Okay, welcome back everybody, Jeff Frick here with theCUBE. We're in the Palo Alto studios having a CUBE conversation. You know, we go out to all the events, we talk to a lot of executives and engineers and developers, et cetera. But what we really like to do when the opportunity is here, is talk to practitioners. People are actually implementing the technology, putting it in play, trying to get a competitive advantage and we're really excited to have our next guest, he's Stephen McKay, he is the senior director in user services and support at the Lending Club. Stephen, great to see you. Thank you. So for people that aren't familiar with the Lending Club, just give us kind of the basic overview. Sure, we've got sort of two halves of the business.  So you could come to us for a personal loan. The other half of our business is for people who have money that they wish to invest in a different kind of vehicle. And so they invest in other people's debt and it gives them a steady cash flow because when the loans get paid back, they get paid. And so we provide a marketplace for those two halves to me. So that's really what makes you different. It's clear there's lots of places that people can go get a loan, but I've never heard kind of that second half of the equation. So what percentage of your capital comes from people participating on the supply side? So almost, well, all of our debt is designed to be sold on the marketplace. We invest almost, well, we invest a little bit of money just mainly as a float, but almost everything is for our investor. That's so cool. And is it done in like a fund or how's it kind of structured or you're kind of buying into a portfolio of loans? When you go on to our platform, you go in and you can see the type of customer that you wish to invest in, certain FICO score, certain geography, certain background in jobs, the reason why they're wanting a loan.  Let's say someone wants $5,000. You're going to invest $25, $50 in that person and you're going to find 100 people like that to invest your money. So that way, if someone does default, that does happen. You're not out your entire... Right, right.  So you have a few days to decide which loans you're going to fund before they disappear because we are going to have to give the people money. How cool, and how can you share the scale of kind of the size of your operation or I don't know what's public or private so you don't say anything. So we actually are the nation's largest personal loan lender in this market space. Wow. So yeah, several billion dollars a year. Very cool. So presumably you have an advantage because you're a modern company, you're looking at different types of data, more data, cutting it different ways than maybe a traditional bank that's just using your FICO score, some of the kind of more traditional scoring methods. Exactly. So big data and data in general, tremendous piece of your guys core business. So what are some of the things, maybe I don't know if you could share that that you look at that maybe people would never think that's a valuable, not the whole portfolio, but are there some funny ones? I can't get into too much details because it is somewhat, you know, so probably it's the secret sauce. Don't tell me any secrets. But we do use a wide variety of the traditional sorts of things that people are familiar with, but we look at things that are a little bit outside the box too, that have a lot more to do with who you are as a person and the type of credit that you've had in the past and things like that. Very cool. All right, and then what do you do? Obviously, Ridge, you're title, but what do you keep busy with all day long? Besides coming to visit us here and follow all the tests? Yeah, I keep busy with making our internal employees happy. So I'm on the corporate technology side of Lending Club and I make sure that our employees have the tools that they need to be able to do their jobs on a daily basis. So I'm running back-end infrastructure or things like our email services and stuff like that, but also just the day-to-day grind of laptops and things like that. Right, keeping the lights on. It's a classic kind of IT. So you're here on behalf of Cohesity. So where does Cohesity play in your world now and then we'll get into it a little bit deeper as to why and how. Sure, so Cohesity is basically our new backup platform. We were a very traditional backup environment, standard software, back-end virtual disk system, very traditional type shop. Honestly, I've been in IT for over 20 years and I put in a system like this, one of my first gigs as a consultant over 20 years ago. So it was time to look outside the box and maybe shake things up a little bit and look at something that's been developed in the last decade. And so that's how we kind of landed at Cohesity. So what appealed to you? What were the kind of top two or three things you were looking for? Well, our huge, our biggest challenge was, I mean, back in when I started in IT I was backing up four gig hard drives and four gigs was awesome, you know. And now my phone is- Four terabytes, four gigs. Yeah, and now my phone is bigger than four gigs, a lot bigger than four gigs. And that backup system couldn't back up my phone. And so we have terabyte file systems and things. And with the traditional backup system that was, if it was successful, it took days, four days or so to actually do a backup. And so that's not tenable. And so going to something that rather than copying every file every single time does it on a block level and is a little more integrated directly into our virtualization layer was the right way to go. Well, I love how you said before we turn on the cameras that when you make a decision to replace something you try very, very hard to actually replace something and not just add something new. Yeah, so I drive my staff a little nuts because they know that when they come to me and say, we're going to do this new exciting thing and we're going to stop doing this over here. I'm like, you're going to stop. That means I'm going to walk in the data center, flip that thing off, right? And they're like, well, but there's that old stuff. I'm like, yeah, well we got to get the old stuff out. And so that was really one of the competitive advantages that Cohesity had for us is because they're not just a backup appliance or whatnot. They do have a file system in there. We could basically replicate all our old backup jobs into the Cohesity. And that way we have to keep the software around, you know, and being able to restore an old job if we had reason to do so, we'd be able to. But at least we can go into the data center and shut that old device off. So were there any particular features that jumped out at the top of the list that, or was it just, you know, you're looking for really a modern architecture with a whole bunch of features? Yeah, it was, it's really, it's a very modern architecture. It has some great capabilities to move data into the cloud and into A to S space to actually use this sort of same technology and the same policies to backup devices in the cloud that you would use on-prem. And so, you know, it has a lot of great features, but to us really the competitive differentiator was that file system. Being able to move our old backups directly into the system and be able to use our old backup software. We didn't have to do, you know, restore and re-back up or anything crazy like that. Right. So all your peers are all probably wondering how hard was it, you know, what was kind of the scope of the effort? What was the scope of moving the old stuff over? Well, so, what would you tell to somebody making, you know, considering this move? Have a good partner. I hired our integrator to do the actual migration. And one of the reasons I chose the integrator I chose is because they were willing to bid on this, knowing that what they're really gonna do is dial into my system for four hours a week for a very long period of time and just scheduling backup jobs to keep the engine humming. And there wasn't a lot of like sit there and there was no value of having one of my people sit there and watch stuff because it's just backup restores. It's not rocket science, but it does take a little bit of hand-holding. So I outsourced the actual migration of all the jobs. The actual setting up of Cohesity is like, you know, a couple of hours.  That was relatively simple. And you mentioned that one of the things that appealed to you was an integration to public cloud environments beyond just the on-prem. Are you using that? And if so, how are you divvying up what goes where? Yeah, so most of our services are on-prem or cloud services. You know, no infrastructure. We're just, you know, the sales forces that workdays, those sorts of services. And so we don't have a ton of stuff in AWS based on the corporate side. My peers in the product side would be very different answer there. But what we're doing is we're doing migration so that we can do our DR in the cloud. So that we can keep stuff on-prem, but if we needed, you know, if we had a problem on-prem, we can do DR. We're also doing replications between our colos, but that's our primary use case. Is to get it off, so it's cool. So do you consider that kind of secondary storage or it's really more just pure backup there if you had a problem? Yeah, so I mean, so we are looking for secondary storage and things, you know, our file servers and things like that. We've had such good performance with the backup migration. And so we're looking at getting off of our file server so we don't even have to back it up so it's just native objects inside. So I'm just curious in terms of kind of the data growth that you have to deal with on a day-to-day basis. Your data growth in terms of the IT shop is probably, your explosive stuff's probably happening more, I would imagine, on the core product or you're smiling and making it a funny face. Well actually, yeah, so it's something we didn't talk about earlier. So one of the things that was very interesting to me, we put in the Cohesity system and we sized it all out and based on our data volumes and things like that. But