Onepod Podcasts

Vikram

In this edition of The Half Life Show, we talk to Dr. Sidharth Mohan, a bio physicist and an expert in protein biochemistry. He is a friend of ours for many years, and we often refer to him interchangeably as Sid, and SidMo throughout the episode, he is an excellent teacher who is able to break down even the most complex topics into what he calls, despicable analogies. We talk about the fundamental building blocks of life, the wonders and secrets that Mother Nature won't reveal to us, and the implications of learning those secrets on the future of humanity. We discussed drug discovery and how the entire scientific community came together as one to develop and deliver the COVID-19 vaccine to the world. Throughout the conversation, you will hear in his voice, the excitement, respect and wonder he has for science and nature, through his lens Subu. And I developed a deeper appreciation for the world around us. And we hope you will feel the same through this episode, we talk about his personal journey, his inspirations and influences, which got him to where he is today. Sidharth is an excellent heavy metal drummer, even though he trained as a classical pianist, and the co founder of What's The Scene India, which showcases and promotes the live music scene in India, and has a dozen editorial teams in various cities all over India. Now, onto the episode. The whole reason we got started about all these things, of what you do, Sid, as you know, biophysicist that you are, is when you visited San Diego, and you came down for the American Chemical Society Conference.

Sidharth

That's right. When you stop by my place, this is what we did, right? We put up a campfire in the backyard, and then poured ourselves a couple of whiskies and got started on talking about biology and physics and all that stuff. So basically, what we're trying to do is recreate that environment, right, and get you to say some of those cool things we spoke about. Because Subu over here, was not privy to that awesome conversation. So that's why we definitely have to get you on the podcast. And now get you to explain those cool things you told me. And probably more. No pressure. Yeah. And so while I aim to, please let it be known that most sequels fail, and so that is the disclaimer, I'm definitely putting up front before we sort of dive into it. And the one one thing I do want to say is, it really helps, I think, I TA'd a lot I was a teaching assistant in graduate school for for many years. And you to explain a lot of concepts, I found myself clutching at straws, with explaining things in an academic way, which is taking the same scientific principle and just sort of rehashing it. So I started boiling things down to terrible, absolutely despicable analogies, but for some reason, they would often work. And one of the things that I explained is this, this idea of this Levinthal's Paradox, which I know you guys have asked me about before, and the analogy actually is not sort of unique to me it is it is derived from conversations in the field of protein folding, and protein science, where these, these chains of proteins are sort of compared to beads on a string. And so the idea of a sort of a long sort of string that has an inherently you know, you can think of it as a, as a two dimensional entity, it's just a bunch of characters on a screen actually has a three dimensional implication and so that that field of protein folding is, is kind of at the core of what I do as, as a scientist and, and what what fascinated me and sort of got me into the field in the first place.

Vikram

We've all heard of proteins, right? What is the importance of proteins? And why is the synthesis of proteins so important to the functioning of the human body? Or any biological system, for example,

Subu

especially because you have a lot of bros right? I mean, especially say in office in the gym, who are smacking themselves with two or three whey protein shakes a day and no, this is very big, you know, there's, there's a big call out for increasing protein, especially in Indian diets, you know, because current diets is not good enough and so on. So yeah, it's that's a good question. Let's start there. Okay, why do we why do we need protein? What's Why do we need protein? Oh, my God. That's that's a that's a jacked question to take your Brotein analogy as far as you want to. But simply said, they are the bio molecular basis of life. It is a it sounds like a very sort of grandstand grandstanding type. Have statement, but it is fundamentally true, you are alive and breathing. And you're able to sit here and hear and see and smell and taste, since we are enjoying some nice beverages because of proteins, they are the sort of biological molecules that that perform and execute pretty much all all of life's functions, right, they break down the sugar, they, they help you flex your arm, they are the fundamental biological basis of life, they mediate all life and all functions.

Vikram

Awesome. So it can make an organism function better, the lack of proteins can kill, and you know, misformed proteins can affect the functioning of various functions of the body, things like that.

Subu

So you know, for all things to be okay, all of these proteins, and these little molecules have to fold and look a certain way and kind of behave in a certain way, if they have to chow down on a sugar or break down fat or whatever it is, right? There's this a lot of lot of functions. And, and exactly, you hit upon the converse, which is that if they are not folded correctly, and folding is a very important word, we will get to that in a bit, then things can go astray. And so disease is born out of that there's issues, health issues. And that kind of segues into I think what you were saying Subu that, you know, why is protein necessary for health? And why is it necessary for building muscle, right, so there's, I don't obviously know exactly the mechanisms of how they are broken down and consumed. But in general, the idea is that your muscles are made of these, these cells that contain a particular protein called myosin. And that is really responsible for the compression of the muscle, and the tissue and, and that's what causes movement. And so the more of that you have, the stronger you are, the faster you are, etc, etc. And so being able to really consume protein and and break it down and really metabolize it, and force the body to take up that protein that you have consumed from outside means you can put on muscle you can you can gain muscle mass, so to speak. So you mentioned protein folding a couple of times, right. And I hear this a lot in our other medicine related podcasts and stuff. And anytime someone says protein folding this picture of like a thin sheet of paper, like paper folding, that's the image that comes to your mind. I mean, am I in the ballpark? Or do I have it completely wrong in my head?

Sidharth

That's one extra dimension added to it. Yeah, in the ballpark. But it's in a different plane, I would reduce that simplify it to beads on a string instead of paper that's been crumpled, right? Think of it as a very long chain of these beads. And I'll dive into that analogy right away. Right, so there are 20 basic amino acids, they are the fundamental amino acids and the building blocks of proteins. So if you want to talk about these in terms of the analogy, there's 20, different colored beads, let's say, okay, and those beads all have unique properties as a result of their little colors. And so you know, you can attach the beads end on end, and you can make a chain that's as long as you want and in any sequence of colors as you want. Right? So if you think about that, from a just a mathematical standpoint, as huge as a huge space, there's a huge coordinate space in which the sequence of these things can exist and and the the, the shapes that you can crumple this string of beads into it's gigantic right? There's virtually limitless numbers of confirmations, if you want to call it that. And so protein folding specifically deals with this field at the at the molecular level. And why is that relevant? Right? Okay, so it deals with it. But why is it relevant? Right? It's so I said that, okay, proteins form the basis of all life, they mediate all these functions. So you know, Pac Man needs to be yellow and have a conical mouth to run around the track. If he was any other color, the DOS would reject, reject the game. And that's exactly the same thing with proteins if you if you have a bead, and it goes blue, blue, red, red, black, yellow, green, yellow, red. And if you and that forms a chain that may be a crumpled little ball of yarn that that looks a certain way. And that can chew down on a sugar, let's say, right, if you change that last bead to a purple, suddenly the whole thing falls apart. It happens. Now the question is, why does it happen? Can I ask you one question right before that? Yeah, this analogy moves fast. So please, yeah.

Vikram

Are there any protein sequences that are purely a straight line? Or do they all have to fold and if they have to fold, why do they actually fold

Subu

so they have to fold so there is no real blanket rule nature, nature has evolved over over time proteins are as long and as folded as they need to be to do a function. So it's driven By need not just by, oh, you know, I have something that's 800,000 sequence amino acids long, therefore it is not folding. Like that's not the way we think about it. So nature has evolved to say, Oh, if I need three amino acids to do this function, I'll bet you money that there's something out there that doesn't. Now, is there something probably not, you need a little more of a three dimensional shape to do certain functions. That is, therefore the prerequisite is that it has to be at least a few more amino acids long, so that it can kind of fold and form a relatively more interesting three dimensional structure. Okay, that is, I think that's the best analogy I've heard of protein folding. Now. It now this protein folding is, in my mind is gonna switch from an image of folded or crumpled paper, to PacMan of different colors. And a different color essentially, represents what kind of sugars or what kind of molecules it can chomp on? Absolutely. I think that's a good analogy to start off, considering, you know, you said there was a five year cut off here. So now, this is very interesting, right? I mean, I have I have this graphic image of how proteins fold my mind now. But do you guys actually see this in under? Is that how they actually look under a microscope? How are you guys seeing this? How do you observe proteins folded.

Sidharth

So there's a lot of inferential work that is done, you don't really see it? See, it does, it has enough information gleaned from freezing, let's call it freezing the folding process. And there's also ways to interrogate it. And when I say interrogate, I mean, you could change those beads one by one by one through by two technological means. And see if the folding rate changes. And then you can sort of take snapshots, instead of instead of requiring a high speed video camera to do it, there are ways to kind of take a good SLR picture of it. And then you can then put those images together in sequence. And you can kind of elucidate the sequence of how these things are happening. And the actual pathway. Now, that being said, that is very, very sort of cutting edge and very challenging work to do. And there are other ways to, to kind of understand it, without really needing to sort of visualize it as a as a video. And nowadays, with molecular dynamics simulations, known as MD simulations, you can render every single atom in three dimensional space in a computer, and put in all of the basic rules of physics and chemistry, like, you know, if there's charged particles next to each other, then they will either attract or repel. And you can do this at scale, right. And then you can kind of visualize the entire protein, as if you were at the scale of the protein. And so the more refined the models get based on the information you get from other science and other experiments, the more informative the simulations themselves become. So it's a little bit of a bootstrap ish process. But the idea is that, you know, we're learning and we're obviously improving the models as we go, and we kind of we, but there is no all encompassing roof for protein folding, which is why we're even doing this research in the first place, which, you know, brings us into these all these sort of paradoxes and, and questions as to well, you know, what, if I change this, what happens if I change this? I think Vikram you were saying earlier, why does it need to fold in this way? Right. So if I change that one bead at that end, or somewhere in the middle, it suddenly changed the fold, and it crumpled into a different ball of yarn. So the question is, why does that even happen? Yes, we've seen that it happens, we know that it happens, but we don't really understand why. And that is the that is that is the most fascinating and frustrating thing about these these molecules.

Vikram

So, it looks like there is a law that nature is following that dictates the folding of these proteins, but it is probably the greatest kept secret of life that we are not privy to yet as human beings.

Subu

You know that that's not an understatement, I think one of the the the holy grail of protein science is to to be able to understand this given any sequence of amino acids basically, if I gave you a string with any number and any sequence of these colored beads, you should be able to tell me exactly what three dimensional structure it will form. We do not know that yet, which is kind of crazy to me at the same time, right? We have all this amazing knowledge we know so much. We have all these orthogonal ways of sort of tapping into the system. And without knowing this per se, we can still make the most amazing medicines, absolutely life changing, you know, technology out there just sort of treating it a little more empirically without really knowing the fundamental mechanisms and that's because you know, these things are reliable. There are some rules like you like you know, like you alluded to, and and we have some understanding of it and sometimes it's enough to actually have these breakthroughs, whether it's vaccines or it's, you know, therapeutics for cancer or anything like, you know, pick any medicine, there's there's so much that we know. And yet there's so much that we don't know. So it's a very interesting place to be as a scientist.

Vikram

So initially, I, you mentioned that there are an infinite number of combinations, given these 20, basic beads, colored beads on a string, the number of possibilities are endless. So it sounds like to me it's a gigantic computational issue. Yes. So we have today, probably the greatest computing in the history of mankind, we have supercomputers and I have come across this other project called Folding at Home, which means that you can even contribute your own personal computer and your own CPU. And it's in the show notes. So we have practically everybody's personal computer at our disposal, if you're willing to contribute to this project. So, us in spite of all this, we are not able to overcome the computational complexity that is involved in predicting protein folding?

