This video is brought to you in part by TrueTechTools. Quality tools, essential support. What's up guys? So we're working on this little row of cooler doors. We've got a little problem today. I went ahead and scouted this out. Well, we've got the system we haven't worked on. So it's kind of new to us, I believe, far as happened to work on it lately. The initial problem was the customer had this particular evaporator freeze up. So what she did was turned off the fans, which these are both together in parallel and the system continued to run. They left it off for a week and here's what happened. So to say the least, it got worse than what it was originally. So what I've done is gone up on the roof, flipped off, pumped the system down by putting it into a defrost and then shut the power off to it. We got one fan out here. We've got, I believe, another fan here. That looked like it wasn't one of these didn't look like they were spinning right. They might be working now, but we're completely froze up inside here. We've got milk, which is their biggest concern right now because they just got a delivery. But we're going to get this milk moved into another cooler.  And so I'm up on the roof and I look at this cooler here and it's turned on pushing straight reports and there's no refrigerant. So this cooler is not going to run. And we've got other problems to fry before we would even consider screen with that and they haven't been using it anyway. So we have a freezer. So flipped on the freezer and the freezer runs. So what we end up doing is going in and flipping the thermostat up to 35 degrees. Moving the defrost clock down to 20 minutes because it's a heated defrost and that's going to be good enough to get them by to hold their milk. And it was holding at 35 and with the door open to hit to 40. So we're going to be fine there from now while we're working to get that going. That's the start of what I'm running into. I couldn't find the key. Luckily, a little piece of duct tape really kills that siren and we got that wedged. We're going to do is we're going to try to break off some of this ice from the bottom because I really don't want it all melting all over the floor. As you can see, it's already starting to melt pretty quick. I'm going to grab a trash can, dump it into the trash can, see if we can knock that loose and then then we'll melt the rest of water. We went ahead and knocked some of the ice off. Knock some of the ice off there. You got to be careful though, because you don't want to puncture anything or create more problems. Still got more to get off there yet. But I'm trying to get that done as quick as possible because it's making a mess. They never put floor drains anywhere near here, which just makes it more difficult. We could have put a tarp on this, but I usually don't run into this that often.  There's no reason they should have had these.  There's no reason to have these on a cooler anyway. I see it sometimes on a freezer, but even then you need to wire it on so it kills the solenoid at the same time.  Instead of just letting it run, run, run, run. Melting it away slowly here. We've got most of it knocked off the bottom. Oops, got most of it knocked off the bottom. Once we get the back down, we'll go ahead and get the inside done.  As soon as I can get it to release from the coil and just kind of snap it loose. And then that way, because I'm not trying to melt it completely, I can bump it a little bit and make it pop out great. Otherwise you can spend all day here. I've been doing this kind of throttle on it and shooting mainly at an angle so it doesn't go too far into the coil. That way most of it hits the outside. I've already gotten one piece to snap off, literally just bumping it. And it doesn't hurt the coil as long as you're going in flat all together. Then I thrown it in the trash. But yeah, we got the back about completely done. And then we got a little bit in the front to do at the bottom there. We'll get that here in a moment.  So we're going to replace that one. And I already got one that's completely dead down there. We'll get that one back all done. Everything's fine. Kind of rinsing off some coil. See a little bit of dirt there. Not horrible, but definitely got a little bit there. I don't think that was enough to slow down air flow to create what we got going on here. I think we got defrost issues. It was set for like, I think 30 minutes total. And if they're trying to make this into a beer cave type style thing, that's not going to be long enough. Usually we got to go somewhere between 45 and 60 minutes of timed off defrost, but meaning it just doesn't run the compressor section. We got that. We're going to pull a couple of these fan shrouds and get in behind there and you got to make sure that there's nothing in between in the coils themselves because it will hide there and you'll think you got it and you don't. Well, you definitely got some ice out of there. The Klein flip bit here. Works out perfect for making that quicker and it's magnetic. So it kind of holds it right there for you. All right. So we were able to get the last bit of it. If I can sneak behind here and not get it on the motor, I get it in here this way. I need to really watch out for the motor. I'll go ahead and yank it out. But literally was able to pull that out without bending it, speeding it up. Now what I did notice here while I was working on that was when I seen this one shut off before it it there. You can see that it's it's sometimes catching and stopping. This one here just an easy turn. Look how long it goes. Just an easy turn. This one here easy turn. This one stops that when they're still going and this one here stuck. So we're going to get these replaced. I'm going to check this coil here. We'll probably spray it off just to be safe. All right, broken loose with some pliers there. Get these out of here while I'm in here. I'm going to look around and see if there's any wires that are getting close to shorten into anything. These were pretty well wire tied so they wouldn't. This was wire tied to this wire right here. The nice big nuts here probably won't fit the new motor. So we'll have to use the small ones that come with the motor replacement motor and a little stand there for us. Okay, got it out. Little Fasco motor looks like it's 115 volt. 20th horsepower. You can see the oil that had leaked out of it. It does not want to turn. So it's definitely got some issues there. This is where I like regular keys like this with the extension. How I can break them loose. It seems like it's easier. There we go. There we go. And this one here would not spin at all.  And since they just got in deliveries, it works out great for a little table. What we use is the Rescu 9662s. Pretty much about the same motor. It's reversible rotation. You carry 230 volt version of it. And like I said, the capacitor might need switch to the front. It's hard to say if I do, I can just undo it from there and move it to the front. It does come with both plugs. I think it takes that wider one. So I'll tape that together so it won't fall out. And rotation will just see what it is when we get in there. All right. What I ended up doing is just trimming off the nubs on the back side. I got plenty of room back here. Okay, I went ahead and taped that together so it doesn't pull apart. Got to leave that adapter on there because it's a wider plug. Then I'll put a wire tie on that so that it can unplug itself. And strap that down to these other wires. Flip it on real quick. See what my rotation is. See what's going on away. They had a do not turn off the fan thing there. Some of them make stickers. And that way, I don't have to unwire it. And then they will at least know because they didn't know any different. So there we go. Switches two wires back in. We should be good to go there. Okay. We was able to wire tie it there and that won't pull out from there. Got that reversed. And now just put our blade on. So we got them all together. Let's get them on. Make sure all the fans are going the right direction at the running. They're all blowing. They're all blowing. We're good to go. I went ahead and washed the grills. Washed, washed, washed, whatever. Clean that up. Those ones are going to need a little bit yet. Those are all got cleaned up because I take them all apart. And so we should be ready to adjust our defrost and check operation. See if our cyclist is full. I'm not sure exactly why this froze up originally other than just one fan motor, which I find that kind of hard to believe. However, we're going to check the thermostat there, which who knows what that's set at. Like it's set for 35 degrees, which is perfect. I'll give us enough for a traditional off cycle defrost naturally. And then the defrost clock will do the rest. Let's go ahead and go upstairs and see what we got going on up there. They had high pressure port on there for some reason. And let's just see how this things feed and see if there's maybe a TXV issue, something like that. Who knows? I went ahead and repositioned the bulb over back behind here. So that way it'll have the air coming through and that would be more accurate than set and dead in the middle. I usually like to see it either up in this corner on the back corner close to it, bulb pointing down with a refrigerant line coming into the top. And I already go horizontal, which is what I did there. Yeah, I'm on the conduit, which normally would not be a good idea, but all this conduit is inside here. It keeps it away from the wall and that's not an outside wall anyhow. So let's go up on the roof. All right. So here's the condenser.  I didn't want to forget the defrost already taking them up to 45 minutes a piece, kicking it on pressure switch kicked in. It should be about ready to release and roll. Seven forty five. Let's set that up. She goes in defrost when I do it. It's a little sticky. You could have a defrost clock going bad, which I think I'm going to replace it. I got to check the contactor yet. You can see a lot of the parking marks there. Really happy with a lot of this stuff. Got wires with like the coating. I think that's just the outside coating that started to crack wire nuts there that's been punched instead of squeezed and the insulation on those is cracking. The sight glass is staying cool so far. Hopefully we don't have to mess with that because like I said, this is not the refrigerator we use. So that's what we're starting with. I'm going to go ahead and put it into a defrost. That way we can go ahead and start checking some of these electrical connections. Like look inside here at the contactor a little closer. All right.  