what we didn't realize is that we had a system that is part of our statistical analysis for our loan modeling, okay? And what we didn't understand is we couldn't back that up. It was too large and we couldn't back it up with our old backup system. And what the statistic guys are doing is they're building a model and going, does this work? And they'll run a ton of data through there and they'll create a model and it'll be two terabytes in size and they'll take one look at it and go, nope, that doesn't work, they'll throw it away, okay? And then a week later they go, well, you know, maybe, let me look at that again. And they call us up and say, I need to restore that two terabytes. Well in the past, they couldn't do that because we couldn't back it up, all right? And so all of a sudden, we can back this stuff up. And so it's getting backed up when we're just starting to do these restores. And so they only had a working size of 20, 30 terabytes or something like that. But what we found out was they generate like 10 terabytes a day and they throw it away. And so our backup volume had nothing to do with the size of the volume that we were giving them. It had to do with how much data they generate. So they generate a ton of data. We had to- So they want to back up Mondays, Tuesdays, Wednesdays, and Thursdays, even though the sum of that is five X, you know, what is their working amount. But they still want it backed up and they still make the call. Well in the past, they wouldn't be able to call us. So they would rerun the job, it would take them a day or two and then they'd have their answer. Now we can expose that old backup job directly to them. It's maybe not high performance because it is secondary storage, but at least they can take a look at it and kind of go, yeah, okay, let's bring that back into our primary storage and continue working with it. And that recreation is not so much a Monday, Tuesday, it's really like a 10 a.m., noon, two, four kind of thing. So has that changed the behavior in kind of the frequency or it's their work environment or now they feel more comfortable having a lot more of those models, a lot more simulations, and ultimately should help their business? Yeah, well and the thing is, is that it gives them the ability to quickly play with the model, throw it away and they can throw it away knowing we can give it back to them quickly rather than having them to completely regenerate the data. So they are able to churn through a lot more models. How many weeks do you keep that stuff? Or how many versions? Well, yeah, there's a lot of debate around that. It's zero to infinite, but there's a lot of debate about that. There's some negotiation around that. I mean, we have multiple different working areas and some of it's like, okay, if you think you might need it and you want to keep it around for a while and we actually may use it, then it goes into one storage area and we keep that for a lot longer. That's funny. That's a really elegant example of something we talk about all the time in theCUBE, which is at what point in time will the value of the data become a balance sheet asset? Whether that's your core data in your product set or I'm sure there's a whole lot of value in all these models that they're building. And before data wasn't necessarily considered an asset. It was a liability because I had to buy all this stuff to store it and keep it in. Like you said, some stuff I couldn't even store. Now people recognize it's a huge value. It's not necessarily in the balance sheet yet. I think it will be at some point down the road, but this is a terrific example of how you can explode the value by exploding the access, the reuse, the capability, without necessarily exploding the budget that you got to take back up to your boss. Yep. Yeah, very much so. And then to be able to drive all these different models, tweak them, customize them, standardize them, target them. Really, they must be loving that. They're very happy. Yeah, yeah.  The data explosions go and bananas. You're in a pretty cool, unique little marketplace. What are some of your priorities? What's next for you? Well, okay, so this is nothing to do with what we've been talking about. But a month ago, I turned my laptop into my desktop support team. And I now run everything off my phone.  My physical, that thing, I don't carry that thing. That's too big, big. So we have a VDI implementation. And I have a Samsung phone that has a dock. So I dock it and I have my monitor and I go in to do VDI. And but I don't have a laptop anymore. I can do everything I can do for my phone. And so I think that is like how to make that something that the business users can do rather than just us techie guys who want to push the boundary and push the envelope. I think that really is the future. And the whole idea of mobile first, it's kind of like mobile only. We really shouldn't be doing mobile first. It's mobile only and how can you make it work? And I like your style. You're just extreme. Like you said, you just turn off the old light switch. If you're going to make the move, make the move. Just rip the Band-Aid off and get on with it. They've got the laptop. I told them to redeploy it. It's a nice laptop. Give it to somebody else. If there's something I can't do, I'll go get one alone. Don't say that too loud. Chuck's looking for a few laptops. All right, Steven. Well, thank you for coming by and sharing the story. I got to dig more into the company. I didn't know that whole kind of backside in terms of the investor opportunity. That looks pretty cool. And again, thanks for stopping by. All right, thanks for having me. All right, he's Steven. I'm Jeff. You're watching theCUBE from the Palo Alto Studios. Keep conversation. Thanks for watching. We'll see you next time.


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Is this, is it true that it was at there? Ah, not really. Now, the first time you guys saw this guy was during Sheryl Gabriella's birthday. Not today in studio, we have him. Nteza kumpa fulsa aize kujintroduz, but if you are new here, Karibusana Ndanya, SPM buzz. My name is Shiko Gitao, Karibusana. Asa nchi. Please introduce yourself. Kwa taza majiwek? Ah, my name is Muya Manuwal. I am Manu on Instagram, TikTok and all other social media platforms. I'm a full-time forex trader. Manager, director, FX champions. Itunadil na two forex. I can say I'm a leader. Ya. Itunivingi. Itunivingi, mambo limingi. Now, of course, what to, or maybe I would say the public mostly knew you during Sheryl's birthday, 23rd birthday, because you guys know whatever happened. Okay, Sheryl said he was dared to kiss someone during her birthday. He is the guy.  Is this, is it true that it was a dare? Ah, not really. It wasn't a dare? No, no, no. What happened? Maybe you can tell us. He is through me. Yes, I'm a tema. Yes, babu, apu even doa tu ali kwenia sana, apu even doa tu ali kujulia sana. Wewe na others. Kidago. Apu even doa muli niya na. Yuna kwenia video ili enda viral? Nili wana. No. Naja palivu kwenia YouTube channelia, Sheryl, ali eza kwenika I was dared to kiss someone during my birthday. On the other hand, Manu, you have stated that it wasn't a dare, but maybe uneza tumbia wewe na Sheryl muli juwana lini. No, kusia kia ili designia ni me say ma, aiko wa dare, but aiko wa dare. Even doa na joy ko wa dare. Enu ili kwa birthday haiki. I remember ili kwa January, Tarre Sabah. Yes. Then, iu period, doa tumu kwa tunna juwana na Sheryl. Kus tu li juwana last year, I guess, November, December. Onaga. Tu kwanza ku vibe ni ni kawida wile. Onezonam say na mrembu. Onaga tia. Ya. Tu juwana like last year. Then, ika fika time ya birthday haiki. Iko January, Tarre Sabah. Wiki ya kwanza. Sosa sii, tu kwa enda birthday tu normal. Jutu li kwa iyo time, doa tumu na juwana. Ni kwa ikato kia like ni kama ni kwa birthday boy. Sii ni kama, I was the birthday boy. Birthday boy yosiku. Sosa iwa nili to kia? Birthday boy by the virtue of she is your friend. She is your girlfriend. By the virtue of what? Kwa time she was my friend. Doa time tumu na juwana. Doa time tumu na try kwanza things. Onaga. Bati ni ni muka icha tisha sa ama. Veni wali tisha tisha ina manisha kwa mba tulia ribu ama. I can say liya. Tulia ribu. Muli chwa ama. Jutu kwa tu me panga iya enda embu evo. Iwi li tukia yosiku ya birthday. Ika tukia tukua cameras ni ni. Ikaw ni kana kitukubo. Na ikua kubo evo. Vili na chachi shiwa. It was just a normal thing kawida. So this means Manu, you and Sheryl or probably on your side you were fallak of a better and easy word. You were in love with Sheryl then? In love. Or maybe attracted to? Attracted to the word. Not in love. Ako kwa in love ikumbaliki dog. And also Juzi li kwa nga birthday. Ako happy belated birthday. Sijuli celebrate Vipi because on the evening of Valentine's Day Sheryl was out there having dinner I think with someone because she was in a corner. You didn't see? No, I didn't see. We actually thought it was you. No, no, no. See what made me. I easy got it made me. So how's your relationship with Sheryl right now? Relationship. She's my friend. No bad blood. But I had to go and get it. Nira fiki twile normal. Ako busy na mambozangu. Ako busy na mambozaka sasa. Iwoto. So this means maybe probably out of the video ambayu li go viral. Ili weekend relationship or you're talking with Sheryl. Now what happened? See, you're back here in Lifanyika in January. Then your time to go to court. Talking stage. Then I got trust. Like I beat there. Kama bad day ake. Na jo ille bad day boy nikopale. Tukafanya bad day. Ikafika time ille sasa si juya keiki. Watu okopale wakai chachi shia. Like isi pe anu ena nakiji ko ame isi pe anu na fok. Isi pe anu ena mkono. Meaning they knew, like you guys were in talking stage? No, no, they knew. Na jo marafikoto likopale. Wengini wakai. Actual whatini wakai but kuna marafiku anu piya. Tuna juwa na watu olikopale, tuna juwa na. Na get siliku ille jokes ni ni ni ni. Eh, but ange kata. Anona. Eh, shed a chance yaku kata. I don't know. What would you tell her probably now, since you are, ama, ama tuna zasema mliatia pale kwa talking stage. What would you tell Sheryl Gabriela