Subu

Sadly, yes, because you cannot brute force this problem that I think that is the fundamental block. And why you can't brute force this is because the sample space is gigantic. So Vikram, instead of what you're telling me is, instead of using my compute resources towards mining Bitcoin, I should rather put it towards protein folding and at least save a life.

Vikram

Yes,

Sidharth

And eventually make an NFT out of it. Whatever that means in this in this space.

Vikram

Bitcoing are only 21 million, right. But, so how much are you going to mine there are only 21 million Bitcoins. But if you can solve protein folding, you have solved the secret of life. So yes, please contribute to that instead.

Subu

And Mo, you made a joke about NFTs. None of, the three of us know nothing about it. But I'm sure What you said makes sense. Because in in our space, we had this protein folding conversation. NFT probably stands for non folding talk. I'll give you a B minus on the joke.

Sidharth

Oh, my god, yeah, I'll take it, I'll take it. But let's let's talk about this. I know, Vicki, you mentioned, you know, we have all these computational resources, you would think that you know, this is something that's really truly within our grasp, and that we should be able to solve this problem. And the reason that we can't is because it's, it's just too large a sample space. And so we let's try and use this beads on a string analogy to kind of break it down, right, so let's talk about maybe a five bead string. So nothing crazy here. And let's assume that these five beads fold into something that looks like a little bit of a knot a small knot, you could hold it in your wrist, okay, it's a reasonably sized, we're about a centimeter, so you have something that crumpled into maybe an inch in diameter. Now, we said that the beads have a distinct relationship with each other, they you know, based on their properties, some basic biochemical properties charge, whether they're sticky, they like water, they don't like water, etc. And so that determines the kind of relationship between the two of them. So if you take two beads, or the first two, let's take number one, and number two, you know, blue and red, if you if you lock the position of one bead, then the red bead, let's just say it can be in one of three angles relative to the blue bead. So it can be either, you know, look at, look, if you're looking down the string, and you have the blue bead, the red bead can either be here, maybe so at zero degrees, maybe at you know 120 degrees, and then maybe at 240 degrees, and then back at zero. So one of three confirmations. Let's call it a confirmation,

Vikram

right? So any three angles, so let's look at it this way. So if you're looking at it, think about it something like a Mercedes symbol, right, you got three places. So you're looking at it, just want to put the second bead at any of the three tips of the Mercedes symbol. So these are your three choices.

Sidharth

That's exactly right. That's exactly right. And now you extend that to the relationship between bead number two and bead number three, beat number three and beat number four, four and five, etc, right? So it becomes an N raised with three number of combinations, right? Where N is the length or N minus one if you want to get into the details because the first one is locked. So let's say for 100 beads string, there are 99 sets so it'll be 99 raised to 3. What I did was I was very generous with the number of confirmations that you can use. I said you can do three. Okay, but there's infinite. If you look at it in three dimensional space, there's the whole 360 degrees so you can either sample it every degree, every half degree it's limitless, right? But even if you lock that down. Now, actually, in nature, there are some very specific angles that you can actually work with the amino acids relative to each other. They're known as the phi and Psi angles. So that's the sort of actual actual limitations of where these things can exist relative to each other. Even if you take those limitations into account. And if you could sample every single combination at a picosecond scale, okay, every picosecond, you can sample one conformation of one bead relative to each other. When you scale it to the length of a protein, given the Phi and Psi angles, it will take longer than the age of the universe to compute the entire chains, entire sequence the entire set of folds. Which is impossible. Okay, so but the paradox that that Cyrus Leventhal talked about, I think, in 1968 1969, was that this is a this is a thought experiment, right? We didn't really, I mean, they aren't really beads on a string, they are amino acids, they exist in real in nature. The thought experiment was, yeah, okay, it would take an really, really, really, really long amount of time to compute all of those folds. And, and map them out and have a piece of paper in which you can draw them out and say, okay, look, the one that is in nature is number 9,451,000,000,000, whatever, okay? But nature. And in nature, this happens on the millisecond timescale, right. So, because proteins are translated, they are read off, you know, the ribosomes are generated off the ribosomes in the cell, and they kind of fold kind of immediately, and boom, they're ready. It's like, it's like animals in the wild, you know, the, the giraffe gives birth, and that the baby giraffe can walk in no time. It's basically that at the molecular level, that's a terrible analogy. We're sticking with it. So the paradox is that it happens in nature at the millisecond timescale, and these proteins are viable, and they work. But if you were to brute force this algorithm, computationally, it would you wouldn't be able to solve the problem. So what gives? What are those rules that we don't know we are very well occluded, we don't really understand fully what's going on. But I obviously have to do you know, there is a disclaimer there in that we are shining more and more light on that every single day. There's a lot of research, and I don't do not want to discredit anyone's effort in doing it, it's a noble problem to solve. But like you alluded to earlier, from the it is the answer to life, if we can figure this out, then I can give you any amino acid sequence, and you would be able to tell it's exact fold.

And that's what nature does, in this scale of milliseconds to a second roughly. Yeah. That's incredible.

Subu

Okay, so now all of the stuff, right, all these experiments that you're running, so what what does an experiment look like? So I'm guessing that a large number of experiments that you run, are trying to take X number of amino acids, put it in the chain and study how it folds? Is that what you're, How do you? what are your experiments Like?

Sidharth

Yeah, that's one that's I think protein folding labs specifically interrogate and try and test different sequences to see if there's any impact on the function of the protein. And, you know, how long does it take to fold and more, some of my research actually had to do with trying to understand the interactions between beads in the sequence and through sort of relying on what evolution has already done. So another sort of twist to the story is that there are a lot of proteins and proteins exist, that they can be called the same name, it doesn't really matter that can execute the same function in different organisms in humans, in maybe an albatross that flies really high in the sky, or in a little bacterium that sits at the bottom of a deep sea event. And these proteins can do exactly the same function in these different evolutionary niches. Right? So you'd expect the structure to be identical, maybe, or close, you know, very, very, very similar. And since the structure is, you know, is derived from or sort of dependent on the sequence, because we've already established that the exact sequence determines the fold, right? That the sequences will also be the same now that's that's the wrench that nature has thrown at us. This these sequences need not be the same and they aren't we've seen in nature that there's families of proteins, they can that can do execute the exact same function break down the same sugar and do the same thing in metabolic pathway for example, but can have slightly varying structures slightly and vastly different sequences. So now that that opens up another can of worms it's like okay, wait, what I thought a sequence determines a fold. And I thought it's gonna be only that. But nature has allowed for some wiggle room. I mean, very literally, there is wiggle room were in very similar sequences. And now you can start quantifying what this word similar means you can start thinking about these similar sequences and the impact on changing those specific positions or on the activity of the protein on on its function.

Subu

So are you saying that if if we go back to your previous example of a protein with five beads, right, for five beads of specific colors, sure. So that could fold in different ways and could behave differently in a bacterium versus an albatross versus a human?

Sidharth

No, it could fold in slightly different ways based on maybe you change the fifth bead, but it will do the same function. Okay. So that's another twist, right? You'd expect the same sequence through one sequence to do one four to do one function, but it is there is some amount of promiscuity, so to speak.

Vikram

So the sequences are different, but that means the physical structure is also different, but they perform the same function,

Sidharth

they will be they will be very, very similar, they may be minor changes in parts of the protein that may not be critical to the execution of the function, but exists there for a reason. Maybe because it helps the albatross metabolize faster or okay, maybe I think the easiest analogy is, suppose this the same protein exists in humans. And in a bacterium that lives at the bottom of a deep sea went deep sea went extremely high pressure, extremely hot. And proteins are biological molecules, they are inherently fragile. And what that means is that they are as stable as they need to be in the niches that they exist, right? You don't need a protein in your body to be able to tolerate boiling temperatures. That is just a waste of evolutionary resources to have to find that sequence. Right? That Oh, you know, your rock solid, great. No use for you. Yeah. So over time, but what has happened is those sequences have changed enough that they can do this exact metabolic function, but they can tolerate that extreme heat and pressure, because it is critical to the organisms survival. And so that is where those slight differences come in, in the sense that maybe the catalytic activity is the same, but some structural component is is different. And that is where you will also begin to see the delineation of smile, delineation of structurally important positions in the protein and catalytically important positions in protein. So it's not all, not all amino acids are equal, but some are more equal than others. So that's a salute to George Orwell's.

Vikram

So it's what is important from this example you just gave is that even slight changes in protein, you know, in the protein, or the sequence of amino acids can create vastly different impacts on an organism like the deep sea event organism versus us, you know, chilling out by the beach kind of an organism, right, but doing the same function, which means conversely, speaking, suppose we figured out nature's secret of how to predict the fall of a given amino acid sequence, which means that we can now engineer life to the way we want it.

Sidharth

That is exactly right. And before we even go to engineering life, which is if you look at a single celled organism, right, that has life that has a complex set of biological functions being executed in real time, it's really, really complicated, even though it looks pretty boring under a microscope. Now, I don't want to, you know, sling mud at any biologists. But before we even get to engineering life, I want to take what you just said, and even dial it down to the level of let's engineer the proteins, right? So protein engineering, as a field itself is, is vast, there's so much that can be done to just make biologically active molecules and engineer them for a specific purpose. I don't know if you remember, but back home in India, we had a new set of these dishwashing liquids and soaps being marketed. And they were like, Oh, it has enzymes for catalysis to break down, you know, grime and stuff. You remember that?

Subu

And they show these these gooey little things running all over it now

Sidharth

unfortunate as the scaling is, in terms of the size, you can you can't see these things that microscopic, the reality is that we were making these proteins and these enzymes and they are exactly what we talked about at the beginning of the conversation. They're they're three dimensional molecules that are very precise and can do very precise functions. Very quickly. I will add that I was a little underwhelmed by those advertisements, because by that time, I had fully understood what enzymes were and I was like facepalming every time those ads would have Well, first of all that is as big as like, you know, a golf ball compared to like the earth and you're showing the earth in the yard. I mean, these enzymes are molecules, little bugs.

Subu

So in the next time one of your uncle's ask you, son, what do you do for a living and tell him, Hey, your shirts are cleaner because of me? You know, because of me.

Sidharth

You know, I had, I was talking, there was one gentleman who was asked this question, I don't I don't remember who he was, but it was a while ago in my field, and someone had asked him, What do you do for a living and he was in, you know, like, therapeutics biopharmaceuticals. And he gave a somewhat dark answer, he said, I help you die better, oh, my god, which while precise is a little disconcerting. And unfortunately, that hasn't left my mind, I'm tempted to often say that, I have to check myself when someone asked me, what do you do for a living? Oh, I make medicines that extend your life that helped you die better? No, no, let's not say that.

Vikram

By the way, now that you mentioned this, you know, how to die better, because we have better drugs, does all this like, you know, protein folding and the discovery of how these things behave, actually help us make better drugs?