Chill power. Get in there and look at this. Points a little better. They definitely are a little pitted like our capacitor here too while we're at it since it's just kind of hanging around three factory wiring with a wire tie. All right. I thought this capacitor felt a little warm. We isolated one leg 5.9. I don't need to do the math. It's that's weak. So I get that changed. Got a new clock ready to go. Got a new contactor and new capacitor. New capacitor here. We'll make sure it's all right. Comes in at 7.7. Got the new contactor mounted up. They had some jumpers there. I went ahead and snipped off the ends and just connected to under one spade terminal there on that. And all we got to do now is just change the clock. But it should be a pretty easy swap out. Just get that popped into place. Got the wire nuts there that we're looking a little ratty got them taped up. Checked all this wiring. It's just that plastic clear coat that's cracking. The main insulator is fine. Everything in here looks to be good. It's all tight and tidy. We're going to go ahead and kick her back on. I've got the clock set up for 345 minute defrost off cycle defrost screens on good. Got the new contactor. Got the capacitor mounted there. Used a heavier wire tie than that traditional one. It's a pretty heavy one. It should last at least a couple of years. But we're going to head into the stropping temp. See where our cyclist is at fan cycle control. Looks like it might be a little out of whack, but I really don't want to try to adjust that today. But for the most part we've got most of all our base is covered. We had like I said that clock here that is it does not want to turn. I mean it you can see the jump there. I mean it's like I'm trying really hard to turn it. It's that thing's junk logical solution. Here's the defrost clock was sticking. Sometimes it probably would work. Sometimes it wouldn't then when they turned the power off that just compounded the problem. We're going to put it in defrost to make sure it times out and shuts down with this new clock. Nothing worse than finding out tonight over time. But it's not running correctly. All right. Good deal set up for the clocks time. You can see how easy this turns. I got just one finger not better. So it'll go into a defrost here on one. We got about an hour or something before it needs to drop down. I'm going to put in mind it's skipping that. You know what? I've actually got four of them here by accident. Let's switch that to three. There we go. Normally two is all we need, but in this case here because they're running the beer and they may decide to try to go colder. We need to try to get as much of a can. This is not a heated defrost. Like I said, it's just timed off. So all you're doing is using the room temperature to fall out that coil. That's why you can't go below 32 and even at 33. I mean, you just not enough heat there to really melt the coil anytime soon. It's just longer. You're going to have to go the colder you're running it. So it's pulling pretty good on that. I mean, could we spend time here? Clean coils and all that stuff. Well, normally we do a lot of PM stuff for these people but I've never been to this store. So I don't know if they're going to have us do a PM on this one or not. But for right now it appears it's all right unless I dig in there a little further and find dirt trapped in there. It's probably going to be let go. So far we're still staying solid here. We ain't at box temp yet and it's not cold out but one of the things where, you know, it may need more but I can't stay here all day. We've been here for a while as it is. Coil and stuff took a while. Going to go down here and watch to see how the box temp is dropping. So far everything looks like it should be good to go. Got to make sure there's no other hidden problems. Went through. Notice dies off by a little bit. My addition was a little off. So like I said, every eight hours we'll be got to set for 45 minutes. I'm going to jump it up an hour. It's not going to hurt nothing. We're going to fall. Well, I don't know if we're going to fall back and fall or not at the time. I thought they voted that out. Not sure. Anyhow, whatever. I don't want this thing going into a 45 minute off cycle here in less than 45 minutes. This thing needs to run nonstop. Get that box down to temp. All right, so hopefully that sticker there which they have writing on that one but they did not have it on this one down here. So got it on both. It still leaves it so we can do our work and kill the fans simple. That way, you know, they've been told they do something that causes that and it's their own problem. So recap we've done thermostat moved the bulb over to here replaced bad fan motor here bad fan motor over there replaced contactor up on the roof defrost clock on the roof and weak capacitor on the roof. Condenser coil could be used probably a wash in wouldn't hurt nothing but I think we're going to try to get that on a PM if not the whole store looks like it needs a PM. I think it's a new store they acquired. So currently right now let's start to drop the temp. All right, so we had a little bit of ice left over from before. Feels like it's feeding pretty good. There's super high humidity in here because all the water on the floor. We got warm coming in getting cold as it goes through which is normal. Feels cold here. Looks like they're all feeding so we're probably fine there. Don't know if it'd be worthwhile to check it over with a super heat or not. So we looked at that one's distributor tube and now we're looking at this one here. Seems to be feeding about the same way. You know, it wouldn't surprise me.  It was originally R22. So, you know, you've got to ask yourself why did they have to change?  Is it a continuous leaker? You know, it's one of them things where I don't have a whole lot of information and I don't think anybody here really knows much about what's going on either. I'm going to just let it go for right now. I mentioned it on the paperwork that we may need to stop back and check that. And that way I'm not spending more time here than what I need to. The thermos thermometer here doesn't look like that works very well. I don't know if it's broke off or what. Yep, it's broke off. So that's bad. That's not very useful. Let's go look at that freezer that I used for a cooler while it's temporarily working on this and we can shut that thing down. They can move the milk back in there. Looks like the walk-in freezer's right here. Let's see if that's the one. Yep, it turned off. It's holding pretty good at temperature. They were just inside here. So, yeah. Just one of them things. So that's, there's that. I want to ask them if they want to use it as a cooler or whatever, which is fine. The thing beside it is a cooler, but it would give them extra room for cooler. But I don't know if they really need it or not. I'm going to let them decide on that. They want to use that freezer for a freezer. So we're going to go over there and turn that back up. I wanted to check the site last a little more time. We're starting to get some bubbles. That's not good. That's what I was afraid of. That's probably other issues. Probably has a leak. It just, everything just piles on. This is what happens when you let everything just go and you don't maintain it. So this is the freeze. We're going to put this back to the 35 minute, 35, almost 40 minute defrost. That way we've got it back to normal and they can turn it down at the thermostat. Right now they still got that milk in there. This just has a regular old traditional clock, honestly, I like better. They work better. They last longer. They don't care about dust and dirt and all that. So set up about 37 minutes area there. It's got a termination on it. Everything in there looks rough. That's on, we'll mark it on the outside so we know it's freezer. It was a little hard to decipher it there with the way that's all mangoided. All right, so it went solid again. I'm going to just tell them that we probably need to do a leak surge possibly on it. I'm going to see if we have any history on this thing.  I don't want to spend a lot of time then they question the bill. The system seems to be working pretty good right now. Those TXVs could be adjusted. I can't really set the super heat right now with the box not being at temperature. It always changes once it gets to temp. At this point, I'm going to wrap this thing up and then we just come back if we need to come back. All right, guys, if you enjoyed the video and you want to see more like it, please give it a thumbs up. Consider subscribing. Check me out on Instagram. And until next time, we'll catch you guys on the next one later.


Hey, you guys, this is Gabrielle with OXMAR Auto Group. I'm here at OXMAR Toyota today and today we are super excited We have the brand-new 2018 Toyota CHR. This is the XLE Premium Edition and we have Chuck here with us And we're gonna actually be going over the exterior features of this car So Chuck if you want to go ahead and show us everything that'd be great. Oh, absolutely This is a very cool looking car We're excited to have this and I want to show you this has a very capable very fun to drive 2.0 liter 4-cylinder engine if I get that there we go Look at this thing with the CVT Very durable You know very fun. I was driving this thing around the parking lot and it's a blast I mean, he's got a little bit of pickups and get up and go. Yeah But then also just the styling of the front. I mean this is a beefy, you know aggressive look It's got LED daytime running lights halogen lamps. This one also has the Fog lamps down there and it has Toyota Safety Sense P2. So you see the glossed-over Toyota emblem Yeah, there's a multimillion-meter wave radar there in dash camera up there I mean, this is just a stylish aggressive look Toyota's doing a really good job of making their cars look sporty It's beautiful and this sits on 18-inch tires These are alloy rims, you know It's a very the very durable last long time and then also this sits up higher than most vehicles in this size, right?  So it's just a unique, you know, very cool interesting Just different car. You don't really see much like this out there at all And then back here you got the hatchback and Gabby if you did those brakes This is one of the coolest. Look at those lights.  Yeah, they're it's beautiful All right, you guys so we're very excited now if you guys have any questions would like to come in take it for a test drive Let us know give us a call at<PHONE_NUMBER> You can also visit our website at oxmore Toyota comm we'd be more than happy to assist you guys Thanks so much for watching this. Bye. Bye


What better way to know if something's going to work on your cancer than actually take the mutations and properties of your cancer and its biology and put them into a living organ and seeing what happens? That's what Lara Tower, the CEO of Vevent Therapeutics does, where basically they take avatars of your tumor and put them into a whole bunch of fruit flies and subjects them to different cocktails. This is actually done at other academic centers in the US. And based on those cocktails of interventions or therapeutics, some of which, believe it or not, are drugs that are like metformin and albuterol that we use in other diseases that are not cancer related that show evidence that they might actually have efficacy with targeted therapies that are cancer directed. Check it out in the podcast Target Cancer, where we talk about this really novel way of kind of treating precision in its most kind of precise form.


