What is electronic design? Okay, all of us have been studying design all these days and teaching design all these days. We must ask ourselves the question, what do you mean by electronic design? What is this activity called electronic design? Okay, there is no book answer or correct answer for this that you write, you know like they say, what is physics? Physics is the science of matter, energy and their interactions, correct 2 marks, none of this. Okay. This is, we have to apply our minds and think what is design? What is electronic design? And as I said there is no only correct answer whatever you feel, this is my answer to this question. Okay, so my answer is this that given specifications, there is no electronic design if you are not given accurate specifications, specifications must be complete. So given specifications, we want to develop a circuit and what is a circuit? Like connecting known electronic devices, okay, such that the circuit meets the given specifications, right. That means what are specifications? Specifications are a description of the behavior of the circuit, okay. So suppose you want somebody to design an 8085, so you will say design a circuit which will read instructions like this clock circuit, it will put ALI, high ALI, low etc., you are describing behavior, okay and then you will say when the instruction or this bit pattern is presented, it will move the contents of A register to B register etc., etc., right. So these specifications are essentially description of the desired behavior of the circuit. That is the starting point. What is the end point? I will say you connect this register to this and this register to this and connect this transistor here and put clock here etc., etc. Then this circuit will have the behavior that you want, correct? Then that has been done, the design has been completed, okay. So this is my definition of what electronic design is. Now there are many keywords here which we now have to understand. What are the keywords? First of all, we are saying known electronic devices, right. Where do you stop the design? You do not have to go down to a single transistor. For example, you are doing a board design. Now I need not go beyond a D flip flop. I will say, okay, use a 7474 here or use a 4013 here. That is where my design ends, right. So where does the design end? Where I reach an interconnection of devices which are already known, right. Now what is already known is a variable that depends on the context. For example, suppose I am doing VLSI design and my company already has a copy of the ARM processor. Then if I am designing an MP3 player, then the ARM processor is a known component. I do not really have to design the ARM processor. I just put it down and that is called IP core-based design. So essentially, if known components become more and more complex, then I can design more and more complex circuits because my design stops when I reach a known component and an entire microprocessor is now for me a known component. Essentially you are doing board design on a chip, right. This is what you would have done on a board design. You would look at it and say I think an 8051 will do it. Therefore when does your board design stop? You do not have to design the innards of the 8051. You say I will put an 8051 here and therefore 8051 is a known component, okay. So once you reach known components and known components only, there is nothing unknown. Then your design is finished. So that is one part there. The other part is specifications. You might make the charge that I have defined something in terms of something else which is as are known, okay. So what are specifications? My definition is not complete unless I have told you what are known devices and what are specifications, right. So known devices, we have now had a discussion, we have a good feeling of what known devices are. We should now know what we mean by specifications, right. So the specifications refer to the description of the desired behaviour of this, right. Specifications must be complete, right. So you must have complete description unless this is complete. I may make something which has those specified behaviour but it may do other things which are quite bad and you may not be acceptable to you. Then my design is not complete, okay. So electronic design is the activity of connecting known devices to produce acceptable behaviour and the starting point is the description of the behaviour, okay. Hardware description languages are designed to meet this requirement, okay. Therefore I need to mix two things which appear very incompatible with each other. One is a behaviour description and the other is a simulation of a known circuit, right. Circuits are described differently, behaviour is described differently. Circuits are diagrams, behaviour is some algorithm, some logic. Now how can I merge these two and come up with something? That is what hardware description languages do, okay. That is the purpose of hardware description languages. So electronic design is the process of converting a behavioural description. What is the behavioural description? Which says what happens when? Kind of description. To a structural description. What is the structural description? What is connected to what, right? And when the behaviour of these two is the same, then my design is complete, okay. That is electronic design. Now what is the problem with this? Now we have understood it. Now electronic design is all complete, we can all go home. The problem is that designs have become very complex, right. So every year new technology comes. In every 18 months, we can put twice the number of devices on a chip and our market expects devices which are twice as complex every 18 months from us. And if we do not meet that expectation, we will be outdated, okay. So new microprocessors come, they work at higher clocks, they have new memories come, they have higher capacity and so on. And if that expectation is not met, then no design is required, okay. You will be out of business. So therefore this complexity is now becoming very huge. The problem is unfortunately that the human mind is not improving at the same rate as the technology, okay and unfortunately it is the human mind which is required to translate this behaviour to circuit. So how do I manage this? That is the major problem. So the major problem of modern design is conquest over complexity, okay. Now one obvious way of conquering complexity is divide and rule, okay. Rather than talking in the air, let us take a specific example. Let us say that somebody was to come and tell us design in 1885. We all know 1885. It is not a big deal, it uses 15,000 transistors. That is nothing compared to modern designs, okay. A Pentium uses something like 10 to the power 8 transistors. About 100 million transistors, tens of millions or 100 million transistors. I am not asking for much, I am asking for 15,000 transistors. So the expectation will be by this afternoon design the 1885 and that should be done. When the original designers of 1885 came together, they took about 15 man years, okay. But at that time all these advanced tools were not there, right. So now with the modern tools, just do it, okay, don't take an afternoon, take a week. We are suddenly afraid, no, no, it may be a very simple circuit. But I can't up and design in 1885 like that. It's too complex, okay. It may be only 13,000 transistors for you, but 13,000 transistors is too many for me. That is because the human mind has a limitation of handling a certain number of objects at a given time. And typically, there may be geniuses. But ordinary human minds, they handle tens of devices at a time, okay, not more than that. If you have more than 10 objects to handle, then it becomes just too complex for you, okay. An example is a game of chess. I don't know how many of you play. So in a game of chess initially, there are 32 pieces there. That's too many for you to comprehend and to keep track of. So therefore initial moves are all stylized. Somebody has worked it out painfully, etc. So you pick one out of 10 or 20 choices, right. So you want 10 or 20 choices, no more. You pick one out of them and then follow it. Till some Mardhar occurs, the number of pieces is now reduced to about say 20. Now you can start applying your mind, okay. So this is the trick. At a given time, handle no more than 10 to 20, right. So this is called hierarchical design. When a complicated design is given to you, break it up into no more than 10 to 20 parts. So, okay, you say 1885, you describe the behavior, all right. It has an ALU, okay, not very complicated. It has a register file. You told me that these are the registers, I need a register file. It has a certain way of reading from the bus, so it has a bus interface unit, okay. It has an interface controller, all right. Now it looks somewhat manageable, okay. So I'll have a register file, I'll have an ALU, I'll have an instruction decoder, I'll have a bus interface unit and now I have to figure out how to connect them, okay. If I do all of this, then I have the design of 1885, right. So already I'll start feeling better that this task can be managed. So the way to conquer complexity is by hierarchy, okay and I'll go top down. Next I say, all right, what is this, all right. I don't worry about other things. My team has been asked to design the ALU. So what does this ALU do? Not much. Just add, subtract, not even multiply, okay. Just add, subtract some logic operations and and or XOR, okay. That I know how to do. I know how to make an adder. I know how to convert an adder to a subtractor. I know how to do