Sidharth

Absolutely, it's undeniable, I mean, a lot of the interactions between molecules, whether it's an enzyme eating its substrate, or it's an antibody binding an antigen, which is the, you know, fundamental basis for a lot of treatments of lots of cancers, and immunological disorders, and whatnot, all of that is informed by that very, very specific site, or sites, or those specific sites of interactions between molecules. So you, they're called, you know, binding sites, or interacting sites or whatever. Now, and remember that all of these surfaces and points of contact between molecules, maybe you're trying to block the function of a cell that's gone haywire. And that can be implicated in cancer, right? So you want to be able to specifically bind one type of cell and you want to do this quickly, you want to do this reliably, you don't want any promiscuous binding and off target binding, because then the drug is, is doing random, crazy things. And you want to be able to also clear the drug from the system. So I'm capturing very, sort of very broadly and very, I've simplified a lot of the prerequisites for what a molecule like that should, should should be, like, what its behavior in the bloodstream should be like, you know, is it something that you have to ingest? Well, if you're going to ingest the molecule, it has to survive the you know, the digestive tract, it has to survive the digestion process, it has to enter your bloodstream, it has to find the cancerous tissue, it has to bind the cancerous cell, it has to maybe trigger another set of events to maybe trigger your immune system to recognize that troublesome cell and and then, you know, then your immune system can attack the cancer cell and then you're sort of treated of that cancer. And so you know, with as far as these mechanisms are concerned, I really liked talking about them from from like a warring government and agents and spies kind of view. Your immune system is basically a highly coordinated network of agents and warriors and informants and, and battle structures and databases, and you know, stuff like that. And viruses and cancers are infiltrators, that's the basically the analogy, right? That's how they override that's how they cheat. And that's how they can stay hidden, you know, cancerous tissue basically evades the immune system. So a lot of new age cancer therapies and treatments are immuno centric, which is we will not directly target the cancer, we will act as flags, we will act as guides, you know, you so you deliver a molecule that can basically specifically it doesn't kill the cancer cell, but it flags the cancer cell, so its only job is to alert your immune system. So it is now you can think of it as as a as a target like your your painting, it's like, you know, you're able to get in there, you're painting the target, and you're and you're leaving that so that way you leverage the body's own system. So it's, you know, quote unquote, a little healthier, a little safer, a little less toxic, at least those are the plans and those are the that's the vision of drugs like this.

Vikram

So these drugs go in and put like Bullseye targets on the problem cells for the immune system to then come and take over. So it's not actually, you know, taking care of the problem cell on its own. It's just putting a target on it and getting out of there. So that the body's immune system can do it on its own. Yes,

Subu

so is protein folding. Just one of the mechanisms of drug development are there like other basis for drug development that are popular?

Sidharth

Protein folding in general is the collective set of phenomena that determine the three dimensional structure from the sequence information. Understanding protein folding has implications on drug development. But it's not like that is the pathway to drug development. Drug development can happen and does happen irrespective of that, right. So at the end of the day, like I said, there are interacting sites, and we can interrogate and change those beads so that those proteins can, you know, either look, you know, like triangles attacking a sphere, or squares attacking a sphere, or you have a concave surface that attacks the sphere, which is ideally, you know, the best match, we can do that without really having to worry too much about protein folding. That, but that isn't to say that it's, you know, fully exclusive. Of course, understanding protein folding means that maybe you can engineer a better concave structure, right to attack a ball like, canceled, for example, I'm really pushing that analogy very, very far. It's about to break, but we can, we can take, we can take a step back. And

Vikram

yeah, let's see if we can take a step back and see, you know, how we can use a simple example, right? We all get these like mild fevers or whatever, and we go to a local grocery store or drugstore and get a little Tylenol, Advil, Crocin, you know, there are so many brand names, but I guess they're all paracetamol ultimately. How do you even make something like that? Let's say, you know, I asked you, Hey, can you make me a drug that reduces the body temperature? So how would you even start on a problem like that?

Sidharth

Well, other than asking for a billion dollars, I would require what is called as a compound library. So, you know, back when we didn't really know that much about biologics based drugs, and protein folding was not very, very well developed as a field, and we didn't really have a very good grasp on bio technological tools, medicinal sort of chemistry was was was the way to go. And so you had compound libraries, you know, organic compounds, and organic chemists would, well, organic chemists would make organic compounds that that spanned a variety of different shapes and sizes, and they had a huge plethora of properties. And and you could make these way, way faster than you can make a biologic, you can make them very rapidly, you can make a very, very large variety of these things. And you would basically have, you know, pharmaceutical companies that owned these compound libraries, and then you would really rely on thorough screening. So the science is not in the generation of the molecules alone, it truly is in the the ability to screen for a valid function, being able to reduce, like you said, a fever or, or, you know, help attenuate some, some trauma, right, some for some reason, whether it's to increase the rate at which your blood clots when you have a cut, or you so all of that is born out of a marriage between chemistry from from an organic chemistry standpoint, and a knowledge of biology as it continues to progress over time. And so being able to see which of those interacting partners can interact, can lead to the development of pharmaceutical compounds.

Vikram

So is that what you mean by screening, like, as in, you got all these compounds in a library? And you have some effects to generate? And it's like a match match one to the other? Is that what screening is?

Sidharth

That screening is basically that the actual experimentation that I know Subu, you were sort of asking earlier, what do we do? I think that is the core of the experimentation. It's one is making the molecules and having a vast array and tools, but then only one wrench really can tighten the bolt, right? And so you have to try the different tools to see what fits and what can do the function the best, what doesn't hurt your hand, what is easy to tune to different varieties of bolts, what has reliable metal, let's say or what has an aesthetically nice, pleasing shape, you know, pick your analogy, but the idea is that screening is where the challenge of finding the right molecule, or the best molecule within within the list that you have. And so a lot of the data analysis and promoting those molecules through a pipeline, so to speak, is where that comes from the idea of screening for the best function and finding the best molecule for that purpose.

Subu

So you have a bunch of scientists whose main job is to show up at work every day and create these molecules. And I guess by trial trial and error, figure out how those molecules behave and if it is something novel then added to the database.

Sidharth

That is a good oversimplification Yes. A bunch of us just turn up we high five each other we run one experiment we find the best one molecule and a billion dollars later we have the drug.

Vikram

So you just do.

Sidharth

Exactly right. I mean, but that's where a lot of empirical work, right, so treating things like an engineer actually really helps, which is where I sometimes find myself like, it's great to have that ability to tap into the fundamental science and see what you're doing. But at the same time, you want to be able to zoom out look at it from a slightly broader perspective treat treat your engine or experiment as a black box and and just Time is money. So at the end of the day, you want to find something quickly, rather than later. So but to to your answer to your questions Subu the experimentation and the data base and the data analysis, yeah, that is promoted down the line. And you have to do a lot of experiments to first of all, repeat those effects. And it could be some random effect, maybe the sun was shining through the window that day and so you know, that heat did it a little extra and something worked. So that kind of scientific or whatever. What is the right word,

Vikram

repeatability.

Sidharth

repeatability and testing the veracity of it is really critical. I think. Now, that being said, before you even you know, just it doesn't just go to the market as a drug, it doesn't run up on the shelf. That's where it needs massive approvals. There's a huge approval process, the FDA is involved drug goes through trials, it has to really be effective, it has to really not kill the patient. I mean, that's probably side effect number one. And yeah, oopsie but it's especially funny when you see drugs, you know, unfortunately, that are advertised as okay. I think Robin Williams had joked about it on his stand up show that he was looking into a drug or to take a drug. I don't know what it was for. And I could be completely wrong here. He joked about oh, side effects might cause anal leakage. problem he had with that was that it was not a side effect that has an effect.

Subu

Oh, God that's incredible. Okay, so two questions. Now. The I mean, you're talking about trial and error, empirically studying molecules and things like that, right. Now, we know that when some popular drugs like penicillin, were discovered through accident, right now. Now, the question is that what percentage of these drugs? What how much of it is accident? And how much of it is like, you know, intentionally drilling down and meticulous experimentation? Or? I mean, no, you know, where I'm going with this. How much is fluke, is what I'm asking is

Sidharth

Very, very, very little. I think the kind of breakthroughs of why penicillin is even rarer in the public consciousness is that because it was revolutionary, right, we had no idea that there was such a biological mechanism, right? That, that there was a set of compounds generated by a fungus that can do this. So it's kind of crazy that? Well, it was, I think it was also a little bit of an eventuality, like, I believe that a lot of discoveries in science. And as we progress as a species in the Great Universe, that we live in our eventual discoveries. So those are the sorts of maybe known I think penicillin, to us at that time was definitely an unknown unknown. So that's, that's why it was so it was a game changer. And the fact that that we could then mimic the the biological processes in a chemistry lab to make that class of molecules, and then change a few of the bells and whistles on those molecules. And I'm definitely oversimplifying it, to, to attack different kinds of infections and fight against different kinds of infections. That was a game changing thing. It was unbelievable and unheard of. Right? This is no idea that we could do this. And then we did it. And then obviously, the effect on you know, like, like the war happened. And so you know, with injuries and infections, it was definitely it had a truly global impact. And we see that across the board over time, whether it's the development of medicines or the development of technology, right? So penicillin is one example. But the polymerase chain reaction, PCR is another example. And some of the vaccine is ridiculous. It's like it's so present right now. But when you fast forward 50 years later, it's going to be looked at as one of the major feats of humankind. I would say it rivals the landing on the moon.

Vikram

You mean the generation of the COVID vaccine in the timespan that we did it as a human species to combat something totally unknown to our kind, and, you know, have a mass produce a vaccine that works to a large percentage on almost the entire population with almost minimal side effect, I would say,

Sidharth

yeah, absolutely. has never been done before. And if it has been done before, if I've gotten my facts wrong, not at this scale. So definitely this while the scale of this and to price is is incredible. The fact is that the mRNA vaccine platform as a as a sort of technology is revolutionary, we always knew that we could do this, I think there were a lot of difficulties and challenges faced by companies who dabbled in mRNA technology. And, you know, thankfully, companies like Moderna had a lot of the platforms in place. So and this is where the sort of empirical testing and screening really is important. Like you can have the best knowledge but if you and you can make the what you think is the best molecule, but there's no way to really truly test its efficaciousness. And its effectiveness if you don't have the right screening, and to be able to do that for millions and millions and millions of molecules very, very rapidly. So I think a company like like Moderna, and this may be like a plug for them and unintentional plug for them, but that's okay. They are they are I think at that, you know, they're on the pinnacle right now. But the fact that they could do that as is, is a big deal. It's a big deal just for the human species, not just for the company.

Vikram

From a biology point of view, why do you view this discovery and implementation of the COVID vaccine and it being mRNA based as some something so significant to humankind? I kind of stated it from a layman's perspective, but from a biology point of view, why do you think is significant

Sidharth

it is significant, because we need a little bit of a primer before we can apply this layer of paint. So Here go the here I go all aboard the analogy train once again. So so the basic sort of paradigm of molecular biology is DNA is transcribed, to form RNA. And RNA is translated to form protein. So we have all lived through the the excitement of the Human Genome Project, right? We all know, that was in the public consciousness for a decade. So many, I think billions of dollars was spent on kind of decoding the human genome. So why was this so important, right? And this is like, really a five step back. So you know, bear with me a little bit, but the, the idea that we will have an end to end knowledge of the alphabet, that makes up a human life was undeniably attractive, it was absolutely essential that we understand this as a species because all proteins come from DNA, right? They it's that sequence that we spoke about, of proteins is, is a photocopy. And here's where the analogy comes in. Right? So let's just talk about this DNA is the blueprint of all life, okay. And I'm again oversimplifying, but if you think about this as the Coca Cola recipe, right, the recipe for Coca Cola is supposedly stored in the locker somewhere, and there's just that one copy, you know, if you take that same analogy, DNA is that one copy is in the cell, but you have to make Coca Cola. So someone has to take and have access to that copy that blueprint, they have to have access to a highly regulated Xerox machine to generate the photocopies that only one FedEx company can ship to the different printing centers at different Coca Cola factories, so that you can make Coca Cola right. So in this analogy, the photocopies the mRNA, the RNA, the mRNA, the messenger RNA, okay. And the factories are the ribosomes that take the transcript, the mRNA transcript, which is the black and white, photocopy the blueprint stays in the locker, okay. And, and then takes all the ingredients, the different beads on the string, and is able to make this, this this coke bottle, which in this case, analogy is the protein. So, to to be able to read that blueprint in its entirety would mean to know everything about COLA to take that analogy to the limit, right? So that was the Human Genome Project. Now in biotechnology, we can sort of interrogate all these amino acids and you know, change these different protein sequences and study protein folding. Because we can change the DNA, we can shove this DNA into a micro organism and the micro organism can act as the photocopier and the factory at the same time. Great. So we can make different kinds of cola, let's say the cherry cola and the orange color based on how what what we tell how we are hacking into the transcript in the in the locker in the or in the DNA in the locker. Okay, so is that an analogy? Okay with us so far?