And now for our featured presentation. The Stephen Padgett Memorial Lecture celebrates the life of Stephen Padgett, who passionately believed that a greater understanding of physiology would lead to better medical science. He was a founder of Research Defense Society, which later became Understanding Animal Research. And this year, we're delighted to celebrate the 86th Padgett Lecture, adding Professor Emerita Alawalia to our long and imminent list of lecturers. Emerita is dean for research in the Faculty of Medicine and Dentistry at the BARTS in London School of Medicine and Dentistry, director of the UK CRC accredited BARTS Cardiovascular Clinical Trials Unit, and professor of Vascular Pharmacology at Queen Mary University of London, where she's principal investigator, leading the Vascular Pharmacology Group. Her research focuses on understanding the role of inflammation in cardiac and vascular disease with a focus on non-canonical pathways for nitrous oxide generation and delineating the mechanisms that underlie sex differences in cardiovascular physiology and disease. Throughout her career, Emerita has persistently raised awareness on the sex bias in experimental work, increased diversity and representation in biomedical science, promoted responsible animal research, and nurtured the next generation of researchers. She was a project license holder for over 15 years, and her seven years as editor-in-chief of the British Journal of Pharmacology, she helped to generate an incredible amount of practical and internationally relevant guidance on animal experimentation. Tonight, I'm delighted to introduce professor Emerita Alawalia in her lecture, Sex and Drugs, My Rock and Roll. Thank you very much for that really very kind introduction. It's a massive honour to be delivering this lecture this evening. I had a look at the website and saw the list of the previous recipients, full of the great and good in biomedical research and in vivo research in particular. It's a massive honour. I'm chuffed to bits, in actual fact, to be the newest name on that list. So thank you very much to those who nominated me, and thank you to the UAR for awarding me this honour. I have a list of, I'm just trying to see the mouse of disclosures at the bottom, I can't see it, at the bottom of my slide. It's the final one on that list that's relevant for my presentation today and you'll understand the relevance as I go through my talk. But before I start talking about Sex and Drugs, I thought I would share with you some observations that I've made when I was doing a little bit of research in preparation for this lecture. Scenes that, despite appearances, that Stephen Padgett and I have rather a lot in common. Both of us actually trained at the same medical school. Stephen Padgett moved to St Bartz Medical College in 1878 to study medicine. I didn't bump into him in the corridors when I was there. I arrived, I know I've got a lot of white hair here, I'm not quite that old, I arrived in 1989 to do my PhD under the supervision of Professor Rod Flower. In doing my research as well, I came across some really interesting facts that I thought I'd share with you that have some relevance to my presentation today. So Stephen Padgett was one of four sons of James Padgett and James Padgett was actually the very first warden of the first residential medical college at Bartz. He took on this role in 1844 and he was obviously a really forward thinker for his time. By 1850 he had recruited the first ever, or allowed the first ever woman to attend training within a medical school in the UK. Sadly, those that followed him didn't really follow in his footsteps. Her name was Elizabeth Blackwell, she was the first and the last woman for some time at Bartz Medical School. She actually became a leading light in women's rights and women's lib. In the United States she set up medical school and she had a very prolific and successful career leading equality for women. I'm sure James Padgett, however, would be very happy with the current circumstances at the Faculty of Medicine and Dentistry. I think I'm a good reflection of the way that the medical school looks today. I'm currently the Dean of Research in the Faculty and I think that's testament to some major changes that have taken place. Secondly, both Stephen Padgett and I have an interest in typhoid vaccine. I'll explain to you during my talk how my interests originated and how I've used typhoid vaccine as an experimental tool in studies in healthy volunteers. But Stephen Padgett was a massive proponent of vaccination to protect against typhoid fever. In his famous book, Experiments on Animals, he published this chapter specifically focused on typhoid vaccination and he actually reported some of the experiments and data that led up to roll-outs of vaccination, particularly at a time when it was needed with the infantry that were fighting in battles. And this just highlights the section that I wanted to bring to your attention. It says, good luck attend all 18 of them. These were the first individuals ever to receive typhoid vaccine. It was an experiment. Then, army personnel, and another interesting fact, actually the majority of them were Indian medical officers. So good luck attend all 18 of them and immunity against typhoid wherever they are.  I think this book was published in 1900 and it is a testament to Stephen Padgett's promotion of the importance of early studies in animals that then are translated into the clinical setting. The importance of how effective they can be in providing us good guidance on what to do next when we go into humans. And I've taken this translational approach to my research in my laboratory and almost everything that I do in the animal experimental setting I take into the clinical setting find ways of simulating the experiments in humans. I'm also interested in supporting best practice animal research. It's a given that Stephen Padgett was interested in that. And in the main, my opportunity to promote good practice came during my tenure as editor-in-chief of the British Journal of Pharmacology. I did that job for seven years. I had an absolutely top notch editorial board and an even better, I mean could it be possible? Yes it could, senior editorial board. And with all of their hard work together with me we generated a number of pragmatic internationally relevant guidelines for biomedical researchers doing pre-clinical research. Many of those guidelines actually focused on animal experimentation and today towards the end of my talk I'm just going to share one of the specific initiatives that we launched during my tenure focused on the subject matter of my lecture today. So sex and drugs. I thought that I should probably declare from the get-go that this is not going to be a racy talk. And my sincerest apologies. The drugs that I'm going to talk about are the ones that I used in my experimental methods in animals, animal tissues to explore signalling pathways that govern cardiovascular physiology but also to identify potential therapeutic targets for cardiovascular disease. The sex that I'm going to talk about is actually the difference between males and females. So apologies again. It's not going to be racy but I hope that it will be at least interesting and illuminating and perhaps even entertaining. So why am I interested in the cardiovascular system? Well it's because cardiovascular disease, CVD, is a major killer in the UK. If you look at that I can't get the mouse to work here. Whoops, here we go. So this is data that is published by the British Heart Foundation on an annual basis. This is the data for 2023. You can see here that the BHF state here that 25% of all mortality in the UK in 2023 will be due to cardiovascular disease. This is a frightening number. It's number two in the major killers in the UK with cancer being the top of the moment just a percent or two more. But I think the number that's probably more frightening is this one. Over seven and a half million people living with cardiovascular disease. And I think this number actually highlights the importance of continued efforts to try and identify novel therapeutics to address this epidemic situation that we have with CVD in the UK. But it's not just in the UK. This is the global burden of disease dataset that was funded by the Gates Foundation published in Lancet. And you can see here that it's not, cardiovascular disease is not just a disease of high income countries but we see very high cause for mortality in middle and low income countries. And whilst it's true that cardiovascular disease mortality is actually decreasing in western countries and in high income countries, it is completely the opposite in low and middle income countries. And we still have to grapple with. This is some additional data that's come from the BHF and it is just to highlight the key aspect of my talk today. This shows the deaths by cause in the United Kingdom in 2019. And the data is disaggregated. It's not often that you see disaggregated data between men and women. And if you look at the top dataset, this is all about cause mortality. And you can see I've highlighted there, there's an interesting phenomenon that's taking place. You can see that there's approximately double the number of men in the early age ranges dying in comparison to women. And what you start to see is we age that the numbers start to converge. There's many men and women dying but then you start to see that the numbers of women eclipse the men. And this is causing this apparent sex difference. What diseases are driving this? Well I can tell you that it's not cancer. So whilst men and women suffer different types of cancer actually the total mortality is pretty much in terms of numbers in the UK the same. So while the actual type of cancer is different mortality is equal between the sexes. That is not the case with cardiovascular disease. I've highlighted the data here. What you can see that as we age that there is almost, well there's double sometimes even triple the numbers of men dying of CVD in comparison to women. That the numbers start to converge as we age and then you start to see the numbers of deaths in women eclipsing those in men. There's been an enormous amount of research epidemiological research in this kind of data and it's clear that women have a 10 year advantage on male counterparts and that this is in pre menopausal women. But once women pass through the menopause that that protection seems to disappear in terms of cardiovascular disease and there's been an enormous amount of preclinical and clinical research that has implicated female sex hormones particularly estrogen in providing the protective effects during the pre menopausal years. In my group we have focused on assessing some of the causes of this sex difference and I'm going to share some of that data with you today. What we know is that cardiovascular disease mortality there is one big risk factor it's hypertension. Hypertension drives the statistics I showed you on the previous slide. 