right now? Nami, si na kitu ya kumambia. Si na kitu ya kumambia. Hauna kitu ya kumambia. Apoj tu kazake vizuri, vilana pushingi. Aji tu metu mkawida. Given a chance, would you probably accept going back and continue from Mali mliatia pale kwa talking stage Mwendele? No, I don't think so. Si umesema kona mutu, ama. We are not sure, because Tuliana tumkono. So we are. Si labda zasahut ni mutu, ama. Ama. Sida ni vibe ilisha. I can say. I don't feel it. Ilisha. Manu labda si Juneza kumambia watu vipiku lenge, because at the time when people were asking you and also Sheryl on Instagram whether the two of you are dating, madam kumnesema you'll talk about it later, not then and things like that. But we had a couple of people who thought or who said that actually since Manu is a forex trader or Manu does forex then these people, this group of people have a tendency of dating non girls ili wa julikani. Si Juneza wambianini, because a lot of people associated whatever happened with you wanting to gain that fame. Uneza wambianini? Sisa ni ono Lisa. Julikani diyo ni nifani ke. Onagat. Nimo oni okitu chenye neza se mani ikotu vice. Nika miko vice versa. Aoma dim, ao ni. Ao famous. Diwa not after east side ya forex traders, kwa ono ili vibe, east side ni kubani, ni kuzuru. Vibe ko top. Onagat. Vibe ko top. Pesa ni ngili liya posas. Vibe ko top kila kitu. Alav si si forex traders, muna si tu neza kwa bad nao famous gas. Aoma mi expecto aditu na ki nanani. Si ni si si? I think the reason why wengu ulkona se maivo, they were trying to associate this situation of Manu and Sheryl with that of Kafuri and the lady ulkona itu nanani, Arian. Ata si danika malikonat after i. Hadja Kafuri is my friend, my brother, close kabusa, si Danika malikonat after kitu kama. But the timing nanani, arian. Timon으로 na uu ni diwa badu na wao. Kwa u failures o hakiili ni 100. Nili 200 ikakuibo, ...mingu. Mungu nakuwa game pangezu na me pangozot. Kwaibu nanasema kwaiteva pipua hafta kwaizu flashes. On the other hand is it like maybe... ...the famous girls are looking for the forex too. Na, ola jalazima hote. Mali kuna kitu. Kuna vibe. Yeh nia na, pelikana na. Naaisahid niku z collar. niku mot. Kuz collar in terms of. Kilaishit. We like it. So Manu, were you hurt with everything that happened? No. You were not? No. I was not. I saw on your Instagram page, of course during your birthday, Sheryl wished you a happy birthday. When was the last time you guys talked? No we don't talk. The last time you guys talked was during your birthday. The last time you guys talked was during your birthday. The last time you guys talked was during your birthday. I think we met at an event in Fulani. We are best friends. We don't talk. We don't talk. We don't talk. We don't have bad blood. Is Manu dating? For now, I can't say. For now, you can't say. What's up? You can't say. We are not dating. I can't say. I can't say. I can't say. I can't say. I can't say. Golds are 2024. I can't say. I can't say. My birthday is in Peter. I can't say. I can't say. I can't say. I can't say. I can't say. I can't say. I can't say. I can't say. I can't say. I can't say. I can't say. I can't say. I can't say. I can't say. I can't say. I can't say. I can't say. Waski. Hot. Ultimately happy. Hot. That is me and the others remaining. Yes, that is it. Can you pouch it in? bikingT It is OK. I am OK.  I can say full time three years two to three years full time nini kwa lani kido kido tu meza kumisha kwa mawatu ingua me pusa forex because forex me kwa a topic of discussion especially here in Nairobi and in our country most of the time the discussion is probably not so good ino fanyanga labdana fila just forex traders does it affect maybe probably your work the trust issues people have with forex? no I don't think so forex ni kwa mabiashara twingi ne na mwenya na takak fanya forex ya kuna interest kama kuna interest uta henda kufanya kutenda kufanya yukitu viloneza funguwa tu salon viloneza funguwa kitu oneza henda kwa forex gifunze, ujue na we invest pese ako so forex ni aki lamtu na forex siyo scam na ita wikua scam na pata forex ni malo ni invest pese ako kama manu labdana especially on your social media platforms you are very active on instagram siju kama ni kitu meko affect guys telling you that probably you guys are faking and things like that ujue kufek vileningum sasa siju sasa tuna fika aji siju kwa kiyu kufek inyo nasema ningum inyo nasema ningum siira isi kufek just go there try kama ni raisi kujia nita kupia lunch ufek, ufek, ufek ufek it's not easy faking siira isi aji ni brokambu gani ama ni ni not brokambu gani ni tu aji ni kampuni gani agari nizakupia agari bure ujue onawapi I think a reason watu alianza kufekirieivo is because to think kidogo kidogo kidogo kidogo amenunu kidogo kidogo amenungeza at a very young age watu amenunu ma shamba ma gari juzipia kufure leza kongeza gari, sindio? mi kitu neneza sama ni aji akuna kitu watu anasa we ni tango God's grace wezi bishangan na mungu mungu akisha chagwa labda alini chagwa kachagwa mungi neneivo evo amsideed through forex onapata nizakama tu ni le God's grace ni onawasi yungi nona na bimagari ni mungutu anawasidea nawewekama manu labda una fukiri kwanini forex traders wana face a lot of this maybe kukemewa inakanti neneza sama watu anawakemi ni nitu labda failures muntumu anawana sorry bro sista kama kupalingi neneza make it usi usiku eni uchuki ju mwenzakwa me make it no piga mamubi ako visuri tafta kitu akufanya forex tafta knowledge tafta investment tenge neneza pesa ako reisivo so are you a student of kufuri not as such but you not really but i can say ok student nuna manche student asha iku lead ana kukuside neneza idea kuforeks 10 asama 10 asama na tumesidea na sis whatu tumeside neneza sayin mefika maalile time tuna discuss asama nili ju kutridkilam tu ame ju anew our group tu wa tuna discuss market vile ina move leo tufanya je tu bayama to sell tufanya je goldi neneza api nasi neneza api witu kama iso tuna discuss ume ju evo neneza bahi nenezi witu kama iso na neneza time neneza foreks neneza land daily mbo na evi ime move mbo na jihenda evi neneza idea idea neneza wakesha tu na unu kwa korek unu kama up na nenezi witu kama strategies so i have my own strategies kujua ina sale ame neneza bahi ina do trick be there foreks nenezi witu a strategies nenezi witu a strategies neneza neneza ni kujia neneza kujua foreks ok neneza neneza idea je kitu mi neneza fanya ofko siyesi kupia knowledge ju mi mi pia i head to invest kwa iyo knowledge kwa iyo knowledge mi mi neme lipa class iya my guy ako gana sayi ako right now neneza kujia efeks na neneza kujia neneza kujia neneza ni pitiya classes kanda neneza wa tu tu nani pa classes neneza lipa 1000 $1000 kwa kwa kwa kwa kwa kwa kwa kwa kwa private class modern $1000 kwa kwa kwa kwa kwa YurWhen a unitinalismu kufanyip mobil okikuji to blamed r mama milaw 5 Ya o okika n ok.) Bora So kuhama So this means probably for me to get into a forex class, I'll probably need over 100,000 for a single class. No, no, no. So this means probably for me to get into a forex class, I'll probably need over 100,000 for a single class.  I invested more than a thousand dollars in classes, different parts. I went to classes because of South Africa. I went to Ghana and Kujo online, one summer. Let's get to hear from you Manu, he had to join you on a good day. I'm talking about a good day here as a forex trader. How much money do you have in Kenyan shillings? It depends on how much money you invest. Forex, it depends on how much money you invest, but you can invest more than. If you want to invest, I advise you to invest less than. You have to earn a lot of money for the economy to go to Kenya. It depends on how much money you invest. On a good day, it depends on how much money you invest.  There is probably a hundred. It depends on how much money you invest, but make sure you invest in Kenyan shillings and forex. If you have enough money, it depends on how much money you invest. It depends on how much money you invest. How much money do you invest as a forex trader? How much profit do you invest in a day? I invest in different traders. I invest in trade news. I invest in trade news like last year. I invest in CPI. I make more than $3,000. More than $3,000. But I make more than $3,000. I make more than $3,000. So this is approximately over $300,000? Yes, over $300,000 in Kenyan shillings. Let me ask you, how much news do you have in your hands? Do you have any trade news in your hands? I have a lot of news in my hands. I have a lot of interest in forex, because I have knowledge. I have a lot of knowledge. I get classes in my hands. What is your business? What is your business? I invest in news. I invest in trade news. I invest in trade news. It depends on what you invest in. You need some basics. You need some knowledge deep in your hands. To support the business, there is no need to be knowledge. I invest in trade. I invest in trade news. I invest in trade news.  Naumamu wajit Wrap I am very active. I am manu. I think Instagram TikTok niko active. Instagram niko active sana. Kila sana niko active. Ipa wunha zan hi reach. Ukini reach kwa Instagram nita reply d antsi yam yako. Of course tu twang. Haa tu dyuwe tu ta fanya aji. So the best way kukuku patan ni via your Instagram account? Yaa Instagram account. Nita apotseza Instagram awlshya waw mahi personal details zi ni raitisi ta kam nanamba tu ta kommunikir raitisi. Office ni? Nya office ni. Wunha ni o office ni wunha zan ni pata daily. Manu. From manu. We are listening to some of the youth from Manu and that from the people that we are with it. Thank you. Thank you for being here today. Thank you. All the people that you have been giving support to were moving forward. Also that we are here today. It was really incredible. It was really incredible! And thank you. All the people who are here today. Thank you. So, thank you. Thank you. This is our fun andności day. Forex trading classes Manu is very active on Instagram Amapalivo kwenyo ofisi as he has told you Thank you so much for watching this video Until next time I've been a host Shiko Gitao on all platforms You can find us at www.espmbaz.ke You can find me too at www.iendesco.com Shiko Gitao