and and or XOR. Not a big deal. Not is also okay. I know how to rotate. Then what else is there? Put all these things together, your ALU is ready, okay. Then somebody says, what about the register file? Not a big deal. I'll take eight D flip flops and connect a gate at the input, gate at the output, connect it to the bus. If this register is to be loaded, then the input should be turned on. If this register is the source, then the output should be turned on. So I'll decode the address of this register, okay. And there are eight of these possibles. So I'll have a three-bit code and depending on whether it is input or output, one of eight switches need to be turned on, register file is not a big deal, okay. Similarly decode or similarly other things. You start doing it at that level. Things start becoming manageable. And that is the only way that we can do complicated design. Therefore, a hardware description language should be one which allows you to work like this, okay. You can't work at a circuitry level, right. Whenever things become complex, then graphical descriptions become too difficult, okay. Remember, how did all these languages, how did scripts evolve? Script evolves like a picture, okay. You drew some picture or some concept and so on that this is a man or this is sky or whatever. But soon it turns out that the complexity is so high that you can't use pictorial representations, okay. Similarly, a circuit diagram is a pictorial representation of the actual circuit. Pictorial descriptions won't do. We need text descriptions. When things become complicated, things become abstract, then you need a language and not pictures. And as a result, the same thing is true of electronic design that you go from circuit diagrams to hardware description languages when the complexity becomes too much. So that is what the hardware description language solves. So these are the things. You need hierarchical design. We have already established that with the ATT5 example. You need modular architecture. So that once you have said ALU, it should not depend, ALU's design of ALU should not change if somebody changes the registered file design, right. So that different people can take on different things and quickly do it. So this design should be modular. It should be text-based rather than pictorial. And you should be able to reuse existing resources, okay. So suppose a register has been made, then tomorrow you want to design an AT51. You should not be required to start from scratch, okay. You are not using the entire AT85 design but your company has been very successful with an AT85.  Now you want to come out with AT51, it should not be required to start from scratch, okay. Register files are similar, ALU has similar functions, some additional functions. You should be able to compose it out of the same building bricks, okay. So therefore each of these entity of design should be reusable, okay. These are the requirements of a hardware description book and we take a page out of the software designer's book because this is what we have learned from software people. Software people have managed high complexity for a long time and this is what they do. They use libraries. So you can reuse functions in different these things. You do hierarchical design. You have programs calling subroutines, calling other subroutines and so on. You do hierarchical design. You have modular architecture. The change of one subroutine should not change another function and so on. So all this is what we learned from software designers. Therefore our description should resemble software. Unfortunately this is taken too far. Many people teach hardware description language as if it is a programming language and that way you are invulking disaster, okay. Because it is anything but. There is something to be learned from software.  It is very different from a software and how is it different is the next thing that we should, okay. We come to this that hardware is actually different from software. Fine we will learn from software but that is not where we stop. There are other things. How is hardware different? Why is hardware description different? First hardware components are concurrent. Software things happen one after the other. Only one thing is happening at a time. Whereas in hardware different blocks are operating all at the same time and interacting with each other, okay. So to describe the behavior of something which is so parallel, requires constructs which are inherently different from the constructs which are required to describe a sequential algorithm, okay. So that is concurrency. Second, the behavior, the timing is a indivisible part of hardware description. Whereas timing is not, you know, you may have a slow computer. You may have a fast computer. The correctness of the program is determined by the result that it produces, okay. May produce it in more time, may produce it in less time, okay. But the programming language is not concerned about it. The hardware on which this language will run, that is separate. But the programming language need not be concerned with time, it never is. On the other hand, hardware crucially depends on time. Which pulse will come before the other can totally alter the behavior of your hardware. And therefore, it is not software. It is different and these things must be incorporated into the very construction of the language. It should not be like a software program. And those who teach hardware description languages like a programming language are producing students who will be unemployable really because they don't understand. They will often commit mistakes and everybody commits mistakes. But the problem is that they will commit mistakes and never realize the kind of mistakes that they are committing. I will take perhaps a few examples of the kind of mistakes which are not understood by those people who teach hardware description like programming languages, okay. So these hardware description languages need to have the ability to behave. Something may be described behaviorally. Let's say that the register file has already been described structurally. I will reduce it down to transistors. But my instruction decoder is described only behaviorally. Now whether the whole thing is working or not, I need to put it all together, right. So therefore, I need to have a language which can put together elements where some are behavioral, some are structural and overall mixed, right. That capability must be inherent and that's why hardware description languages always have behavioral and structural parts. I already said that timing is integral to hardware description, right. This is, by the way, there is one kind of software where timing is important. That's called real-time software, okay. In fact, if you read any book on real-time software, they say it is like hardware because timing is important, okay. For example, suppose you have a microprocessor in the anti-locking brakes of a system, of a car, right. Now this program determines the force to be applied to the brake pads given the current situations. Now it may arrive at the correct value, but not arrive at the correct value fast enough and the car may crash. It doesn't help you very much that it finally found out how much pressure should have been applied to the pads if by that time the car has crashed. So when you have a system like that, then that's called a real-time system, okay. And hardware is always a real-time system. Hardware design is always a real-time system. So we must understand time, okay, or delay. Let's see how do you describe delays? So here is the simplest possible example of a delay. You have a pulse at the input and the output should be a delayed version of this pulse by 30 microseconds. Now essentially this is an example in VHDL, but don't worry about the actual structure almost in any language. This is what happens. I am saying assigned to out the same waveform as in, but with a delay of 30 microseconds, right. What do we understand from this in hardware terms? If we stopped it here, then we will really not understand hardware. Then we would be treating it like a programming language, okay. So let us see. Suppose I open out this black box into its hardware constituents. And this is one possible way that I can put down the hardware, okay. Let's say there is this RC delay with a delay of 30 microseconds and a buffer, right. This RC delay will lose the TTL logics or whatever logic levels because of the RC, right. The output will not be sharp anymore. Therefore I need to put a buffer, okay. And RC will provide the delay. Now this will meet my specifications of this circuit, right. The buffer is non-inverting. So output is the same as the input with a delay of 30 microseconds or is it? Let's look at it. Let's say that I give it two pulses. The first pulse is much narrower than 30 microseconds. The second pulse is much wider than 30 microseconds, okay. Now let's see what happens. This is an integrator. So when this pulse goes high, it starts integrating up. However, before it has gone much further, this pulse goes low again, right. As a result, this point being low, this capacitor now starts discharging through this, right. So as a result, in 30 microseconds time, this will go up a little bit but since the pulse did not last a full 30 microseconds, it will come down, okay. And therefore this guy will treat it as if it has been zero throughout, right. So at the output, this little wiggle will actually disappear. On the other hand, if the pulse is much wider, then with a 30 microsecond time constant, this will charge up, stay there and when the pulse