we actually saw was active also in human cells. So it was something that we could translate potentially to patients. So this was something that was very interesting for us. Now, how do we bring about doing this? And so as we're doing our mouse-based studies, we're also looking at human-based model systems. These are always in cells. We are not yet at the stage of bringing this to patients, of course, to make sure it's working in the right environment. And it seems to be working, which is very exciting for us. Now, in terms of how are we going to bring about this to patients? So this is where, for those of you who know Nancy Wexler, you know that when she wants you to get something done, it's going to get done. And so very much thanks to her, she got me to be able to kind of ask people. And thanks to the Scientific Advisory Board and members of the Davidson, Frank Bennett, we're actually looking at ways by which we might be able to increase Alfie levels in patient brains. And whether or not we use an ASO-based approach or kind of a more typical medicine-based approach is something that is left to be seen. So we can add Alfie to human cells. And we can also put this into human cells. We're studying Alfie in the context of other diseases. So one question is, Alfie being studied in other diseases yet, another related question, hi there. I'm curious, Dr. Yamamoto, is there an Alfie for Alzheimer's disease? Another question that was very similar. Do I saw somewhere along the way? I'll answer these two questions. Is that we are currently actually looking at Alfie in the context of mouse models of Parkinson's disease, mouse models of Alzheimer's disease, and mouse models of frontotemporal dementia and ALS. The reason we can look at all of these different diseases is because they all share a feature, which is the formation of those clumps. And so if we can direct Alfie to those clumps as well, maybe we can bring about the same kind of promising outcomes that we're so far seeing for Huntington's disease. Now, there was a question about how do you get Alfie to go to the aggregate? And that is a really good question. This is something that we are trying to understand. Basically, what we believe happens is that the clumps get labeled by the cell, kind of like a postage stamp equivalent. And Alfie can find that postage stamp. That actually is a whole other set of machines that we have to kind of bring into play. And what we were hoping by just focusing on Alfie was that Alfie was the part that was missing. That is to say that these clumps are getting stamped, stuck all over them, saying, please get rid of me, please get rid of me. But we thought maybe Alfie was the one that was kind of slow to respond. And that's why we thought boosting Alfie levels will actually help. So what is the current research emphasis from Gloria? Well, the current research emphasis is really about trying to understand increasing these Alfie levels can indeed drive not only the change in behavior, the positive change in behavior we see in Huntington's disease, but we want to understand that more deeply. There's always a question, how much of the mouse really mimics what's happening in patients? And so we really want to understand that translation better. So that's really one of the big emphasis right now in our work. Let me see, the questions are starting to come in. So for those of you who can't, it's like the scroll, it's so hard, it's hard to read. I have a good one here. Comes from an anonymous attendee saying, how is the research with Alfie applicable to those with HD now? Do you think this research will be useful to them within the next 10 years? Wow, within the next 10 years. That's a very good question. Thanks Megan, that was really hard. Okay, let's see. So it's always hard for me to know how long these things take, right? I mean, so first of all, I guess one of the things that I can definitely say is that we all have Alfie in ourselves and we know that the Alfie that we have currently is working as hard as it can to do the best it can. That is to say that we did experiments where we actually removed Alfie from otherwise seemingly healthy animals and it made them sicker. So we know that Alfie, the Alfie is there is helping to sustain whatever movement we can. Right now, there is nothing that we know of that can boost Alfie levels with any kind of known medicine or known vitamin or something like that. So I would say that right now it's still very much in the experimental phases. We are working very hard to see how quickly we can apply what we're finding in these animals in patients. As I mentioned before, it is a current focus of ours. And so I would hope that in the next 10 years we will be somewhere where we're actually talking about what is happening potentially in patients. And given the breadth of Alfie's function, hopefully we'll be able to get there soon. But I have to say that this is really outside actually bringing this kind of experimental work to the lapse of patients is very a bit removed from my expertise. And so it's difficult for me to give a real timeline on this, unfortunately. I following on from that is an interesting question from another anonymous can be, are these experiments being done in a rescue model where the Alfie are gone after the mice have developed symptoms as would be the case in a human treatment? Yes, so those experiments are currently ongoing. So Catherine Croce and the lab who's leading these experiments, who's wonderful by the way, she is currently doing experiments where she's taking mice and waiting for them to become, to have clumps in their brains especially. And then turning on Alfie, right? And using our genetic tricks. This is not with a medicine, but with a genetic trick to see whether or not it can affect change. Of course, these are the kinds of experiments we would like to do because of course, this is how we would envision a therapeutic being used. And so this is how we're trying to address these questions in a way that we know best. Okay, and another question from Christopher Pearson who is on our scientific advisory board and is going to be presenting our next research spotlight webinar in June. Christopher writes, does Alfie alter levels of pelvis in the brain? If so, is there a specific cell type that is affected? Okay, so, and for those of you listening, I'm gonna go into a little bit of jargon here because I'm just gonna take advantage of the fact that Chris should understand. So