50% of all heart attacks somewhere between 50 to 75% of all strokes are due to high blood pressure. These BHF statistics in England in 2019 you can see the percent of the population with hypertension and hypertension is high blood pressure with blood pressures of 140 over 80 and above. And you can see the pattern between men and women is identical to the cardiovascular disease mortality statistics. Hypertension is driving the cardiovascular disease mortality and that means that there must be some specific differences between in blood pressure regulation between men and women that we haven't quite got to agree with that is driving this. It's important because the medicines that are available to treat hypertension have been largely tested in men. So it means that we have not been serving women well for these many, many years. Well this is some data actually that was generated by my colleague Vikas Kapil. He was doing a PhD with me at the time. He's a clinician, a clinical pharmacologist. He's now a senior lecturer in clinical pharmacology at the faculty of medicine in dentistry and a consultant in cardiovascular internal medicine. This data, Vik, generated when we were exploring the possible blood pressure lowering effects of inorganic nitrate. It was mentioned in my introduction. It's one of the areas of my lab. This just shows baseline characteristics of the cohort that he recruited. We recruited young people. I have to tell you the truth. They're all medical students. They were willing and happy to do the study. You can see there are some obvious differences in their BMI but there are massive differences in blood pressure here. This is histolic blood pressure and diastolic blood pressure. These individuals are healthy volunteers yet already we're seeing that the men have much higher healthy blood pressure than women. Indeed, these young men are very, very dangerously close to the levels of blood pressure that have been identified as predictive of moving into hypertension in the later years. Close to 130 over 80. So worrying but importantly highlighting that there are big differences in blood pressure regulation between men and women. Well, we know that blood pressure is determined by two key factors, cardiac output that's the work of the heart and peripheral resistance and that's the resistance of blood flow as it's flowing through the circulation and resistance is largely determined by the resistance vasculature. These are the smaller arteries that actually penetrate our organs and perfuse them and provide them with oxygen and nutrients. The vascular tone of resistance arteries is primarily determined by the sympathetic innovation that penetrates the walls of those resistance arteries. These neurons release catecholamines that cause an increase in intracellular calcium of the smooth muscle that they're innovating and this causes smooth muscle to contract and therefore the blood vessel to vasoconstrict. When it vasoconstricts, this then creates a resistance to blood flowing through the blood vessel. This contractile activity of the sympathetic nervous system is constantly buffered by an endothelium-derived vasodilator influence. The endothelium is activated by blood flowing through the blood vessel. It creates a shear stress, a frictional force on the intraluminal side of the endothelium and this triggers the endothelium to release three different vasodilators that act to constantly buffer this vasoconstricter nature of the sympathetic nervous system. This plays an important role in maintaining healthy open blood vessels that keep essentially our blood pressure down. Well in my group we've been very interested in the nature of those factors and how they operate and function and we're interested because what we know about cardiovascular disease that one of the earliest signs of impending disease is a dysfunction of the endothelium and that dysfunction is due to a loss of the protective activity of endothelial mediators. These two prostacycline and nitric oxide are two of those three endothelium-derived vasodilators. A great deal is known about these I'm not going to talk much about them except to say that prostacycline is generated in the endothelium by the activity of cyclo-oxygenase 1 and nitric oxide is generated by the activity of the endothelial form of nitric oxide synthase. These factors pass through to the underlying smooth muscle interact with their cognate receptors IP for prostacycline soluble guanalan cyclase also known as GC1 for nitric oxide they elevate cyclic nucleotides and lead to smooth muscle relaxation that's how they provide that opposing influence to the sympathetic innovation. But in this talk I'm going to talk about the third vasodilator and that is EDHF EDHF stands for endothelium-derived hyperpolarizing factor and it's still and the fact that we're using an acronym and a very generic term tells you something about the controversy in the field with regards to its identification. The controversy exists but actually there have been some excellent researchers who've worked using a range of tools that we have available to us to delineate the signaling pathway for EDHF activity. People like Arthur Weston, Chris Garland, Tudor Griffiths, Paul Van Hoot, these individuals have helped us to understand the exact signaling pathway for EDHF even if we can't agree on what it actually is. What we know is that when the endothelium is triggered by severe stress or by circulating hormones, the EDHF is released and this is associated with potassium e-flux that emanates from these two calcium dependent potassium channels, the SKCA and IKCA. There are specific drugs that have confirmed that they're involved that we use in vitro that there is an increase in the potassium levels within the space in between the endothelium and the smooth muscle and this is associated with activation of the sodium potassium ATPase and an inwardly rectifying potassium channel that leads to hyperpolarization of the smooth muscle and further relaxation. Again, these tools have been critical but actually all of the experiments that have helped us to understand the EDHF signaling pathway have been done by eliminating nitric oxide and prostacycline from the experimental environment and this has been done using non-selective inhibitors of nitric oxide synthase and cyclooxygenase. The vast majority of the research that has led to this very clear understanding of the EDHF signaling pathways has come from in vitro experimentation. You simply can't take toxins in vivo and certainly these drugs are not selective for the endothelial isoforms of the enzymes. It's impossible in vivo to tell whether the effects that one sees are due to EDHF or some other unrelated effect on isoforms expressed in other regions of the body. So despite the fact that we know the signaling pathway well, the true in vivo significance with regards to blood pressure was largely unknown. Until I collaborated with a good colleague of mine is actually sitting in the audience Adrian Hobbs, we decided that we would create what we called the EDHF mouse and this was a mouse that we felt would provide us with an environment where we could look at the activity of EDHF in vivo. And what we did is capitalized on the work of those individuals that had generated the ENOS deficient mouse and the COX-1 deficient mouse, that's Huang and Oliver Smithies and Langenbach. We crossed those mice and did repeat crossing of the progeny until we generated mice that were doubly deficient in both ENOS and COX-1. Now I have to admit a terrible, terrible fact at this moment. So this was early 2000, so please don't hold this against me. We did what everybody else did at the time. We didn't think about the female animals we took the male animals and we started off by doing the classical in vitro experiments that everyone did to study EDHF. And actually it was a massive disappointment. We thought that what we were going to do is create mice where there was a substantial EDHF response that we could then start interrogating. These are resistance, these are data from resistance arteries taken from the mesenteric resistance vasculature. And if you look at the first two bars of the wild type mice these are arteries that have been pre-contracted. And then acetylcholine which is an endothelium dependent vaso relaxant agent applied. And you can see that in these male arteries there's a prominent relaxation response as we would have expected. However, when we looked at ENOS knockout mice you can see that this response is profoundly reduced implicating nitric oxide as an important mediator of this response. Some reduction when we used the cox one knockout mice, but the response was almost completely absent in our mice that were doubly deficient of both those isoforms. So it meant that we had a piddling response that we really just didn't make studying EDHF very easy. And you can see that in the different genotypes that if we used either endomethacin or L name endomethacin in the ENOS knockouts the remaining response was completely blocked. If we took the cox one knockouts used L name the NOS inhibitor we completely blocked the response. So that told us that in males, EDHF has practically no role to play in endothelium dependent relaxation. And I suppose you could call it an act of desperation or an opportunity or a fortuitous opportunity but it was also because we were wasting. I mean this breeding process was necessary to generate the doubly deficient animals but not using the female mice was actually really a scandal. And so we decided that we would repeat the experiment in the same type of artery in females and I know this is a busy slide but just focus on the open bar and the grey shaded bar. You can see here immediately that in these female arteries if we added the L name endomethacin cocktail we only suppressed the acetylcholin induced relaxation by about 20%. Hooray! It meant the EDHF was the likely mediator of that response and then when we looked in each of the different deleted mice you can see that the cocktail actually didn't really affect the response. It was slightly reduced but it wasn't inhibited any further. This doubly deficient animal, this relaxation is due entirely to EDHF. We confirmed this by using those drugs that others have identified were effective inhibitors of the EDHF signaling pathway. Raising extracellular potassium also blocks EDHF and you can say they see that they profoundly suppressed the response. So what we had discovered in this fortuitous experiment is that EDHF was an important endothelium dependent vasodilator in the resistance arteries of females but certainly not in males. What we wanted to confirm in vivo that this made a difference to blood pressure responses and we used bradykinin which is an endothelium dependent vasodilator to look at blood pressure responses. This is a the top is a typical trace from a female double knockout mouse and this is a male double knockout mouse. You can see there's a dose dependent decrease in blood pressure. These are anesthetized mice and instrumented and absolutely no response in the males and that's the summary data. Okay, so EDHF is important for mediating blood pressure lowering responses to an endothelium dependent vasodilator. What does that mean for baseline blood pressure? Does it make any difference? And the answer to that of course, I wouldn't be telling you this story if it wasn't, is that yes of course it is. This shows each of the different knockouts female and open bars male in their filled bars and you can see that for each genotype, whilst the females are completely norm intensive the males when they're deficient in ENOS and when they're doubly deficient in both isoforms are certainly hypertensive. Slightly more in the doubly deficient animal. I know you may not agree with me but just humor me. Okay, well actually Adrian has his own separate research program. He was interested in EDHF because of this thing here, actually C-type lateratic peptide and we helped him publish a few papers that have demonstrated without doubt and with the use of a number of different genetically transgenic mouse models that CMP, C-type lateratic peptide from the endothelium is binding to an NPRC receptor to result in hyperpolarization that mediates EDHF responses in females but not males and plays an absolutely critical role in regulating blood pressure. So there are many that argue about the identity of EDHF but there is no question that CMP plays an important role. I had a PhD student funded by the Barts Charity, Melissa Chan, she's now a senior scientist at the Framingham Institute in the US and actually Melissa's PhD was all about trying to understand what was causing the regulation of EDHF in comparison to males in the females and what she discovered is that females expressed substantially more of the endothelial pathways for EDHF, the SKCA and ICCA and that this was driven by the activity actually of estrogen. I don't have the time to show you the data here today but great paper published in British Journal of Pharmacology. But what's also true about these endothelial factors is that they're not just simply vasodilators. As I mentioned these factors, at least certainly for prostacycline and nitric oxide there is a wealth of literature that shows that these factors are critical not just for keeping blood pressure down but actually maintaining the patency of our blood vessels by preventing platelet activation and by preventing inflammatory responses. And so, of course we wondered could EDHF possibly be responsible for this influence in females. So, we decided to focus on the I'm going to show you the data with regards to looking at inflammation and we used mouse models to look at inflammatory responses in mice. This was work done by Conchie Villar who is funded by BHF Project Grant in my lab. She's actually now a senior in vivo researcher at AstraZeneca. And Conchie used this model that was actually developed by a colleague of mine at Barts, Professor Mauro Peretti where one injects air under the skin of the mouse and it creates this rather cool little pouch into which one can inject inflammatory stimuli that lead to an influx of fluid and leukocytes. And what one can do is wash out that pouch and actually analyze the pouch contents. And that's exactly what Conchie did. We used interleukin 1 beta that plays an absolutely critical role in the acute inflammatory response but has also been substantially implicated in the progression of cardiovascular disease. We