Subu

Nice.

Vikram

So far, so good

Sidharth

So far, so good. Okay, so this is a bit of a story. The only problem is those mRNA copies, right? They are like the glasses that Tom Cruise wears in Mission Impossible. They self destruct in five seconds after the message is read, which is to say that this class of molecules is inherently unstable. And that has to do with the fundamental sort of molecular structure. There's, you know, there's an extra oxygen somewhere and I'm not going to get into those details.

Vikram

Like nature's see, you know, layer of, you know, security like security, because you don't want these messengers to hang around with copies of important information. So once you deliver the stuff, you have to, you know, eradicate yourself. Right?

Sidharth

Exactly. It's a very efficient also, more than the security, it's scalability, okay? One factory needs maybe one copy, okay. And then you can have 10,000 factories. And and so you can, and the moment the factory uses the copy, it breaks down, right? It's eradicated. But if I kept sending a stream of copies to the factory, I can really make this a highly scalable system. And that's exactly what happens in the cell, you essentially have the one main, you know, blueprint kept in the nucleus of the cell for eukaryotic organisms that have a nucleus, and that DNA is transcribed. And so there's again regulated Xerox machines that forms the mRNA. And the mRNA, then gets translated in a ribosomes, where the amino acids are kind of put together based on the mRNA sequence. So literally, the DNA really informs what the protein sequences going to be. That's, that's the basic paradigm. Now the problem is that you think, Well, why do we have to keep messing around with the DNA? Why can't we just change the mRNA and hack the system, and then you know, we'd be set, right. But that's because the mRNA is unstable. So that's one of the main reasons so to be able to go and vaccines in general, when we deliver vaccines and older sort of vaccines. And that sort of typical vaccine, if you think about it, is a protein or a killed virus, right and attenuated virus, like the flu vaccine that you normally get every year. And these vaccines, basically, you can think of them as ghosts, that you introduce into the cell, or into sorry, into the human body. And when you introduce these ghosts, or the vaccines into the human body, your immune system remember it's that informant network, and that's, that's fine at work, they see they're lacking this, this thing looks suspicious, maybe I should call home guard and tell him to prepare, prepare some troops and keep them ready. And that's exactly what it is you're training the body to recognize. So when or if you are unfortunate enough to get infected by a virus that that the ghost was derived from, then guess what your immune system can go to red alert immediately before that virus, and that infiltrator really infiltrates your system and does its job, which is to make you unhealthy? So that's what so that's sort of we segwayed very organically into what a vaccine is,

Vikram

but that's making a vaccine that is Whoa, those are the kinds of vaccines that

Sidharth

`vaccine Okay. Exactly. So, so there you think of them as viruses that are attenuated, they look exactly like the real virus, except that they have no properties. They're just a shell of its former existence. Okay, now, how we make that I'm not gonna go into that. But the idea with the mRNA vaccine is, remember that these viruses are proteins themselves, they have protein on a protein coat, it's a shell of proteins, right? So let's not forget the basics here. Those proteins were derived from mRNA, which came from DNA. So ideally, you might want to be able to introduce DNA and then your body doesn't discriminate. It says, yeah, it's DNA, I guess we you know, we'll send it to the photocopiers we make some mRNA, they'll make some cola, whatever, that's what you that's what you think the body will do. body's not that naive, right? The body is also a little regulated. Second, this looks suspiciously different. I don't trust this document. Maybe the spies are trying to out spy us can get really messy. So unfortunately, this analogy is being pushed to its limit. But hey, that's that's what we do. So the idea with mRNA vaccines was let's introduce the mRNA circumvent that security in the main locker. Let's just go straight to the Xerox machines. That's kind of what kind of what we're trying to do here. So you're giving the body enough information. It's useful, you're not you're not really, you know, trying to destroy the body through the vaccine. Like that defeats the purpose. So you're trying to give the body it's think of it as a benevolent third party. Okay. A benevolent third party has infiltrated both sides of the war, and has said I favor one side. I want to give the body the team on the left, which is you know, the few the humans that want to survive. They seem like good people, some some information that might benefit them. And so suddenly, the body's like, Whoa, there's this photocopy out here. It's it's like, Oh, my God, it's sugar free Cola, I guess we can make.

Vikram

So I guess what we're doing is we're telling that the messenger RNA, don't go to the, to the nucleus of the cell where you normally get your instructions. Why don't you use this instead to make some photocopies?

Sidharth

Yeah, you're telling the ribosome. Yeah, the mRNA is the photocopy Oh, coming in with a fresh photocopy saying, hey, guess what, like, okay, so you kind of I mean, there's, there's a little bit of a back end

Subu

this is like that, you know, in your in your exam in your high school, you have a little chit with the answers, right? A little help on the side. So this looks like that.

Sidharth

The things I've had to resort to I'm kidding.

Subu

So we basically gave a cheat sheet or one of those cheats to our body. And that's essentially what the code vaccine is saying here you go.

Sidharth

Oh, actually, let's do it. Let's take a different analogy. So they're not say the teachers have figured out that the students are passing chits, okay. Okay. So you start writing them in Morse code. Now they can read it. Everyone in class gets it but the teacher's can't.

Vikram

So. So that's how you fool the body into accepting this new set of instructions and to trick the factories into making more proteins based on these instructions you just gave?

Sidharth

And what is the protein that that the body is going to make? It's going to make the COVID Like, Spike protein? Yeah, so then you're making the ghosts. You're not just delivering the ghosts to the body, you gave the body the blueprint to make the ghosts, which is like some next level alteration.

Subu

Yeah, this is such an enlightening episode.

Vikram

So this is why it is like a feat. This is never been done to this level of skill, where you tell the body to make the ghost instead of giving it the ghost, which is what we've done for decades, right? Giving the ghost now you make it yourself.

Sidharth

And and speaking to the scale of the operation, you know, people had concerns about oh, it happened so quickly. emergency use authorization this that. So without me getting into the sort of geopolitical landscape of this whole thing. I think the idea here that what what organizations did, and kudos to everyone who worked on all of these teams, literally, from the truck drivers to the scientists, like it doesn't matter. Every single person here knew that there was a worldwide crisis that needed to be solved and stepped up to the plate. So first of all, it's a, it's human spirit. Aside from the science, like that, to me is unbelievable. That is a group or there's groups of people out there that just got it and did something about it. So that to me is just like chilling. Like, right now I'm getting chills just thinking about. And and the second thing is a lot of these processes typically in science and in pharmaceutical development are, are contiguous, right, so they're a buttered enter to end and they can be, and that is why, you know, it takes time because you know, there are checkpoints or milestones that need to be hit. And there's a lot of money and effort that goes into these things. So you obviously want to minimize the risk. As the process progresses, in fact, the closer you are to making a drug, the costlier the risk of failure, right? If you think about just from an economics perspective, but in this in this particular case, I know that I mean, there's a lot of articles in the New York Times had a few articles, where, you know, suppose there were three or four lead vaccines or drugs that, you know, had maybe very competing data, very similar data, but we didn't really know maybe the effect on the body yet, I'm just giving one example as a potential route for elimination of one of them. So So you have four leads, but you can only make one, right, you only can make one at scale, and share that information with the world and deploy those resources to make it you can't have a variety of the same. It just doesn't make sense to do that. Because then you have heterogeneous effects, you know, maybe someone takes one and the other like, it can be crazy. It can be an absolute storm that you do not want to weather. So but but in the spirit of reducing the timelines, the manufacturing side, for example, would make all four at scale, ready to deploy. And then when the one was promoted, they would kill off the three Oh, my God. So we willingly took on having to waste resources, precious resources, because we needed to save time. So the groups of people who made those decisions, I mean, I salute them. It's incredible that we had that forethought. And it's so unfortunate that we had to waste some money and time and resources, but we needed the vaccine yesterday,

Subu

especially especially in the beginning, especially the beginning of the whole COVID pandemic, right in the first month. I remember I mean, we all didn't know what we were up against as a species. And it was, the roads were deserted. And like in the first few days, a lot of people including me, we stocked up on groceries and stocked up our pantries. But three weeks later, or a month later, we still were in the same spot, we didn't know what we were up against, because the death toll is rising and hospitals were inundated. And then I had to restock my groceries. So I go to this local big box store. And it was, you know, a scene from one of those movies like contagion where they allow, it's a huge big box store. And they are allowing only 10 people to enter at a time. And it's in everyone's like, boy wearing these yellow suits and masks and shields and I went out to get some advice, stock of groceries. And I'm like, Man, I don't know if I'm, I might just catch this bug and come back. But there's no food at home. What a terrible time that was.

Sidharth

Yeah. Oh, my God. You know, actually, it's very interesting that you mentioned contagion. Contagion was, I think, somewhat more scientifically accurate, rooted than most other sort of sci fi apocalyptic movies with viruses. It's kind of crazy. I mean, and then the reality of the human condition sort of mixed into that, you know, with, whether it's bureaucracy or a power struggle, I mean, those those are real things. And so those are challenges. I think that I'm sure people in like all of the companies that made these drugs faced, it's just what what do you do? You can't it's we don't live in an ideal world. And in spite of that, we succeeded. So to me, this is a human feat, like I said, that rivals, the moon landing, I feel.

Vikram

it's a good explanation, I think, really clear that up, in my mind, at least as to why this is so amazing, is Yeah, yeah, I

Subu

didn't know. I mean, I have a newfound respect for this. This type of a vaccine. I didn't know. After all those countless podcasts and other stuff that news articles I've read, I didn't have this kind of an understanding, as you explained. So thank you for that. So now, Vikram and I are both electrical engineers. I mean, we are hardware engineers. So from a from one question, what one thing I was curious about is, from a medical electronic standpoint, you spoke about all these developments, right? And all these experiments that you run all this stuff that happens day to day in your labs? Are you is most of the stuff most of your equipment that you use, is it still test tubes and pipettes and centrifuges? Or can you talk about some groundbreaking electronic equipment that has been that you use and things like that?

Sidharth

Oh, yeah. Oh, my God. So to answer the first part of the question, no, I think that is definitely a vestige of your time at esteemed University at undergrad school.

Subu

And every time I use it, I mean, I was hoping the answer is no, because every time we had to titrate and you know, find these things. And every time I used the pipette, I drank half solution.

Sidharth

Oh, my God, this mouth pipetting thing. You know, when I when I oh my god, it's so crazy that you say this, because when I've talked to my peers, or even juniors and uneven students hear that yeah, you know, I had to mouth pipette. And you know, we really, we really understood how differences in viscosity of the liquid that you were pipetting affected the, your mouth, because if you if it was too thin, and you thought it was like a Ghee like thing when you're tasting like I can veritably say I know exactly what potassium dichromate tastes like,

Vikram

you have to admit that actually forms a sentence in your life. I mean, I'm serious.