For more videos on people's struggles, please subscribe to our YouTube channel. Welcome everyone to today's discussion. We are going to discuss today COVID-19 related death estimates in India. It has been under a lot of controversy. And what we have with us is Professor Christof Gulmoto. Professor Gulmoto is a senior fellow in Demography at the French Institute, IRD. He has joined the Center for Science and Humanities in Delhi in the year 2021 to work on demography of inequalities. Recently he has contributed a paper looking at critically reviewing India's death estimates and providing alternative estimates. Welcome Professor Gulmoto. It's a great pleasure to have you here. So as we discussed in and this is very much in the news currently in India that India's COVID-related death estimates are being debated a lot in the country. Bodies were floating on the ganges. We saw scenes of crematoriums overflowing with people and that shook our conscience, the nation's conscience in a great way. There have been reports that the government estimates of deaths are much underreported compared to what could have happened to the population. It's very important that we record deaths and we have proper death registration. Professor Gulmoto's own estimate suggests that we had 2.2 million deaths in India till May whereas official figures are only 0.4 million. Professor, your initial reactions, why should we have a better estimation of deaths, why death registration becomes very critical to a health system or country, not only in the context of pandemic but in general, your initial thoughts on the current situation? Well, I think the first issue is to count people and to give recognition to people's lives and that is mostly about their births and their deaths. And as you probably know, there's been a tradition in Western countries related to the Church of keeping record of deaths and births and marriages as well for more than 500 years. Whereas in India, it's pretty new. It dates back to the colonial period. Of course, death is taking but also birth and death registration which was introduced in the 1870s in some cities like Mumbai and Kolkata. But strangely, this process of registration of death and birth has moved on very slowly including during the last 50 years because there is still serious under registration of birth and death and especially of death. So birth as we know are more often registered because a certificate of birth is useful for many purposes especially for schooling the children. But death is considered of lesser importance to register even though it is supposed to be useful to sort out inner-reference issues and many others. But first of all, death is locally recognized in communities and that's enough to decide about sharing of property and or residential change. But otherwise, the under registration of death is still very important. It's still very significant in India because a lot of people do not consider it necessary to go and report death. So it varies in some areas and the registration of death is minimal. It can be for example in some cities or in Kerala or in Punjab. But elsewhere like in North Central India, about half of deaths are actually reported. So that's the first environmental feature to take into account. And then there are sudden deaths due to a catastrophe and that is to some extent something new in India because there's never been a catastrophe of death volume in India for the last few years maybe since the Bengal famine. So now there was a need to quantify this catastrophe and to quantify it just to let the country, to less citizens understand what has happened. And I think one thing that is very important and it has probably impacted a lot of us resident in India is to see and to hear about people dying around us. So that's new to me. You know, I'm coming from France. We had two serious waves. But to tell you the truth, I had nobody close friends or relatives who died. Whereas in India, after two months, it turned out that had several friends who died. So I realized that finally this crisis was for real beyond numbers. And then I asked for some of the friends here in India and I don't have so many. So that's why I was surprised that some of them, some of my own friends died. Sorry, but also surprised that it could happen. Then I asked other people and everybody knew of someone. So sometimes it was somebody close enough among their relatives. Or anyway, it was somebody they knew, at least one person. And I said, then how is it possible, you know, from a statistical point of view, how is it possible that everybody knows at least one or sometimes more people affected. And here I'm not talking about somebody who got COVID, but somebody who really passed away. And that's, I think the beginning for everyone in this country is that it's not only about what newspapers are saying, it's not only what numbers are being published. It's something that is affecting us, you know, be it in South India or in North India everywhere. This seems to have happened. And I think for most of us, it's something entirely new. We've never heard of something we've really heard during the last 50 years of people who might have died of, say, road accidents, but not within a year. So this became something considerable in our lives. And so it should be, at some point, be translated into numbers. Definitely. I think you made a very profound point. And that's very core to our understanding. Everyone in our family, every one of us have someone in the families, relatives, friends dying in this period of time. And definitely the government estimates currently are not able to reflect the magnitude of the problem. So coming back to our issue of estimating COVID-related mortality, there are some systemic issues of underreporting because of weak system, weak registration system. But also there are some, I think that there are systematic ways of undercounting COVID-related deaths. But how can we, how can we have better estimates, better methods of estimating COVID-related mortality if we do not have good registration system? Or, say, our health system is not equipped to capture what could be the other ways if you can enlighten us on the methodology of it? First, I guess we need to understand what's happening after a death. So it may be a COVID death, COVID-related death. It may not be. So if it is not a COVID-related death, let's forget it. It can be, for example, road accidents. So there is no doubt. But then there is a huge category of deaths which were of unknown reasons, especially, I would say, in rural India. And again, even local people had no clear idea whether it was a sudden fever or something like that. So there are no words to describe it. They might have heard about COVID, of course, but it didn't relate to their past experience of morbidity, of disease, of bad health, and finally of, and so a lot of these unknown deaths were just ignored. And some of them were due to COVID. And at times, even people had the fake feeling that this death may be concealed because they were, the family could be looked down upon as a family with the disease. And so they better attribute the disease to something else like all the age, because mostly all people were affected by COVID. So all the age is a blanket answer for death. And then you had deaths that were, in fact, clearly identified or could have been identified as COVID deaths. Now, a lot of these deaths are simply not registered. And so even if they were properly identified by the relatives or even doctors, but anyway, they were not registered. And so among those who were registered, there was even another difficulty is that they needed formally to be tested for COVID. But a lot of people, I mean, we've heard stories of people dying before getting even tested, and that in cities. So you can imagine that in rural areas, people were just, there was no way for them to test them in a primary health center or still they were dying. But then they would not count as COVID deaths because, and then finally, among those who were tested for COVID, some of them, and we don't know what proportion, in fact, some of these deaths were attributed to co-morbidity to some other thing that people would die of, say, diabetes. We know that diabetes patients were extremely vulnerable to COVID. So the cause of death would be attributed to something else. And this is definitely not what it should have been, what should have been done, because right from the beginning, the WHO requested practitioners and registers to use a very broad definition of COVID deaths. So whenever there was a suspicion of the deaths being caused by COVID, it should be registered at COVID, by COVID, and not under some other category. So here we are talking finally about the minor parts of deaths which were identified as COVID. A lot of them were just ignored. We, nobody had an exact clue or wanted to come forward and say it must be COVID. Among those few that identified, then they had to be tested and most of those recorded had to corresponded to persons who died in a hospital. And we're finally there. Somebody could say, yes, it is COVID and it is not another source. So if you look at the long line of the different steps whereby deaths may be ultimately registered as COVID deaths, they are, I mean, every time you go from one step to another one, being identified, being registered, being tested, blah, blah, blah, you lose 20% or 30%. We have no clear idea. But what we discovered soon that beyond our experience, which told us that it must be more than the official numbers, beyond that experience, we've seen local numbers that did not match with what people were observing on the field and be it registered deaths, be it deaths of all kinds. And then later on, incinerations or other dead bodies identified the number of which would not match official numbers, which I wouldn't call by the way government numbers. I would say just official numbers because they were counted by different local health authorities. And more or less everywhere we had the feeling that there was severe underestimation or say under registration.