basically when we did a loss of function experiment, it's almost like talking in a different language. I feel like I'm talking in Japanese all of a sudden. When we did the loss of function experiment, so that is to say when we decreased the levels of Alfie, we actually did not seem to affect cell death. That is while the patients, the mice got sicker, we did not accelerate cell loss. Now, and this has a lot to do with the fact that the mouse models we are using do not experience cell loss. So it's very difficult for us to study. For those experiments that I was showing today where there were fewer inclusions and then the onset of the symptoms in the mice were slowed. Here we, again, don't know if we're affecting cell loss because at least the ages that we've looked at thus far do not show cell loss. So this kind of question is something we would like to answer in the cell-based and the human-based models. These are the direct conversion, so where we take fibroblast skin cells from patient with patients and turn them into neurons. That model actually shows cell death and we are trying to figure out a way to administer Alfie to those cells to see whether or not we can affect cell death. So I can't answer a question about cell type specificity but hopefully we'll be able to with the direct conversion-based model. Okay, I have another somewhat scientific question for you to answer from another anonymous attendee. Is there any evidence from the human GWA studies that variation in Alfie affects the age of motor onset? Yes, so this is the very exciting work by David Hausman, Nancy Wexler, Jean-Paul Vonsetel in collaboration with the New York Genome Center, turning to one of our most precious, precious resources, which is of course the Venezuelan cohort. And there, I mean, it's like unbelievably enough, but really what really galvanized our work and gave us kind of the courage to really go after this full throttle, had to do with the fact that it was found that in patients, Alfie, like all proteins in everyone's body can be slightly different. They're called variants, they're slightly different from one person to the next. And there was a group of patients in the Venezuelan cohort that actually showed a variant of Alfie, and Alfie that was slightly different. I mean, super, super slightly different that actually had a delay of onset. So instead of say at 40 years of age had the disease onset at 50 years of age, average age of onset delay of 10 years, the maximum delay age of onset was 23 years. So this is what really got us excited because we were like, oh my goodness, sometimes when we do experiment at a bench and play with mice all the time, we start losing track of what's real and because we hope so much of what we're seeing as real. This was what we call independent confirmation, a study outside of our realm telling us that we're onto something. It looks like, and Catherine's also exploring this, how does that little change boost the way Alfie can work is something that we're exploring currently because this is really going to give us insight into how we can potentially increase Alfie in patients. So yes, the GWAS studies were very valuable to us. Okay, thank you, Matt. I hear a couple more related questions from two anonymous attendees. If we already have Alfie to clear the aggregates, why do they form to begin with? And related to that is, are clumps the only thing Pac-Man eats in ourselves? Yes, so for the first question, yes. So basically what we know is that if we didn't have Alfie, the clumps would form a lot sooner and we would experience patients, experience symptoms we would argue sooner. That is to say that the Alfie we currently have working in our brain works for as long and as hard as it possibly can, but somehow Huntington's disease is stronger than Alfie. That's why we need to kind of give Alfie a boost. I don't know, a Superman cape, something, something to help it work better. But what's promising is that there's something in there that's already working and we can potentially take advantage of it. Now, of course, Alfie shuttles the aggregates to Pac-Man. So does Pac-Man eat me? Well, we have to tell Pac-Man to eat the aggregates, the chocolate macaroons, because actually Pac-Man likes strawberry macaroons, lemon macaroons, all sorts of macaroons. That is to say Pac-Man goes around because it is important in keeping ourselves nice and clean and healthy. It has to clear out all sorts of other things. And so in essence, what we've been trying, what Pac-Man can do is eat other things. And that's why we have to boost Alfie and say actually you have to like the chocolate macaroons a little bit more. We of course don't want to distract Pac-Man. Pac-Man needs to do its other things as well because that's also very important. But we know that for some reason those aggregates are much of a distraction for our brain cells. And so we're trying by maybe just making them like chocolate a little bit more than strawberry. Maybe we're hoping that that's really what's affecting change in these brains. Okay. Thanks for that. I, another quick question. I'm calling on an anonymous attendee. What was the HT mouse use in these studies? Oh, wow. We used a couple of mouse models. So for the slowing, so for the elimination of the aggregates and the slowing on the behavior that we showed in the video, we use the mouse model known as the Borschelt model, the N171 model, which is a fragment model which uses only a portion of Alfie. And then we also use the CAG 140 knock-in model. That is to say it is a mouse uses mouse Huntington where the Huntington's disease mutation is put inside. There are experimental subtleties as to why we use both models. But what's very exciting is that Alfie seems to be able to work in both situations, which was very important because of course, in patients, one could argue we could see one, if not both of those forms of the protein. So we wanted to make sure that Alfie can act on both. Okay, thank you. And then another question, a bit on the more scientific side from a participant named Boone Mabam Malou. I'm sorry, I probably got that totally wrong, but his question or her question is, could the targeting system used be applied to epitopes presented on other inclusion body disorders? And I think you may have touched a little bit on this already. Yes, so we think so. So what we do know is that, like what we've tested explicitly is we've looked at Huntington's disease like polyglutamine inclusions, and