know that systemic inflammation precedes disease and IL-1 beta is one of the key inflammatory cytokines in that setting. And you can see here she took male and female litter match mice and you can see while I asked IL-1 beta caused a profound leukocyte infiltrate into the pouch that the females were actually unresponsive and this was at a 4 hour time point. What she also did in this experiment is actually took half of the females and ovariectomized them and you can see that the removal of the ovaries resulted in a phenotype that almost was identical to the male mice. So this is ovarian hormones driving this difference in the inflammatory response. Well we wanted to look a little bit more closely at what step of the leukocyte recruitment paradigm might be different between the sexes. And this is a famous cartoon from a Nature Immunology review published by Klaus Lay. It shows that the leukocyte roles adheres and then once adherent it commits its fate to margenation. And we can use the in vivo method of intervital microscopy to focus in on venules and look at this paradigm in real time in an esthetized mice. And that's exactly what we did. We took male and female mice, we injected them intraperitoneally with IL-1 again and we followed leukocyte rolling and adherence and this is just one of the images that we generated you can see here leukocyte rolling along here this one is actually starting to commit its adherence and you can see some that have emigrated here. But what's clearly obvious is that there are way fewer cells rolling along the endothelium of venules in females compared to males and that this leads on to a knock on effect of reduced adherence. So the difference is taking place at the very earliest stages of leukocyte recruitment. Well, Conchy then actually took a few of the mesenteries did some simple H&E staining and here again this is a venule here and you can see that these leukocytes are polymorphin nuclear cells. This is one that's actually starting to adhere here another one that's flat and you can see here I don't know how visible this is but this is a PMN in the extravascular space. We know that these PMNs are neutrophils because Conchy assayed for myeloperoxidase which is a neutrophil specific peroxidase and you can see that in supernatants of the mesenteries she showed that there was a time-dependent increase in MPO activity that essentially matches this leukocyte rolling and adherence time course and we saw a very much reduced response in the females. Well, another postdoc of mine again funded by a separate grant from the British Heart Foundation project grant helped us to look at these responses a little bit further. We took sham and ovariectomized female mice and explored the response to interleukin-1 beta again and you can see that in the absence of the ovaries there was a response to interleukin-1 that was very similar to the response that we had achieved in the males here at the same time point at the four hour time point about 30, 32, 33 cells per minute rolling along the endothelium. I want to highlight an important point here which shows the importance of conducting when you're using a surgical model of conducting a sham control. If you look at the sham controls here, the sham's no response to aisle 1, but look at this the control animal. Here there are somewhere in the region about 14, 15 cells rolling per minute. If you go back here I know it doesn't sound like a lot but it is actually. If this is the healthy female mouse and it's sitting somewhere around 8. So the surgery itself did change the environment so it was important to do sham control in that setting. What we did, what Rayman next did is actually we used osmotic mini pumps from which estrogen was released estradiol was released and you can see that by restoring estrogen levels in the ovary ectomyse mice we substantially attenuated that leukocyte rolling response to interleukin 1 beta. And finally, we wanted to make sure that this wasn't just some freaky deaky response to aisle 1 that actually it was something that was pertinent for other inflammatory stimuli and we repeated the experiments looking at male and female mice using tumor necrosis factor alpha another cytokine that has been implicated as a very important mediator of cardiovascular disease and you can see that there is an identical phenotype to the one that we saw between the sexes interleukin 1 beta. Well, what about EDHF? Does it have any role to play in this response? Well, we went back to our different genetic deletion mice and you can see that in the Cox 1 knockout mice the females don't respond so it's certainly not prostocycline that is attenuating that response. In the ENOS knockout mice there was a small response but it certainly wasn't at the magnitude of the response to interleukin 1 beta in the males so the females were still protected and you can see that in each case in the males there's a prominent response to the cytokine in the males but also a raised leukocyte rolling here in the ENOS knockout mice but in the doubly deficient mice you can see that the females still maintain their protection. We did a time course just to be sure here in these experiments while the males are responding to interleukin 1 the females are not. It is EDHF that is inhibiting that leukocyte recruitment. I can tell you again that Adrian and his group have gone on to demonstrate that the EDHF doing this in part is C type naturitic peptide. But as I said, whenever I do this kind of pre-clinical experimental work I seek opportunities to translate to the clinical setting and that's exactly what we did next. My colleague Vic Capil went and learnt two separate models of inflammation that we could construct in healthy volunteers and we looked at the differences between the sexes in this response and this is just the paper that we ultimately published. Krishna Raj was a PhD student cardiologist doing a PhD with me funded by the NIHR at the time. He did all of the work that I'm going to show you. He's now actually after getting an NIHR ACL has got his permanent post and as a senior lecturer only last month at Barts in the London and a consultant cardiologist at the Barts trust. So the first model that we used was the Cantharadin induced blister model. This was a model developed by Tony Siegel at UCL many years ago. Cantharadin is a protein that's found in the blister beetle and it's actually used clinically. It's used to treat warts and varucas and what Tony Siegel did is he used Cantharadin applied it to the skin of healthy volunteers and it creates a blister and from that blister we can collect the blister fluid and analyse it for cell numbers but also for inflammatory mediators. And the protocol involves two time points 24-hour time point after Cantharadin application which shows you the big, strong acute inflammatory response and a 72-hour time point where the inflammatory response is starting to wane and the inflammation is recovering. We designed the study based upon our pre-clinical experimental work. The response in females was 50% less than the response in males and that is exactly how we did our power calculations expecting 50% less neutrophils in the blisters in females compared to males. When we collected the 24-hour blisters what was clear almost immediately was that both men and women could mount a strong inflammatory response but actually it turned out that it was identical. There was no difference again massively disappointing. This seems to be part of my research life that the first experiments the hypothesis we're testing we're wrong. I'll show you the data that confirms that in just a moment but the absolutely brilliant finding in this study was that the 72-hour blisters in 60% of the women there was no blister at 72 hours. It had completely resolved there was no sign of it. It hadn't popped the males pesky males not very good at looking after their blisters you can clearly identify a popped blister but there were no resolved blisters in the males whatsoever. When we looked at the we interrogated the blister a little bit more closely this confirms what I've just said to you for our blister. No difference in blister fluid volume. No difference in total cell number within the blister. No difference in neutrophil numbers and absolutely shocking finding and certainly not what we were expecting. You can also see reduced blister volume at 72 hours a hint that maybe there are lower cell numbers in the 72 hours in the females here but again no change in neutrophil numbers. What was different were the number of inflammatory monocytes and inflammatory monocytes perpetuate the acute inflammatory response. They're key in responding to the inflammatory trigger and in the second cell they're really after neutrophils. We did a whole host of other experiments mechanistic studies to try and tease apart exactly what was going on here and what we found is that with the reduced inflammatory monocytes there was a protected number of the anti inflammatory monocytes. They have a different phenotype and we can separate them out but what we also found is that whilst the neutrophil numbers were identical at the 24 hour time point in the females the neutrophils were distinctly and significantly less activated.  We measured a number of different mediators in the blister fluid. This was an unbiased array externally conducted and we found a number of anti-inflammatory cytokines and chemokines that were raised in the female blisters at 24 hours versus the male blisters. What does this mean for the cardiovascular system? Well actually there's a second model that we set up and that's systemic inflammation induced vascular dysfunction and this is where the typhoid vaccine comes in. This is a model that was developed by Arun Hingarani and Patrick Valance at UCL many years ago and essentially what they did is used brachial artery ultrasound measurements to measure the brachial artery diameter before and then after the release of a cuff of the lower arm when that cuff is inflated it stops flow when it is released what happens is that there is an increase in flow through the brachial artery I'm just at the cartoon here now brachial artery and the increase in sheer stress that is thought to trigger the release of nitric oxide that then causes a vasodilation and that's what you see here. This is the flow mediated dilation response and then what Professor Hingarani next showed is that after eight hours, eight hours after the administration of the typhoid vaccine this induced a systemic inflammation which was associated with a significant reduction in that FMD response. We know that this relates to endothelial dysfunction and not smooth muscle cell dysfunction because he conducted some controls this is GTN it's an organic nitrate and an N-O donor that delivers N-O to the smooth muscle independent of endothelial cell activation and you can see no change in the response here. So we designed our study to have equal numbers of both men and women subjected to FMD and typhoid vaccine. We also had a 32 hour time point. This is a transient systemic inflammation and the individuals do recover and that's reflected in this data which shows the blood differentials after typhoid vaccination and CRP levels. You can see it's not a serious systemic inflammation but in both sexes after typhoid vaccine there's an increase in circulating white blood cell count. We used two way analysis of variants with sex as a factor here both we saw an increase in white blood cell count in both the men and women. No difference between the sexes and this increase was driven almost entirely by an increase in neutrophil circulating numbers and again no difference in numbers quite different to the mouse air pouch and the intravital microscopy but those studies were the precursor to doing these studies in healthy volunteers. So what about FMD? So the flow mediated dilatation response in the two sexes was very similar at baseline after eight hours you can see that the males suffered vascular dysfunction the females in contrast surprisingly showed an improvement in their vascular function which was a definite surprise and this just shows the change in the FMD response from baseline comparing the two sexes which is how we powered our study. We know that this is endothelial dysfunction because this is the response to GTN in the males and the females in the vaccine. We then went on to do a whole host of mechanistic studies biochemical analyses of the blood and we found that whilst the neutrophil numbers were elevated in both sexes actually the activation state of the neutrophil was reduced in the female at the eight hour time point in comparison to the males and that this was associated with an increase in the anti-inflammatory monocyte expression associated