Subu

20 years later, I remember what industrial wastewater tastes like,

Vikram

oh my god, this gets worse.

Sidharth

That and that's a complex mixture, I had a mono phasic one entity kind of thing. And that was how bad enough I felt like my teeth are gonna follow goodness for you. I'm glad that you're here. industrial waste is probably like, you know, I'm grateful but also kind of sad that you're not like a Teenage Mutant Ninja Turtles.

Vikram

I'll take potassium dichromate any day over Subu's stuff.

Sidharth

I think so too. So that's the first part of the question. No, we don't use that that kind of glassware and stuff, but not on a regular basis. Yes, to handle a few mixtures and compounds here. And there. We occasionally use plastics and pipettes that are, you know, mechanical, and they have variable volume adjustments, and you know, they can be multichannel. So because you can do work in a matrix format in plates right now. So that allows you to increase throughput. And that word is particularly sort of close to my heart. Because I work in a high throughput lab, I do a lot of screening work. And we look at multiple proteins and biologics at once instead of one at a time. And so with the interests of experiments and hardware, I can give you an example of one device so there's I don't know the physics of it. So a lot of the devices and experimentation and experiments and instruments are based on the fact that the molecules we work with are not viewable under an ordinary microscope, right so biologists in a pure bio biology lab could see a warmth and a cell with basic with a very basic optical microscope, we can't really do that with what we have and what we work with. So you need more advanced technology. And it's not just oh, just oh, we need an electron microscope. But there are properties displayed by these molecules that you can sort of track through light scattering, through ultraviolet absorbance, through fluorescence, through you know, there's all these kinds of properties that really are derived from the fundamental makeup of these molecules. And so to that effect, you know, I can give an example we use something called a multi angle light scattering device. Great. Sounds amazing. What the hell does it do? So it turns out that physicists figured out that you take any object that is maybe a certain size scale, and if you have a light being shone on it, the pattern or the way the light scatters as it hits the object is indicative of the size of the molecule. Great. Fantastic. So and it's also dependent on the concentration of that molecule in the solution. So So you have sort of a three variable system vaguely speaking, right, you have the scattering effect dependent on the size of the molecule and the concentrate. So now if you knew the concentration of your molecule, from some other means, you can open up Pandora's Box, clearly, you can now tell what the average sizes and I can use that information to tell to tell me if there's a aggregation of the molecule in question to form larger super structures, like suppose they're all very sticky, and they just, they do not like being suspended in solution alone, for example, then you'd expect the light scattering signal to correspond to its molecular weight, oh, it's supposed to be 150 grams, and 1000 grams per mole or Dalton's. But it's behaving like it's something that 600. 600 is a multiple of 150, which means probably they're sitting in groups of four. So you have to establish a lot of standards and controls to really make sense of this data. But you know, we've able, we've been able to bootstrap over the course of history to kind of make sense of what are the fundamental behaviors and sets of behaviors, and then you can kind of glean from that what your molecule looks like, or how that behaves. So that's one example.

Subu

Nice. You briefly mentioned, what PCR is, right? You we spoke about the COVID vaccine and how that was discovered and what exactly that does. And but then I've been to a PCR test site, like a dozen times in the last year. So when I go to one of these sites, and I get my nose swab the throat swab, or whatever it is, what exactly is going on there? What what the swab and what are they looking for, to see if I have COVID?

Vikram

Who and I guess what does PCR in this context mean? We all use the acronym now freely. Polymerase Chain Reaction is a case in my limited understanding is a significant cornerstone of scientific development. So I guess you can help us understand why that is so and how it has helped us in this COVID journey.

Sidharth

Yep, if I have my historical facts, right, and I should, as a scientist in the field, with that use has, has used and uses PCR every day. It was a Nobel Prize winning discovery by Kary Mullis. And it really, I think, Snowball set the ball rolling or snowball, the biotech failed into what it is today, it was it was a fundamental sort of control over DNA sequences. You could you could alter and mess with DNA and change the sequence of DNA, which therefore means you can change the sequence of proteins as we now know, to your to your benefit to whether it is to explore a function or to make a drug or or to test something, right. So the idea of taking some some some known DNA, changing it shoving that into an organism in like a E coli bacteria. And then having those E. coli bacteria pumped out that protein at a relevant concentration in relevant amounts for you to do some biophysical tests meant that we could start doing protein engineering work. Really that's just the long story short in terms of what PCR is, and why is it called polymerase chain reaction. And every time I say the full fledged, sort of the expansion of PCR, I remember this rather terrible movie with Keanu Reeves and Morgan Freeman called chain reaction. I don't know if you guys have seen it, but it had something to do with this fusion reactors and the runaway chain reaction and you know, obviously everything goes goes to shit. Thankfully, that's the inverse event with PCR. Now that being said, it is a nuanced experiment and work that you can do in the lab, and it's highly tunable sort of experimental what it means is you can make vast amounts of quantity you it's it's a chain reaction because That is multiplicity involved. So if you think of DNA as a double helix that, you know, we're very familiar with. What you don't see in those images and in those beautiful video renderings is that actually DNA is an is a set of inverted strands, right. And there's a directionality to a strand. So it's it's not a real ladder, it's not a parallel ladder. It's an anti parallel ladder, that has implications on how sort of some of the molecular mechanisms happen and they go left to right, if you're looking at a frame of reference, if you lock your frame of reference, then then some some mechanisms happen left to right on one chain, and then some mechanical happens right to left on the other chain. That's just just just for general knowledge. The idea of polymerase chain reaction, I think came about from the story is that he kind of that Kary Mullis thought about it on the on the way he was away for a vacation with his wife. And he conceived the idea in his head and then left the vacation spot, straight to the lab. So I don't know what happened with the with the marriage, but he definitely got a Nobel for that balance, I guess. But, but he sort of conceived the multiplicity, he sort of thought of it as a thought experiment. And then they had some of those sort of, I think molecules in place, they just didn't put those building blocks together, so to speak. And so the idea is that you can rip apart the DNA strand. So when you're doing that, you're exposing, let's say, the printheads of a printer. Okay, so now you can shelve paper and on this side and get a photocopy of paper and on the side and get a photocopy. But by doing that with a molecule that essentially acts as a photocopier, you can now you have a sort of an exponential rise, right, so one becomes two, two molecules, because you have the strands separated, then you make a matching strand on on one, right. So if you have a photocopy of the one strand, and then the other strand will now resemble this photocopy right or other the other strand is exactly the same as the as the photocopy or not the photocopy, but it's called a compliment. So it's called a complementary sequence. So the two strands already complement each other, you rip them apart, then you make a complementary strand on the one that you ripped up on the left, and the one that you lift up on the right, so now one became two, guess what happens two becomes four, four becomes eight, right? And then so that becomes really multiplicative. And I'm clearly demonstrating my lack of geometrical progressions. But the idea is that this becomes an exponential increase in the amount of DNA that you have in place. Okay. This is for doing blank photocopies, I gave you the extremely naive sort of zero level PCR reaction, right. And you can do this by by by regulating the heat and the way it works. And so DNA is denatured to expose the anti parallel strands, and then you have what is called as annealing of the primers. So the photocopying machine is called a polymerase, hence the name polymerase chain reaction. The polymerase is just an engine, it photocopies, it can read the DNA base here, and it can put in the complementary base on the other side. So it's the photocopier machine.

Vikram

So DNA is basically taking the two strands and denaturing means breaking the DNA strands, you know, into two strand instead of like, a double helix, you make it like two single helix.

Subu

And then you basically take some clay and create mold, you stick it to each of the each end of the strand, and you get essentially like a like a mold of what it looks

Sidharth

It's like It's like fission and cells, right? If you if you've seen cellular reproduction from under a microscope, they just like cleaved in half, and then they like. know that that gets, but actually what's happening at the molecular level in the nucleus is the replication of DNA. And it looks exactly the same here, except that we're doing this in the lab, and we can control the way it happens, which is crazy. If you think about it, you've mimicked a biological process with temperature. Now, why is temperature important? Because the way it works with this actually broad steps, so you denature the DNA, you really boil the crap out of it. And then you anneal the primers. So remember that the little it needs a little bit of clay, like you said to at the very beginning of the of the strand, so that the photocopier can start doing its work, okay. And so the annealing happens at a certain temperature, the reading happens at another temperature, and then they kind of seal so now you have two copies, like we said, and then the two becomes four, four becomes eight, etc, etc. Now the really cool thing that you can do with PCR and why it's so revolutionary is like not only does it the zeroeth level is powerful and just making so much so much more DNA that you that you need. Is that isn't that a annealing stage. The primer that you put that allows the photocopier to start can have a forced error. Okay, that forced error, or a mismatch means there's some some length of that complementary that sequence that you put in the little bit of DNA that you put in, that doesn't match at every single base to the parent strand. Okay, that's it. parents tend, you introduced one primer, you thought it will be exactly ATCC, it will be the matching sequence and then they'll continue, everyone is happy. No, you forced the error. Because you want to make an amino acid sequence of your choice. So you're going to make sure that the DNA sequence matches that amino acid sequence. But based off some parents DNA, right. So through forcing that error, you will now and if you do the math, and we're not going to get into it, you're going to get only the predominant species in the population as this explodes exponentially, yeah, the predominant species in the population is going to be the DNA of your choice with that forced error, because you wanted to make that. So now you can take that error, error DNA, or rather your manipulated, clone the DNA, right, and put that into a cell and make the protein that you want it to make. So you went from intent to execution, because of PCR, you could never do that before or you could but it was very, very, very, very hard, very laborious, very, very trivial, very difficult to do at scale. You can do this in a, in a school lab, it is taught at the school level nowadays, which is ridiculous cover given our backgrounds, where we came from and where we are, it was like, Oh, we had to take you know, get a degree in the field to kind of understand it and test it in the lab. Now it's an everyday thing.

Subu

What year was this Nobel Prize? Given? What year was this PCR?

Sidharth

That was a good question. I'm gonna try a very sort of Joe Rogan approach here and try and pull it up for you. 1993

Subu

Okay, that's pretty recent

Vikram

that that late huh?

Sidharth

Yeah. It's think about it, right. So in from 1993, to now we've, we've got mRNA vaccine technology, that is solving a pandemic level problem. That's ridiculous.

Vikram

So in the PCR tests that we do, by swabbing our noses for COVID, what happens there

Sidharth

they are. So we know the sequence of the spike protein. And so we know what its DNA can look alike. And so I believe what they're looking for is the presence of that genetic information in the swab, which means that the virus is present. So you are directly probing for the DNA sequence that the virus has. So if you think about the virus has its genetic information, you're sending in the known photocopy, saying, if the photocopy happens correctly, and 25 repetitions later, I have two to the 25th, the number of copies, holy crap, you have the virus, that's basically it. So you're using the photocopy machine to increase your signal to noise ratio to tell you if there is actually that genetic information, which is indicative of the virus or the presence of the virus.

Vikram

So the forced error here is the virus's sequence of DNA.

Sidharth

It'll be viral related sequence.

Vikram

Yeah. And when you introduce that goes into the tube. Yeah. And if the dominant process, the dominant sequence that arises when you do this exponential multiplication of DNA, if that matches the original sequence, you have COVID. Because what multiplies was the COVID virus sequence DNA see exactly. And not?