we've looked at alpha-synuclein-based inclusions. And Alfie seems to be able to work in both in cell-based systems. For synuclein, we are now looking in the adult brain, so in the mouse brain, so we're doing this in mice. In theory, we think that it should work generally. The way protein clumps are viewed by Pac-Man, it doesn't care what is in the clump. So in theory, we believe it should work more broadly. We're currently looking now at TDP 43 inclusions and we're also starting to look, which we haven't fully started yet, but we're starting to look at intranu- intracibata that's still inside of the cell. So we think it should, but we have to actually test to make sure that it does work inside the brain as well as it does inside the cells in a dish, which has been a lot of, where a lot of our experiments had been done thus far. Okay, we have another question about the mice test. Did you use any other bolder cognitive tests to evaluate the mouse symptoms? And if so, what were the results? So the tests we showed are thus far, I believe the only tests we have run are the two motor tests. We have not done cognitive testing on the mice yet. That would be a good idea. That would be a good idea. And here's another interesting one from an anonymous attendee. Are there other proteins like alfie that are also in our cells that aid in clearance of aggregates? I mean, there is a whole lot of proteins inside the cells that kind of all play into what alfie does. However, in terms of actually working like alfie, people have looked at, there are a couple of proteins that might work like alfie. And at least for a couple of, at least one of them in cell-based systems, it doesn't seem to work like alfie in the sense that if you eliminate the proteins, they all cause more aggregates to form and accumulate inside the cells, just like alfie. But what set alfie apart in our very early studies is that if you increase levels of other proteins, it doesn't seem to affect the turnover. That is to say, it doesn't seem to drive Pac-Man to eat the aggregates faster or more. It doesn't increase its preference for chocolate macaroons, as I like to say. So, alfie was unique in that increasing its levels is what actually drove the elimination of the clumps. And that's why we focused on it. But I've always, you know, never want to kind of rule out the possibility that there's like, you know, more than one way to shoot a duck or whatever is the metaphor. Maybe not a duck. I don't mean to kill any ducks, but whatever is the expression. So, but as far as we know, we've only, in the screens we use, only identify alfie. It's what, it was a factor that made it stand out. Okay, thanks for that. We have another question from Casey Fox asking, is alfie a large molecule and therefore difficult to increase its concentration in the brain? Is that one of the reasons that it isn't closer to being used as a drug therapy at this time? Oh, hello, Casey. Thank you for a very, very good question. In fact, alfie is very big. This is one of the reasons why, you know, we're trying to think of alternative ways to increase the levels of alfie in the brain. We can't just put it in because it is very big.  It's just that I guess I was a little shy. But, you know, with the help of doctors, Bev Davidson, Frank Bennett, I get to use their brain power to help, you know, figure out how it is that we might be able to tell the cells themselves to actually make more alfie. And this might be one of the more effective ways to actually boost the level of this very large protein. I do want to address a question. I think it just happened to be floating in front of me by Sandra Fienko, which I think is an interesting question, as many of the others have been, is alfie labeling other slash physical proteins for autophagy? And could this contribute to possible adverse effects, right? We've only talked about, you know, how increasing alfie can be good, you know, could increasing alfie be bad? And, you know, this is something that, of course, we are very concerned about, right? If Pac-Man goes around eating a lot of other things, and this is where I could think it's especially, it could potentially be bad, that first alfie might distract Pac-Man from doing what it needs to do. And that could be adverse. It could cause, you know, just garbage to start piling up in the cells and then affecting it, affecting cell function. The other thing, though, is, of course, alfie might call other things chocolate macaroons, right? Call things that the cell actually needs and make Pac-Man eat those things by accident. So these experiments that we're doing in mice, where we're increasing alfie levels in mice, I think is the most telling. So so far in Catherine's experiments, where she's increased the level of alfie in the entire body from the beginning of time, she's also doing it in later in life. So far, these animals of just alfie, higher alfie levels are not showing any issues. They are not showing any signs of stress, discomfort. They are aging apparently normally. They're breeding normally. They're behaving normally. So it looks like while the possibility is always there, at least in the approaches we have used thus far, we are not seeing any detriment in the presence of increasing alfie levels. But in all fairness, you know, understanding how alfie works, what alfie does inside the cell is still actually quite early science. It's not very well studied. And so there's still a lot to be learned in that regard. So thank you for the question. Okay. I had another couple of questions about CRISPR. Here we go. One from an anonymous handy who writes, I know research is slow on this disease as my mother passed away from it 21 years ago. How does CRISPR factor into experiments? And then related to that. Is one again from Booney, my bomb balloon as a CHE expansion disorder with anticipation becoming significant around 40 years. As you said, would it be possible to utilize embryogenic CRISPR to delay presentation of this glutamate channel disorder? Okay. So for the first question, can CRISPR somehow play a role in this? So I guess, you know, the way I would answer this question is that it will go to those experiments looking at what I was referring to earlier when someone asked about the GWAS GWAS studies about alfie. And I was talking about how in the Venezuelan cohort, there were a group of patients that had a change in alfie, a small change, but that a single nucleotide change, as small as you can be. And