comes down, it will start discharging. Now because this acts as a discriminator of a low level and a high level, the output will be a 9 square pulse somewhere depending on where the discrimination level is. And now you will get a delayed pulse, pulse delayed by 30 microseconds, okay. So the pulse much narrower than the delay has vanished. Now this is how hardware behaves. Should we say that all delays are like this? Answer is no. Let us look at opening out that black box in a different way, okay. This was my black box, delay of 30 microseconds. Now instead of implementing my delay by that RC circuit, I implement it like this, okay. I give it to an LED here. I have a long optical fiber whose length corresponds to a 30 microsecond delay and then I convert this light back to digital level at the output, right. This black box also should be described the same way. That output equal to in after 30 microseconds, correct. Now let us see what is the behavior of this black box to the same input, okay. Now of course this pulse will also appear at the output because there will be a flash of light which is brief. This flash of light will eventually appear 30 microseconds later at the other end of the fiber and this longer flash will also appear as a longer flash at the end of the optical fiber. If that did not happen, you would never be able to send messages from here to Delhi. The delay is so long, hundreds of kilometers, the delay is really long there, right. And yet pulses which last nanoseconds are sent. So in this case, the narrow pulse does not vanish, okay. So therefore there are delays and there are delays. We must describe these delays differently, okay. So hardware is a strange object, should not be treated. If you just thought of it like software, you would have said that assign to this variable this value after so much time, okay but that would not do. In fact the thing has changed qualitatively because of the way that the delay is implemented, okay. You must understand time very properly and in fact these delays are called inertial delay and transport delay accordingly and you must specify it the same. Now very often you are not worried about timing at the start of the design, okay. You just want to know the correctness of your logic. So then if you do not specify a delay, what should the hardware take it, okay. Now by default the delay is taken as 0, okay the default value of delay if you do not specify is taken as 0 but that cannot be physically right, okay because there may be race conditions and if pulses arrive in different order that will cause problems. So what the hardware description language actually does is that it takes 0 delay to be a delta delay, okay. The assumption is no hardware can update its output in 0 time, okay there is a definite amount of time of delay from input to output. If you are not worried about it then I agree to play along with you and I will call it 0 delay but as far as my internal operations are concerned I will define a special unit of time called a delta delay, okay. So this delta delay is something whose value is reported to be 0 when the final timings are reported but as far as my internal operation is concerned I will assume that all things which are a result of something that the delay between cause and effect is always a delta delay at least. So if you say 0 I will put it as a delta delay and keep detailed accounting that the time is 30 microseconds and 5 deltas, okay and then when you ask when did this happen I will say it happened at 30 microseconds at that time I will put delta equal to 0 but for internal operation I know that something which happened at 30 microseconds plus 2 delta occurred much earlier than 30 microseconds plus 5 delta and there was a 3 delta time in between where one was true and the other was false, okay and if that has specific consequences then I should take into account those kinds of consequences. For example the one input is coming to the clock the other is going to reset. Did clock come first or did the reset come first, okay did clock go away first did reset go away first correspondingly the output could be a 1 or 0, okay the overall delay can be 0 but the output result itself will become wrong if I do not take this scheduling into account therefore I must have the concept of delta delay which is special to hardware description languages. This by the way is the source of much headache to all the students who learn HDL as if it is a programming language, okay because what happens is that you say assign 0 to A and if A equal to 0 to this now because they are so used to software they feel that A equal to 0 is correct indeed it is not what when you say assign 0 to A A has not become 0 a 0 is scheduled to be given to A in the future after a delta time if you test the value of A before that delta has occurred then you will find that it is actually non-zero, okay so these students just like used to be used to software writing say why is it misbehaving A I am saying assign 0 to A and if A equal to 0 to this and it is not doing it, okay that is because you have learnt hardware description like software whereas it is not software you have to take that into account and test the value of A only after the assignment is complete only in the next delta. So therefore the language has this responsibility now to remember to assign values in the future when the assignment is encountered it is not immediately assigned, right so it keeps what is called a transaction register a transaction queue what is a transaction it is like saying that I place an order but the equipment arrives later, okay so that is a transaction somebody has to keep track that this order has been placed this money has been paid whether that thing has finally arrived or not, okay so this whole thing constitutes a transaction now the hardware description language keeps a time ordered list of transactions, okay and things are executed in the order of their time so for example if you have let us say AND gate one input is 1 the other input is 0 now at some time the zero input goes to 1 then the output should become 1 AND gate the other input is 1 this has gone from 0 to 1 output should go from 0 to 1 you do not directly assign a 1 to the output you have to take down a note that after a gate delay make the output equal to 1 so during that gate delay many other things may be happening all around the circuit you have to do those first and then make the output of this AND gate 1 otherwise you will get wrong results right so therefore you need to have the capability of storing these pending events all the time okay and that is called a transaction a list of transactions and you must understand it in terms of transactions and assignment is not immediate it is not like assignment to a variable it is a transaction okay so it is like a little black diary you have so when somebody says give 10 bucks so you say and you write down in a diary give 10 bucks at this time but your wallet still contains those 10 bucks when the time comes then you will give that 10 bucks so if somebody says if your wallet has more than this much amount do this or less than this much amount then do this then if it is treated like a programming language you may come to the wrong result okay so that that that is the difference between programming languages and hardware description languages how do we handle concurrency that many things are happening at the same time because after all the program can do only one thing at a time we are simulating it on a computer so the computer can do only one thing at a time whereas you want everything to happen at the same time so the answer is that we have a separate variable called time and we keep the value of this variable frozen now we can do all things and only when everything is completed we advance the value of this variable as a result even though we did that sequentially as far as the time tracking is concerned they would all have occurred at the same value of time because the value of this time variable was kept frozen okay so this is how this is even driven software always works like this we discussed this for IRCM also so you have a time variable you hold the time variable constant you place a transaction when something comes and then advance the time to the earliest next time when something needs to be done okay so this is called event sensitive handling so let us take this practical example okay what do I have I have a NAND gate here there is one input which is direct and the other input which goes through an inverter the inverter has a delay of 8 units the NAND has a delay of 6 units okay so I have a slow inverter a fast NAND let us see what happens now if you were to analyze it without time then you will come to the wrong result right why because at least one of the inputs must be 1 and the other must be 0 okay if A is 1 then the other input is 0 if A is 0 then A is 0 so at least one of the inputs must be 0 at a given time therefore the output this being a NAND output should always be 1 okay in reality that is not so what happens is that when A goes from 0 to 1 this becomes 1 immediately but there is a delay of 8 units of time before this becomes 0 therefore for 8 units of time both inputs are 1 and the output has a 0 going glitch okay this example will illustrate how that glitch is correctly reproduced during simulation okay otherwise if we treated it like a programming language you will always come up as a logic simulator we will always come up with the wrong result okay so that is why this example is illustrative so I have a time ordered transaction list what is a transaction