Now I have to behave and fabric disease. In the lab, we have several projects which all come together and they're under the umbrella of inflammation and fiber and formation. Even fabric disease has some inflammatory stuff going on after one second to me is the one I'm going to talk about today. I go in the clinic, I see patients and then I go back to the lab, and I look at some of the basic science behind what I see in the clinic, and sometimes some of the things I see in the lab, go back to the patients as well. Now this doesn't work and goes back to patients as well. So I'm always thinking on both sides of the fence and hopefully creating things and finding new information that can help patients down the road in the future. That's my favorite disease rat. So I'll speak about cataract and cataract surgery today. It's the most common procedure in the United States that affects all ages, and some are at a higher risk of complications, particularly children and patients with EVitis to the point where it changes our surgical plan and management. This is something that really got to me when I was a fellow where I had these little babies and they had cataracts and we had to operate right away. But then it was really just really complicated afterwards and that's why people didn't want to do it. So the key here is children are not just small adults. It's not just actually their eyes are almost the same size as an adult at birth that grows a little bit in that first year, but pretty much the same. But they behave differently. One big thing that's different about children is amblyopia. So grandma and grandpa has a pretty gnarly cataract. They don't need it out ASAP because they aren't going to lose their vision and have physiologic and anatomic changes that could be permanent. Children yes. So that's why when a child is born and they have a dense cataract in one eye, we want to do it before six weeks or both eyes. We want to do it before two to three months to prevent nystagmus because the longer you let that vision be low in that eye, the more solidified that poor vision is going to be. Then you have to deal with the amblyopia. So that's half of the challenge, probably more than half. That's because children have a critical period of visual development. If you don't correct the problem and rehabilitate the vision by about eight years old, you're never going to get it back. And that's something that parents don't understand in the post-op period. And the younger the age, the faster it changes. So if you have a two-year-old with a globe trauma and one eye has reduced vision, you're actually racing to rehabilitate that vision. If they have a cataract or something, because the longer that sits there, the faster those changes are going to happen. So same with pediatric intraocular surgery. Cataract is pretty common for intraocular. And if you don't treat it, they get amblyopia and you're fighting against that.  The problem is you're trading time and the physiologic and anatomic changes for amblyopia with that exaggerated post-operative response. So that's why we try to wait till six weeks to operate on a child, even with being a lateral cataract. Because if you do it earlier, you could have more complications. And in fact, we don't, I don't know why this keeps going randomly, but in fact, we don't even put in a lens in the eye at MCW, we don't even think about it before the age of one. And between the age of one and two, we think hard about it depending on the compliance of the family. So it would be nice to have an intraocular lens in the eye, but we're left with two options with these newborn cataracts. One, we take the lens out and we leave them a fake it and give them a contact lens for unilateral. At this point, we've been giving glasses for bilateral more often, partly cause it's difficult with the lens fitting and lens loss. There's a risk of infection. It could be completely impractical in rural areas of the world or certain situations where they can't come back and forth very easily. And if you don't use that contact lens or some sort of refractive correction, they get amblyopia. So you're trading off one for the other. The other option is to put in an IOL which we don't do under the age of one because they could get severe inflammation. They could literally have a fibering cocoon around the lens. The membrane conform and change the location of the pupil where it's not in the center at all and give them cartopia. And one thing we learned during COVID is that the pandemic really highlighted the need to address this and be able to put IOLs and babies. I swear to God, I don't know why this is happening. Maybe it's from the light. Oh, okay. I'll just keep doing it. But it highlighted that we do need to work towards getting IOLs in younger ages cause the soul soft lenses for the A-fakes were not manufactured for quite some time. And we had a lot of trouble getting contact lenses for babies even now it's still difficult to get them. So we had to go to RGPs and that's a little harder to teach parents to use. So lots of issues in the pandemic has even highlighted even more that we need to get this taken care of. So postoperative issues that we see are that membrane formation. This is actually a picture from a 10-year-old child. So it's not just babies. The synechia and pupil displacement can be highly relevant. We have several that we try to re-op and then they just scar again and all sorts of issues and intraocular pressure could go up. And we watch that for their entire lifetime. In the infant apacheal treatment study where they looked at kids who had an IOL put in versus no IOL, they found the IOL group under six months old had a five times higher rate of needing follow-up surgery. And that's just not a good thing. So that's why we lean towards not putting in an IOL under a year of age but it would be nice to not need to do that. So the rationale for our lab is if intraocular surgery is so high-risk due to this inflammation and fibrosis making us delay surgery, sometimes changing the surgical management not putting in an IOL. Maybe we could find a way to not have that happen. And in order to do that you have to understand why it's different. It doesn't make sense that a newborn baby with no immune system has an overactive immune system in the eye. It just doesn't logically make sense. So we really wanted to understand the scientific reason for these differences and see if we could target new therapies to prevent or treat these complications. And this is highly applicable to children and patients with EVitis which is what this picture is from. So the nice part about IOLs and kids would be constant refractive correction, reduced risk of corneal infection which actually happens quite frequently. Even with the infant AFAKIA treatment study that was an ideal situation where they had access to lenses all the time, lots of teaching. That's not the real world. The family is highly burdened and hopefully in the end it could improve their vision. So we looked for a good animal model to look at the post-operative response and we chose New Zealand white rabbits. They're nice to handle and they're often used in industry to evaluate new intraocular lenses. So they already have that mechanism to look at how things do after implanting an IOL. And the enter chamber is similar to humans. So we use the same instruments, we use the same materials actually. Everything's all the same. We use the fake machine, even the same IOLs. They have a seriously exaggerated post-operative response or our thought was if we could get a rabbit to have a good post-op outcome, children should be almost a slam dunk and it's a very controlled environment. You sending a child home with whatever you're doing, you don't know what's being done. Rabbits, animal models are great because it's a controlled environment. We don't give post-op steroids and we look at their post-op response and it creates a lot of scientific rigor which makes it easier to translate to people down the road. So the way we did this was we had the hypothesis that some proteins go up after surgery and there are probably some that go down or become undetectable after surgery. And we did this by sampling the aqueous humor before and after a lensectomy and looking at clinical exams. So just keep in mind that the aqueous, the enter chamber pretty much holds 200 microliters on a good day if you could get that much. So that limits what we could do. So this is actually a video of our rabbit surgery. So we do a paracentesis before to get the fluid and then we do use an MBR to make the incision. Oops, why is it doing this? Oh no. So we just make a small incision because we expand it later. So we put a viscoelastic and then we open up the enter capsule with the cystitone needle and micro forceps. And then I actually use Simcoe irrigation aspiration to remove the lens because it's so soft and easy in little baby rabbits. This is actually my old grad student. We pre-placed the suture because it could get... There's a lot of posterior pressure. We put in the IOL and then we take out the viscoelastic and close the wound and bury it. And if we put in medications, we put it in once the suture is buried and everything's stable. And then we do slant exams post-operatively. So that's a nice... I had to borrow this from the web but that's a nice example of the kind of sewing flare we could see in our bunnies. But these are bunny eyes. This is a normal eye without surgery. And this is one of our rabbits post-op and you see that huge chunk of stuff. We used to call it a clot because we weren't sure what it was made of. So that's pretty dramatic. And so we could literally look with the slip beam and do the exams just like you do in clinic. Same slip lamp. So we just first wanted to make sure it was an appropriate model. So we looked at older rabbits and younger rabbits and the older rabbits were much less responsive post-op than the young rabbits. So here on the left is a young rabbit with a huge chunk in the anterior chamber and on the right is an older rabbit with the same surgery. We did it with FACO and it looked pretty good comparatively. So it's a good model. First, our first question was, what are the different surgical stages do to the proteins in the anterior chamber? So we actually broke it down step by step. We did just paracentesis, then just the incision.  