that was sufficient to potentially boost alfie function. So one place where CRISPR could play a role is to make that little change in alfie, right? And if, in fact, that change is sufficient to drive alfie, increase the way alfie works, that's somewhere where CRISPR could play a role. Now as to the second point in CRISPR and kind of like eliminating the CAG and embryogenesis, because this is where I guess, I think that's what the question was asking. I mean, I guess in theory, I think embryogenic CRISPR, there are a lot of other concerns and ethical concerns that are related. And this is really, I have to say outside of my wheelhouse, because this is a technology that we actually are not using regularly. We kind of use more traditional approaches. So as we are not developing CRISPR-based approaches. So as I said, you know, we just put the information out there. And so if there's someone that can do it, please, please go do it. Because we can't do everything ourselves. Okay. I have Thomas both, the HDI, who asked the question. Hi, Tom. In addition to your histopath and behavioral studies, have you performed any molecular comparisons by RNA-seq, SCN RNA-seq, and proteomics and controls versus alfie over expression mouse studies? So thank you for this question. So we are collaborating actually. So the reason we're using the CAG 140 mouse model has to do with the fact that we're collaborating with William Yang, who has of course done a huge study characterizing the impact of the trinucleotide repeat expansion in Huntington mouse models and using the full panel of omics-based approaches to characterize the impact of potential variants and modifiers on the transcriptomics and proteomics of HD. And we are going, we are basically using his design, much in consultation with active consultation with William, where we're doing both a loss of function, partial loss of function of alfie, as well as gain of function of alfie and looking, doing RNA-seq. Now, one of the things that we are, we have some hints that the impact that we are having on the HD phenotype in these mice is beyond just the histopathology and the behavior, but those studies are really early going and we would like to do kind of the full blown RNA-seq. And I believe Catherine has finished collecting the sample so that we can do so. Or we're almost there. I don't want to, I don't want, if Catherine's listening, I don't want her to like dive a heart attack because I guess she did more than I claim. Sorry, Catherine. I think we're in process. That's for sure. And so we should be getting the RNA-seq data soon. Okay. I think we're going to need to wrap it up. We've got about five more minutes. Do you want to just take a quick look and see if there were any questions that you would like to address before we wrap it up? Sure. There was just one quick question. Because I do like to say, you know, there are possible issues. There is a question by an anonymous attendee, were there any mouse models where Alfie did not work? Of course, we always focus on the positive data. But so far, no. But that's because we've only done it on the models that I showed. So there's still plenty for failure, I guess. Those scientists, we kind of prepare ourselves for failure. It's basically, it's something where we train. And then there was another kind of technical question. What kind of cells, for example, astrocytes and microglia are important for the Alfie mice, clear, misfolded proteins. And I would say that it's within neurons. It can also be through microglial engulfment or potentially autophagy and microglia, as well as potential autophagy and astrocytes. We still don't know. But Alfie is expressed in all of the cells. And our experiments are also targeting all neural cell types and not just neurons. There are a lot more questions, but I think it's just like lots of them. I'm happy to answer them offline if people would like. So if there's a mechanism by which that is possible, I'm happy to do so and address any of those questions as well. Yes. If anybody would like to send additional questions, you can send them to me, Megan, M E G H A N, Donaldson at hdfoundation.org. And I can pass them along to Dr. Yamamoto. So just one last question that I'd like to address, especially given the news from Rose, Roche and Genentech. I just want to put it out there. It's from Robert Burgess. Robert, thank you very much for your question. If a trial has risk, but the risk is greater to live with Huntington's without any treatment. Shouldn't a patient be able to choose to get treatment now? I think that's an excellent question. And thank you so much for asking it. It's a very difficult question for us to address. And in light of everything, the news last night, I think we just have to wait and see what happens with all of the data from that trial. And there are lots of other trials that are continuing to go forward. And let's just hope that those have good results and that we'll find some good results from the, the Roche, um, Genentech trial. I think that's something that, um, I would recommend you discuss with your family and your, your doctor and your genetic counselor. I did you want to add anything? No, no, I mean, that sounds great. Yeah. I'm also anxious to understand what happened. I think we all are and hopefully we'll get some more, um, information soon. But to wrap up, I, um, the predatory disease foundation would like to thank, um, triplet therapeutics for underwriting our research spotlight. Webinar series and also Novartis for sponsoring today's webinar. We'd especially like to thank I am a moto for her incredible research and the time that she's graciously spent with us today. Thank you also to eyes colleagues, especially Catherine crochet, who has performed much of this work that I discussed today. With the valuable insights that we gain across the medical and biological professions, we're better able to serve victims and other families of Huntington's disease and so many other brain disorders. Thank you also to all of today's attendees for meeting, learning and sharing with us to continue our research spotlight webinar. We will be hosting our next one on June 15th, noon Eastern featuring Dr. Christopher Pearson.  Registration is open. www.hdfoundation.org slash webinars. Thank you again. I for this incredible presentation and thank all of you for joining us and for asking your excellent questions. Great. Yeah. Thanks for having me. This was a lot of fun, everybody. Thank you.