Sidharth

That is correct. You're not looking for your sequence? We mean, that kind of information is, first of all irrelevant in this context. We were checking to see if you have COVID. And secondly, can you imagine if every single testing facility, I mean, the implications of storing an individual's genetic information is it's this huge implications. And you know, there's ethics and, and regulation and regulatory stuff. So that's obviously much more challenging. So what you're really looking for is, is there viral genetic information? At some level in your nose? If you if there is, then it is we're not just saying, oh, you know, there's you have the virus, it's very, very likely that you? Yeah,

Vikram

got it.

Sidharth

And that we could do because of us. So here are PCR was used as a diagnostic. So you see, the flavor of applications is virtually is, like limitless. There's biotechnology for making something that you want, you know, you have quantitative PCR where you can make a set amount of DNA, there's diagnostics like this. I mean, it's and then it's gotten more and more sophisticated in terms of what those primers are. And you know, it's expanded the repertoire of molecules and photocopying machines that do that photocopying has also increased, which means that there's there's probably uses of PCR out there that even I'm not familiar with.

Vikram

Amazing. Amazing. Awesome, so we should refresh our drinks. And yeah,

Subu

So let's get let's get into it methodically now. You Yeah, you are. You are an accomplished scientist in your own right, right. And you're doing important work work that you enjoy, but How did it start? I mean, why biotechnology? Why this line of science? And have you thought about, you know, did you get into all of this by accident? And by just circumstances of life and decisions back then? Or was there a knot? Do you feel like it was an obvious trajectory that led you to this.

Sidharth

So there was nothing, I think obvious about it. And while one can argue that Hindsight is 2020, and you know, you look back and you say, of course, that this was inevitable at any given point in time, you're always trying to operate from the perspective of I'm going to make the best decision I can now with the knowledge I have hoping for the best in the future. That's it. But there were some I think he milestones, right, and this trajectory, so to speak, one of them actually overlaps with you guys, which is kind of insane. I think on a previous episode you talked about surely you're joking, Mr. Feynman. That was mind blowing ly formative for me too, which is, I was like jumping with joy. When I heard that on one of your previous episodes, I couldn't believe that you guys want I remember calling Vicki and telling him immediately like, Dude, this is insane! Like, and what that did for me, let's drill into that, like. To me, the free spirited sort of nature of the man was was one thing, but his fearlessness in research. He didn't care about the, you know, the cost or whatever, he was not bound by those rules. Not to say that he was, you know, terrible as a person and did all and blew things up within reason within his social constructs. He, he operated freely. And he was curious. And he was unrelenting, right? That tenacity of questioning, it didn't stem from an insecurity, it didn't stem from at least my understanding, right from a need to be validated. It was genuine curiosity. And I think he gave me some permission to ask those questions. I therefore, I became the one of the most annoying people in my family, like, there was, what can you say what this kid will not shut up asking why. And this is a 19 year old, not a five year old, but the idea was just that, okay, you know, you say we do this, and there are customers, and maybe it was but why? Like, I acknowledge it, I'm not questioning the, you know, the power structure or whatever. Right. It never came across atleast from from my perspective, when I asked the question, why I think that was when the researcher took root. In my being, I think. I think all kids are curious. I mean, you guys both have kids, you know how they operate. Right? And and I think, maybe for a while that curiosity of mine was dormant. And you know, you're in survival mode as you go through school. Where are you going to be curious, just getting through our education system was a task in and of itself. Do you guys know that?

Vikram

Oh, yeah. We had a whole episode on that too.

Sidharth

Exactly. pedagogy is insane. Big, big topic. So what Yeah, so the permission to ask why and never really stop, right? That the frame of reference keeps changing when you ask that question. And then thankfully, having the tools to kind of adapt and let your brain kind of wrap itself around those concepts and and be satisfied with the answer that you are given them. But probe the next level probe to the next level. I think that that was where the research was really too root, like I said, so. So the book was one line. And that's just one of those random time points. And it was a crazy story, how I got that book, too. We had some distant relatives visit us for the first time when we were living as a joint family. And I think they didn't expect me to be there as one of the cousins they had my other cousin in mind. And so they got two books, okay. And at the time, my cousin was interested in cricket, and the other book was like something about Rahul Dravid or Sunil Gavaskar. I'm not really sure what. And they just happened to have bought, surely you're joking, Mr. Feynman, in thinking that they would give it to my cousin, but the Fluke is in the house. So they gave one to me. And it happened to be that otherwise, I don't know, maybe my cousin would have been like the scientists and I would have been like a CPA or something just crazy that that happened.

Vikram

Crazy thing, and it's such a great book to get at a certain point in your life. Surely, you're joking, Mr. Feynman, because it's all these stories about his, you know, adventures about lock picking. And you know, how he did all his various staff like physics in strip clubs, and it's so so crazy. It's amazing. Yeah,

Sidharth

I know.

Subu

In my case, right, I think the reason why this book, this specific book, you know, Vikram was the one who ended up getting it I don't remember how Vikram got that book, but he told me Look, you gotta read this, and that's how and we were in 12. And we were preparing for those IIT exams. Right. And I think that was very formative for me, simply because in 12, while I was preparing for IIT, along with Vikram, and a couple of other really good friends, I was not good at all, I didn't live up to the mark, right. And if my education in physics or chemistry and math was limited to the instruction we were receiving in class, and to those demoralizing exams, where I pretty much flunk each and every one of those, so if my, if that if my exposure to physics was just that right instruction in class plus exams, then I would have totally ditched science, and gone to economics or done some MA or something like that, right. And I would have concluded that it's not for me, but because of books like this, which invoke your natural curiosity, even though I was flunking these physics exams, in my mind, I imagined myself to be a it it primed, primed my mind to be curious, and start loving, you know, space exploration, and all of those other stuff that humanity is attempting. And it wasn't too hard for me to I didn't get I didn't get discouraged from these poor marks. And I went on to do some engineering and I like my life right now.

Sidharth

That's that's very self effacing went on to do some engineering pursued physics and physics like field in spite of not doing that speaks to more of your ability and sort of tuning to that research Well, research mentality, so to speak, right? But research mentality research has as a word, I think, has gotten such a, such an unfortunate negative connotation of people just being very, you know, just droning on and being very wearing, you know, plaid coats and having elbow patches, I think that's the very unfortunate image that it can construct. But but research is exciting, because you truly are at the edge of the unknown. I think one of I saw a very interesting graphical analogy. So when and this is like drawn on a paper, and I think I may have told you this in the past, but we'll just talk about this again. So human knowledge, as explained on a two dimensional canvas, okay. So when you're born, you are a dot, you know, nothing, your consciousness has just emerged, so to speak. And so you're just learning to speak, walk, eat, sleep, fart, poop, whatever. And then as you grow your, that circle expands and becomes a centimeter in diameter on a scale of say, all human knowledge being maybe the size of a parking lot, let's just say that your circle is maybe one centimeter and as you grow, that circle keeps widening, you know, you get a degree, it becomes really quite broad and you have a good understanding, you know, you're in your 20s, you you've gotten through a probably a very critical phase in life, as we know it right? To be able to get through your 20s is a parenting win, it's a genetic win, right? As far as the biology is concerned, you are primed to now pass on those genes in this social construct to the next generation and procreate and etc. Now, when you do something like like an engineering or a master's and you start really specializing, that circle starts becoming more egg shaped. So you're there is now directionality. And there's a vector to your knowledge on this canvas, okay. And when you get a PhD, or an advanced degree, or even a master's in that sense, it that oblong shape, and now it's actually expanded into an arrow that goes right to the very edge of this parking lot. Okay, which means you are now literally the world expert in this like, there is nobody else who has done what you have done and knows what you know about this very, very, very narrow, narrow topic. So that's why it went from being like a shape that had some area to it to being a lion to the very edge of human knowledge. And then to do something pathbreaking is to take that edge of the parking lot and they show this zoom into the kind of line in this next graphic in the window. And it creates a small bump, it extends the boundary by a millimeter, thereby broadening the entirety of human knowledge. Yes, isn't that a brilliant analogy I used to see this visually it was so amazing.

Vikram

So that little bump in human knowledge is is a plenty of a lifetime achievement. For most people even.

Sidharth

Yeah, to be able to do that means you know, and and like, you know, the framework of regret, as I like to call it is like, Oh, if and when, when, when the time comes for you to to move on, you know, maybe you maybe the bucket of regrets you have is is tiny. And so that framework of what your regret looks like when when it's time for you to die, as however morbid that may sound needs to be needs to be healthy. needs to be something that you know, you don't care about your overall career progression is that have you lived a good life? Have you done something good? Maybe, if you, you know, minimize the number of regrets you have. And if you've done all of this while doing pathbreaking work and making money and being well loved, man, that's that your heroes like to dream? Yeah. That's the dream. Yeah, exactly. And then, but then that's also not I think, truly an unachievable goal for folks like us who've had the backgrounds that we've had. And when when I speak to students back home in the same colleges that we went to, I tell them this, I think the first question is born out of insecurity when it comes from the audience. Oh, what about jobs? Sure, that is a valid question. But not to, not to sort of decry that that line of reasoning and thinking, you guys are going to have degrees from these universities, you are going to be able to find those jobs, you will work hard and you will survive. So trust yourself trust a little bit in the process. But that's the baseline, go beyond that, right? Push for your interest push forward you want to do, you will if you are insecure about it now, that's, that's healthy, and that's good. But it has to kind of you have to contain it and hold on to it and be inspired. And holding on to that inspiration and seeing it through and eventually achieving your goals. That is the accomplishment. I think the the preconceived notion of I will be successful from off the bat. That's it, you know, I'm going to graduate when we get the best job in the world. I have achieved my life goal. That doesn't happen. That's unrealistic. Right,

Vikram

Right. Yeah. I think a lot of times, you have to define for yourself what success means to you, like he's pushing the edge of human knowledge. one millimeter, is a major form of success most people can only dream about. You know, you can earn billions of dollars and not do that. But you know, there are people who, you know, the guy who invented the zipper didn't make a penny. So you know, I mean, but we all use it everywhere. And I do consider that a bump in one one mm of human, you know, knowledge, because it makes life so much better for all of us. But so, you know, to each person has to define what success means. And where inspiration comes from for that, right? I mean, like we all spoke about, surely you're joking, Mr. Feynman, such a good book, we all have common roots, that book, I suppose. Yep. But I'm pretty sure that we will come across people in our lives who have driven us towards these things. Who are some of the people that drove you know, you towards being inspired? In the field of biotechnology to get you where you are today?

Sidharth

Absolutely. That's a great sort of cornerstone or a pillar I think for for anyone who does any kind of research. There's either one or more individuals who, who've done that for sure that it's there's no single uninspired researcher, right. And or, or musician or whoever, I think. Yeah, exactly. So the for me, there was there were a few people, I think there was, there was a Mr. Vasu, who was my 11th and 12th, standard teacher. So this is high school here, who taught chemistry and he taught us physical chemistry. So this was some of the quantum stuff that was really exciting, like, electron and a box and stuff like that, it was just like, I could not get my head around the fuzzy logic of, you know, the wave particle duality, and things were bad, which is insane, and you just have to accept it. And then later on you fast forward 10 years later, when you listen to Feynman, in these YouTube videos, and he explains, you know, a rubberband and atoms and, and he says that it would be unfaithful, to explain something in the example, in the context or through an analogy of something else, because that would also reduce to what you are trying to explain now. So so he says that he'd be cheating you if he did that. And so he says, at some level are at that level, when you drill down to that subatomic level, you have to just accept the reality for what it is. And that to me hearing, you know, Mr. Vasu say that in like 2001 and then fast forwarding to like, I don't know, 2020 2021 say 20 years later, that is an immovable truth that just is and that there's some beauty in that I think, I think there's getting that validation when my brain is obviously changed and is less neuroplastic I would say is blowing. To me, that is a reality that just persists irrespective of your belief system, being able to see that you can engage in dialogue and argue the science and just bash your head against the wall say I don't understand it. Like I understand the premises I understand the building blocks but the concept doesn't materialize in my head and Mr. balsa wood would smile and and he knew what we were going through like that level of scientific empathy. Let's call it that was now when I look back was That's it that became a cornerstone. And let's rewind the clock a few years before that in middle school, middle to high school. So let's let's talk about seventh to 10th grade, there was another Mr. Hart, who taught us physics and math, what really made it work for me it was the clarity with which he taught things. So it was not just shoved down the throat, it was you were baited enough to, to walk down that path and let your brain wrap itself around physical concepts. And he did that through structure, right? It was not an unguided situation, it may have seemed very academic, and no, this has to be done by the syllabus. But he was irreverential enough to the academic syllabus of the school wherein he could get that done, you could do well in the exams. But you also had understanding which to us, you guys know, it was mind blowing, if you had something that really you could connect with.