Good morning. I hope everyone had a good weekend. My name is Henry Lester. TAs, do we have any announcements? If there will by any chance be a quiz this week, and the average is one quiz per week, it would also cover the material in the second half of the first lecture about deriving the ions in and near a cell from first principles. I think we have to stop deriving things from first principles today, though, and actually start some tough work. I do want to remind you about the Buy Neurobiology CNS 150 homepage. Please note my office hours. They will also be on the final slide of this lecture, and please read the book. Now, we exist here at Caltech in two worlds. We have the world of research and education, and we have the world of compliance. Many people who work in the world of compliance have a tenuous arrangement with the rest of the Institute, and the registrar in particular is usually a couple of weeks late with regard to what goes on in a course. And because of this, if you drop the course or if you want to register late, email the head TA, Geron, don't just talk with the registrar, who is an extremely nice person but has lots of other things to do. Now, this morning, ion channels in the news. The Nobel Prize in Medicine or Physiology was announced this morning, and it went to William Campbell, Satoshi Omura, and UU2 for therapies that have revolutionized the treatment of some of the most devastating parasitic diseases. The first two winners in alphabetical order, Campbell and Omura, discovered the ivermectins, including the drug ivermectin, and I will tell you about that drug in a moment. UU2 discovered an anti-parasitic drug that works on malaria. It's always fun to teach by 150 in the fall because you always need to step up and announce the neuroscientific connection with the Nobel Prize that gets announced at the beginning of October. So the ivermectins are the world's largest selling anti-parasitic drugs, the world's largest selling drugs for agriculture. Ivermectin and its analogs irreversibly activate an ion channel. It's a chloride channel found among many invertebrates, but not among vertebrates. However, its closest vertebrate homologue is the glycine and GABA receptors, which we will discuss at great length in this course. Right, I'm going to show you ivermectin in its channel. Many, as you know, there are various courses at Caltech about protein structure and about biochemistry. This is a molecular viewer called Paimo, which is one of the most common channel, which is the target for ivermectin, is called the Glucel channel. I will tell you a little bit about that in a moment. Here is an x-ray crystallographic structure of ivermectin down to the channel. The channel actually consists of five identical subunits, each with a different color here, and we are now looking down the axis of the channel. So that's where ions flow when the channel is open. Ivermectin would actually bind to all five subunits. I've only shown one of them here in this position, and it is an irreversible activator of the chloride channel. So we would see only chloride ions going through here, and this has the physiological consequences that I will describe in a moment. Does anybody have any questions about this structure, what it represents, or any other aspect? I'm going to shut down Paimo because having too many things at once does tend to crash my computer. So today's news very much exemplifies how contributions to neuroscience come from many other fields. In this case, from parasitology, but some of you are math majors, cns majors, chemistry, biology, bioengineering, other fields that have contributed to neuroscience are that, choreography, public health, biochemistry, molecular biology, statistics, physics. I was actually majored in physics and chemistry in college. So it is not surprising, it is not surprising, typical that parasitology should contribute to neuroscience and vice versa. So when Ivermectin binds to the invertebrate glutamated chloride, gated chloride channel called Glucel, this forces the cell, clamps the cell, if you would like, to go to the nernst potential for chloride. So the dominant ion whose permeability is in the plasma membrane becomes chloride. This is around minus 70 millivolts, and as we will learn in today's lecture and mostly in the next lecture, this prevents the cell from firing action potentials. And so the parasite, the invertebrate, cannot feed and it dies. And here's an experiment that we published a couple of years ago, taking the Glucel channel and making it into a tool for neuroscience. But you would see the same sort of records in a native invertebrate expressing the Glucel channel. Here we have transfected a modified version of the Glucel channel into vertebrate neurons in culture. Am I using any words that you would like explained, such as transfect? So this is an empty DNA, it's the control, and you inject current into the neuron as though you were a synapse, and the neuron starts firing without ivermectin present. Now, we first tried this technique a few years ago with an older version of the Glucel channel, and it did inhibit firing when we added ivermectin, but not enough. And so the optimized version of the Glucel channel, which we mutated and put GFP on so we could find it and all that, inhibits nearly all of the firing of the neuron. And so if you think you have a neuron whose function you want to understand, then you can use various techniques to engineer silencing into that neuron, or as is done in other labs around Caltech to activate that neuron. So they allow the experiment to silence the neuron by applying ivermectin. So very important drug, world's largest selling agricultural drug, ivermectin, and it has also become a tool for neuroscience. Same action, activating your chloride channel. Today's Nobel Prize. All right, moving on now to the usual substance of the lecture. You may remember that we said at lecture one that ion channels have a selectivity filter and that potassium ions lose their waters of hydration. Well actually, ion channels have two major functional characteristics. They have an ion selectivity filter, which determines which kinds of ions will permeate through them, and they have a gate, which determines what opens and closes the ion channel. Very few ion channels are constitutively open, although some are, and that's what causes the resting potential. For instance, nature has tricked today's Nobel Prize winning channel into being opened by ivermectin. Ivermectin gates the channel open. It's not usually open. Usually glutamate gates it open. So we are going to be talking then this week about two major types of ion channels. We're going to be talking about electrical transmission in axons, and here the gating function is accomplished by an electric field, actually by a change in the electric field. And in future lectures we're going to be talking about chemical synapses, chemical transmission at synapses, and there the ions are gated by neurotransmitters. Or in the case of today's ion channel by an external drug. Could anybody else name an external drug that activates an ion channel in the vertebrate central nervous system? This is an experiment that has done around the world 150 billion times a day. Yes. Okay. Very good. Thank you. Nicotine. Okay. I've told you that there's that electrically gated channels are gated by the electric field across the biological membrane. That is surprising because we don't usually think of cells have a resting potential of only the nudged potential minus 70 millivolts. Let's calculate from first principles the electric field across a biological membrane. So if we look that way toward Mount Wilson and toward the St. Gabriel Hills, and incidentally today is a beautiful day finally, we see high voltage transmission lines that carry current at about a megavolt, 10 to the sixth volts. The ceramic insulators which prevent the lines from shorting through ground to ground are about a meter in length. And so the field across those ceramic insulators is about one volt per meter. Engineers like to play it safe. And so that's presumably much less than the dielectric breakdown voltage of that insulator. And in fact, if we do look at dielectric breakdown fields in the real world, ceramic has a dielectric breakdown voltage of about eight times 10 to the seventh volts per meter, silicone rubber a little less, PVC a lot less. Right now, let's calculate the field across a biological membrane. The resting potential here, we're saying it's about minus 60 millivolts. The membrane thickness, however, is really small. It's about 30 angstroms. And so we do the division and we come up with an electric field of two times 10 to the seventh volts per meter across the membrane of nearly every cell in our body. Now, this is pretty close to dielectric breakdown for many materials. Well, what does dielectric breakdown mean? It means that dipoles in molecules get extended and sometimes they get extended so much that they break. Just so, in a biological membrane, the dipoles in a channel protein get extended and distorted by changes in the membrane field until those extensions and distortions make a conformational change that's great enough to open the channel. And I can tell you from experience that if we accidentally apply a membrane field only twice as much as usual, those ion channels will undergo dielectric breakdown. The membrane will break. We have to stop the experiment and try again on another cell. So, biological membranes exist very close to dielectric breakdown and instead the proteins are moving around under the influence of an electric field. Any questions? So, here, another simple electrical fact that I believe we talked about in lecture one is that we can think of an open channel with its complex alpha helices and beta sheets and fluid conducting pathway. We can treat it most of the time in neurobiology as a conductor, a little resistor with a finite resistance. Now, I prefer to call ion channels conductors rather than resistors, although they mean the same thing. And that's because treating them as conductors will allow us, as you'll see in the next lecture, to add very nicely in parallel. But they are little conductors. Any questions about that? Okay, let's go back to 1973 when I first came to Caltech.  