Привет. Вы меня хорошо работаете? Да. Окей. Меня зовут Дмитрий Пелевский. Я говорю о истории, о продавании русских криптовых алгоритм в OpenSSL Kit. Хотя это содержит некоторые детали русских, это будет полезно для тех, кто хочет продать новую крипту. Я думаю, не только для OpenSSL, но и для любой функциональной криптовой криптовой. Итак, кто я? Как я сказал, я Митрий Пелевский. Я читаю OpenSSL Code с 2005 года. Иногда я понимаю, что происходит, иногда я не понимаю, особенно в тех, что касается постоянного времени. Моя главная работа currently is Russian Cryptographical Company, Cryptocom Limited. Я директор, и я тоже participated in various OpenSource projects, mostly related either to cryptography or to domain registration stuff. Итак, first of all, what is Russian Ghost itself? It's a set of cryptographical algorithms. The eldest one was approved in 1989 at Soviet Union time. At that time it treated as a bit paranoidal with its very long K, but it's still alive and the concurrence who will have gone. Russian standards include both first class algorithms, full stack of them, such as block ciphers, message authentication, of course digits, of course digital signature. But these algorithms themselves are almost not usable with our supplementary algorithms. We need a pseudo-random function, we need the derivation of the shared K, we need various functions related to K-Rap, and so on and so forth. So the history for me begins in 2005, when the company I worked it was a Cryptocom Limited time too. It was my prepreviews work, not only current decided that we should provide support of Russian Ghost Crypto and various, in this or that worldwide use toolkit because self-designed formats weren't convenient and we wanted to use worldwide ones. At that moment we studied, in fact, three variants, OpenSSL, GNUT LS, which was not major enough at that moment, and NSS. We've chosen OpenSSL, I don't remember the exact reasons, and then we decided to look into the code. So the first we've seen was many, many case switching in the parts of code related to digital signature. It looked like case RSA without this, case DSA without that. Sometimes there were some cases where something was done when we had elliptic curves cryptography. It was very long ago. We thought whether we have to add the case Ghost, decided not to do it, and took a more closer look at the engine subsystem. It was a way to provide plugins that could carry the implementation of ciphers, of message digest, and of custom, for example, hardware of load, implementation of worldwide algorithms or random number of generations. And the last but not least, we've seen that the main body of code contained hard-coded algorithm clearing. Shawan was the king at that moment, and well, okay, you can use any digital signature algorithm if your digest is Shawan. It didn't fit our purposes because our standards required that only Russian Ghost function should be used with Russian Ghost signature. And in the study we decided that the best way will be writing a custom patch that will implement callback structures for digital signature algorithms as it was already done for ciphers and digest. And then we thought when we write it, we provide it to upstream, they will accept our patch and everything will get brilliant. So here is a very brief history of Ghost implementation in OpenSSL. In the branch 098 we implemented a separate patch, which has broken binary compatibility, but we expected that it will be a good starting point to discuss. Then, for some time, Ghost Engine was a part of OpenSSL bundle. Then it moved to a separate project. For now, in the main stable branch 1.1.1, we have a chain of breaks and fixes. It's a normal process for not widely used functions. And there is a branch for walking progress, where I am implementing the current Russian Taylor standards. So when we have finished the first version of patch, for the 098 version, we were happy that we did it. Everything was OK with OpenSSL self-test. And we published our patch at the OpenSSL web mailing list and waited, well, not immediately accepted, but at least we were waiting for this or that reaction. As far as I remember, the only comment we got was that we should switch comments from C++ style to C style. It was rather discouraging at that moment. So then it took us about half a year to implement the patch. And then for about four or five months we did not know what to do with it. Because there was no discussion, there was no public interest. We made some presentation at various developer conference, but it was not our aim, because we had to adopt this patch to the changing versions of OpenSSL itself. And it was rather time consuming. So we needed to push this patch to the main codebase of OpenSSL. At that moment I noted that the signature of one of the OpenSSL team members, Dr. Steven Hansen, contained the line, we need funding and we will discuss the suggestion of work. Our bosses came into contact with OpenSSL team directly with Steven, if I'm not mistaken. And began the process of the integration of the library to upstream. It was finally added to version 1.0. And this version contained a lot of changes, but I will say about those that were the most important for us. There appeared a concept of public key context, pkcpx. There appeared two code structures, one to implement the crypto operations themselves. And the other two with ISM.1 encoding, decoding, and so on and so forth. And the reference implementation of Russian ghost cryptography, which became part of engine subdirectory. Well, and mention two points, which are the lower part of the slide, because they will be repeated again and again and again during the rest of my presentation. There are a lot of fixed internal lists in OpenSSL, enumeration of algorithms, enumeration of aides, enumeration of cypher models, on and on and on. And in fact, every changes in stand-ups requires changing in this list. And OpenSSL team treats it as a breaking compatibility, and I'm afraid they are right. I'm afraid they are more right than I am. And the other problem was, we tried to implement code structures in the LibSSL, which was a TLSSSL stack. But in fact, we didn't succeed because of some support of legacy, Cypher's use, and because of, well, at that moment, more irregular structure of the LibSSL itself. LibCrypto was more regular at that time. So, it was okay till 2014, when the HardLead vulnerability was published. At that moment, the OpenSSL team was significantly underfunded. So, when HardLead occurred, the community understood that, guys, we have a very important product, which is developed without regular procedures, underfunded people. So, it means that such incidents will repeat again and again. So, something should be done. And from OpenSSL team, OpenSSL management team was reformatted. The processes were set up. Now there are strict processes of reviewing the patches. Now there are public roadmaps. So, in fact, it means that, after HardLead occurred, the modern history of OpenSSL