list this says make the input 0 at 0 time make the input 1 at 20 this is also a transaction and make the input 0 at 50 right these are the three transactions which are pending so I enter them here in this transaction list and order them by time at 0 make A equal to 0 I do not know what A was before that is unknown okay so at 0 make A equal to 0 at 20 make A equal to 1 and at 50 make A equal to 0 okay these are the only transactions that I know what I do I first make a sensitivity list okay this is what we had discussed for IRCM also we simulate only partial circuits only a circuit whose inputs have changed should be resimulated right so we notice that the any event on A will cause resimulation of the inverter as well as the NAND any change on A then the output of the inverter and NAND must be recalculated after recalculation it may come out the same that does not matter but recalculation must be done on the other hand any change on B only the NAND needs to be recalculated okay so we say it by saying that inverter as well as the NAND are sensitive to A whereas the NAND is also sensitive to B okay and now we keep our time order here next we advance the time to 0 okay now at 0 what is the pending transaction make input equal to 0 okay so you update and make A equal to 0 now the old value of A B C were unknown new value of A is now 0 therefore A has had an event okay A value has changed earlier it was unknown now it is 0 therefore I must re-evaluate all gates which are connected to A who is sensitive to A both okay so therefore if when I do the inverter its delay is 8 so it says make B1 at 8 units of time current time is 0 delay is 8 input has become 0 therefore I calculate and say make B1 at 8 units of time correct and when I evaluate the NAND I do not care what the other input is if one input has become 0 then the output must be 1 right I already know the output so I say make C equal to 1 at 6 units of time right these two I insert into my transaction queue right and take out the transaction that I have already handled so make A equal to 0 I take out see this is the transaction queue before and this is after so I take out 0 and insert at 6 make C equal to 1 and at 8 make B equal to 1 and the remaining transactions remain what that then I am through with 0 what is the next earliest time at which something happens 6 right this is the earliest time in my transaction queue so at 6 what should I do I should make C equal to 1 right that is the diary that I my diary says at when the time says 6 makes equal to 1 so that is what I do the current time is 6 A was previously 0 now remains 0 B was previously X remains X right but C should now be made 1 at 6 okay and that nobody sensitive to C so no recalculation needs to be done so I removed this transaction at 6 right I have made C equal to 1 right so notice this C has gone to 1 here I am constructing the waveform here A is 0 and remains 0 B is still X and C has gone to 1 here okay at 8 at 6 next what is the earliest thing remaining at 8 what should I do at 8 I should make B equal to 1 right so at 8 B goes from X to 1 therefore B has had an event who is sensitive to B only the NAND okay so I must recalculate the NAND so A is 0 B is 1 right so what should be the output of the NAND it should be 1 if either input is 0 output should be 1 so I scheduled the value 1 at current time plus 6 right the delay is 6 what is the current time 8 so 8 plus 6 that means at 14 makes equal to 1 okay I do not care if C is already 1 so I insert at 14 makes equal to 1 notice that C is already 1 but I do not care I insert that at 14 makes equal to 1 and then I am done I can remove this transaction right so this is my remaining transaction list now when is the next earliest time when something happens 14 right and what do I have to do makes equal to 1 so that is what I do I have arrived at time 14 the old value of C is 1 and the new value of C is 1 there is no event nothing has changed even if there was an event nobody was sensitive to C right so no re-evaluation is required okay so C remains 1 okay notice A is 0 B remains 1 now and C remains 1 now what is the next transaction at 20 at this time at 20 make A equal to 1 right so you make A equal to 1 at 20 so these are the old values at 14 0 1 1 and at 20 you make A equal to 1 so A has an event correct A has changed who is sensitive to A both inverter and the NAND so I must evaluate both of them so what is the value of B because this is 0 B must become 1 at current time plus 8 right what is the current time 20 so I must schedule at 28 the value of B becoming 0 right input has become 1 input has become 1 from 0 at 20 so the output should become 0 at 28 that is the behavior of the inverter right so therefore at 28 make B equal to 0 when I do the NAND what happens A is 1 and B is 1 so the output should be 0 this is an NAND at what time at 26 right current time is 20 and the delay is 6 so at 26 makes equal to 0 correct these two transactions I insert here and take off this 20 because I have already handled the 20 right so at 26 makes equal to 0 at 28 make B equal to 0 and this 50 A equal to 0 remains correct alright these are the waveforms at A we have had this transaction and now my C has become 0 at 26 right that is the next transaction so I advance the time to 26 and makes equal to 0 C has an event but nobody is sensitive to C therefore all I do is that makes equal to 0 and then advance to the next time what is the next time B should become 0 right so I come to next time at 28 B become 0 26 B was 1 at 28 B become 0 so B has an event if B has an event who cares the NAND right so if B is 0 then 6 times of unit later make it 1 so therefore makes equal to 1 at 34 right so we insert this in the time ordered queue next time is 34 I make C equal to 1 C has had an event but nobody is sensitive to C so therefore C which had gone to 0 now becomes 1 this is that glitch we have correctly reproduced it okay next event is 50 A become 0 right that is the only thing left when A becomes 0 both must be re-evaluated therefore that means make B equal to 1 at 58 and makes C equal to 1 at 56 right and take off this 50 transaction right both and inverter and NAND must be re-evaluated so then at 56 C equal to 1 at 58 B equal to 1 then I advance the time to 56 do the same thing at 58 make B equal to 1 and this is my final at when my transaction list is empty then I have finished my work okay so therefore this hardware description language behaves differently if you had just use assignments to C at the same time without this delay you would have never got this glitch and that is indeed the behavior of this hardware you would not do it if you treated it like a programming language okay and as a student you would never understand why this glitch is coming in a hardware description language if you wrote the corresponding C program the C does not never shows it always shows that the output to be 1 but it is only the hardware description language which will show this glitch okay I think we will stop here


but I'm here to say work is important. There's no doubt about it. Children are important. There is no doubt about that. They're important.  Let me say that equally as important. Okay. If you're up to recognize when two people begin a physical relationship with each other and an emotional relationship with one another, you can get rather attached to this person. And as a way for some men to detach is to use their children and to use their professional life as a way to detach. Now I want to be candid with you. And I'm not proud of what I'm about to say, but I did this, I did this myself. I used my


My name is Brian Wesson-Williams. I am a professional track and field sprinter. I am also a fitness enthusiast. I do group training, one-on-ones, personal training, and I'm also a fitness model here in Los Angeles. So when I was in middle school, I was always clean to be the fastest kid in class, and I took a lot of pride to that, because that was pretty much the only thing I was really good at at the time as far as being a group go. But when I got to high school, I got to football. Football season ended, so I needed to stay in shape, and sure enough, I joined a track team, and I got into a track team, and I started beating all the fast kids in the team and started going to track me, start winning races, start being very, very good at it, and I kind of made a commitment towards it. So the following season, football season came, you know, I was the fastest kid in the team. I was put as tail-back, corner wide receiver. I was always, always the fastest guy on the field and whatnot, and very, very prestige means like Arcadia and whatnot. My fastest time became a 10-58. I was 10th in the state, two years in a row, and I got recruited to play football and run track at Arizona State, so I got to Arizona State. Coach got fired, got a new spring coach that got to the program. They didn't know each other, they didn't know me, so I came in here with a partial track scholarship, and when I got there, you know what I'm saying, I was for being me tall, I was going to play football there, but they were like, no, you're not. So I ended up sticking with track at that time, and I'm kind of glad that it turned out that way because, you know, I'm not, I haven't really experienced concussions, shoulder problems, knee problems, you know, common football injuries and whatnot, and I do know that guys who are my age, which I'm 28 right now, guys who are still into football and NFL or some other type of league are, they start a football at a younger age in a professional league, to this day they're dealing with