And then what happens when you take the lens out and put it in an IOL? And we wanted to know what the protein changes were before and after. So clinically, just a paracentesis, really no difference. It's a nice clear lens. Incision with suture, also nice clear lens dilates well. After lens activity without an IOL in the juvenile rabbits, there's some fibrin in the anterior chamber, but not nearly as much as when you put in an IOL. And it's really, really reactive across the board. We see this so often that our number of rabbits to achieve statistical significance is only six, so it's pretty dramatic. So your question is the fibrin on the lens, is it on the posterior capsule? Yeah. It's where the lens was. So it's in that, I didn't do a posterior capsulotomy and a vitrectomy in the e-fake it, because I just wanted to know the lens removal. So that's in that capsule area. So it's not on it, it's just kind of in there. And we did remove the lens a lot. There weren't lens particles remaining, so that's just the inflammation. In the lensectomy with the IOL rabbit, you could actually see turbidity of the anterior chamber fluid before we were done, because it's so reactive. Yeah. Yep. Yeah. So the, so the age of the rabbits were six weeks, exactly six weeks, because that's when they could be weaned. And the weight was usually, it was under a kilogram, usually it's around 700 grams or so, 600, some of them were smaller, it was variable. So, and then the heparin in the irrigation fluid, we didn't use it, and you'll see my NFD slides. It actually, I know that they use it in the veterinary world, and that was part of the reason why I wasn't surprised at some of the results we got. Stay tuned, right here. So one thing that we did see, which would be a surprise to some clinicians, but not to Dr. Werner, is that there is a large increase in coagulation factors, but there's no blood. It's not like it's, I call it a vascular coagulation in the anterior chamber. So there's a large increase in coagulation factors and fibrin. So that goes to the heparin in the irrigating fluid because one of our targets is using for preventive therapy is using low molecular weight heparin. So, but we didn't use heparin itself, we used a different one. The other thing we saw was complement components also increased dramatically, especially in the Lensectomy with Iowa Lensectomy groups. So we knew there was inflammatory components, that there's a blood coagulation cascade activation. And then another thing we saw was some immunosuppressive proteins really dropped in these young rabbits, even just a parasyntesis. So that told us that perhaps the young rabbits or the young children also, we still have to do some of those studies, perhaps the reason they're so overactive isn't because their immune response is hyperactive, it's more that they haven't developed inhibition or immune privilege. So maybe that explains why an immature immune system results in these overactions, it's like a double negative. So they haven't developed immune privilege is our gas, but we're still working on that. So we actually mapped out with the coagulation cascade all the things that increased. And so everything in red is increased postoperatively and that gave us an idea of where we could target our therapies. Also in the inflammatory cascade side, we saw that the classical lexin pathway was pretty activated in using the proteomics. So, and some of them talk to each other, which makes sense with COVID too, where you have a high inflammatory response and you also have a lot of coagulopathies going on in these patients. So once COVID hit, I just said, oh, they have to be anti-coagulated too. And look at what we do now. So the question for us was, now we know all these things increase, but where is it coming from? Is it from the structures in the eye or is it from the blood? And we did that with RNA sequencing and I won't go into too much detail because this is a clinical talk, but we've looked at the RNA sequencing changes in the iris and ciliary body after surgical intervention. And it turned out that all these pathways increased as well. So perhaps it comes from the iris and ciliary body, but we also looked at the cornea and these RNA transcripts also increased. So that tells us that the eye is a very dynamic system and that perhaps a lot of this development of the post-operative immune and scarring response may come from the eye itself and maybe some of the coagulation activation may also be from within the eye. So in conclusion with our surgical step-by-approach, the fibrin formation goes up at each surgical step clinically, but really after lens removal and especially with increased implantation of the IOL and the juvenile rabbits. The proteins detected, we have some that increase, but also some interesting ones that decreased following the surgical intervention at all steps.  We tried ELISA and it was just, we would have had to use so many rabbits that it wouldn't have been a good way to do it. And that the ocular structures likely contribute to these changes. Yes. Hi, I can't see anything because it's all bright. Some aspects of rabbits, kids are like rabbits in some ways, not all. Do you use heparin in the kits? Oh, okay, yeah, yeah. Yes, they do. That would be great. Yeah, I actually, a lot of our animal model development came out of scientists from Utah. So I used a lot of your papers. I was walking around and was like, I know who that is. So thanks for your pre-work, helping us develop this model. And so you'll be happy to hear about the prevention side is to, so knowing the coagulation cascade and the complement cascades are activated, we wanted to inject candidate medications at the end of surgery and see if we could prevent these responses. Right now, all these in the real world is steroids, right? Try and send a loan intraocularly if we get really worried systemic steroids, topical steroids. And that's really not ideal in kids because kids tend to have steroid response glaucoma. So I had a child, actually one of the first pictures is the patient that inspired me. And he had increased an IO, he came to us for surgery. The attending didn't want to do the surgery. So we did the surgery, sent him back but she wanted to handle the post up. He was on steroids four times a day, which I usually do a lot more than that. And then his pressure started to climb. And so she stopped the steroids because she thought of a steroid response. And then it really went up. And then he ended up with surgery. And then he had glaucoma surgery because it was all just stuck down. And then he had a retinal detachment and he had all sorts of complications after that when if we could have just prevented this, the inflammation and scarring post up, perhaps he would have done a lot better. Maybe there wouldn't be this confusion about whether a steroid response glaucoma or a fake glaucoma. So he's my inspiration. So now looking at these cascades, instead of heparin, I thought, let's put something in the eye that's gonna stick around post up and stick around for as long as we can. So I chose anoxaparin or lovinox. It's already used in children. So I love that part of it. And we still use trial and send alone. So I use that as one of our controls or standards of care. And then I actually put them in combinations to see if they have a synergistic effect. So this is our untreated as you have seen multiple times again and again, it's just a big chunk in the anterior chamber. This is with anoxaparin. So it acts on the coagulation cascade and prevents that fibrin clots. And we actually did some immunohistochemistry and proved that it was fibrin. So now I call it a fibrin clock because I know it has fibrin in it. So that's really nice. It doesn't have that nice red reflex that we like to see but it's definitely a huge improvement. Eight milligrams is a huge dose. We didn't see bleeding post up because we did it when the anterior chamber was stable. So I think by not having those changes in the IOP we didn't have bleeding events. So that was, and it was pretty consistent. Trinocinolone, I used a very low dose intentionally because I wanted to see just with a low dose and then in combination, that white stuff is the trinocinolone painting, the inflammatory stuff in the anterior chamber. That's not vitreous like you see in the OR. So that's just fibrin that's being highlighted. So we actually got a little confused because the cell was more in the trinocinolone but that's because we were seeing trinocinolone. So I'm not gonna present that data. And then when we put them in combination that's when you see that nice red reflex. So we were pretty excited. So I wish someone would put this in a kid on tube chicken to do it. So we need more dose response curves and a little more science behind it