We had one already. Now we're going to get another couple. I went to see Max Delbrook, who was a biophysicist and had indented molecular biology and molecular genetics. He died in around 1980 and had won the Nobel Prize for essentially indenting part of molecular biology. Max said to me, well, Henry, what are you going to do? And I said, well, gee, Max, I don't know. But, you know, I like ion channels and maybe I could measure individual ion channels. He said, well, I don't know how to do that. But it's getting to be noon. I have lunch with Feynman every day over at Chandler. Let's go there and ask him how to do it. So Delbrook and I went over to Chandler and Delbrook said, Feynman, this is Lester. Lester, this is Feynman. Dick, Henry wants to measure single channels. Tell him how to do it. Now, also present that day was Carver Mead, who is now an emeritus professor of electrical engineering at Caltech. Poor Carver was the only person in that group who did not eventually have a Nobel Prize. But he does have the Lemley Award from MIT and the National Medal of Technology and a couple of other distinguished recognitions. And probably among his more famous accomplishments is that he invented the touchpad on your computer that his company called Synaptics did that. And just recently, the New York Times, sorry, the New York Times had an article about the future of Moore's law and actually Carver is recognized in the Times and other places as the person who actually told Gordon Moore about Moore's law. Okay, and then there was me in 1973. So Feynman asked me, well, these ion channels that you're trying to measure, how are you trying to measure them? I said, well, we have these sharp glass electrodes and we stick them inside cells and then the potassium channels are open, sodium channels might open. And so currents obey Kirchhoff's law and flow in a complete circuit and that changes the membrane potential across the membrane. And if we can get low enough noise, we measure those ion channels individually. He said to me, no, I don't think you're going to do it that way. He said, the problem is that the membrane has a resistance and a capacitance. And he asked me what the resistance and capacitance were. I said, well, the capacitance is about one microfarad per square centimeter and the resistance is about 10 to the fourth ohm centimeter squared. And then there's the Nernst potentials in the membrane. He said, yeah, I was afraid of that because this capacitance, this RC circuit is going to act like a filter, which you all learned about in PHIS 1B practical. How many of you took PHIS 1 analytic? Oh, good. Oh, just one. You'll have a little trouble in this course, but the rest of you should do fine. So Feynman said now the RC circuit is going to filter out the noise, filter out the signal due to the ion channel and you won't see it. You need to think of a better way. The time constant is going to be about 10 milliseconds if you multiply this times that. Think of a better way. And he suggests, why don't you come up to the cell with a device that allows you to measure the currents directly rather than the voltage and just put in an electronic ammeter here in the circuit. And so the currents that flow will be forced to flow through the electronic ammeter. And the whole key though, said Feynman, is to make sure that the currents don't get shorted underneath the pipet and go back on their own. So he said to me, how close can you get? I said, well, I don't know. Never thought about it, but maybe a tenth of a micron. And he thought for a minute and he said, no, that's not close enough. Come back when you can get closer. So, of course, the field never stops and other people figured out how to get much closer. It's called the patch clamp or the gig ohm seal. And so now with the patch clamp and the gig ohm seal, all of the current that flows through a channel has to flow through the electronic ammeter. And here are the results, a single voltage gated sodium channel. So this is five picohamperes, which equals 10 to the fourth ions per millisecond, roughly, in 20 milliseconds. Now, the patch clamp is a wonderful device. For one thing, it has a tremendous dynamic range. One can record a single channel, even if it opens for only 10 microseconds. And even if it does this only once every 20 minutes, which is a dynamic range of 10 to the eighth, almost as good as the human genome. And the patch clamp can record currents ranging in size from two picolamps, which is about the size of this sodium channel to 100 nanoamps. And there are roughly 50,000 channels per cell. So we will come back to stories about Feynman and Mead next time. But meanwhile, the people who actually discovered the patch clamp, oh, let me show you a better version of that. So this was, my goodness, 25, almost 25 years ago, 1991 Nobel Prize. So it was sort of 24 years ago today. Erwin Nair and Bert Sackmann, function of single ion channels in cells. So the story that I just told you had got around. The morning that the Nobel Prize was announced, the editor of Nature called me and he said, listen, there's a rumor that Richard Feynman told you that you could never record ion channels. I actually said, no, it's not the way it went. He told me how to do it. I just didn't pay attention to him. So the Nobel Prize announcement said, this new knowledge and this new analytical tool has, during the past 10 years, revolutionized molecular biology, facilitated research and contributed to the understanding of the cellular mechanisms underlying several diseases, including diabetes and cystic fibrosis. So here we're back to this annoying concept today that actually electrophysiology and ion channels is more than simply neuroscience. Neither diabetes nor cystic fibrosis are neural diseases. And here's a simple diagram of a patch clamp as it existed in those days. Nair and Sackmann's scientific achievements have radically changed our views on the function of the cell and the contents of textbooks of cell biology. When they say textbooks of cell biology, they actually mean the early editions of the alberts, which you used in by nine or by eight. Both, okay. And finally, toward the end, here's a little bit about neuroscience. Drugs against anxiety act on certain inhibitory ion channels in the brain, alcohol, nicotine, and other poisons act on yet other sets of ion channels. So we have brought matters up to near the present. We're going to talk today about shaker, which is a very well studied, voltage gated potassium channel. Shaker, as Ralph explained, shaker is the name of a channel, but it also is the name of a gene that encodes the channel. It's a Drosophila gene. It was first studied in the late Seymour Benzer's lab by graduate students Lily and you knowing Jen, who are now very famous professors at UCSF. And it was isolated simultaneously by their lab and by Mark Tenoway, who was both a Benzer postdoc and a Caltech professor now at Berkeley, still doing wonderful science. And if I have my wits about me, we are going to simulate the gating of shaker, and we're going to use Francisco Betsenia's simulation programs at the University of Chicago. So here goes. This is Java, and you have to convince Java that it's not going to rob you or steal your identity. Yes. Okay. So we are going to talk about the so-called Hodgkin-Huxley formulation of a neuron membrane. Hodgkin and Huxley made this formulation in the early 50s, and they emphasized the gating of channels. But actually, they never mentioned channels or they ever measured a single channel. If you've been following the lecture, you know that single channels were not measured routinely until the 80s. And so Hodgkin and Huxley talked about macroscopic properties, permeability of membrane, which we now know is the sum of all of these little conductors in parallel. And Hodgkin and Huxley measured these macroscopic properties as functions of voltage, not of time. And so the conformational changes that I'm going to show you today in individual proteins, microscopic conformational changes that underlie the macroscopic properties are mark of processes. Would anyone like me to define what I mean by macroscopic? Okay, macroscopic means much larger than an individual molecule or an individual event, but characteristics of the population itself, macroeconomics as opposed to microeconomics. And so these macroscopic processes are typically sums of individual microscopic processes. And a Markov process is a process that involves a transition between two states of a molecule or a state or a system that has a defined simple probability of occurring with time. That probability does not depend on the history of the system. So ion channel gating depends instantaneously on the electric field across the membrane on the voltage, not on the history of the voltage. But it's easier to show you this in a simulation. Okay, now here is the simulation of a membrane. And we're going to show you the following events. The channel is actually shaker that's been simplified. It's been mutated to remove inactivation. And I will tell you about a little about activation later. The model is based on several types of evidence. It's based on biochemistry, electrophysiology, lots of site-directed mutagenesis, x-ray crystallography more recently in fluorescence. The model shows only two of the four subunits. So there are four subunits. They are not color-coded by subunit. They are color-coded by transmembrane helix within the subunit. And so in front of the screen and behind the screen, there are two other subunits. There are several states in this model which would be associated with conformational changes of the protein. So you can put the protein in one state or another. Here is closed number one. Here is closed number two. You see there is a rather small change, but that's enough of the change to open the pore when it occurs to open the channel, when it occurs simultaneously in all four subunits. And if we now make the change further, that's the one that actually opens the channel in all four subunits. So let's open this one and open that one. And now the green arrows go all the way through. Those are potassium ions leaving the cell. So we don't need to think about current, we just need to think about the flux of ions here. Is anybody lost or requiring a further explanation? Tell the Java app change the confirmation of the protein into states that do not allow the channel to be open. The question is, when you press the buttons, what happens? And the answer is we're modifying the confirmation of the protein to go into states' conformational descriptions that are not associated with the open channel. So here we've opened one. The channel doesn't open with only one of the four subunits in the active state. It needs all four of them, but only two are pictured here. So here we are activating the channel manually. Now we're going to do what, yes, why do the white and yellow, the question is why do the white and yellow not move at all? These are, the white and yellow are actually helices within a subunit. They are not by themselves subunits, show me. And so you may remember that the pictures we showed of ion channels have big complex proteins, and in the middle of that complex protein is the conducting pathway. You could think in this case of the white and yellow helices as insulators that hold of the rest of the protein in place to respond to the voltage, and also interface with the lipids in the membrane. So they are the, if you like, if we made the analogy to a wheel on an axle, that would be what holds the bearing in place. Any other questions? You know, I've been explaining this for so long that I think everybody knows what I know, but now is the time to say, well, Professor Lester, actually you need to step back. Right? So now we are going to let the program do a voltage step. If all, oh dear, start. Okay, now that all this is very complicated, what the program is doing here is repeating a voltage step. Now you remember the Markov process, the Hodgkin-Huxley formulation says that the rate constants for opening and closing channels are not functions of the history, they are only functions of the voltage. And