also with the expression of a number of anti-inflammatory cytokines that accelerate the recovery to an inflammatory response. There is a paradigm now where the resolution of inflammation is recognised as an active process this is a lovely cartoon that came from a review published in Cirque Research a few years ago but essentially you have your inflammatory stress and trigger it causes neutrophils to get to the site where that stress is taking place. They start to attack and deal with the cause of the recruitment of those cells. Some of these cells become apoptotic macrophages start to engulf those apoptotic cells but this results in further signals that recruit more pro-inflammatory monocytes but at a certain point that aphorosis actually triggers a switching of the macrophages from a pro-inflammatory to a pro-resolving phenocyte. What we think is happening is that in females there is an up-regulation of the expression of these macrophages that these lead to an increase in anti-inflammatory cytokines and chemokines and that this then results in the reduced activation of PMNs and the accelerated recovery of the inflammatory response. We are currently exploring this in studies in patients with coronary artery disease and comparing that to age-matched healthy volunteers and hopefully I'll have some more ideas on the actual targets possibly for estrogen in this setting. Finally, in the last few minutes of my presentation I focus on the work that we did at the British Journal of Pharmacology trying to address that issue that I mentioned to you of the sex bias in research. We know that pharmacology also there's a great deal of sex bias in the published literature but at the Journal we decided that we would take a look at what the issues were and then think about what we might be able to do to support the research community in changing their ways. So what are the issues? Well, actually when we were exploring all of the issues driving sex bias it became clear that there are a number of issues specifically pertaining to preclinical research but that there is a very prominent sex bias in clinical research that in some way has influenced what we do at the preclinical setting and this is driven in large part drives in large part the reduced number of women that are recruited into clinical trials and the reduced number of women relates to the concerns about risks the risks of harm to women but also to the potential unborn fetus. I mean the experience of the Thalidomide scandal is something that we are still living with and actually even though over the past 10 years or so there's definitely raised awareness that there is a sex bias in clinical research and we absolutely have to do something about it the underrepresentation of women in trials persists and it persists because people are concerned but also we think it persists because actually the design of clinical trials that includes sex as a factor is not well enough developed yet. There are lots of people that are on the job but we're waiting to hear outcome for clinical trials how that's going to run. The preclinical researchers have some good ideas and I think the clinical world could learn from them and I think this is in part underlies the clinical setting and it is a big problem and it's something that we absolutely have to do something about and this data here these numbers give you a good reason for why we need to do something about it in the period between 1997 and 2000 10 medications were withdrawn by the FDA from the US market. 8 of them were because of safety issues in women and the reason those safety issues occurred and weren't suspected is because women weren't involved in the trials and females weren't involved in the preclinical research that preceded it. So what about the preclinical research setting well I've said already the majority of preclinical research there is a sex bias and this is largely due to this perception this impression that there is greater variability in females this is actually controversial in fact has now been refuted by a number of prospective and robust studies it seems that the neuroscientists and the psychopharmacologists are way ahead of the rest of us but they have already done a great deal of work in this area showing that this just simply isn't true and we do need to do something about it because if anything I hope that my presentation so far has told you that there are major differences in the biology between the sexes and also the molecular mechanisms of drug action. So we cannot rely on studies just in male animals. Of course there is also that terrible serious ethical problem that hit us when we were doing those studies in the doubly deficient mice. The surplus breeding and waste of female animals is simply unconscionable and now we have been woken up to it we cannot continue in that way. But one of the big problems that is holding people back is understanding how sex as a factor can be included in experimental design. So at BJP we wanted to take a look at what we had been doing in the past and determine whether pharmacology was plagued with the sex bias as much as the perceived view in other disciplines. So we did an audit. Now we assumed the worst because one of the great pharmacology leaders of the past Lewis in his book had the textbook of pharmacology told us all in 1960 that we must make sure that the animals that we use in our studies are from the same strain of similar age, weight and sex and I have to tell you that we were obliged in the pharmacology research community. This is just some data from the very first issue in BJP and an issue that was from BJP in 2019. You can see that in 1946 they didn't much care what sex they were using for their experiments. There was absolutely no mention and if they did mention the sex it was predominantly that the animal was male. In 2019 we were doing a little bit better thankfully. At least we were saying the sex and it turns out that all we're doing is using males for our studies certainly in the material published in the British Journal of Pharmacology over 50% of the papers. And in fact this is a terrible indictment from a group that have actually done quite a lot to raise the alarm for sex bias in biomedical research. As occur of Prendergast and Beery they did an analysis published just in 2022 of historical trends globally in biomedical pre-clinical research and you can see that in the early 1900s nobody really bothered saying anything about sex that by the 1970s that that had decreased thankfully but it turns out that all that people were doing were using male animals in their studies and this has largely persisted. Perhaps the most frightening bit of data from this is that it turns out that the pharmacologists are the ones who are doing the worst out of all of us. The neuroscientists are pretty good. In 2009 you can see that about 25% of their published material used both sexes in their studies. That has massively increased more than doubled in 2019. The pharmacology community actually has decided to resist and if anything we've decided that we absolutely will not work on female animals.  What we did at BJP is try to change the rhetoric led by two of those wonderful senior editors, Jim Dockety and the wonderful Claire Stanford who is a previous recipient of the Paddock Lecture and is sitting in the audience. Jim and Claire led an editorial that we put together, the senior editorial board and myself and dating a new change to publication of articles in BJP where we required that sex needed to be considered as an experimental variable where we actually gave some good advice on how one might be able to do that by using factorial design and we also insisted that there must be some discussion of the relevance of findings for both sexes at the end of the manuscript. We took a pragmatic approach we recognized that publications or articles that were being submitted to the journal were the work of the previous could be two, three, four years we would accept papers with only a single sex but would require full justification and discussion and this was a mandate that we published in 2019. Now what we did during my tenure as ERC at BJP is we introduced guidelines and then we assessed how well we were doing so a few years later 2022 we decided to determine whether had we really eliminated sex bias with these new mandated guidelines we wanted to know whether researchers were more aware of the importance of considering sex whether they were actually incorporating sex as a factor in their experimental design with a view to providing a change in the published material and improved reproducibility and of course transparency. Claire led the publication which came out earlier this year in 2023 you can take a look at it we discuss our results and now for the results oh goodness Claire I'm going to show everyone in fact the story is good and then not so good so what Claire did is audited the journal back to 2014 there's an enormous amount of work and you can see that what we were doing in the years from 2014 is largely doing experiments in males only when we introduced the change in 2019 we actually saw this really rapid change in the numbers a decrease in males only and you can see here increase in the use of females and these were being used with both sexes largely in our publications it improved a bit more in 2020 and then what must have happened is we took our foot on the brakes and we stopped accelerating and we're starting to see a decrease in 2021 and the 2022 and a reversion to type it does seem so what this highlights is that it's absolutely essential to remain vigilant that when we introduce guidelines the audit is really important publication of the audit and sharing of our data absolutely essential and hopefully when they do the next audit in another two or three years time the data will look much, much better fingers crossed actually an important aspect is of course teaching and training and I've had the wonderful pleasure of working with this individual researcher here Natasha Karp she's a leading light internationally in highlighting the issues around sex bias Natasha is a senior statistician at AZ and Natasha together with her colleague another statistician Ben Phillips have created a training module at the request of the UKRI working with the MRC to support researchers in introducing sex as an experimental variable and we trialled that training at a recent international conference for pharmacologists that I was secretary general for earlier in the year and what we also did at the same time with the help of Breanna Gaskill is actually look to see what the impact of that training in fact was on the individuals who attended it we did a survey before and after the training as well as surveying conference participants we are actually repeating the workshop in London at Barts in the new year so that we have a sufficient sample size and we will be publishing the outcome of this test case in 2024 it's been wonderful working with this group of individuals we actually also got some PhD students at the conference to help us and they did a fantastic job in collecting the surveys too so thank you to all of them it's not all bad news the research community in the UK is waking up there is absolutely no question the NIH were the first they mandated a change to their grant submission process in 2015 where they insisted on sex being considered as a, they say biological variable but Claire very rightly highlighted that it's experimental variable in the design of their experiments they did survey the panel members a few years later after the introduction of the mandate and there was a small improvement in consideration of sex and experimental variable again highlighting the need that we have to keep pressing on the accelerator, keep pushing if we want to bring about change it'll be slow but it will eventually happen and the same is actually happening in the UK the UKRI just earlier this year published their mandate that sex has to be part of experimental design the preclinical level the clinical level and this includes in vitro, cell, experimental work too the cardiovascular world has also woken up that this is an important issue those numbers that I showed you from the BHF statistics clearly women are dying from cardiovascular disease eventually and we don't have treatments currently that is that caters directly for their disease the medications that we have have only been tested largely essentially in men and this commission organised by the Lancet published a number of recommendations there are loads, you can't read those but the fact is that there are so many I had to make it tiny so it got onto the slide but there are two main areas that I think I feel that I've contributed I hope to contribute to