before we translate it to patients. But this is really exciting because if we could prevent things an ounce of prevention is worth a pound of cure. So we also looked at some of the inflammatory and coagulation cascade changes post op with targeted mass spectrometry. And this is just a heat map. So you don't get into the weeds of the details. But if you look at the combination, black is closer to zero, red is increased. Some things don't look any different or better with the combo treatment but there's a lot more dark. So the compliment, some coagulation factors are significantly closer to pre-op compared to untreated or an oxaparin alone or even just try and sit alone alone. So that shows that there are definitely proteomic reasons for the clinical exam we see. And we could actually map it out and quantify it with the way we did our mass spectrometry. And it kind of correlates with what we see that the combination is closer to the preoperative proteomics in the anterior chamber of the eye. So we're pretty hopeful that this may be something to target in the future to help our kids. Now, the question is what happens further down the road and you do well in that first week close up in the rabbits but what does that mean in a child? Does that prevent it way down the road months, years from now? So there are a lot of questions between the rabbit and children. So the other thing we thought about is treatment. Cause we have a lot of patients who have scarring or UVA just patients with a big fiber and membrane and we don't know what to do. So we thought let's target the coagulation cascade again and look at our clot buster tissue plasminogen activator. So we injected it at the peak of that postoperative response and examined it 24 hours later and we used a dose of 25 micrograms. So what it does is it targets plasminogen to plasmin and increases the degradation products and breaks it up. So this is our classic chunky, chunky interchamber of the eyes before. Our controller's just balanced salt solution says you'd expect it really doesn't do much. Maybe it clears up a little bit but that's just the natural way things happen. Pretty nasty looking. And then one day after tissue plasminogen activator injection there's a nice breakup of that fiber and clot. Didn't always go away but I'd be pretty happy if I had a patient with a nice cocoon around their lens and lots of problems and it looked like that. So we're also pretty hopeful with that. We did look at the cell and flare and in the TPA it looked to go up but that could be because those fiber and chunks are breaking up into little pieces and that's what we're seeing. So we did look at coagulation cascade and compliment changes as well because we didn't want something that would make it even worse and make it more inflammatory because it looks good but who knows? Maybe the eye is gonna get bad CME or something like that. So we did look and it doesn't look nice the way the preventative therapies does where it goes back closer to normal but it wasn't much worse. So we're still looking at this and seeing if it's a viable target. So it doesn't look like it reduces the inflammatory or coagulation cascade components but it doesn't increase them significantly but that might be just because there's kind of high to begin with. So still more work to be done on that but also promising. So in conclusion, anoxaparin is a hopeful use for preventing this postoperative complications especially with trienceanolone. It prevents and fiber and formation and then the trienceanolone kind of works in synergy to decrease some inflammation. TPA is also hopeful to improve the clarity of the visual axis after scarring has already occurred and one of my fellowship mentors has actually used this in desperation in one of her UVitis patients and she said it worked pretty well but the guy was blind already. So she just didn't want him to have complications but I'm not that bold. Future directions are looking at these eyes longer term. Really these experiments are done in one to two weeks. I wanna look months out and see other things especially the PCO. That's the poster capsular classification is totally different field that I do not have the time to attack. So we could always talk and see if there's other things that you could work on based on this research and all our data is publicly available. So if you wanna look at other proteins it's on our public data sharing website. And then our next goal. So we did the rabbits and this is a clinical talk and I haven't talked about a patient really very much. Our next goal is what about humans? So rabbits, we have a nice story with the rabbits but who cares if humans aren't the same? It's hard to do human subjects because there's so much variability in the population genetically, environmentally. Pediatric surgery is pretty rare overall. We see it a fair amount but that's just selection bias. And then the pre and post-op regimens are different depending on the surgeon and the compliance. So it's really hard to gather data in just an end of sex in humans. And we need a lot more numbers. So this is where we're still trying to work out a lot of the methods. And our main question is are we similar in animals versus humans? And this is where Dr. Huang comes in is what about the vitreous? The aqueous is not an isolated system. The vitreous is also very dynamic. And how can that help with PVR and post-operative fibrosis in kids? Because in an adult, they get a retinal detachment. There's some standard if it's mac-off you could wait a little bit. But in kids, even if they have mac-off their entire retina is just gonna be one big vibrant folded mess. So maybe children can, we can take advantage of some things in the posterior chamber as well. So what we're doing now is we have a relatively large sample bank with about a hundred aqueous and vitreous samples of all ages, some surgically naive, some not surgically naive. It's hard we can't get pre and post-op samples because our post-op samples and the rabbits are three days after that's not ethical in human subjects. So we really have to take advantage of surgically naive and patients who are re-ops for whatever reason. So it's a little statistically more complicated but we are building the sample bank at this point. And we did do some preliminary runs. We really have to alter our method because humans are so variable and very different from each other. So one is there's just a pediatric trauma patient here. So it's hard to parse out whether it's a pediatric or the trauma, but their complement was way higher and some of their coagulation proteins were elevated as well, particularly the fibrinogen so that we're still trying to get more pediatric samples to run together because we need a higher end but it was just a method run to see if it works. But definitely there are some proteins that go down and some that are lower and some that are higher because we don't know if it goes down or up because we don't have a pre sample. So we're looking to collect a lot more samples for more analysis and also the vitreous as well. So in another interesting patient we had a pan-UV-itis patient and looked at sample versus the pool. So we had to look at controls which were just adults that were having cataract surgery and nothing else going on. And it looked compared to our controls that the pan-UV-itis had elevated fibrinogen which is that coagulation fibrotic cascade, RBP3 which may be an immune suppression protein was decreased, which makes sense in UV-itis because they're very immune activated. So maybe that's an immune privilege issue and their immune inflammatory proteins were really high, really high compared to our controls. So we're still looking at this and just trying to get more samples and run them together. So lots of interesting things with our pan-UV-itis sample with our pediatric sample, which I pointed out already the fibrinogen was really high and the immune suppressor protein was low even though it was just a trauma patient we, they, she hadn't had surgery. The surgery was for cataracts. So the other thing that was interesting about her or about the sample was that it was a trauma patient we didn't see a frank tear in the eye in the lens capsule or even during surgery we didn't see a tear in the lens capsule but there are lens capsule proteins were super high. So that tells me that there's probably a micro tear that created the hydration of the lens and created the cataract. So why does this matter is pretty obvious but we do have patients that have had gene therapy and in pediatric patients who have had lexterna they are known to be at a very high risk of cataract and we're not sure if it's an inflammatory thing or if it's related to lens protein release. So this would be really helpful in those patients or those subjects where they develop a cataract if we'd go do the cataract surgery perhaps we could parse out if it's truly inflammatory or if maybe there are micro tears in the capsule. So lots of potential here. So we wanna increase sample size use our surgically naive in different age groups as the control we kind of throw them all together and look at the differences as a whole group. We realize that's not good to do in humans that are in rabbits and we really need to focus on reoperative samples because that's what we're looking for is why they have post-op issues. So that's requiring collaborative sites. So Dr. Huang is a collaborative site, Duke and Iowa might also join in as well and we're expanding our vitreous. So I just wanna say most of this work is not done by me directly. I was very hands-on but Jonathan Young depended his thesis based on most of this work. So I have to give kudos to him as well as Joe Bogard who's now doing his retina fellowship and the collaborators I have are straight-up biochemists and other really important people in my world. And this is John's cover of one of the journal articles he published that on the cover of a journal. So this is really pretty, I thought he did a great job. And then just in terms of hobbies, I love traveling. So this is my family and the reason why I have so many pets. Any questions? I see one from, should I read it? So Marissa LaRochelle said, we routinely inject TPA in the AC after you've had a cataract surgery cases at the end, 25 micrograms. So sounds about right. I would be curious to know if that really prevents the post-operative scarring that you see especially if you use IOLs. And I would be curious to see if anoxaparin would help as well. So anoxaparin, why do we use anoxaparin for prevention and TPA for treatment? We actually use several different drugs for the preventative side because I wasn't sure what would work. So Heparin was just too short. We did lobenox because it's really cheap and already approved in kids.   