began. And now it's significantly much more secure than it was before, both in process and in code. In 2015, I met some people from the OpenSSL team during one of ITF meetings. I met Rich Salz, who is a well-known person in ITF cryptography community. And he was very in agreement. How should we deal with Ghost cryptography in OpenSSL? At first, the problem is that nobody in main OpenSSL team is interested enough in Ghost support. So, nobody was interested either to provide new algorithms or to fix something. And at that moment, we came to agreement that the engine should be moved to a separate project. But patches, which are not algorithm-specific but Russian protocol-specific, will go to the main tree and so it was a process that took three months to make support of Russian Ghost matching the new OpenSSL standards. It made me significantly rewrite the patch, in fact. I was very happy when the patch was accepted to main tree. At last, now I'm sure it compiles every changes. Of course, as new Russian Crypto standards were developed since 2005-2015, there were again changes in internal list. It was a more clear design of the Libre-SSL to that moment, so various support of Russian specific was moved to separate callbacks. And I hope that there will be a day when these callbacks will be fully encapsulated in engine, so it will significantly remove the process of negotiation of what to do with it. So, it was accepted and Russian TLS became first class variations of protocol at 1.1.0 branch. Currently, if you download OpenSSL you get support of standards that were penalized in Russia until approximately 2014. As I mentioned before, there are some work-in-progress branches. Ghost Engine became a separate package and it's available at least in Fedora, in Debian, in Russian widely used distribution as Linux and some other distributions less popular. There is a problem that as Russian crypto-algorithms are not worldwide standardized. We have a lack of packages containing certificates of trusted CAs. It's a really big problem. And last but not least we have a problem with Red Hat specifics, with Cryptopodes. Because I understand why Red Hat people don't want to have anything to do with Ghost. I think sooner or later we'll get an agreement how it can be without breaking the compatibility and system ideology. And maybe Ghost specific crypto policy will become a part of Red Hat package of Ghost Engine. Oh no! There are a lot of problems and interactions because of course customization has separate timeline which has nothing to do with OpenSSL process. So the best solution we have is distribution patches to main code of OpenSSL together with Engine for those who really wants to support brand new variations they will have to rebuild OpenSSL sometimes. But most of these patches are usually accepted to main code base. Of course Ghost has low priority for the world outside of Russia. We understand it. Yes. Of course we still have a problem of extending hard calling lease. And we will have it forever. It's okay. And we have a relatively poor state of international civilization of Ghost Algorithms though a lot of documents are published as informational RFCs so we are working on it. Here is the list of Algorithms which were supported in various versions of the Engine. I don't think it's interesting, it's just for information so you see that deprecator standards are fading out and new standards are implementing. It's okay. We have Russian national standards for X509 profile PXS10 We have national standards for PXS8 and PXS12 Of course we have standards for SMIME CMS Of course we have standards for TLS It's the main purpose for our clients but till the last versions it was impossible to implement DTLS using Ghost because some extra protection was not DTLS friendly it requires fixed order of packets to be delivered New TLS specification allow using DTLS There are walking progress specifications which are partly finalized in Russian partly implemented in code so I hope that sooner or later they will be part of Open Selling at least in TLS part and some additions in CMS and X509 And of course we have standard set of supplementary algorithm HMAC, BRF Many of them The next one Ghost or any in fact any non-standard crypto in OpenSSL Well, you still need to support it in the application that use crypto libraries For the most cases if the application uses the modern API you can just load configuration file ensure that engine providing the algorithms is loaded with the configuration file and everything will be more or less ok There are still now some legacy applications I marked them we know about RSA It means that in most cases you have to rewrite from this or that algorithm specific API to universal API to EVP API And sometimes for example the product of Cryptocom Open VPN Ghost we had to look into the protocol and carefully replace basic primitives to Russian Ghost specific and it's relatively rare case but you should take it into account Well Problem which stays unsolved till now is the support of Russian crypto in Java There are at least three Java implementations of Russian crypto There is a Wildfly OpenSSL Java library which uses OpenSSL and crypto provider I tried to provide to provide the ghost algorithms, ghost cyclers use ads in this library but I did not have any success First of all, because I almost don't know Java Loading the configuration the file did not help me so I hope that somewhere this work will be done and we will have support of in fact any as I told before any national cryptography in this script provider So, ok we took a wildwiple kit We used a lot of did not want write ourselves, such as I send out one library and many many other So, what do we give to community? First of all I think that Russian ghost engine using it is the most comprehensive testing of the engine subsystem in OpenSSL Then I should mention I should mention 2 bugs found during implementation of new standards The first was a very nasty bag that elliptic curves matter was changed in shortly before 1.1.1 version was released we found that it works with sub curves used in ghost because points on elliptic curves people who did this change they agreed that it was a bug and fixed bug I think relatively significant was stacker's exhaustion during TLS can shake though I don't think it was acceptable outside but anyway it's better to have it fixed As Russia is where non-latin base script is used of course we had a lot of problems related to internalization so some patches providing more support of non-latin scripts was fixed and now I have a side project related to support email address international in X509 I hope it will be finalized Yes, many people don't like email address international I don't like them too but as there are people in such countries as China Thailand and many others where alphabet has nothing common with Latin people will need these scripts to deal with it that way then I hope that sooner or later we will move some national specific structures to we will