so the essential trick of the voltage clamp experiment is that the voltage is always constant, then it's jumped to another voltage, and then it's constant there and jumped to another voltage. That's what clamping means. Now cells do not do this, their voltages vary all the time, but in order to understand how ion channels work, experimenters have developed the voltage clamp paradigm. The patch clamp is one example of a voltage clamp. The old-fashioned intracellular electrodes that Feynman told me to throw away are another example of a voltage clamp. So we are repeating the voltage pulse every trial for many times, and that's because each trial which concentrates on only one channel is a microscopic event. Now by doing the experiment many times on that one channel with its statistical stochastic behavior, we sum that behavior until we get a macroscopic signal, but actually this cell is doing it all at once during a single trial by summing simultaneously the microscopic behavior of a lot of channels, in some cases 50,000. Okay, so far? All right, so there's a lot of stuff in these traces, and you can see that the traces differ from one to the next. That's because the Markov process is inherently a stochastic process that is governed by statistical transitions from one state to another. So the left shot and the right shot really mean the states that we previously controlled manually and now that voltage is controlling them, going from C1 to C2 to A. That's simply what the shots mean. Eye gait is something that we won't discuss at the moment, although it's really cute. The main current that we want to talk about, the main phenomenon we want to talk about is eye single. Now, if you look monomaniacally, you'll note that when the green arrow goes all the way through, the current trace is up, and when the green arrow stops in the inside the cell and can't get through the trace is down. So this is simply the ionic flux through the channel. When the channel opens, current flows. When it closes, it doesn't. And you'll note that there are fluctuations in the opening. That's because of the stochastic processes that keep the channel more or less open, but sometimes closed, and more or less closed, but sometimes open. At the resting potential, by definition, the cell is at rest, and the channels do not have a high probability of being open. They are gated by voltage, actually by changes in voltage, actually by changes in the electric field across the membrane, which pushes and pulls on the dipole moment in the protein until these conformational changes occur and open the channel. Now, down here at the bottom is what we are used to seeing. At now after 105 summed trials, we actually have a fairly smooth macroscopic action average of all of these individual guys here. So this channel takes a couple of milliseconds to open all the way, but then it stays open. Any questions about the individual stuff here? You can run this yourself, and it's a pretty good simulation of the actions of a particular type of channel. Remember, the channel is the Shaker potassium channel, and you will come to understand this code. The code is who or where, and there's only one correct answer usually. So where was this channel discovered? Which institution hosted the discovery of this channel? Caltech. All right. Now, there are a couple of interesting little electrical characteristics here. Remember, I told you not to worry about this current called iGate. Actually, this current called iGate is a capacitive current, and it has to do with the fact that the charges here in the membrane that move in response to the voltage actually move in or out of the membrane. And if you think about physics one, the charge moving in a dielectric is a little bit of a capacitive current. And so sure enough, since the charges don't leave the membrane, the time integral of the charge movements is equal at the beginning and the end of the pulse, because those charges don't leave the membrane. And another interesting point is you see this little blip down here, pause, this little blip right here, and this little blip right here. When we step back to the resting potential, the channels don't like to be open, but they don't close instantaneously. They close according to stochastic processes. And so therefore, the currents stay on for a fraction of a millisecond. So, electrophysiologists have talked about all of these characteristics, and they give great insights into the way that ion channels actually make information in a cell and conduct information in a cell. Now, the charges here are not surprisingly positive charged side chains on the protein. They are lysines and arginines, and typically they occur every third residue in a helix. In the helix, there are great arguments about whether the helix twists or moves up and down still. Green stuff in the middle here, without those insulating helices that Shaman asked about. Roughly corresponds to the ion channels I've shown you before, the very simple ones, that have only two transmembrane helices per channel. What is the counter? Now, because where is the voltage, actually? Well, we're in the membrane here, and the voltage is somewhere around, across somewhere around here, probably toward the top of the channel here. This is the extracellular solution, this is the intracellular solution. Now, note that active state, a closed state, the gate is near and prevents ions from flowing. And so, this helix actually consists of a break and then another helix, and this is not surprisingly to those who've studied protein chemistry. It's a glycine, which does not have a side chain and is often associated with breaks in proteins. And so, if there's a hinge, it's right here in this glycine. All right, I think we have done enough, any questions before we turn this simulation off? Question is whether modeling ion channels as a marker process is relevant to processes that occur in the nervous system? Absolutely. It is a very good approximation, but sometimes unexpected environmental influences can change the transition probabilities, and sometimes those transition probabilities do change with time and with other events such as phosphorylations, proteolysis, etc. I'll get to it on the next slide. There are many voltage dedicated potassium channels, and each has its own voltage sensitivity in kinetics, and so one needs to understand the details of the voltage sensitivity in the kinetics for every potassium channel. Now, we're going to move on from potassium channels to sodium channels, and you remember that sodium channels, there it is, sodium channels are selective to sodium, and their side chains make a sodium ion feel more comfortable than a potassium ion. Sodium channels have a water-like pathway that trick the ion into being comfortable in the channel and make it a little conductor for sodium, but returning to the question that was just asked, how good is a Markov process? Well, the way you make a better Markov process, a more realistic Markov process, is that you keep adding states to the model, and so in order to understand the gating of sodium channels, you need to add another state, and that is the inactivation state. Ion channels open, and most of the time they don't stay open, they close again, even though the stimulus may still be there, even though the voltage may still be there, they close again, and so typically, back in the form of the years, people added another state called the inactivated state, and the more we know about sodium channels and other channels, the more we understand that typically the inactivated state looks like a flap that can cover the pathway and prevent ions from flowing in a different way. Now, not only are there flaps in ion channels that can inactivate those ion channels, and as you know, sodium action potentials are rather brief. They last only a millisecond or so. That's because sodium channels inactivate after a millisecond. We'll talk about this in the next talk. Not only are there parts of the channel protein that can come in as a flap and inactivate the sodium channel, but there are lots of external chemicals that can come in and inactivate channels or activate them. We talked about ivermectin. This one is tetrodotoxin, the puffer fish toxin. So in order to do a fair analysis of the sodium channels, which are after all the basis for the action potential, we need to look at another simulation again from Betsania's website. This model is a bit more venerable than the potassium channel, and you'll see the graphics are not so great. The potassium channels have four identical subunits. Sodium channels have four nearly identical subunits, but they're all actually hooked together in one polypeptide chain. So you could call it a pseudo tetramer. The individual repeats resemble an individual subunit. The part that actually selects for ions, the so-called P or poor region, differs. We are going to see now that the single channel events in this simulation, sodium channel simulation, there we are. So the sodium channels, the graphics are a little cruder, consist of these four homologous domains. Here's the inactivation flap down at the bottom. We're looking from outside the cell. So now we go to the simulation. We're going to start the simulation, and we're going to start it. So this green stuff around here is the membrane. The orange balls are sodium ions, which want to enter the cell, and as in the previous simulation, they only enter when the gate is open, but also they can be prevented from entering when the inactivation flap is closed. And so we have an additional so-called Markov state. I don't know why it should be running all the time. Here the channel is open, ions are flowing. You see little plus signs on the helices. Those are responding to the electric field. They're being driven one way or the other to produce conformational changes that open and close the gate. And then after the channel has been open for a few milliseconds, there's a statistical probability that the inactivation flap will come in, stop conduction, and terminate the action potential. Did I answer your question? Any other questions? The question was both the activation gate needs to be open and the inactivation flap needs to be out of the way. Those are two separate conditions, and they need to be met. Yes. In the Hodgkin-Hurksley equations, which are macroscopic, there are two parameters called brilliantly enough, M and N. M describes the activation gates, and since there are three of them, you have to cube them. N and M runs from zero to one. N describes the inactivation gate. It runs from one to zero. Since there's only one inactivation gate, it is there in the first power. So Hodgkin and Huxley, 40 years before the gene for sodium channels, was known. Guest from the kinetics of how they had to model the channel, that there were three subunits that could open it, and one subunit that closed it, even before they knew what subunits were. Okay, our next lecture does employ electrical circuits. Please review your material from This 1b Practical, Huxley-Ohm's Law, and you can also see Appendix A in Candel. And my office hours are Mondays, not Wednesdays and Fridays, entering near the red door from 1.15 to 2. See you Wednesday.