So we abandoned that really early. So we did try several based on the science. And so we kept going with anoxaparin because it has a pretty good safety profile too. You don't, you inject it at home. You know, you don't have to have it infused. And so I don't think systemically it would be a good idea to use it but intraocularly it's really isolated. And then TPA that was pretty much the only clot breaking drug that I knew was available. And we were very careful with our choice of the TPA. We use rabbits recombinant TPA because we didn't want an inflammatory response from like recombinant human TPA. So we were very careful in what we chose. Yeah, so anoxaparin acts in several sites to inhibit that end production of fibrin and TPA breaks up the fibrin which is probably why we saw more cell in the TPA. We didn't do it immediately post-op. I didn't even think of that. We just wanted to see if it would work in that huge fibrin trunk. So that's a good thought to use that immediately post-op and see. That looks pretty good. Yeah. Yeah. The half-life of TPA I think is not terribly long. So it's good. I do think there is a lot of value to preventing or to not having this fibrin scaffold in the beginning. And I think it has long-term benefits. So I think whatever method you use to not allow fibrin to form even microscopically because we don't always see it at the beginning. I think it's probably gonna have benefits months and years down the road. That's why we need to do longer-term studies. Yeah. Yeah, so good control prior to surgery. We do have like two or three patients that have been stuck who are in that amblyogenic range and have new diagnoses you guys can't see anything. And we're just in this horrible cycle of when do we intervene? What do we do?  It's better than nothing. Yeah, absolutely. I agree. But that's why if we could have a couple of more things that are non-steroidal in our armamentarium, I think it would be super helpful. So I'm excited to hear that TPA is being used at the end. And we are in the rabbits, we use it the same way right at the end of surgery, everything stable.  So, and then is there any concern with retinal toxicity with lobenox? I did. So that's a great question. And we had the same question in the beginning and we actually did ERGs on the rabbits after the lobenox and the steroid and the control and the ERGs were completely normal. So we didn't see anything, any signs of retinal toxicity but that the more studies would need to be done. Yes. So your question is if we saw Hyfema post-op TPA or lobenox, because you saw it in the TPA in that study, we actually didn't. I think TPA is a fibrin breaking drug. So it's very possible that it's opening up some of the blood vessels. And some of that has already happened because you're using iris hooks and whatnot. So that's I say potential for health and UVitis because it's a much more complicated surgery and you're definitely touching the iris and tearing it. We didn't see it with either one but we were very careful when we did it. Not to, we didn't have as much trauma because they're not UVatic rabbits. So, but I'd be curious to see what lobenox if you get the same problem. Was the Hyfema significant to create problems for you? Yeah. So that's helpful. Yeah. Yeah, I think we could deal with Hyfema pretty well. Yes. Thanks. Yes. So in children today, what would I suggest to minimize the inflammation with or without what has already been done? I think in general, I think surgery is getting better in a lot of our instruments are better. I feel like the more you touch the iris, the worse the post office.  So we actually are parsing out our dark-eyed versus light-eyed subjects for the human samples. So I think the less manipulation, the better. We actually recruited an amazing adult surgeon to do the surgery itself. And then we deal with the post-op because he doesn't touch anything and his post-ops are amazing. But for children, this is not even close to ready for prime time. You have to do things in adults first and then march back. I wouldn't use lobenox or TPA in any child right now. My chair, old chair at some point, we had a UBI just child who had a post-op mass. So can you inject TPA? I was like, no way. And it turned out they had an ophthalmitis. So I was glad I did it because I would have been in a lot of trouble. So I think there have to be those finding studies, a lot more work until you even put it into adults and then march it back to kids. The adults have to prove that it's good enough. It's just huge, pediatric IRBs are really tough. Yeah, sorry. Yeah, it's vague. I'm very vague with the slide. It's not that specific for trying similar. I'm being vague. Yes, post-operatively is upregulated. Yes, so because of the, I'm gonna go back to the pathway. Can we take advantage of the classical pathway activation to find something that is non-steroidal? Yes, I'm very interested in that. The more new drugs come out, the more excited I get. You just have to be careful. We actually targeted prothrombin inhibition and it was a mess also. There's a lot more to a drug than just the drug. There's the vehicle, there's a preservative free. So you can't just go willy nilly. I think that's what animal models are great for because you can kind of shoot a little bit in the dark and hope something sticks. But yeah, I would be really interested to see some of the new anti-inflammatories. We have to get off of the steroids. It's too vague, especially in children. If they have it systemically, it could hurt bone growth. So there are lots of issues with steroids. Yep. I think there's a lot of crossover with Andy as well. I think a lot of stuff in the eye is immune dysregulation. And just to be clear, I know there's a lot of cytokines and chemokines that are really heavily researched, especially in uveitis and other inflammatory eye disorders. We did not detect any, but that's most likely because we can't detect it because it's such a low concentration. So we haven't been able to get at that really well. We do see some changes in the RNA sequencing with the chemokines and cytokines, but we just can't detect it. And that doesn't mean anything. We even looked at doing human protein analysis using those antibodies. And it's not worth doing that because the crossover isn't good enough. So we haven't chased that lately. There are so many things to do. Yes. Yeah. So your question is, have we taken advantage of the immune privilege? Am I interpreting that right? Investigate. Yes. So we are really trying to get more in the specifics of the test of the immune privilege question, but that's a really broad thing and it's pretty messy. So grant funding is a little difficult when you go for something that's very new and unknown. So we're trying to get some more baseline data before we really go after it. But right now it's so fruitful going after the coagulation cascade and the inflammatory cascade that seems to be the closest immediate low hanging fruit. And that's also something that we do kind of on the side. That's where grateful patients come in really important because then I could explore quite a bit. Great. Thank you. Again, at noon to the research faculty and personnel in person here. Thank you.