in the future that I think are absolutely critical firstly we need to develop educational programmes and hopefully the work that I'm doing with Natasha will lead to something tangible that leads to benefit secondly I hope that the work that I've shown you today has left you with the view that I have contributed in some small way to understanding the differences between the sexes but absolutely it's the funders that need to fund this research and it is true that historically they have rejected these grants but there has been a change there is a change there are calls coming from charities and from the UKRI to submit grants focused on this particular issue so hopefully we'll see funding for this really important area of work just to acknowledge all of the funders of the work and the support that I've received for the work that I've presented today thank you very much everybody for listening I hope that you're not disappointed that it wasn't racy but it will be useful in my presentation thank you thank you for allawalia for an outstanding presentation just highlighting the importance of the interaction between human and non-human animal research and the benefits that can lead and also just this blatant bias I think you are overly flattering on the neuroscientific field a lot of folks are still not reporting so I think they also have a lot to do we've got time for some questions please if you have for the speaker I couldn't help but notice in the analysis done by Clair that the moment you put out that idea of you've got to report the sex and you should be including females as well as males what you actually got was a lot of papers that didn't report the sex yeah we did get more papers we did see an increase in the total number the proportion of the total number that didn't report sex but the upside of that was that we did actually see more less that we're using males only and an increase in the numbers of females being used you know you put in guidelines and it requires the team I did say it was an excellent editorial board and they are but you know the editorial board certainly of the journal that I was editor in chief of they do that work voluntarily and it's in addition to everything else that they do and so sometimes I think there are articles that slip through the net but the audit is really important because it highlights those papers and highlights that we need to be extra vigilant to make sure that that isn't happening but I mean it's shocking that there was no reported sex so I'm a lay member of a work of Manchester and I'm just wondering if what role you think Aweb has in sort of dealing with this inherent sex bias that seems to be so prevalent, shockingly prevalent actually and if any I'm just wondering what your thoughts are on that I mean you guys should be pushing it I mean it's absolutely we should, the baseline position should be the equal numbers of males and females are used in all experiments for the first question you need to be doing that that should be your blanket starting point if when you've done that it's clear that there are no sex differences then you probably need to continue with both sexes in your studies if there are prominent sex differences then that is the rationale for doing a sex specific study if you really want to interrogate pathways but the Aweb is critical you have an opportunity to make a difference here at the university if when people are replying for their project licenses they're told this is simply unacceptable you need to include both sexes well they'll do it because you won't approve there you guys in the committees within the ethics committees you're in a position to actually influence that outcome so you have to use your position of power to steer researchers in the right direction yeah I mean I often point that out as a lay member and the answer that comes back is there's always an answer and there's always an excuse and I don't have the skills I don't have a scientific background at all so I can't actually press them on it so I just think Awebs and lay members in particular need some sort of support something some data just a way to actually press this boy because I think it's really critical I always ask this question well the excuses generally are incorrect they say that it costs more and that it's not relevant and that there's increased variability in females actually a lot of those issues have been proven incorrect or debunked now and actually what could be done is some summary information that supports individuals like yourself in making that case I mean I think universities in particular have to move their position they have to say that it's unacceptable not to be considering not to be balancing your experimental work at the early stages it simply has to move into that position thank you can you plug for the NC3R's experimental design assistant terrible echo can I just shout it if you could use the microphone it's really echoing I wanted to plug the NC3R's experimental design assistant that has the functionality to allow robust experiments using both sexes you're absolutely right and in fact in our guidance we did we have highlighted the EDA excellent online tool that can be used to help design experiments address the question from the A-werbs this is what we should be using the A-werbs should be pressing that the EDAs use both things like this while Clive is getting the microphone I think you hand your hand up thanks I'm going to ask you to extrapolate wildly from your research sounds like fun the question is does the difference in the way that the blood vessels respond between the sexes means that there's a difference in the way they respond to exercise to prevent us from getting heart disease and does that mean that as a male I can do less exercise than my wife absolutely not it means that your wife probably can do less exercise than you because she has EDA check that's what I would say right because I'm a female I think the women are protected in the pre-menopausal years and it does seem that endothelium dependent vasodilator influence over their resistance arteries is driven in females by EDHF we need to work a little bit harder to confirm this in humans but the pre-clinical experimental data is very strong and does support that position when the endothelium is activated by sheer stress it causes an increase in activation of the endothelial cell and that will release NO, EDHF and prostacycline at the same time what is true about studies exploring endothelial dysfunction in experimental models both actually in animals and in humans is that endothelial dysfunction seems to have some preferences over the endothelial mediators it seems that NO and the pathways for NO generation are a main target in that disease process so wild extrapolation since you need ENOS to protect resistance arteries and I need EDHF and the factors that trigger endothelial dysfunction actually seem to target ENOS and NO generation I would say that you're the one who needs to do more exercise than me we have a question from Clay I'm raised a really excellent lecture I just wanted to ask you a more technical question so you showed that endothelial derived mediators like nitric oxide and prostacycline don't just influence vascular tone but also can affect platelet leukocyte function and the lay picture that everyone shows about how leukocytes get out we now know it's slightly more complicated that platelets are required for that and I just wondered whether you or anybody else has done anything to understand whether there's a sex difference between how platelet leukocyte interactions occur to lead to those changes in leukocyte infiltration that you saw in your preclinical model so in preclinical I'm not sure I've got the answer to that what's for sure platelet reactivity is different between the sexes Adrian's just raised his eyebrows at me of course in his CMP deleted models endothelial CMP deletion and the NPRC knockouts it's quite clear that platelet leukocyte interactions are enhanced in females in the absence of CMP and that this is something prominently evident in the females versus the males so there is some evidence that that is happening as well it's true that NO plays a really important part in interfering with platelet leukocyte interactions as well it regulates the expression of key adhesion molecules and so you will see an impact of endothelial dysfunction on platelet reactivity in males and platelet leukocyte aggregate formation too I'm not sure if I've completely answered your question there but I hope I've done it in a radical way thank you please help me to thank Professor Elawalia for a wonderful lecture so I'd like to thank again the openness awardees congratulations please consider nominations and nominating yourselves or others and the next openness meeting is going to be at the Francis Crick Institute next December 2nd so please mark your calendars finally we'd like to invite you for some nibbles and drinks out there in the foyer so thank you very much have a lovely evening


All right today we're here with our one of our new recruits this is Sammy he's 12 weeks old golden doodle he is as hypoallergenic as you can get so for all the families we sell to everybody wants if they don't want the golden hair we have the golden doodles or labored doodles for people that somebody in the family has allergies and this is as good as it gets when it comes to being hypoallergenic so he is a sweetie he's got a lot of personality he's very loving very playful he will be fantastic with a family we just got them yesterday and he is a great great dog so come here buddy let's put you up here for a second he's a lover he loves to cuddle if you walk around he'll just hang on to you and look for you all day the cuddle so he'll start his training today tomorrow yeah all right come here buddy all right let me he's a lover lover so his training will start today tomorrow and depending on the level of training somebody's looking for our level one he'll be ready in about three weeks level two four to five weeks so we've had a lot of inquiries already about the the ones we've gotten in right now so if you're interested they go fast so give us a call and we'll talk to you about whatever you want to know about having one these guys go to your home and be part of your family so this is Sammy 12 week old golden doodle that's okay let's see what happens all right and now we have sissy also same breeder 12 weeks old and also very loving very affectionate very playful sweetheart this is just great dogs and our staff is wanting to keep them themselves so it's a big problem but yeah also as you know the highest level of hypoallergenic you can get so again anybody who has a family that you know allergies are concerned these are the best of the best when it comes to that hair type and very low maintenance these guys very smart breed this both of these are golden doodles both 12 weeks old difference between them is size she's gonna be the small side and he's gonna be the large size so if that's a concern for people a lot of people they have a preference some don't want the small ones they want the big ones want the small ones don't want the size but either way both very low maintenance no hair in your house you know you don't have any of that kind of maintenance the coats are very easy to upkeep so also very sweet this dog so if you're interested we have both of these golden doodles 12 weeks old and both same thing she's gonna start her training today tomorrow and three weeks she'll be ready for level one and four to five weeks they'll be ready for level two if you need a service dog give us a call we sell tons of service dogs whatever your needs are if you have special needs we have to add anything special to the training you'll let us know and we customize the training for you as well so if you're interested give us a call they go quick so that's sissy ready