Stuck in the rewind haunted by shadows of the past feels like yesterday's on repeat crushing your today Stop hit fast forward. You ain't stuck.  What lights your fire? What makes your heart skip a beat?  Tell your body. This is what's coming. Get ready clear mind Pumped heart. That's the recipe for change


Hi everybody it's Sam at Mixed Up Girl thank you for watching my tutorial today I'm going to be showing you how to make this really nice rather large size gift box it's all reinforced it's got a flip lid you can see inside there some of you if you've been following me for a long time you'll recognize this bow it's a slightly smaller version of my big bow that's what I call it my big bow topper I made this oh I probably want to say about two years ago I reckon and what I'm going to do because I won't be showing you how to make this because I've already done it I'm just going to link you to that one there so if you click on that one it will show you how I made this bow I will give the measurements for this one and the quantities because obviously it's a little bit smaller but also the box that I put this on in that video is another nice 6x6 I think that one was it's really lovely but yeah I thought I'd show you this one actually reminds me a little bit of like a carriage clock it's got that kind of look about it you could maybe cut a little aperture window here put some acetate behind it so it's really really fun like I said all reinforced and yeah let's crack on and I'll show you how to make it okay so there is the bow for today's all right so that one's all done again like I said I'll list everything below and I'll give you the measurements to that topper later on now I've done one base because you do two of these so this is one already done and I'm going for this blue today because the papers I'm using are from the new paper addicts magical stories so this is a Christmas paper pack however you know there's so many patterns here that you don't have to just use for Christmas I mean with this one here if you just remove that it will make a lovely box for a birthday so you know that I did say that as well when I done the unboxing that you know you can use these papers that a lot of the reverse size sides as well so that's the paper pad I will link all of that below but the one I've pulled out is this one here with the Christmas baubles on and it's got this nice kind of just very light blue colour and I just thought this card worked really quite in the well with it so you're going to need I'm going to go through all those measurements in a moment so it's a deconstructed box I've made this side this style box before and I've done it for the teacup and saucer it's that one with like the hinges that's how we're going with this one today so for the base and the lid you want two pieces that are eight by nine and a half and on all four sides you're going to score it one and two inches okay rotate one and two rotate one and two and again one and two these are both reinforced so you want to do that on both pieces you'll have two like I said I've already done this one and I'll show you how to do that one in a moment then for your side so what I've done is I've reinforced these by sticking them to the the blue card stock so this is actually a strong I would say it's it's borderline paper and card it's a really good quality so it works great on its own and it would also work as a you know a card blank you could have that you know as the front of a card and then do your mats and layers and so on but because the way I'm doing this box and because this is reinforced I've decided to stick these on another piece of card stock and I'm using the Kalau glue because this just dries completely you know stiff so it just creates more of a chipboard feel to your projects so it's entirely up to you if you do that your card stock your pattern card that you're using might be even thicker than mine and you might not be using pattern you might be using a coloured you know just plain card stock and in that case I'd recommend like a 300 350 gsm you wouldn't need to do this but if you do then you're going to need two pieces of pattern paper that are three and three quarters by eight and then two pieces of a coloured card you know mine's all matching so it's just the same card from this it's actually the little card stock if anybody's wondering you want them the same size and just stick them together you can hear there they're just solid pieces now much much stronger than that on its own then you'll also want two pieces so this is five and three eighths by eight inches you want two pieces of coloured and two pieces of pattern so I'm just going to show you because I'm going to stick this one over the top now I did do this long you don't need to ignore that I folded that over really you don't need to do that at all I was just trying something else out at that point but I'm going to stick them together in a moment then for your hinges you want four pieces that are three quarters of an inch by eight and then along the three quarters of an inch you want to score it three eighths of an inch right down through the middle and just fold and burnish those you might want to do thicker you might want to do two by eight and then score at one inch or you might want to do one inch by eight and score at half an inch it depends I just wanted it quite thin my kind of hinge so again those of you that have made these because I know so many of you have made like loads of the teacup and saucer box then you know yeah you you know what you're doing then you'll need four pieces of three quarters of an inch by five and a quarter that's the mirrored cardstock so four pieces and then four pieces of I think I just dropped this down yeah half an inch by five okay and then again you'll want four inches of three quarters of an inch by three and three quarters and you'll want four again of half an inch by three and a half inches and these are to decorate the bottom here so that's completely optional and I've also done it on foam so the blue is raised so stuck them all down you'll see they've got my foam in between so I've got all those ready to go as well and then for the topper the mirrored cardstock is three and three quarters by five and a quarter so the white piece on top will be three and a half by five okay so that's all of my sides already so then we'll do this so you'll have two pieces now you will end up cutting one of them slightly different but I'm going to show you both ways on this one so first of all just fold and burnish all of those score lines okay and then we just need to do a bit of cutting so along the shorter side you will have four squares here you want to cut down pass the first score line down to the seconds there's your first down to the second and again do the same with that one okay so there's still those four squares these two outer ones you want to remove completely and then that one as well so then all you're left with is just this one which is going to create a tab so do that on all four corners okay so this is what you should now have and all of these ones should all be free apart from one side and then we just want to go and take little wedges off of all of the sides of everything okay so off of there just kind of mitering it all this one here every edge that you have apart from these actual outer side pieces just all of the all of these bits here okay so you're going to do this the same for every single one but because this one now is going to become the lid I am going to do this one slightly different so what I want to do first of all is show you how you will glue all of it down for the base okay so I'm going to glue on one of the tabs and then just bring one side right the way up like this okay and then at the other end of the long side because I'm doing the long side here you're going to get again going to pop you will end up putting glue on all four okay so if you're doing this as your base you'll you will stick all four of the corners in like this so this one this one as well but I don't want to do those ones because this is going to be the lid I've already done the base so you see there how it's going to look okay then if you imagine I've already stuck these in you're now going to fold in all of those sides with glue and you would do that on that one as well and that will give you this okay but because this is like I said going to be the lid the difference you need to do with this so although the scoring is all the same and we cut it to that point you then want to cut away so what we want to do here is just cut away this completely and this one here and then cut all the way down this one here okay so then all you're left with is that hinge these pieces then you'll just stick in as normal all right just get your bone folded and just go in really burnish that side and just make sure that glue is spread out and you'll do this whether you're doing the lid or the base it's all exactly the same the only difference is you're cutting that piece away on the long side on one of them all right okay so you'll have one that has all folded in and then you'll have one that has this piece just flapping out over the side okay so now you want your hinges and one of your front pieces you'll have your front and back of the bigger ones and then these two are your side pieces and we're just going to start putting them all together so you're going to have a piece of the hinge stuck on the front like that just half of it like so okay so it's up to you whether you want to add the glue in there first or on here I'm actually going to add a very thin bead onto here and then you can just kind of fold this over like and just kind of push down both sides because the glue is only actually going to stick to one side but it just means that you can really stick it into place then okay so there's now one of my hinges on so I'm going to do the same on this side okay and do the same with the other large piece okay so I've got my two front and back pieces there and then you want to grab your sides and grab any of them and you're now going to stick one of the sides on so again I prefer to put my glue onto this piece and obviously make sure you've got your directional paper if you are using it up the right way and again just go around sticking it all down okay and then you want to attach this onto there so again I'm still going to stick my glue on here okay so that's that piece now while that's still just drying a little bit I'm going to stick these on the side of this piece they're still going on all four sides even though that bit's slightly disconnected so I'm just going to stick all them down okay so stuck my top on as well so that's my whole lid all ready to go with the hinge there at the back and then this here the easiest way to do it is if you push in the sides push everything in and then you'll get it into the lid or sorry the base like so and then once you stick everything down it should all fold completely flush which it does okay so what I do again is I put my glue this is liquid glue is going to be better for this project because it gives you that time to move things so I'm just going to cover about an inch because that's the depth of the the base and I do have time with this glue as well so again just going to squeeze it all in get all your points in and then it would just push itself out pop it on its side and you can go around get your hand in and really push down make sure you're pushing it right to the base there okay so you keep everything straight and there you go really lovely very very strong this this really is a super strong box and if you want to reinforce it even more you could add another you know you could add some chipboard on that base there as well if you wanted to but this is going to hold you know hold definitely a bottle of something so yeah it's very very strong and then all you got to do is pop your lid on so what's going to happen is that's going to go over the front and then this back bit is going to stick onto the back of this so that then when you lift it up you've got the hinge okay so again I'm just going to pop some glue in here side if you've got a preference to the front or back my designs are pretty straightforward and if you put it on at the front first push it then right back and then bring that down just hold it there for a second and once it starts to tack then you can lift it up and just make sure it's all but that way you know your lids on perfectly whereas if you go to put it on lying it down lifting it up slightly you might not get it in place but you can see already now everything's nice and flush and yeah it's a really special box I like this one a lot okay so there you have it a really lovely very very strong nice size gift box I didn't actually give the overall dimensions for this one at the beginning but it will end up being five and a half by eight by three and three quarters so it's a great size I love them I love that they're not your traditional Christmas which for me is a little bit of a newish thing because I tend to always stick with my greens and reds but I'm enjoying playing with these papers I need to put a top on there actually I'll do that nearer the time but you can see what I done there that was just using a one of the papers in the same pack what you can also do is if you want to have it as a closure then there's no reason why you can't I've got some scrap card here you could stick a piece of cardstock underneath there then put a nice topper on here like a circle something said Merry Christmas and then put some Velcro underneath here or a magnetic closure and then it would close and it would keep the lid if you want to do that way you guys you always amaze me with what you share over on the Facebook groups so I'm sure some of you will come up with something you can also turn it into a gift bag there's no reason why you can't put hole punches here and have a handle on it I think that would look really really nice as well put some feet on it you know make it look like an actual clock there's just so many ideas with this but it's just a lovely size box so I hope I've given you lots of inspiration I hope you've enjoyed today's tutorial as always if you have please give me a thumbs up I do really appreciate it and subscribe to the channel so you get to see more because I've got so many great things coming yeah until next time thanks for watching bye