In this second lecture called flow classification, I will explain the purpose of the lecture and then tell you something about the vast scope of convective heat transfer through the flow types and how we are going to select from this vast canvas only a few situations that are of still practical interest. So in convective heat in mass transfer, we recall that we are in concern with bulk fluid motion as such everything that affects bulk flow influences heat transfer coefficient H and the mass transfer coefficient G. All flows are governed by three dimensional time dependent partial differential equations of mass momentum and energy transfer, but not all flows can be elegantly treated by analytical methods and hence we require numerical methods to solve these equations. The complete equations under all types of boundary conditions and complexities of flow domains can only be solved by the technique called the computational fluid dynamics. It will solve the entire three dimensional time dependent partial differential equations and therefore, the scope of the subject is very vast both in terms of its physics, in terms of applications and in terms of mathematical complexity. We need to reduce this complexity so as to make it tractable through about 40 lectures in a classroom situation and therefore, what I am going to do is to classify the kind of flow situations that are of interest and from this I will give you the selection made for this particular course. The first type of flow classification is well known to you called the forced and free convection. So, consider a hot cylinder say a tube or anything like that across which the cold fluid is flowing and if the fluid motion is caused by external means such as pump, blower etcetera we call it forced convection heat transfer. In this case the temperature differences are such that density differences due to temperature differences induce very little or no motion at all. The entire motion is really driven only by the power provided by the pump or a blower as a case may be such a situation we call heat transfer by force convection. On the other hand if this cylinder was kept in stagnant air you would see the fluid motion would still be caused, but it will be upwards acting against gravity G which is downwards since the cylinder is hot the density near the cylinder is low and the temperature of the stagnant air is very low and therefore, its density is high as a result of the density difference lighter fluid simply climbs up against the action of gravity we call natural convection. In moving past the cylinder in this manner it picks up heat and transfers it to the ambient. So, if the fluid motion is induced by density differences arising from temperature differences we say the situation is completely free or natural convection situation. But many a times the external flow is not high enough to suppress all motion due to density differences and in such a case then we would have the two motions are comparable and such a situation would result in the net flow downstream of the cylinder going somewhere to the northeast. It is a combination of northward flow in natural convection and eastward flow in post convection and therefore, it would climb somewhere northeast such a situation we call mixed convection. Now, of course, you have come across experimental correlations of this type N u from the cylinder the Nusselt number from the cylinder would be constant into Reynolds number raised to power m Prandtl number to the power n and this we say is in this situation the Grashof number divided by Reynolds number square is very very small in other words all natural convection motions are suppressed and we have essentially forced convection. In this situation we have G R by R e square is very very much greater than 1. In fact, the forced convection does not even exist and therefore, when the Grashof number divided by Reynolds square is very very large we say it is a natural convection situation and if Grashof by Reynolds square is of the order of 1 then we have a mixed convection situation. In natural convection then Reynolds number gets replaced by Grashof number whereas, in mixed convection both Grashof and Reynolds are present Prandtl number of course, is present in all cases. The Grashof number G R is the ratio of the buoyancy forces and viscous forces. Most often we are interested in forced convection heat transfer and Reynolds number is an important criterion with Reynolds number is the ratio of inertia forces to viscous forces. As you all know when the Reynolds number is less than critical Reynolds number the flow in the tube will be laminar and the friction factor versus Reynolds number relationship on a log-log plot would be linear. It is F equal to 16 divided by Reynolds number is a very well known solution to laminar flow inside a tube. If Reynolds number is greater than Reynolds critical then we have turbulent flow and the friction factor would be doing that. For these two cases the heat transfer would look something like this. The Nusselt number would equal 4.36 if constant heat flux was applied at the wall and it would be 3.67 if wall temperature was kept constant. All this is known to you in laminar flow. Notice that the Nusselt number is independent of Reynolds and Prandtl number and it is essentially a constant. But as soon as the Reynolds number exceeds the critical Reynolds number the Nusselt number becomes function both of the Reynolds number as well as of the Prandtl number. What about the in-between region we call the critical region or the transitional region in which the laminar flow essentially is converted to a turbulent flow. For ducted flows as you all know Reynolds critical is about 2200 in the range says from about 2200 to 3000 would be the critical Reynolds number range. Like critical Reynolds number there is also critical Grashof number in natural convection. So, in all kinds of situation that we will be studying critical Reynolds and Grashof numbers have been experimentally studied. Theories exist to estimate them but usually the theories fall far short of what is expected from experimental results. It is extremely difficult to identify all causes of transition to turbulence and theories can only capture some of the causes of transition to turbulence. So, laminar and turbulent flows as well as the transitional flows are important classifications from the point of view of convective heat transfer. In fact, many of our equipments are deliberately run in the transitional Reynolds number range simply because it is in this range that there is a very sudden increase in heat transfer rate. A third classification is incompressible and compressible flows. Now of course incompressible flows routinely occur in liquids. We all know liquids are incompressible. The distinction is really applicable to gaseous flows. In gases we say the gas flow is incompressible if the Mach number defined as the velocity of the fluid divided by the velocity of the sound in that fluid is less than 0.3. Since you know that in at ambient temperatures the velocity of sound in air is about 300 meters per second what we are saying is if the under ambient conditions if the velocity was less than about 100 meters per second the gas flows could be considered as being incompressible. Now of course it so turns out in majority of our heat exchangers and other equipment gaseous flows are indeed at low velocity and having velocities much lower than 100 meters per second and therefore gaseous flows can also be considered as incompressible as far as practical convective heat transfer equipment is concerned. There are applications like in gas turbines combustion chambers and in turbines you do have situations where Mach number could exceed 0.3 but then we are not at the moment considered with such extreme situation. What is implied in an incompressible flow is that the density is either constant or it is function only of temperature. If it was a force convection situation of course the density variation with temperature would be very small but if it is a natural convection then the density variations would be significant. In compressible flows which occur mainly in gaseous usually Mach number is greater than 0.3. Mach number would be equal to 1 for sonic compressible flow it would be greater than for supersonic flows and the main distinguishing feature of a compressible flow is that the density in this case is a function of pressure and temperature both not only temperature but both pressure and temperature. Remember compressible flows occur only in gaseous. Another important consideration is wall flows and jet and free flows. Remember we are interested in determining H and G at the interface between a surface and a fluid flowing past it. The surface may be solid or liquid and the fluid flowing past it may be liquid or a gas. So naturally we are only interested in flows which are bounded by walls because it is only there where heat transfer coefficient is defined. But in fluid mechanics one also is interested in what are called free flows such as those that are formed in the form of a jet say air issuing or a gas issuing from a nozzle or a wake that is a flow behind a ship would be a wake flow. These flows of course are not bounded by walls but nonetheless one can identify an imaginary surface in which significant velocity variations occur and therefore it is of interest in fluid mechanics but not so much in convective heat transfer simply because there is no bounding wall and our interest is in determining H and G which is defined only at the bounding wall. The flows with interfaces are termed as wall flows and internal duct flows, external flow over a tube, wind flow over a lake etcetera are examples of wall flows and our interest is in wall flows as I said but not in free flows such as jets and wakes because there are no interfaces. The examples are jet discharge of hot water into water body, flow behind a ship etcetera. These are not of interest. Then there is the very important classification called the boundary layer flow and a recirculating flow. Boundary layer flows are flows that are long and thin that means if you consider a surface past which a fluid is flowing then the viscosity affected region would be very thin of the order of delta and the dimension x would be far greater than delta. So, we call such flows as long and thin flows. The velocity u also would be considerably bigger than the velocity v because of the predominantly unidirectional flow they are also sometimes called one-blay influence flows. By this we mean in the predominant direction of the flow the conditions at a cross section would be influenced completely by the conditions upstream of that cross section. Conditions downstream