a lot of issues as far as injuries go. So when I go to the track, the smell that, when the sun hits the track and the smell of that rubber, that turns me on. I love the smell of it, I love, they make me want to go work out more and whatnot. When I go to environments like Arizona State Track, UCLA, USC, Track and whatnot, people get in and whatnot, that motivates me in order for me to go out there and do my thing. And I love being around people who are faster than me or better at me in some aspects of sprinting because that makes me want to adapt, be more resilient and produce myself in order for me to be better than them and whatnot. And as far as the gym go, I just love the sound of weights clamping, grunting, weights falling down. I love training at gym with no air conditioning because I love the sweat and whatnot. I just love the environment. And when it's something that I crave, when I was working for that aerospace company, I'd be sitting in my office just banging my head on the wall because I just can't wait to be there because I just crave it and whatnot. So that's what I love about those facilities. So it's all about, as an athlete, you got to have your priorities in order. You know, when you go into a training facility, you're going there for a purpose. My purpose is to run fast. So I need to do the necessary things I need to do in order for me to run fast. And when I go into a training facility, I go in there to make sure that I have the proper exercises to maximize the function in order for me to go and do my sprints. So that being said, sprinting is an anaerobic movement. It's a very dynamic connective tissue movement.  And what that's going to do is it's going to burn a lot of fat and it's going to, and then you've got to have a lot of power so you've got to do some heavy lifting in order for you to build more mass. And that creates aesthetics. That being said, aesthetics just came along. It's something that I don't focus on. It's just something that has just happened to just be there. So in order for you to run faster, you've got to understand how the body moves. You've got to understand posture. You've got to understand how, you know, how your legs move, how your arms swings. And that being said and whatnot, you've got to be able to apply a lot of power into your sprinting because sprinting is a connective tissue movement. And when you're using every single muscle fiber in your body in order for you to do a specific movement. So that being said, if you understand power, you're able to go to the gym, do your deadlifts, do your squats, do your power cleans, hand cleans. You've got to have strength in order to apply power. So that means that you've got to do a lot of strength exercises to make your core strong and whatnot in order for you to do this movement. And if you're able to exceed in those two emphasis, you're able to continue to run faster. And on top of the power and strength movements, you've got to maintain mobility integrity. You've got to have the proper range of motion. You've got to stretch. You've got to foam roll. You've got to have the correct body posture in order for you to maximize your sprint so you don't hit plateaus. So again, you're going to hit plateaus as an athlete. In order for you to break those plateaus, you've got to train smart. In order for you to train smart, you've got to look at your weaknesses.  So I'm born a skinny guy. I've got small joints. I'm not naturally big at all or anything like that. So that being said, I don't produce power and strength naturally. I've got to really work for it. I've got to eat a lot of food, drink a lot of protein, eat a lot of protein, get some carbs and me and whatnot. And at the same time, I've got to go to the gym and actually push some heavy weight in order for me to produce more mass in my body. So the first thing that, the first focus in a gym, obviously, you get on a bike, reactivate the muscles, reactivate the core, get the body sweating again in order for you to do your workout and then foam roll. And the first movement, the first type of exercise that I focus on in a gym is power moves. Connect the tissue work and whatnot, which today was hand clean and power clean. Hand clean and power clean is a good exercise in order for you to teach your body how to be fast with big, big, heavy weight. And you've got to use your speed and you've got to use technique and form in order for you to get underneath it and whatnot. Second to that will be strength training. So I'm a big advocate to a 5 on 5 reps and set ratio. So you can focus on strength and be able to have connective, in order for you to work on every single muscle fiber, whatever body part that you're working on. And the reason why I change chest is because your chest is one of the largest muscles in your upper body, along with the lats. You can use your chest to swing your arms and swing it up because the faster you move your arms, the faster your legs will react to it. And then afterwards I train during high reps, a lot of sets, ratios, reps of 20 to retrieve more fibers as they finish the touch. So supplements is an asset to nutrition. You eat the right foods that you need to do in order for you to get what you need. Supplements take it to a higher level as far as like the things that you need to go. So the first type of supplements that I make a focus on is vitamins. So I need fish oil, vitamins for fertility and make sure I get the proper body function in order for me to let my body do what it needs to do in order for it to stay healthy. The second focus I attack as far as like supplements go is protein. I need a certain amount of protein a day in order for me to recover, in order for me to go back the next day from my body to build and for me to keep on doing the things I need to do in order for me to keep growing. So the third would be BCA's amino acids which is an asset to the protein in order for the process inside your body. Fourth is pretty much glutamine and five is pretty much whatever vitamin or so that I'm lacking inside my body that I don't produce enough. What's important is you fulfill yourself from within. Like kind of like what Arnold Schwarzenegger said like when you wake up in the morning what's the first thing you think about? What's the first thing that you crave to do when you get out of bed?  If it's something that you crave to do and you're very very passionate about it make a life out of that as opposed to what society should tell you what to make a life out of.


So, this week we are discussing about protein dynamics which can be proved by NMR spectroscopy. So, in last lectures I discussed why dynamics, why dynamics is important, what all methods are there to measure dynamics and why NMR based dynamics. So, we will continue our discussion for the NMR based dynamics how we can do the experiments. So, typically we discussed that we measure the spin dynamics spin relaxation rate by carrying out two dimensional heteronuclear experiments such as HSQC heteronuclear single quantum coherence or heteronuclear multiple quantum coherence experiment. And then we can like measure the 13C or N15 relaxation, proton relaxation are rather complicated. So, they are not directly measured in protein NMR. So, typical scheme for 2D heteronuclear relaxation measurement is something like this. We have to start with a preparation state where we start preparing a desirable coherence like we start with a proton transfer to N15. And there we introduce the variable delay of various length which actually captures the auto cross relaxation rate. After this we introduce a T1 period which indirectly encodes the frequency such as N15 or 13C. So, that is a T1 period after T1 period we transfer the magnetization to proton because we are going to detect on proton. So, finally, transferring the magnetization back to proton and acquiring on proton while decoupling the heteronuclear like a 13C or N15. And then after acquisition there is a delay period that D1 period and this D1 period ensures that our magnetization are again back to the Z direction before we start the next scan. So, that then we go back and start the next scan. So, typically this is the experiment that is done preparation T1 delay here we are encoding that like a in case of say T1 experiment we will be encoding here longitudinal relaxation rate. So, delay T then followed by a T1 period indirectly frequency encoding transferring the magnetization on proton and finally, acquiring it. That is a typical acquisition period that we have in the 2D. So, if you look at the pulse sequence of a longitudinal relaxation which is like a T1 relaxation T1 or R1 relaxation rate longitudinal relaxation. This is the kind of pulse sequence that we used this is water flip back sensitivity enhanced T1 HSQC experiment. So, like water flip back is required for suppressing water as you know in protein the concentration of protein is typical of few hundred micromolar or maximum 1 or 2 millimolar, but the water concentration is very high. So, we need to kill the water water is general 55 molar. So, we have to kill the water and that is how all the protein experiments in encodes like has this water flip back or water suppression schemes here is also water flip back experiment. So, if you look at typically we are starting here with a inept transfer we are coming on the nitrogen then here we are encoding the T1 right that is a T1 variable and here is frequency encoding and after that we are transferring back to proton and then we are doing the sensitivity enhanced finally, we are acquiring on proton while decoupling on nitrogen.  