Subu

And that's a good word. I mean, this, the scientific empathy is a nice way to frame it, because a lot of teachers, they just got to get these kids who are maybe say, in eighth grade past the finish line, that's the main duty, and then the finish line being that exam, okay, let's just do everything we can to just get the students to the finish line. But in that whole exercise, you've completely forgotten about this curious child who actually wants to know the science. And instead, you're just trying to, you know, sort of teach what's in the textbook or the syllabus. So in your case, you had like a couple of teachers in middle school, and then one in high school, you had those teachers who had the scientific empathy, to go beyond just the syllabus and satisfy the curiosity of the students.

Sidharth

But the thing that where it got really, I think, where it assaulted my senses was in graduate school, because now what is the idea of graduate school, right, like thinking going back to the analogy of pushing the boundary of human knowledge, you've come in with a certain set of skills and knowledge. So you're competent enough, not competent, competent enough to do a certain level of research to push the boundary of what you do and learn something, discover something. But you have to get very comfortable with being uncomfortable. To me, that was very difficult, I think, because over time, okay, you've got the scientific accomplishments, you've understood the science you've gotten through school, you've gone through life a little bit, you feel like you got this and you know, your curiosity is well satiated by by I think the internet as it expanded and bloomed in, you know, as we had access that eventually became Unlimited, right, being able to wiki, anything, Google anything meant that you felt like maybe you had access to that knowledge, or you had that knowledge at your fingertips, literally, right. But being faced with that ambiguity and uncertainty, when a molecule misbehaves or something, and there is no handbook for that. There is no Wikipedia page that tells you what that means getting comfortable with that, and, and defining a framework of reference of knowledge, and then pushing that really pushing that boundary that we talked about earlier. That was I think that was it, right? That was to me was my PhD, like just figuring that out and instilling that behavioral aspect into what I do and, and then doing that work took a solid six years, it was like, my brain just didn't get it, you'd get frustrated, why is this thing happening? I'm trying this, I've done everything, exactly the same, something failed. Well, there was an unknown parameter that you had no idea, it was an unknown, unknown to you at the time. So then you render it as a known unknown, and then you shine light on it, and then it becomes known. And then that is how that boundary gets pushed, because you're accumulating that knowledge of that experience into the model or whatever that you're trying to build. And it's not a model that you're trying to build for the sake of it, you really are trying to wrap around a reality that exists. Again, whether you believe it or not, whether you know it or not, it's there. Those molecules do what they do, but there's a reason for what they do. And so you have to wrap your brain around it. Yeah. So I think graduate school, obviously, was the most recent formative experience in that sense. And then, you know, the two teachers in middle school, and then at the very beginning, I think, as early as I can remember, it was my grandfather, so I actually dedicated my thesis to him. Oh, that's awesome. Yeah, so it actually says, I think my mom was not particularly happy about it, because my mom you know, okay, middle class, Indian enslaved beta padega types of situations and, and that, I think, no, of course it was you I have a chronology doing like a letter that is four pages long, but let's leave the blank sheet. The man's name, so about that. This is really crazy, right? So my grandparents lived in Chennai, and we had moved abroad and we would visit every summer and those and I never could understand why his name was Doctor Parameshwar. And I was like, first of all, he doesn't work. Secondly, he's not a doctor. He knows. That's the one thing. And so that curiosity I would ask truly, very innocently, but I would be shocked by you know, you know, my dad or mom I don't remember or it wasn't addressed enough to make me happy,

Vikram

right? Just be like, be disrespectful

Sidharth

Oh no, not even that I think they were okay with the but he would catch me he was I was the, the first one on the you know, whatever, dad's side of England so he was very attached to me and I was very attached to him. And he would say, he would just smile and laugh, it tickled him to know and that I was blown away by the fact that like, here's like a six year old asking why are you a doctor but not. And then we would spend essentially the whole day together, you know, running around playing with the neighbor's kids, whatever. And then in the afternoons, I had a routine, you know, there was lunch, and I was very, very fussy eater. So after Mega lunch, we would get, I would watch The Jungle Book, the animated Disney cartoon. And so you know, song and dance, maybe that's the artistic influence. I don't know, we'll decide. We'll we'll cut that up later. And then he had a collection of hardbound books that either my dad had, you know, brought and, and saved over time. And they were on like sci fi. So sci fi from the context of 1989 1990. So this is completely outdated sci fi by today's standards. Okay. But can you imagine what that does to a child's imagination you open you see spaceships that don't exist, renderings of planets that don't exist. theories on propulsion that are still vaporware as far as today's technology is concerned. So at that age of like, I don't know, maybe 6789, I was exposed to the concept of solar propulsion, the Daedelus project. All this rendered by him by his words, I can't read that stuff. I don't know what it means. And he would just laugh and he was like, Ah, let's see what these aliens are doing today. And you know, and we would look through this, this sci fi book, Think about that. What a crazy fit. My parents had no clue this was happening, right? They are busy with work, whatever. And one random afternoon, grandfather's sits down, this becomes a routine. We're opening these random books. And I was not allowed to touch these books. They were on a shelf somewhere. So maybe the engineering mindset came from there. When everyone kind of goes to sleep and the cartoon is finished. I would sneak into his room and quietly try and pull that wooden chair to climb it to climb the table to reach the shelf to pull the book. Many scoldings later, the book is in my lap. Achievement unlocked. I think that was it for me. And no, I remember this one thing. So he read by here as of I know this guy, Richard Feynman talks about opening up a radio, right? Oh, yeah, yes chapter. I did that to a to a torch. Okay, it was, I don't know, you remember, we used to get those flashlights that were orange plastic with the black switch and the black rim signature like this is just like your run of the mill 80s flashlight. It ran those big ever ready batteries. And one day, I don't know, I was inspired. Let's call it that. Okay, I take apart the whole thing. I figured out basic electric circuitry on vacation during power cuts you need when you need the flashlight. So and I think probably where I think this I was validated. I think for the first time that I remember why my parents were having the scientific curiosity was i i had i figured out which screwdriver to use to open up one of the two screws or whatever, I'm taking up the whole thing of the whole thing. And remember, there was a spring that will also hold the batteries in place. And I knew how to take out the spring right? So small fingers very dexterous. So I think you know Vicki you'll relate to you know your kids having that ability with that with those fingers. It's really an advantage being that small. I think so. I told him no, no, trust me, I will put it back together and it will work on the first try you know raising the stake.

Vikram

Challenge accepted

Sidharth

Challenge accepted whatever they let me be they will probably like what we have other fish to fry. Let's just let this kid do all those things. Lo and behold, it worked. Put the whole thing back. We know what feels like trivial thing like now I mean can you imagine not knowing anything about positive negative electricity making sure the polarity is correct making sure the connection with the bulb is correct figuring out where on the screw type the is the negative and the positive on the bottom empirically tested no idea right and no fundamental concept of electricity as we know it today. So that was I think and then and then being validated by the parents to say okay, good job, whatever, you know, yeah, there's gonna be a power cut so we need it. Yeah. Done. That's it. Yeah.

Vikram

The next time that low power the power went out like yes, this light is on because of me and my science because

Sidharth

of the soon to be you know, doctors. 30 years later.

Vikram

That's nice. You So is Jungle Book where you got all your grooves from your rhythm bones? Because

Sidharth

Rhythm bones huh?

Vikram

why I have to say the bones to people who don't know you already is because you're a damn fine drummer as a Subu. So I'm feeling left out in this conversation because there is a distinct drummer who once told me like, Dude, you play all this guitar, but you really lack a sense of rhythm. I'm like, You're right. You're absolutely right. No offense taken. Agree.

Subu

The thing is when SidMo and I first met SidMo, right, in the in our college Indian Band, he wasn't even the drummer. He was hired to play the keyboards

Sidharth

and the bass. Yeah.

Vikram

He's a man of many faces.

Sidharth

Oh, no, no, I was I was belligerent, I think with the whole band thing, because I had to follow in the wake of the great blacker than for first time listeners here, you know, Vicki, and Subu are powerhouse musicians who achieved a lot. So it's, it's only unfortunate that you guys are not still a part of a touring band, I will say, and I say that I am a scientist because I'm a failed musician. So I'm the other way around. But yeah, I think the idea of grooves more so the Jungle Book was my dad. So my we had, and we still have to this day, the original vinyl records from you know, like, I think, going back to the 70s, of whether it was AC DC, or Pink Floyd or dire straits. And for example, like my dad, I don't know whether he was he is into music, and but he never, it never really got into metal it was he was a little more eclectic in his tastes. And it was just a thing that every weekend in the morning, we would play music and I knew about dire straits, I think from the time when I was born, which is kind of crazy, because that really shaped the you know, my aesthetics, right. And so some very little classical a lot, a lot of rock a lot of what we call classic rock now, and I remember being fascinated by thunderstruck. AC DC And it's like, oh, you know, this is loud, screaming metal and we will play it and I'll be like, what is but that is dad playing music early on. Again, very eclectic, and it kind of died out. I think I pursued music in Russia. So we lived in Russia for a little while. And I was taught the piano by a very strict Russian lady teacher who would whack my knuckles if I played, you know,

Subu

Stereotypical.

Sidharth

I'm not joking. And then you see movie like whiplash? And I was like, Hey, that's

Vikram

not that's my life. My life.

Subu

How old were you?

Sidharth

I think I was close to maybe eight? Nine? Yeah,

Subu

Not that young

Sidharth

eight, nine. So and it was traumatic because she insisted on keeping like the door close. So it was just, you know, the, the sort of overall strong pedigree Russian pedagogical, I don't influence whether if that if that was it or not. And it was traumatizing. I think I I began to hate playing the keyboard it was, it was really something I didn't want to do. I would I had a ear from using in the sense that I could, you know, listen to tunes and replicate it. And I would do that for the longest time. So 1015 years go by, I'm in high school, and you know, yeah, okay, I'll get back to the classes here. And they're very broken, and then call it happens. And then there's this expansion lens explosion of freedom to do what you want, in some sense. And I see this drumkit. And then that's where a lot of the magic for Black Earth also happened. Right. And I remember Subu was like, and I we still, I still call him my drummer brother from another mother. But like, I was very much in an exploratory mindset. mindspace, right. And I remember sitting behind it just being fascinated by it before I could even play a four by fours. It took me months to get a four by I just could not believe how awkward the basic, you know, four - four timing is where your dominant hand does 1234 your bass drum, your kick goes on one. Yes, there goes on three, and it should feel like a beat instead of just you're just clunky on it, you know.