cannot influence the conditions at this cross section x. Therefore, we call them one-way influence flow but for example consider now a case of a surface on which a rib has been mounted. Then although the predominant direction of the flow is this way close to the surface where heat transfer is taking place you would have recirculating regions. In these regions of course, as you can see the influences would travel both from downstream as well as from upstream. Therefore, recirculating flows are often called the two-way influence flows. As we go along we shall note that in during mathematical treatment that boundary layer flows because of their one-way influence are governed by parabolic partial differential equations whereas, the recirculating flows are governed by elliptic partial differential equations. By and large it would be fair to say that it is extremely difficult to obtain exact analytical solutions to elliptic equations whereas, at least some progress can be made in obtaining analytical solutions to parabolic equation. Therefore, we would largely concentrate on situations that are governed by parabolic equations. Then of course, the single and two-phase flow. This of course of great interest in mechanical engineering particularly in boilers. What happens in boilers is that you have say single phase water entering at the bottom of a tube and heat transferred by radiation and convection as we saw in a PF furnace heat flux is falling on the surface. As a result of this first nucleation sites would be established when the temperature of the wall increases and small bubbles of vapor would begin to be formed and they would penetrate inwards into the core of the flow. Such a zone is called the bubbly flow regime. As we go downstream the temperature of the wall continues to rise and the number density of bubbles also increases so much so that some of the bubbles actually coalesce with each other and form large bubbles which are sort of confined and surrounded by smaller bubbles. The large bubbles move like slugs and therefore, such a region is called the slug flow region. If you go further down the tube you will see the slugs get elongated and bubbles continue to be formed and at the water surface. So, we have inner core of vapor long inner core of vapor and a thin outer core of liquid interspersed with tiny bubbles. Such a flow is called annular flow without entrainment that means there is no fluid entrained in the core. But if you go further down you will find the interface between the liquid water and the water vapor becomes very unstable and it causes rupture of the interface sputtering out water droplets into the core of the flow. So, water vapor and water droplets sort of coexist in the core of the flow while the water still surrounds the inner surface of the tube. Such a situation is called annular flow with entrainment where the water droplets are getting entrained into them. If you continue to heat further you will see a situation is reached where nowhere in the cross section any liquid water is found and you essentially have the tubes in contact with water vapor only. Now, as a rule the heat transfer coefficient the magnitude of the heat transfer coefficient in gases and vapors is much lower than the magnitude of heat transfer when in liquids. As a result of this there is a sudden drop in heat transfer coefficient near what is called as a dry out point. As a result of that the temperature of the surface of the tube would rise very suddenly to a very high value. So much so that sometimes even a tube might well melt. So, the objective is of course to make sure that such a situation does not arise in practical equipment and that the temperature is maintained well below the melt down temperature of the surface. So, post dry out you essentially have very very tiny droplets which appear like a mist in an essentially vapor flow and still further downstream you will see all these droplets would be converted to water vapor and the dryness fraction of steam would be 1 whereas it was 0 at the bottom of the tube. So, essentially then you have flow with phase transition from liquid water to complete vapor, but in between there are regions of two phases water and water vapor. In the bubbly flow the dense phase the water phase dominates over the vapor phase. As you move towards the black flow the lighter phase the vapor phase begins to dominate the flow. When you go to this annular flows vapor flow vapor phase occupies much bigger volume than the liquid flow and then when you come to the mist flow region of course the vapor phase almost completely dominates over the liquid phase and beyond x equal to 1 of course you have the purely water phase. So, such two phase flows are extremely complex in their physics because there is a continuous change in the structure of the flow. There is a simultaneous heat and mass transfer accompanied by phase change, but then there are also situations like fluidized bed dryers or pulverized fuel combustors and so on and so forth in which phase change takes place particles from solid phase move into gas phase coupled with chemical reaction. You have situations of cyclone evaporators, you have situations of evaporators, cyclone separators, situations of evaporators, boiling water reactors, fluidized bed dryers, all these are situations involving simultaneous heat and mass transfer with or without change. Extremely complex physics and mathematics are involved whereas in single phase flows the physics are relatively much more simple. Yet another classification is internal flow and external flow. I said we are interested in wall flows, but there are variety of wall flows. Internal flows are those we call ducted flows. So, for example, here is a case of a spin fin heat exchanger or plate fin heat exchanger. In this there is a plate at the top, plate in the middle, plate at the bottom and between the bottom two plates the flow is from southeast whereas in the top two plates the flow is from southwest. There are also the plates are separated by means of fins which are forming here triangular cross sections whereas here they are square cross section. I show this because normally our understanding of a duct is that it is a circular tube as you do in shell and tube heat exchanger, but you can get ducts of variety of shapes, triangular or square or rectangular or elliptical and so on and so forth. Such flows are called internal flows because they are bounded on all sides by a solid wall, but as I said if we consider wind flow over a lake then clearly the interface is between water surface and the wind and we have a situation of a flow bounded by wall on one side that is the water surface, but on the other side it is a completely free expense. Such a flow we consider as external flow. These two are very easy to understand, but in mathematical modeling sometimes we actually treat a confined ducted flow if you like also as an external flow. Where does that happen? For example, if you consider a turbine, gas turbine then the turbine blades are mounted on a disk at prescribed pitch. Such a situation is called a cascade of blades and the oncoming flow will flow through the passage created by the pitch. So, the flow would enter saw and would leave the turbine blade from near the trailing edge. Of course, from heat transfer point of view what we are interested is the heat transfer at the wall and it turns out that in such flows in the core of the flow the temperature would almost be uniform and so would the velocity be more or less uniform whereas the greater part of variations of velocity and temperature would be confined to an extremely thin region close to the surface of the blade. The top surface of a blade is called the suction surface, the bottom surface is called the pressure surface and therefore this blade disk would move in that direction. You will notice because these regions of where temperature gradients exist are very very small compared to the total distance between the suction side and the pressure side. So, as so much so that for all practical purposes we could treat this boundary layer development on the suction surface as an external flow which is bounded on one side by the solid surface the suction surface, but on the other side is a completely free expansion. Likewise, we could treat the pressure surface in the similar manner where the boundary layer is growing on the pressure surface. The surface is the interface where the region below the boundary layer would be a free expansion. Although truly this is a confined flow we can simplify and concentrate our attention on a very thin region. Incidentally it is in this thin region where velocity and temperature variations take place and as a result these variations essentially imply that there is a resistance to heat transfer and momentum transfer is also confined to regions very very close to the wall. Therefore, it is worth considering only those regions to obtain practical information which is also mathematically much more easy to track. So, the classification of internal and external flows is an important one because we often call certain external flows which actually occur in ducted or confined situations. Finally, the dimensionality of the flow this is very very important. The dimensionality of a flow is concerned with the number of independent variables associated with the flow. That means, number of independent variables with which fluid properties such as pressure, velocity and temperature vary. So, in effect there can only be the maximum number of dimensionality of a flow can be 3 x, y and z. So, you can only have maximum 3 dimensional flows. So, if I consider for example, flow in the entrance length of a tube then you will see boundary layer development will take place, but after a certain length the boundary layers will merge and the velocity profile which was in this case function of both x and radius r is no longer function of x at all because the flow has become fully developed and the velocity is a function of radius r only. You are all familiar with these terminologies development length and fully developed length, but notice that in this case the in the development length flow variables such as pressure, velocity and temperatures are functions of independent variables x and r whereas, in the fully developed region they are functions of x. I emphasize this connection of dimensionality with independent variables because very often people think of dimensionality of the flow as being concerned with number of velocity components which is not true.  