So, this is the T1 period this is being introduced to measure the trans longitudinal relaxation rate in T1 pulse sequence. So, what we have typically 180 degree inversion pulse right. So, that inverts the magnetization from SZ direction to minus SZ direction SZ direction to minus SZ that is a inversion pulse right and then this T period is allowed that is given magnetization to come back to the Z direction. So, we first we invert it and then allow magnetization. So, once we invert like this it does the precessional motion come back to Z direction. So, this T period is basically varied and that is what measures. So, you start with a minus Z direction and then with a different T period we are measuring how the intensity changing and coming back to equilibrium. So, that is what you measured MZ at any point T is equal to MZ of equilibrium 1 minus 2 into e to the power T divided by T1. So, this is the rate and this T which was there if I use the eraser here. So, if you look at here we have a T period that T period essentially this T period is the time period that we are giving and T1 rate is the rate like this is the T time delay and T1 is the rate right. So, a 90 degree pulse creates the observable transverse magnetization for detection that is why we apply a 90 degree and then 41 measurement this relaxation block is inserted in the sensitivity enhance sensitivity enhance HSQC experiment ok. So, so that is that is how we do the experiment. So, let me summarize again we start with proton magnetization transfer to nitrogen then we invert it and then allow a T1 period then T1 period to create an observer sorry to spin to relax and then we apply a 90 degree pulse which creates the observable transfer magnetization transfer back to proton detect the proton that is what essentially we are doing in this T1 experiment. So, in terms of product operator if you look at we start with a z direction H z which with a 90 degree pulse converted to H minus H y then we are evolving under coupling and finally, transferring back magnetization to the nitrogen n y. Then we introduce the T1 time period here and finally, it goes to some eta of n z. Now, this factor eta is a T dependent time dependent of magnetization signal amplitude. So, you can write this factor is equal to 1 minus 2 e to the power T that time we are giving and T1 rate right or in simplified version 1 minus 2 e to the power minus T r 1. So, 1 by T1 1 by T1 is r 1 that is the rate and this is the time ok. So, this gives is so, the intensity that has decayed essentially gives us how much the magnetization is changing and that is how we measure it. So, this is the conceptual framework for doing the T1 experiment. Let us go how we set up the experiment and how we process the data in this case. So, we start with a usual stuff right we take a protein sample whatever concentration you can afford take it about 500 microlitre at 10 percent of D2O this is for locking. Then you do the sample height adjustment. So, like in a typical NMR to you, you have to have something around 2.1 cm or around 20 or 21 mm height adjustment. Then you tune the magnet, shim the magnet, set the one frequency one setting that usually stop in any protein NMR experiment that we do we are going to do it. Then we determine the 90 degree pulse there we calibrate it for transmitter and decoupler and we choose the spectral window how much we want to keep for our typical protein that we have. So, here generally you want to 0 to 12 ppm and here say 100 to 130 ppm should be fine for recording an N15 HSQC experiment. Now, important point because we are doing relaxation experiment, so typically we have to set a D1 which will be 5 times of T1. So, typically setup of D1 is recycle delay should be 1.5 to 2 second and generally we have to signal average good right. So, we have like a certainly number of a scan should be 4 or 8. So, in the experiment different T point we are recording right different time where the spin is relaxing. So, we need typically of 8 to 12 T delay which you can range from 5 millisecond to 1.5 second for N15. So, first 2D experiment we do with 5 millisecond, then 50 millisecond, 100 millisecond, 500 millisecond, 800 millisecond something like that at least 8 to 12 we need to do. And since these are inept based sequence, so we have to also look at what is the typical inept sequence. So, you can say you have set the time delay which is here tau it is at 2.7 millisecond right. So, that is what we set essentially and then we set the carrier frequency. So, in proton dimension it should be set at 4.7 ppm or something around that at water. The N15 dimension you have to set it somewhere in between. So, if you are doing that 100, 230 we typically set at 115 or something like that, 118, 115. Now you acquire the data, so like minimum should be 128 complex point in T1 and for proton it can be 1 k like 1024 T2 points. You can also record with this as a 256, here you can record at 256 and this is fine or you can record at 2 k right. So, that is a typical parameters that we are using. So, as we said we are recording 8 to 12 2D points. So, here is a representation of 2D, we are recording and with a different time point different T1 delay the signal is going to change right. So, essentially we start with a minus inverted signal then little bit less, little bit less and that is how it is in 1D that is what we see right, it goes something like this. But in 2D the signal is going to decay as we increase the T1 point. So, we record this data the 2D data and then do the two Fourier transform for each of these 2D data. And before doing Fourier transform we just do the usual stop like multiplying with a 90 degree shift square, sine bell function or cosine bell function or Gaussian whatever you fit it, then you 0 field at least twice of the digital resolution. So, you can 0 field up to 2 k and 1 k or 512 and 2 k, 2 k in in direct dimension 512. So, that digital resolution is typically of 2 hertz per point and then you can apply a 90 degree pulse on proton dimension or nitrogen dimension, 0 field is required before Fourier transform. If required you can do linear prediction that improves the digital resolution, but all the time it may not be required. So, you can take a call whether you want to have a linear prediction or not. So, these are cosmetics or processing data processing. Once you have processed data, remember all 2D like whatever 8 or 10 2D we are recording they should be identically processed right. So, for measuring the intensity, so once we process identically using these stops right. So, typically if you open a broker top spin that is what I have taken from this is the 0 filling you can do up to 5 to 48, 512. This is the frequency that you can for like spectrometer frequency, the SR values, then window function what we want cosine or sine, line broadening what is the SSV, you can choose all of these parameter that you need, phase correction value, now BC mode right all of these you can choose it whatever is needed for data processing getting a nicer well separated peaks. So, because you cannot measure confidently intensity of the peaks which are merged something like this right. So, you need to process so that all peaks are very well separated and one can only take well separated peak for data analysis right. So, we create this series of 8 or 10 or 12 2D, then for each peak now we are going to measure the intensity. So, series of the each series of the spectra is phase corrected right and then proton dimension adjusted according to the first FID, phase or dimension like a phase of the N15 phase is here that phase is corrected here. So, phase you have to correct for the both dimension, phase is corrected. Now essentially we are measuring the amplitude of cross peaks or we can measure the volume integral of each peaks and signal should not be a overlap right. So, you need to have that is what I was mentioning, you need to have all separated signal in the 2D for the data analysis. We measure the volume integral or intensity integral say ij at any point any t point right say 2 millisecond, 5 millisecond, 500 millisecond so all those intensity are measured and that is fitted in a equation to measure the longitudinal relaxation time constant by fitting the equation. So, what is the equation that we are fitting essentially this is the equation that we had discussed earlier right. So, this equation essentially we are fitting to get the T 1 parameter. So, this equation 1 minus 2 e to the power t divided by T 1 or multiplied with R 1. So, rate or time we can measure it. So, if you measure it for say various amino acid that I am showing from one of the protein work. So, here for leucine sorry leucine 24 phenylalanine 36 lysine 46 you see with the T 1 time which is varying from 0 100 to say 1.2 millisecond intensity is plotted and you can see with a time this intensity decrease. And that is what we are going to fit into this equation that we were saying T by T 1. So, we fit this equation and find the T 1 time or the R 1 rate that is coming right. So, this fitting we can do residue a specific manner all non-overlapping residues should be used for this fitting. Once we fit it we can get the T 1 relaxation rate for each of the gemino acid and now one can plot it in a residue a specific manner. So, here I am showing you a protein which is called human sumo that I had worked on. Now, this protein has a globular domain which is here it has a similar fold like a ubiquitin beta beta alpha beta beta alpha fold and it has a long end terminal stale of about 20 amino acid and short C terminal stale of about 4 to 5 amino acid there are loops