What do you mean? What did you say about the Noorish? I said that Noorish is the one who makes me happy. Oh, okay. Let's go and have a cup of tea. If you don't know what I'm talking about, let's have a drink. Hey everyone, I hope you're well. Hey. It's so cute. Welcome back to my channel. If you're new here, my name is Sepi. And today I'm joined by my mom. Hi, mom. Hello. In today's video, mom, what are we doing? I don't know. She has no idea what we're doing, you guys, but as you can guess by the title of this video, I'm going to make my Persian mom guess the meaning of slang words. Mom, do you know what slang is? Yes, of course I know it's slang. Slang is like when one word has like a meaning, you know? Okay. You know, like, for example, javad is slang for dohati. Yes, for deshi. No. But for example, harfi ke manish is like, how do you explain slang in Persian? Let me Google translate this. I don't know what it is, but I don't know in Persian. No, because slang is like, for example, words that the kids use, that the adults don't use. I know. So, for example, like I said, no, babe is not slang. Babe is normal. So, she doesn't even know what slang is, so this is going to be... I know what is slang. Okay, she knows what is slang, okay? So, I'm going to make... So, I'm going to tell you slang words, okay? And you have to guess what do these slang words mean. Look like test, for example.  I think I understand what is a slang. This is going to be so fun. Okay, so this is helpful. So, my mom is saying that she doesn't even understand Farsi slang, so how is she going to guess English slang? I don't know. I don't know. I don't know. I don't know. I don't know. I don't know. I don't know. I don't know. I don't know. I don't know. I don't know. I don't know. I don't know. It's English slang. Okay, so I'm going to tell you words and you have to guess what does this mean? Okay? Okay. What does... goat mean? For example... You know in like... His wife, what was her name? Hoprahim Sultan. Hoprahim Sultan is such a goat. She's like... There. She's the goat of Queens. yes what does it mean I mean she's brave she's so if someone is a goat do you think it means they're brave it's not just only brave I don't want you love that no one's laughing you're so cute no one's laughing okay and means and she believes she can do everything okay but you're just thinking about Sultan like what I'm told I mean if anyone could be a goat okay first they can do anything they want they can reach anything they want okay okay that's close should I tell you what it means yeah goat is G-O-A-T stands for greatest of all time okay next one basic so then you'd be like oh that girl she's so basic yes she don't know that much anything about those things I talk about it that's actually kind of close so basic is like you put someone down like they're not they're not very interesting why another slang term is bye Felicia like bye like let's say like someone asks me to go to lunch and I'd be like bye Felicia I mean that means you're dreaming exactly well done I will not go out with you like bye okay bye seppy she's mean the next one is on point like that it will come literally like like that like your makeup is on point I'm gonna say in force see how I can sit in English you mean your makeup is on point your outfit is on point you're not saying it's exactly what you want no no no no no it's not you can't change your answer say one answer and I'll tell you if it's right or wrong okay what does on point mean on point means on the on the border so your outfit is on the border yes not very good not no on points means perfect amazing outstanding first I said no you said no you can listen first I said it's very good and you said no and I said this perfect very good you said no she's getting so angry she's getting vexed what is vexed to me I didn't I literally just said she is vexing yeah I mean cameo idea saying English it's perfect no vexing is not tough she's vexing and so we're vexing challenge me no it means like really angry like few make next one you have to know this what is why I should sit for this kind of you're lucky it's you otherwise I was gonna be that I know every day no one said you know it but this is what it's fun everyone wants to see if it mums know what like stuff means of course Iranian mums you couldn't compare all Iranian's mine and with me probably they have few boys at home and I didn't see you say a slang that's because you don't listen to me I listen to you what did you say can you can you repeat it it's because you don't let me speak English at home does anyone else's mom yes because they are not allowed they're speaking okay it's true maybe other Iranian mums do know slang better than you but maybe you speak slang with your friends when you go out with your friends and we don't know about it okay so the next one you have to know this okay it's bae like oh imagine like my name is like Sheila okay I'm Sheila and I like fancy guy called like more than I'm like more than is such a bae awesome he's very handsome bae means before anyone else so like my baby I love you before anyone else like more sense my bae my more sense my baby but how somebody you see them first time and you say it's bae don't worry I'm not Sheila I haven't actually met my more son okay so next point salty because salty actually means something totally different in Iranian like my mom said salty in Iranian translates to banana like which means you're very funny like if you're salty you're really funny and and no salty does not mean funny salty means like you're very angry and you're very bitter like why are you being so salty for why if you say salty means angry or bitter yeah why you don't say bitter and angry why did you say funny instead what do you say salty I thought it is like sweet but somebody is salty means really funny you're so salty in Iranian not in English you are last one that we're gonna do okay I think this one's quite easy actually so you should be able to get this fan fan mm-hmm what's up fan chilling with the fan say in English and somebody you like it yeah okay what do you mean someone you like and sit relax and enjoy it okay what's a better word to describe someone who you relax with and you enjoy who you're really comfortable with what do you yes like what what are they like if you're that comfortable they're like you're what do you mean fam you're chill with somebody you like it you're so you're chill with somebody okay that's close fam means family like you're my family like have you heard people like a slang say like blood like oh blood it means like you're like my bloodline miss a whole namby and in camp yeah so fam is the same it means like you're my family so so that being said me and my fam are done with this video yeah my fam me and my fam she literally copies everything I say I love you so much and so we are done with this video I hope that you guys enjoyed that and thank you so much for all to tell everyone thank you for their comments thank you very much for all the comment but you didn't like this video no because it was too much as the rest for me okay if you can think of any more fun videos as to be honest I enjoyed it was really nice good really nice if you can think of any other fun videos you want me to film with my mom or my dad or both of them and then just let us know in the comments down below and thank you so much for all the nice comments you left us last time please leave us some more we love reading them and replying to them and please tell her next time when you won't have a video with your mom at least half an hour before let her know not my hair like okay thank you so cute okay we love you guys so so much don't forget to check out my socials they'll be linked down below but my Instagram is Persian bunny my Twitter is the Persian bunny and my snapchat is seppy Samoy we love you so much love you and thank you very much for all the comment bye


I want you to stay tuned. Okay guys, what's going on here? Put this in here. Sometimes it works What a fail past few times it wouldn't work at all. You had to fiddle with it to get it to There it'll jam up Just like that. So what's going on is this key See the ridges here the teeth are almost non-existent. Here is a new key. Yeah big the Dammit the teeth are and These are worn down to almost nubs So we're going to put this new Lock cylinder in it. All right What we're going to do first now if If if you can't get your key to turn at all You're more than likely going to have to get a slide hammer and yank that out there because This is a little tab on top of here a little tab You have to turn the key in a certain position Right there for that tab to come down and see you can push the tab in It's actually going to go like this. I think it's gonna be on the bottom. Yeah, so you can push that Push that tab up to slide the cylinder out the same way with to push it in So that's why you're gonna need the key for that now Let me show you some things here guys Down under here. Okay. You've got some screws. This has got to come off here. Yeah, sorry This's got to come off here. So you're gonna have 10 millimeter screws You're gonna have one right above your emergency brake handle. This is a 2003 2004 Toyota Tacoma You're gonna have a 10 millimeter there.  I don't know but that's what that's gonna be now I've already got these taken out to save a little on time. It's kind of boring So what we're gonna do is we'll want to pull this down And out. Whoops. Sorry Just like that. Okay Now be careful This is your head latch. You got to be careful with that Now up in here.  I think I'm looking for right There's a little hole. You'll push something up in there and get this tumbler to come out. So first let's Sorry guys See if this will pop out. Yeah See that'll pop out of there Okay, just like a dust boot. So I think all I'm gonna have to do guys is push that in With a little small screwdriver or a pick or something But first we got to put the key in and it worked and you should be able to get up in that little tiny old right there or yeah, right there You know what, hang on a second. I'm wrong with my lens.  There's a little hole right there. See it? You got to push up on Now you're gonna have to turn that key to a certain place Until you can feel that push up. I can't find you guys Okay, there it is Let me show you. See how far up in that goes? Well, maybe not because I'm gonna push that up in and Nice to have a camera man right about now. I'm gonna try to pull this out And now you'll feel it See it coming out and pull your And this thing should slide out. There you go That's what you end up with right there Now check it make sure it looks the same and it does. Whoops. Sorry guys Now this one Remember we've already got it ready to get pushed and go back in So we should just be able to push this there's There's slots right in there. See right there the slot for top slot for the bottom. Okay, so you just line them slots up There you go. All right, once you get them lined up Down here at the bottom See that little a little tab right that little Dot there you got to push it up in With a screwdriver and push in on ignition I'm gonna try to do this one hand or there They say I'll push this up in and right there.  Let me see if I can And now I got a blood pressure cuff Something up here. Just like that push it in till it locks in there you go. I never condition her belt Let's put them on here about a year ago, but She was like you just put them on her last year and I'm like, yeah I know but the belts have like about a hundred Over a hundred and twenty thousand on these belts. I'm like, yeah You can't go by how long something is it's how many miles and so there you go Just like that. I don't know with my left hand And we're good to go Everything works good Good and the steering Never locked on this truck before she's gonna be happy about that But now as you can see It locks. Let's check the other key make sure it works. Like I said the steering never locked She's gonna be happy about that. That's all there is to it now For this to put this back up in there. I already show you guys that I mean it's pretty boring But you just get it up in there just backwards, you know, like I always say just reverse what you did. I Don't know if you can see anything because a little bit Like I said, just be careful of your hood latch cable. It's up in here Over here on this side. There's a couple little tabs tabs right here going in holes That's all you got to do. That's a little crooked, but without a camera man or lady make sure Everything's lined up and your little of course a little tatter there like that once you get your screws in And everything will be held into place. I'm gonna put them screws back in Get this all lined back up the way it's supposed to be. I'm gonna put my little deal back in there So anyway, you get the idea and There you guys go How you change a lock cylinder in a 2003 Toyota Tacoma 2003 2004 I Forget exactly what years but it's Pretty sure it's all the way up to 2004 from 2000. I'm pretty sure about that so But if not, there's little variances, but it's all on the same basis. So there you guys go So thanks for watching and sorry about the movement of the camera But it's kind of hard to do by yourself in a little tiny truck and get everything filmed so I'll try to Get most of that shaky and wobbling this out of it during editing, but you know, hopefully you'll get the idea So there you guys go. Thanks for watching check out monkey 1000s channel, and I'm Shae bear Shae burn 1000 the met the man the legend Shae bear. I'm gone for now guys. Bye. Bye. Thanks for watching