The third dimension is not involved because we are talking about an axis symmetric flow. So, the development length flow is two dimensional but the fully developed flow is only one dimensional in the flow in a tube. But if you now consider a flow in a non-circular section tube like let us say square section tube you will have development length in which boundary layers will glow on all four surfaces and the flow in this region would be three dimensional. But once you reach fully developed flow then the flow velocity would be function only of the cross sectional coordinates y and z and therefore the flow will be essentially two dimensional. So, dimensionality of the flow changes within the same tube or a duct and it is very important that we appreciate the differences between flows in association with the dimensionality. By and large one and two dimensional flows are relatively easy to track than three dimensional flows in classroom situations. Scope of the present lectures then so all the things marked in blue are the ones that I shall be concentrating on while making very brief references to those which are marked in black. The first classification is between forced and free convection. So, I will be largely dealing with forced convection. There are laminar and turbulent flows including transitional flows. I will consider all those types. There are incompressible flows and compressible flows. I will be largely confined to incompressible flows because as I said in most of the equipments particularly in mechanical engineering the flow velocities are essentially less than about 100 meters per second. We will only be concerned with boundary layer type flows essentially flows with predominantly one way influences simply because they are mathematically much easy to track in a classroom situation. Recirculating flows require numerical methods and use of computer and I will therefore not be dealing with recirculating flows in the classroom. Wall flows by definition since we are interested in heat and mass transfer coefficient we will only be concerned with wall flows and not with free flows. And then as I said two phase flows are very complex to handle analytically and therefore I would deal essentially with single phase flows. Finally again one and two dimensional flows will be emphasized because they are simpler to handle analytically. Three D's flows will be ignored. In the next lecture I will continue with the laws of convection.


So, this is the last session that how to publish your application on a Google Play Store. So, in this session I will tell you how to what is the published process and how to how to get ready for a successful product launch. So, before you publish your application on a Google Play Store and distribute it to the users you need to get your application ready and test it. So, the preparation and publishing task are numbered so that it will give you the rough idea. So, the first step is what understand the publishing process. So before you begin your application, so before you begin this steps sorry before you begin the steps in the checklist you should take a moment to read and understand the overall publishing workflow and become a familiar with the process work. Second test for app quality. So, before you publish your application on a Google Play Store it is important to make sure that it meets your basic quality for all the Android applications. Third one determine the country distribution. Google Play lets you control a different territories for your application and therefore we can determine the country distribution. Fourth one confirm the application overall size. So, the overall size of your application can affect its design and how you publish it on a Google Play. So, currently the maximum size for your application that is for an APK on a Google Play is 50 MB. If your application exceeds that size or if you want to offer a secondary download you can use that APK expansion files. Fifth one is what confirm the applications platform and screen compatibility ranges. So, before publishing it is important to make sure that your application is designed to run properly on the all Android platform version and device screen sizes. So, you need to determine what platform your application are going to run. Sixth one decide whether your application will be free or priced. So, there are two types of application which will be either free or priced. So, on a Google Play you can publish your application as a free or price. So, free application can be downloaded by any Android users, but the paid application can be downloaded by only users who have registered form of payment such as a credit card. Seventh prepare your promotional graphics. So, when you publish on a Google Play you can supply a variety of high quality graphic assets such as image, icons to showcase your application or a brand. So, after you publish this will appear on your product details page or in a store listing wherever you will search your application. Eighth one build and upload the release ready APK. So, when you are satisfied that your application meets the user interface or compatibility or quality requirement you can build the release ready version of your application. Second last final checks and publishing. When you think that you are ready to publish then take a few moments for a final checks such as do you have the right version of the application uploaded or do you have set your application pricing free or priced and the last one is support the users after launch. So, after you publish your application it is crucial for you to support your customer which can provide a better experience for the users such as better ratings or any feedback or any kind of problem that may have regarding the applications and such as. Now, these are the four phases of the publishing of an application and we will see one by one. First one is the aiming at the right device. So, first thing you need to consider the Android version. So, let me go to the Eclipse. So, this is your Eclipse IDE and in the left pane I have project name Android app. So, let me open the XML file. So, this is my layout. So, this is a simple layout which has Akash wallpaper and I will tell you the settings of how to adjust the screen sizes, what are the different form factors and how to package your APK and all. So, first thing you need to consider is the Android version. So, for that right click on the project, go to the properties and in the left pane you can see the Android category and there you have a different Android version. So, Akash tablet uses the Android 4.0.3 so I am selecting Android 4.0.3 version which has API level 15 click on ok. So, second thing to consider is the minimum API level. So, double click for that double click on Android manifest file. So, in the left pane I have Android manifest file then go to the manifest tab and in the uses SDK select the uses SDK and there I have minimum SDK version that is already been set to 8 then I have a target SDK version that is 14 and the maximum SDK version. So, what are these 3 versions? That means my application will run will definitely run on API level 8 that is Android 2.2. Now the one more thing to look at the feature set so for that so what is the feature set? So, suppose your application relies on some features such as a camera, Bluetooth. So, you have to clarify this in the manifest file. So, click on add select the uses features click on ok and here you have a name field in which you have to name your feature set. So, suppose this application relies on camera so I will write android.hardware.camera and I need to set the required field to true so that means after you publish this application this application will only be visible to the devices that has a camera facility. Since I do not need this features I will simply remove by selecting the hardware the android.hardware.camera in the manifest extra and I will click on remove yes. So, if you want to learn more about this topic just go to the developers site click on developers go to the API guides. So, in the left pane in the left pane you can find you will find a android manifest click on it then go down sorry go down. And here you have a uses feature category select it go down uses features so these are the different uses features so these are the hardware features that have so these are the hardware features that is bluetooth camera you can also incorporate the software features. So, these are the software features such as a live wallpaper or SIP and all. So, now we will move to the second phase of publishing an application so the second phase is form factor and screen