Subu

And you know, that's the beauty of is putting a bunch of kids who don't know, drumming and keep putting around, we didn't have any teachers, right? And we would just listen to the same kind of music Metallica, and we would just show up the next day to college, and one of us would play and then we would say, you know, I think this is how it goes. I think I figured it out. So there's no teacher no one. And there's no real internet in the way. I mean, there's no real there was no YouTube, so no video, so it's pretty much you had a toy and we just figure it out ourselves. And that was our pastime essentially.

Sidharth

More so you than me. I think I was I was lacking in that sense. But I think Subu was formative in that sense. I think you were because not only the You teach me a few beats here and there. But first of all the positive energy, if you know people ever meet Subu in real life, they don't know what I'm talking about, right? The positive energy is one thing, the lack of barriers was another. And then you started gifting me stuff. I mean, you had that extra pair of the Lars Ulrich signature sticks, which I was like, what? Okay. And then you had that that double pedal kick, double pedal, which I still have, by the way, it's in the cupboard right there. Can you imagine how old is that it must be at least 25 years old now

Subu

Easily because I got it. It was a hand me down from another accomplished drummer. Her name was Durga. And I don't really don't know what she's doing right now. But it was a hand me down from another drummer. And then it came to you.

Sidharth

I think you put the iron Cobra beaters on it, and you sold it to me for a nominal price. Yeah. Which by the way, I had to like crowd a crowd fund. But we made all the money back because we started winning competition. So I ended up paying all the battles back for that. I was just saying that, that that the ultimate validation, I think in terms of drumming skills, and I think the high point for me, we've just before you guys left was you were late for practice or something. And we just had you guys were doing dream theaters, YTSE Jam at the time. And I remember distinctly the one part where you decided to chop 1 part of the bar. The band could just could not coordinate it. And I remember it was a group call you guys made and decided to cut it. And it irritated me to no end that you guys did that. So I made sure to play through it with you when you were late. But it was like Yeah, well, this guy. Yeah, replacement

Subu

almost took my job.

Sidharth

Oh, my God. So

Subu

wait a minute, I want to I want to go back to you casually just mentioned Russia. So what's the story there? How did you end up growing up there? Were you born there? I mean, if you if you're a that was probably just a freshly broken up Soviet Union at that point.

Sidharth

Yeah, so this is crazy. So my parents actually studied in Ukraine. And as life would have it, you know, they worked for companies that then eventually had like offices in in Russia, and they were under sort of retail and whatnot. So, so I spent five and a half years there 1989, to about 1995, around six years. And we are we saw the breakup of the USSR, crazily, we lived on one of the main streets when there was a siege on the White House. And so here, you know, it's like, again, maybe six, seven years old. And my father was traveling at the time. So he was working for like an Oil Corporation. And so he was actually close to the North Pole, like really far away, buried in snow. And then there's the seige happening on the White House and the tanks rolled down our street. Okay, So picture this. So it's like the middle of the night and earthquake is like sort of experience and because the tracks really are very, very loud, and they do a lot of vibrations of the tanks on the road. And it's dark is dimly lit. And so my mom's obviously panicking, and you're like, What is going on? So look outside the window, and I see the tanks, and being the age I am I'm like, oh my god, this is so cool.

Subu

That' the best thing that could happen to you.

Sidharth

Can you imagine? Wow, no concept of war, politics, nothing. And my mom on the other hand, of course was panicked so we had to take shelter in the far side of the house, kind of hoping and praying that nothing and having desperately calling people wondering what's going to happen and and that's it and I just remember being just fascinated as a kid by that just can you so that descendants and that kind of oil and water is sort of a mix of a child's, I don't know, innocence, if you want to call it that. Overlaid in a deeply, deeply dark part of human history, you know, the fallout USSR and where things are today. It's just like, I can't believe that I was a part of those those experiences. It's blow it blows my mind. It's pretty crazy.

Vikram

Going back to your music or, you know, endeavors in life, you have a pretty awesome project with your wife called What's the scene about? Fill us in?

Sidharth

So, as you know, and as I've said a few times, I am a scientist because I'm a failed musician. So I moved to Mysore for my first job as an IT engineer. I land up in Mysore. I frequent Bangalore a lot. I mean, a bunch of other people beasts. I didn't start on the band, but I was playing with another man with another friend. And we were doing gigs locally in Mysore and Bangalore and whatnot. And eventually I landed up in Bangalore because I had malaria. mosquito bit me and I literally died two times. Oh my god, and because of that, I met my wife. So in our context, Cupid was a mosquito. Okay, literally, like it's the only reason we met because I had and in Banglore she was, I think, fascinated by how fascinated I was with music and really was into the scene and was trying to do all this. And she couldn't believe that it was this hard. And we know there are all these challenges, access to equipment, access to gigs, there's no promotional framework, nothing. There's just no real representation. And she is an accomplished Well, now she's a writer, director, actor, playwright and super, super sort of artistic has a super artistic output. But at the time, she was a very, very avid writer, she used to blog like crazy, and she won an All India award and whatnot for like, just being an incredible writer. So that's a whole new, whole new thing podcast you'll have to do with her. But eloquence, yeah, her eloquence, I think, really spilled over into the scene. And so by that, I mean, she wanted to get to the bottom of this and figure out what it was that really makes the scene tick. So we started off this sort of initiative as a newsletter within the company, right? We had a weekly release, we said, We got to get a bunch of enthusiastic people to talk about the scene. What does that mean? It means we have a gear enthusiast to talk about gear, what's the cutting edge, maybe make it advertised bubble and, you know, try and sink our teeth into some collaborations with with import export for, you know, the music stores and whatnot. Or we can also talk about the latest albums that our favorite bands are releasing. And we could talk about bands locally, and kind of put them on the same page. I mean, the minute very literally, and say, and so do these reviews, and we wanted them to be objective. We wanted them to be academic and really, you know, someone has to we have to we wanted to show that we were putting a lot of thought into the output of your average Indian musician and do the same for like an Opeth or Dream Theater. And that's the kind that was our approach to kind of level the playing field. Okay, from a listeners and consumers stem. Yeah, so that was the newsletter now it's suddenly it snowballed. It really grew into something like if we were late by a few hours, not even a day, right? If we did not release it at 3pm on a Friday, yeah, people would email us why is this? What's going on guys? We were really excited about this. I was like, wow, that is interesting. This we did not think that this would happen. So what started off as I think as a journalistic interest project for for Priyanka, primarily. And for me as Yeah, you know, I have the scene experience, I'll be the cool consultant or whatever, we sort of started this together and it really really picked up I mean by we saw that it would be limited by being restricted to a bulletin board in the company. And we decided to go outside we were like we're gonna go Facebook the internet is exploding. There's all these social media platforms This is crazy. And we did we and then we started you know, in Bangalore, let's go attend a gig. Let's write about that gig. Let's photograph it, let's put it up on the website, put it up on the social media and see if people are interested in this. It was yeah, of course, oh my god. So we had a logo built there was a brand built around it, we started the company, you know, just like like like a small company at the time, right whatever, it's like a limited liability partnership or a sole proprietorship or whatever. And we started documenting the scene. Now, word got around, we started getting all access passes to all these venues. And at that time, it was burgeoning. It was really something that was you know, you had all these like Kairos and B flats and it was like amazing to see the scene in Bangalore. Why district to Bangalore we are operating on a digital platform we can start you know if we communicate the value system and the and the model and have a hub and spokes approach across different cities we can have different teams so at one point we were operating out of 10 cities, that is ridiculous.

Subu

yeah that's pretty that's real nice

Sidharth

yeah hailing it so we became known as the the mirror of the scene if you want to call it that, and you know people like Vir Das in US endorsed us 10 years ago. Who is is crazy. He probably doesn't remember us today. Vir Das is a fantastic Indian comedian who is based out of India and has an Indian flavour to his comedy so to speak, but has made it internationally is and I think he was in this kind of music I hope he still is and here endorsed us way back then saying yeah, what's the scene is doing legit work. We have photographers and reviewers on all these cities, we have editors who coordinate all of this and you know, get make sure that the content is delivered on time and delivered to the right place. And you know, we have fact checking and error checking and it was polished content that was out there. Yeah, it's amazing.

Vikram

So it's what's the scene selective?

Sidharth

It is. So as life would have it, you know, my wife and I we were still just only dating friends. And then dating at the time. And so it was a tumultuous ride through life. And you know, I moved abroad and it was difficult to kind of do things remotely and then eventually moved here. And so we kind of went through a bit of a hiatus, and I think it was for good measure, because the scene changed on us, I mean, literally changed. For the worse, I would start with think, you know, there's all these restrictions that exist right now, the way music is consumed in India has also changed all these venues that had sort of supported the independent music scene, I think evolved or devolved, or closed down. And so we went through, I would say, a bit of an identity, I wouldn't say crisis, but change. And we kind of revamped the model in the last three, four years. And we went, again, virtual, but we have a much more focused approach where we have aggregated data, aggregated reviews, you know, the social media platform focus has changed. But we're still essentially trying to do the same thing, which I do. Now, I'm not just sort of restricted to reviews and documenting the scene, we want to connect and allow musicians to kind of leverage our platform to to promote and collaborate and do stuff like that and do and do things in a way that is the representative of a well oiled framework or structure, the scene has no shape, so to speak, if you ask an average Indian Band, or band member, what does it take to succeed, you're going to get a variety of answers. Now, while diversity of thought and execution is great, there isn't really any one clear path. So we're hoping that by doing what we do, that we can help define that and crystallize that, and really tap into the same I mean, we are a country of over a billion, there are brilliant musicians in our country. And so there's all this talent that just does not get access, no discipline while we attend. Yeah, so so we want exactly so we want to be a discovery engine without being an online radio. So you see, like it there's semantics and sort of nuances there that I think we're continuously figuring out and learning but what's the scene is just that it is we want to be those seem, this is the scene, it's our scene, we are the scene. In fact, all the members, you know, it almost became like a I wouldn't say a cult, but it was a great sense of solidarity. When we were the broad older model. It was we are the same WTS we are the scene, what's the scene so there was a great sense of enthusiasm. People did it out of passion, it we didn't make any money off this. In fact, it sucks. Like, you know, we spent a lot of money on making sure the website was was built from scratch, we paid designers or we just went through a revamp process. But the really nice thing is that we the support that we've gotten from consumers, other pillars in the industry, and the bands and the musicians themselves. You can't plan for the stuff. I mean, it's amazing,

Vikram

unbelievable stuff. What you do when you start a project, we will do this, I know that the Instagram handle is WTS, India, for watch the scene India. And for any additional links, it will be in the show notes. So you should definitely check out all the What's The Scene stuff, there's some really good music and stuff on the Indian music scene that is awesome. It's good for the bands. It's good for diversity. So we'll definitely have links to that stuff.

Subu

So you know, SidMo So thanks for joining us on this journey. I mean, educating us about the science that you work on the talking to us about your personal life. And it was obviously always fun digging into our past and making a record of it. I think that's like, that's one of the main reasons why we even started this podcast in a in a way it was for a selfish reason of making a record of all the fun that we have had in our past. Right. And I think that's exactly what we did today. So thanks for that. And I hope listeners are also going to enjoy this episode we'll have links to all of the stuff that we spoke about in the part one of the science section and also all the Indian music that we spoke about. And that's it for this episode. Thank you!

13: Dr. Sidharth Mohan: Discovering New Drugs