here. So, now, we are like I am going to discuss the T 1 data that we have recorded for this protein. So, here is the residue a specific manner plotting of this data. What we see here right so, initially the T 1 of these 20 amino acid shows higher value. Now, for all the well folded region alpha helix beta sheet or so, we are seeing the sorry this is the R 1 value 1 by T 1 so, 1 by T 1 is R 1. So, relaxation rate for the flexible domain is higher for well folded domain is lower that means, the T 1 value for this is going to be shorter and for this is going to be longer. So, T 1 and R 1 has an inverse relation. If you look at all the flexible portion has a high relaxation rate longitudinal relaxation rate again loops here which connects the 2 beta from here and some of the C terminal again have high here loops are high. So, all the loops you can see shows high longitudinal relaxation rate and all well structured domain in this protein whether it is alpha helix or beta sheet shows relatively less relaxation like longitudinal relaxation rate. Now, suppose I put this protein in urea where all measure of the secondary structure is removed. So, suppose I am doing the same experiment in 8 molar urea which denatures the protein. So, now the typically alpha helix beta sheets all the are all of those are gone and now you see what the rates are changing. So, if you look at here residue specific manner 0 to 100 what we see here that more or less it becomes flat. So, whatever you see sequence wise variation here when the protein is folded if you denature the relaxation rate is gone. So, if we measure the T 1 relaxation rate of a protein it tells the motion in a residue specific manner the longitudinal relaxation rate in a residue specific manner and you see if we remove the structural elements from the protein this T 1 relaxation rate becomes majorally flat. That means, all the relaxation sequence wise variation that we were getting is becoming absolutely similar. Good. So, that was about longitudinal relaxation rate which is called spin lattice relaxation rate. Now, coming back to another relaxation rate which is called transverse relaxation rate the T 2 relaxation rate T 2 or T 1 row the T 1 in a rotating frame are more or less similar I will just discuss the T 1. So, what is how we can measure the sorry we will just discuss the T 2 how we measure the transverse relaxation rate ok. So, the sequence of T 2 was initially developed by Faroe et al that I am going to show you in the next slide. So, it is similar to T 1 essentially it is a similar to T 1 instead of creating a minus z direction right by applying of 180 degree pulse here we create a coherence. So, here we inversion scheme is replaced by a CPMG or spin lock sequence CPMG in case of R 2 and spin lock in case of T 1 row. So, this rather than like here in T 1 what we saw our magnetization started with minus z direction and it went back to the plus z direction. Now, in this case we are starting with a transverse relaxation rate. So, rather than decaying along the longitudinal relaxation the heteronuclear magnetization relax in the transverse plane during this time T in the T 2 pulse sequence. So, here we are not going like this here just by applying a 90 degree pulse or CPMG pulse we are coming in the transverse plane and from here now our spin is going back to the equilibrium state right. So, what contributes so, in your addition to spin-spin interaction why because this is transverse relaxation rate is spin-spin relaxation rate. So, other than the spin-spin interaction the field homogeneity magnetic field in homogeneity also contributes to transverse relaxation rate. So, to remove this contribution coming from field homogeneity the CPMG spin echo sequence which was developed by Karl, Pulsall, Meeboom and Gill was introduced in the T 2 scheme. So, CPMG scheme of heteronuclear magnetization what happens here we started with a S z and by applying a 90 degree pulse we came to S x. So, S x evolved during this period of say epsilon under the interaction of chemical shift and field in homogeneity. Then we apply a 90 degree pulse which reverse the direction of process. So, we started with here we went back here and now it is slowly diffusing and then we apply a 180 degree pulse. So, it goes like this and then slowly it defuses and then finally, it refocussed during the second period of eta right. So, first it defuses then you apply it and then it come back. So, the provided the spin being refocused remain in the identical magnetic field during this both period of epsilon right. So, then we can measure the rate. So, resulting transfers magnetization at the end of the even echo period of the CPMG CPMG pulse strain has an amplitude something like this I is a intensity at any time point and I 0 is the initial intensity. Now, T is the time period with which we have like waited for and R2 is the spin relaxation rate ok. So, that is what we measure basically for the T. So, now T is 2n 2 eta that time that we are discussing pulse width of 180 degree pulse. So, that is a CPMG pulse strain that we are measuring and essentially we are measuring again intensity during the T2 period. So, this is our pulse sequence that we are using we have started in a with a inept block we are coming back to here nitrogen then you can see here we are introducing this CPMG pulse and during this the T2 relaxation is happening then we are frequency encoding going back to proton detecting on proton while decoupling the nitrogen. So, this is the T period during which transfers relaxation is happening and that is happening on nitrogen nuclei. So, that is what we measure. So, again we record from 8 to 12 of 2D with different T2 time the Fourier transform them process them and then we measure the intensity in a residue specific manner as we did for T1 and you can get the T2 rate. So, here typically the duration that we keep is 0 to about 200 millisecond. So, here you can see for different residue like Q53, 25 K that lysine and K25 that is lysine and I solution 88 here the signal intensity decay are measured ok. So, now each of this fitting gives us a rate of a particular N15 nuclei. So, data processing is done in a similar manner typically 8 to 12 relaxation delay ranging from 5 millisecond to 150 millisecond is recorded and data processing similar like T1. So, if you do that and plot in a residue specific manner. Now, what we get is this one. So, here this is the again rate. So, here the for the flexible portion we are getting the less R2 value whereas, all the structured portion we are getting more R2 value this you remember this is spin-spin relaxation. Now, this was slightly different in case of T1 here if you look at this is reverse here for the flexible portion we are getting the higher R1 rate lower R2 value. For the structured portion we are getting typically lower R1 value, but higher R2 value. So, this is the typical signature we get again at the end you see the lower R2 values are there. Now, some of these peaks are also showing the high value. The one contribution because R2 consist of R2 intrinsic plus rex. Now, this is exchange phenomena. So, R2 also encodes the exchange chemical exchange that is happening for doing that you require like a relaxation dispersion experiment, but looking at this you can say that this protein is geared to show some chemical exchange which needs to be further probed. So, I am not going in detail of the rex at the moment, but that information is hidden there. Now, similarly if you look at the denatured protein again if you put this sumo protein in it molar urea you can see similar like R1 the R2 becomes also flat. So, all exchange are suppressed all the sequence wise variation is suppressed. So, here is the folded protein in zero molar urea R2 and for ready reference I am also showing in denatured state. So, everything is suppressed now protein behaves quite uniformly because earlier it was quite folded and when you put in 8 molar urea it become like a disorder right. So, 8 molar is zero molar. So, just to show that the dynamics that we are measuring also tells a lot about structural compaction where is the flexibility where is the rigidity. So, not only structure, but also NMR can probe dynamic in a residue specific manner. Now, you can even use this concept for protein folding study. So, here I show you how like a urea concentration wise R2 changing.  So, all the central portion where a structure was supposed to be there basically building up and that is how measuring the R2 dynamics also tells about what is the folding hot spot where the structure is forming and that is what you can measure it in a residue specific manner to understand the protein folding. So, with this I just want to summarize today R1 and R2 are two basic experiment that are done for the protein NMR. Next class I am also going to discuss about heteronuclear NOE, R1 measures the longitudinal relaxation rate, R2 measures the spin relaxation rate and they are for the folded region and there is a like for the folded region there will be low R1 high R2 for like a disordered region or the flexible portion it will have a high R1 low R2 right. So, and R2 can be used to monitor the structure that could be formed during the protein folding that is what I am showing you here. So, with this I am going to close it today and in next class we can take heteronuclear NOE and how we can combine all these basic experiment of protein NMR, T1, T2 and NOE, R1, R2 or NOE to reduce spectral density function or even modal free ok. So, with this I am closing it here for today see you in the next class. Thank you very much.