convert some national specific structures to form that will be managed via engine and it will allow to simplify the main code base of crypto library Of course a lot of memory leaks a lot of small bugs were fixed I want to mention a project we made by one of participants of ghost engine development who provided support of Russian hash Stribog in the Linux kernel if I'm not mistaken version 4.21 will continue Here are the relevant links to the presentation link to ghost engine itself then for those who based it and doesn't know where open cell code lives here the link to open cell well this may be the most important link in this presentation it's a brilliant paper describing writing custom engine in open cell and everyone who wants to write hardware acceleration or something else must read this article not should but must then here is the link to Russian crypto standard body enumeration of RFCs covering Russian crypto algorithms here is the document describing new Russian TLS I actively participate in this work and here are two mailing list dedicated first to Russian cryptography in open source products and the second is dedicated to ghost engine itself well the language currently is Russian but if you write in English of course you will be answered last but not least for those who cooperated with us during the well almost 15 years of development of Russian ghost first of all I should mention people from Open cell team Dr. Stevie Henson mentioned before Rich Salz mentioned before Richard Levitt and Matt Caswell they were very cooperative and did a lot to make possible to provide Russian ghost crypto in the main code base then I want to thank people from Red Hat team Thomas Mraz Alexander Bakavoy many thanks without the efforts it was almost impossible to implement ghost engine as a package in Fedora I want to thank some people from Russian cryptography community Vasily Dolmatov Vartan Khacheturov Igor Ustinov, Stanislav Smyshev who actively participate in national standardization of Russian algorithms and protocols and they provide help when it's necessary I want to thank people from Alexey Novodvorsky, Dmitry Levin Gleb Malinovsky, Vitaly Chikunov for the efforts in improving and development of the engine and of course I want to thank to those who contributed in the engine code itself and Artyom Tufrin who worked in Crypto.com and provided a basic version of the engine Alexey Diktyrov who implemented who provided a very fast implementation of modern Russian hash functions Max Tishkov who implemented Kuznetchik Russian modern cypher algorithms and many other people who I don't remember all of them unfortunately but of course they the efforts were significant for providing Russian ghost in PLS Thank you very much if you have questions please I'm here I'm ready to answer them I don't think more things into the comebacks and engine but on the other hand Chinese got their SM Cypher's directly into the open SSL code and I heard that at least it was like some email or some discussion I don't know if it was on GitHub open SSL or something that there is some interest establishment in the standard at least the RSE that they could into the actual the main code Well, it's a very good question and unfortunately I don't know the answer the question is that why these algorithms became a part of the core open SSL and ghost left aside So the Russian regulation in sphere of cryptography requires certification of implementations and it's much more simple to provide certified version crypto as separate engine then then publish it as a core it's impossible to publish certified version as a part of open source project so of course we can deal with code in the main code base replacing just replacing the code base by engine it's allowed by open SSL API but as the ghost algorithms were first set of third party algorithms appeared outside nobody wanted to build it in the main code base and as far as I know from China who became part of open SSL team or at least contributors and they support the algorithms at least part I made mistake about the status but I've got something like this I don't know yes I have a dream to become a part of open SSL team and support case maybe they will agree that ghost algorithm can become a part of open SSL main code base but well it works as part of engine and it's no problem Do you work in SSL 3? Lessonization has begun but not yet There is a draft describing new authenticated encryption mode which differs from Galois counter mode and CCM when it's finalized I think we will sooner or later provide the specification of using Russian ghost to TLS 1.3 What about Oh. It's a pain for OpenSSL There are no browsers except maybe links who use crypto from OpenSSL Most of them use crypto from NSS As far as I know There are browsers which support ghost but they are not widely used Первый — Ямпикс-браузер. Это форк хромиумов, я не ошибаюсь, и он имеет поддержку российских госталгаристов. The other one is a so-called Sputnik-браузер. It's developed in one of government agencies. It supports Ghost Algorithm 2, and, well, it has an Easter egg when you establish a connection, a secure connection using Ghosts. It shows not a lock, but a two-headed eagle indication that it's not just secure, it's Russian secure. It's offensive, it's offensive-efficient, but, well, if a Ямпикс-браузер is more or less in use, the Sputnik-браузер is almost not in use. Do you use Firefox? Yes, there is a commercial implementation of Firefox, implemented by a Russian company named Lissy. You can try to buy it and use it. Sorry? You said about the certificate of politics for one year. Yes, it's a very big problem, because, first of all, it's a political problem. From time to time there appears a project of standardizing, of creating a rather big Russian CE, and standardizing and making it conforming the CA-browser form requirements. And after that, the Ghost certificate can be added to various operation system bundles, and Ghost will start work out of books. But, unfortunately, now this project is not actively developed. You cannot care for Ghost, you cannot care for Ghost out of rush, except very limited cases. Some Russian sites require Ghost to be connected. And, well, I should mention about some projects of people who don't trust to MSA and decided that if they replace IS with Ghost, they get better security. Well, it all depends on the authorit models. Sorry, I don't... Yes, it's more or less used in Russia for, let's say, communication to government agencies, mostly, but there are a lot of projects of more spreading Ghost to... Если нет вопросов, я должен сказать, что национальная гипнотипография нет в России, а в России уже. Конечно, есть национальная гипнотипография в Японии, если я не ошибаюсь в Корее. И в некоторых репабликах, как в США, у Украины есть калина алгоритм, в Беларусе есть все алгоритмы. В США есть алгоритмы. Таким образом, проблема в использовании не очень worldwide standard crypto для больше людей, чем вы можете думать в первые часы. Спасибо большое!