sizes now this topic will give you a few hints of how to best prepare your application for a different screen sizes on or a form factors. So, if you look at your application in a graphical layout and on a different screen configuration by selecting one of the devices from this list then you will then it will look quite different on a different screen sizes. So, let me take the 7.0 WSVGA screen and you now you can see that actually information of this application on such a big screen looks very small we can also change the android version from here that is from 4.0.3 to android 2.3.3 or 2.2 so when I select the API level 8 that is android 2.2 and now you can see my content is looking some older kind of so for that to handle this we have two possible solution first restrict your application either to specific android version so for that click on manifest so here I have manifest select the manifest tab select the uses SDK and here I have already set the minimum SDK version to 8. So, what is the minimum SDK versions that means your application will definitely run on android 2.2 and the target SDK version is the version on which your application has developed and now I will set the maximum SDK version to 15 so that means my application will definitely run between the android 2.2 and android 4.0.3 that means between API level 8 and 15. After selecting this my application will not run on a higher version that is 17 or 16 that is jelly bean you could also add support screen features so what is the support screen feature so for that click on add select the support screen features click on ok and there you can see I have small screen normal screen large screen so if you want to support the small screen set it to true normal screen if you want set it to true but if you do not want to set your application on a X large screen set it to false since I do not need this restriction I will simply remove this now the second thing you can do is providing a different layout so what is the different layout so for example you may want to provide some additional content for a larger screen or for a larger sizes you simply create a new android XML so for that click on wizard give the name of XML so main click on finish sorry click on next and here you can see the available qualifier and the chosen qualifier so now I will select the size element and if I want to move this size element into the chosen qualifier box I just need to press this button and set the size to the X large click on finish and now you can see I have created a new XML file for a larger screen so this is about the form factor and screen sizes now we will move to the third phase that is packaging a release build so in this topic we will make our application as a production ready so the app so for that we firstly need to have projects that is I already have the project here that is android app so if you take a look at your application folder so let me switch to the eclipse so firstly I need to find the location of this project where I have stored this project so for that right click on android app go to the properties in the resource category you can find the location of this project so I have to go to this folder see you are documenting each studio desktop android app so this is my android app this is my android app and if you open this you can see there are some other folders and inside the bean folder you have some generic classes so we do not need to touch them except the android app dot apk so your apk resides in the bean folder so what is the apk so apk it is a application bundle ready for the installation on android device like akash tablet and this is the file which creates as soon as we hit the run in eclipse so here I have run button so as soon as I click the run button it gets created that is apk file gets created so to create an application package ready to be deployed on a google play store right click on a project click on export select the android category select the export android application click on next so here I got the indication that no error found that means my application does not have any error and it will successfully run on a google play so here the project name is already prefilled click next then here we can enter the existing key store so then what is the key store so key store is the authentication if anyone ask you so it is it is a authentication for your application so if you do not have one you can create a new key store by selecting this radio button give the location of your choice where you want to save the key store so click the browse so I am I will store my key store in a desktop give the file name so I will write the name as key store click on save make up the password we should be sufficiently long click on next give the alize name application application again password then validity years 20 first name so it it will be in it will be your company name or your personal name so IITB so what is the indication that a 25 year certificate validity is recommended so I cannot put the 20 a 25 year certificate validity is recommended so I will change it to 25 and now the indication has gone and these are some optional fields if you want to put you can put click on next and select the destination for where you want to store the sign in apk so go to the browser and again I will save into the desktop only so it is already pre-filled android app apk click on save and here I got the message that certificate expires in 25 years that means my application has a validity up to 25 years click on finish and now I will show you the two files so these are the files key store and my android app dot apk and these two files I am going to use when I when I will publish onto the Google Play Store so this is the last phase of publishing an application on a Google Play Store so before that I will show you a different ways of publishing an application or different ways of distributing an application so there are 3 ways to distributing an app but before you publish your application on a Google Play we have a 3 methods so as an open platform android offers a choices so you can distribute your android application to the users any way you want so using any distribution approach or a combination of the approaches that meets your needs so these are some alternatives for distributing an application so first one is what through an application marketplace like Google Play so Google Play is the premier marketplace for android application and is particularly useful if you want to distribute your application to a large global audience however you can distribute your application through any app marketplace you want the second one is the through email this is the easiest and quickest way to release your application is to send it to the users through the email so to do this you prepare your application for release and then attach it to the email so here you can see this is a gmail and here I got the mail and with the attach file apk that is android.apk and when you click on this attachment it will pop up with the 2 buttons install and cancel and when you click on install it will automatically install it on your device so the distribution distributing the application through the email is convenient if you are sending your application to only few trusted users but it needs a few protection from piracy and unauthorized distribution because anyone can simply forward it to someone else now the third one is what the through a website suppose if we have some website we can also distribute the apk through that website so if you do not want to release your application on a marketplace like Google Play or if you do not want to send it through email you can make up your application available for a download on your own website so to do this you need to provide a download link download link of the release ready apk and then android protects the user from inadvertent download and install the application from the location other than the Google Play so you need to first select you need to first set settings then security and then the unknown resources so if you check this it will allow us the installation from the other resources so this was about the distribution of apk now I will show yeah now I will show you how to now what are the steps to publish your application so first one so first one is go to the play dot Google dot com slash app slash publish there you already have dot apk file so you have to upload it and as well as the icons and images of your application the third step then you will find the listing detail where you can set the language as English Spanish any language depending on your country then the title and description so title will be the title of your application and the description will be the description of your application then what is the application type so it can be the entertainment or related to education business and there are many more categories then set your application either to be free or priced now the first step is what publishing option so in that you have to set the copy protection either to on or off then set the content rating to all set your location to all location and in the fifth contact information suppose if you want the user rating or any feedback then you have to provide the information to the user the that is any website name or your email ID or phone number and finally check for the acknowledge and click on publish button and then your application will upload or will publish on a Google play so now I think you clearly got the idea of how to publish your application and I have explained thoroughly so thank you