Welcome to the 27th meeting of the Education, Children and Young People Committee in 2023. We have received apologies this morning from Ben Macpherson, MSP, and Ross Greer MSP. The first item on our agenda today is an evidence session with the Minister for Children, Young People and Keeping the Promise on the Children, Care and Social... Children, Care and Justice Scotland Bill. The minister is delayed, so our minister ein teidwar o'r rhaid iawn i gynnwyr bwysigau ar gilyddiaeth y gynnwyr y Deilgrifchesteriaidd yng Nghyrchau i'r blaenau. Felly, rhaid iawn i gynnwyr y gynnwyr y Deilgrifchesteriaidd yng nghyrchau ar gilyddiaeth ynghyrchau ar gilyddiaeth ynghyrchau ar gill yn Sgotland, a i'r rhagolau 2023. Galler rheinellu ar gilyddiaeth ynghyrchau yn gyfyrddfer i'r rhagolau, erbynnol will revoke a previous amendment made to the nutritional requirements for food and drink in schools Scotland regulations 2020 during the pandemic, which as a temporary measure allowed education authorities or grant aided schools to provide food or drink which did not comply with the 2020 regulations, where they were unable to procure a product given the supply issues at that time. The regulations will also correct a calculation formula in the nutritional requirements to ensure that nutrient requirement for school meals is calculated as intended by the 2020 regulations. Do members have any comments to make about the subordinate legislation? No. Is the committee agreed that it does not wish to make any recommendations in relation to the instrument? Agreed. I will now suspend the meeting. That concludes the public park for the moment and we will wait for the minister to arrive.


maybe what we should do is we should say what we're thankful for do you know what being thankful for me well you know what well you know what being thankful for like what are you thankful for Aiden? I'm thankful for people. Oh nice. What are people's answers? Like what do you mean? Like when they say something to me I do it. Oh okay. Okay. What are you thankful for? It's so sweet. What are you thankful for daddy? I'm thankful for the three of you. Seriously it sounds simple but it is. Thank you. You guys make everything a better day for me. I'm thankful for family as well. I'm thankful for Aria, Aiden. Can I have a kiss please? Daddy. Oh, more kisses. Thank you and I'm thankful for you guys. Thank you so much for your love, your support. Watching our craziness. Control craziness. I think it's fun craziness. It's real. It's real. That's the truth. So on the count of three we have to say happy Thanksgiving okay? Ready? One, two, three. Happy Thanksgiving from the housing life. Good job guys. Good job.


With the Super Bowl right around the corner and the Patriots in it, I went to the streets of the best sports city to see just how much these fans know. Who are the starting quarterbacks in this year's Super Bowl? Tom Brady and Russell Wilson. Tom Brady. Tom Brady, obviously, and then... Oh, um... Something Russell. Yeah. I don't know what's the first name. Russell Wilson and Tom Brady. Russell Russell. Russell Wilson and Aron Tom Brady. Brady. And we're playing the Seahawks, right? How many pounds of guacamole is consumed on Super Bowl Sunday? Three million. Five hundred thousand. Like six thousand. Also ten thousand pounds. It's eight million. Eight million. Yeah. It's a lot of avocados. What? That's crazy. I just got so much guacamole. So back in 2012, Richard Sherman tweeted a picture to Tom Brady. What did it say in the picture? Uh, I don't know. He's the best at whatever he does. I forgot what he plays though. I imagine he was bed-mattling something. I'm the best. He tweeted you mad, bro. You mad, bro? You mad, bro? What's your Super Bowl prediction? Seattle, 24, New England, 17. Let's go 32, Patriots, and Seahawks, 17. It's going to be a neck-and-neck game though. I think it's going to be a fourth quarter game. Can you give me your best Rob Gronkowski spike? Everybody seems to know how much guacamole they eat on Super Bowl Sunday. One thing's for sure, everybody knows that Tom Brady's coming to play. For Good Morning for You, I'm Kiko Dally.