Good morning everybody another day is here. It's here right now, and we're going to call it Tuesday We've got a delivery this morning into Winnipeg got to be there for 8 a.m. So I'm just warming up the truck right now and in the meantime. I came to check it out. Here's trailer 603 It's all tied down Which I'm glad Usually these roll tight trailers we leave all the equipment with the trailer whereas a flat bed or Stepdeck like this the driver uses his own equipment and takes it with him when he goes And then I would put my own equipment and tie it down with my equipment. I Like that a lot better because then all of the the trailers when they get brought to the yard They stay secured and I don't have to worry about you know finding equipment to tie it down, but It's just my own personal opinion. I like pulling these roll tights a lot I like them I find them fascinating trailers because I've never pulled them in the last while I've been driving since 2006 And I just started pulling them in the last month or two And it's like a flatbed trailer but with a Cover so you never have to tarp you just roll it closed and hit the road But you still have to secure the load like you would on one of those so That's why I came here quick in my pickup to check. I'm here in the loaded lineup It's all ready to go my trucks over on the other side of the yard just warming up right now I figured I'd make myself useful instead of just sitting around twiddling my thumbs while it warms up I don't like to start it up and just give her and hit the road right away So it's a diesel engine you should do that with your gas engine too But it's it's more so with the diesel engine It's even more important I should say because a diesel engine needs to get the coolant warmed up it needs to get the engine warmed up before it starts pulling So I like to give it at least 15 minutes to idle in the morning when it's been cold at night Enduring the summer as well too usually just to get the fluid running actually in winter time It's sometimes it takes even longer than that to warm up But at least a good 15 minute idle when you first start it up in the morning and then she's usually Ready to go out there and kick some butt So let's go say hi to her shall we I left her over there all by herself kind of man am I Today is gonna be a good day I don't know why because it's a day that ends in Y every day that ends in Y is a good day, right? right just waiting for this guy off to my left to Make his little loop. He's trying to make a u-turn and I don't want to be in his way We're trying to get out of his spot or something The main gate fell off its track this morning So that's gonna cause chaos in the yard all day They're gonna have to get that thing fixed pretty quick. There's gonna be a big build up I was one of them I came up to the gate and just sat there. What's going on? Why won't it open Brendan had to come over and tell me how she fell off her tracks There's our girl right there. I'm just gonna do a little loop Oh, hope there she is There she was Back her in here right beside her Be careful not to let my tires slip into those ruts that haven't been filled up yet There we go They'll wait till the ground is finished being confused Right now it's springtime. So one day. It's soft the next day. It's all hard the next day. It's soft the next day It's all hard Shife and crooked here, but whatever It'll work. So the truck should be just about ready to go And I'm sure they'll have someone there working on that gate right away It happens everything breaks everything needs maintenance. So Uh, at least we have a second gate here, right? So it's not like we're all locked out of the yard All right, let's go. Let's tell what she's warmed up. She's an old truck and when she's cold she runs a little bit rough But as soon as she warms up she smooths right out. So that's my gauge of How I know when she's good to go. She's not quite there yet. There'll probably be another like She doesn't like to think she's very much like me, you know, we're good friends co-workers work buddies We both don't like working It's just the getting up member in your head that it's going to feel good once you're working It's going to feel good once you're being productive Getting out of bed And getting moving is sometimes a little difficult. Not every day but sometimes it'd be Nice to get an extra I don't know six hours to sleep All right, but there's too much work to do even on the weekends Oh man, we're working at the land every weekend pretty much on Sundays. We've got stuff going on And once we have kids I'm going to completely forget what it's like to sleep in so I may as well get used to it now All hooked up airing up the system Signals are working Air in those tires air in those tires Air in these tires. Yes signals working mudflaps in place Oh Oh, yeah, these mudflaps are new. We got those mudflaps there freak me out. I just saw this bar here. I was like, oh shoot Missing a mudflap Nope, they just put them back there. That way they don't get ripped off when if you accidentally bump into a curb when you're parking I like that Bonus they're all filled with air nice Signals are all working fantastical.  I guess they were waiting for this stuff We just uh picked it up yesterday. I believe yesterday afternoon Now this came down from arborg from up in uh the interlake of manitoba. So about two hours north of here And uh another driver went to pick it up. I was supposed to go pick this up yesterday But uh my morning because of that tractor that we couldn't get started It took too long so they had to send another driver up there to pick this up But he did a good job with it secured it all nicely So I just hooked up to it this morning and brought the rest of the way to the happy customer here And uh, I think my truck is uh Freaking out here. Look at this Apparently according to my truck It's 100 minus 120 degrees outside and very dusty inside. Yeah, so look at this What is going on with me? I was in a bit of a rush this morning. I didn't have a Chance to dust everything. I just hooked up and booked it out the door and look at us kicking butt kicking Tuesday's butt right here With my back where it belongs Yeah, so uh this thermometer doesn't work I was gonna show you at some time uh every once in a while doesn't do it every time So it's hard to catch it But I I turned the truck on and it freaks out for like five minutes saying those minus 20. No, it's plus 140 No, it's minus 80. No, it's only plus one. No, I think it's zero. No, it's minus 120 again I don't think it's quite that cold outside I mean it's we still got some snow over there But uh, I'm pretty sure it's not negative 120 Just going out on a limb here and guessing Down on this trailer. I don't think it needed to be but again, I was in uh, I was in a bit of a rush To get going this morning. So I just left it down. It's not Illegal or bad if it is down tri axels a tri axle It just wears out the trailer a little further So I'm gonna lift it up Since we're now empty and there's no reason to have it on the ground So that we got to go all the way back up here.  Maybe uh, you're not paying for the tires. Maybe that's why But uh, I always put them up. It has something for us when we get back It takes about 45 minutes 45 to 50 minutes to get home from here or not home But get back get this truck home. This is the exact same place. We were at yesterday Not the same drop-off at the exact same street a little bit of a blind corner here Because those trees block what's coming that way, but it gives us enough time But over here you got the road curving around that way So we're gonna go now and slowly watch this nobody's coming from there Looked like two cops part two of today pulling this box up to too long Let's roll those up before we forget signal's on first So we can check that they're working marker light and signal working Tires filled with air filled with air mudflap in place Marker lights working signals working for light signal light working over there all filled with air I love it. Okay. Look up. Don't forget always want to follow the rules How's it looking? Not too bad not too bad A quick sweeping will be on our way. I've seen a lot worse This means to look up at the latches there to make sure that they're latched in Because if you don't double check very often they don't actually latch in And then the trailer doors aren't secured going down the highway That's how I do Might get you ticket from one truck to the next Don't be mad at me. Okay I forgot you guys here I got over halfway home and I had to turn around and come back I thought I had you guys with the rest of my stuff and suddenly I figured you know what I had a feeling Maybe I should check it out So I pull over and I go through my stuff in the back Right here with my bag and everything My lunch kit and everything else and I could find everything except the camera I knew I had it in my hand.  Looks like they fixed the gate Yep I wonder if they fixed it themselves or if they had to get like the gate doctor to come in and Fix her up It's not the first time this gate is broken It's a big gate and you know, it gets used quite often. We have to keep it closed all day We can't just leave it open. We have to keep our yards secure So it's opening and closing opening closing opening closing every day day after day after day Tomorrow my whole day is marked out for me already There's a load of shingles. I have to deliver first thing in the morning into Winnipeg And then after that, there's two pickups. I got to go and get One up in too long one in our Berg again, that'll be on a roll tight trailer That'll probably take yeah, that'll take the whole day mostly I may have time for another like local delivery when I get back, but I'm guessing that's going to be the whole day So I like that. I like knowing what I'm doing the next day It's not that it matters. I mean I can find out in the morning too doesn't really matter At least now I know Where I got to be and when So let's go home I got everything this time for real At least I better If I forgot anything else, we'll get it in the morning. I'm not going back again


In late 2020, it was reported that basketball superstar Candace Parker was making real estate moves. The five-time WNBA All-Star purchased a $4 million modern farmhouse-style mansion in Tarzana, LA. Prior to this, the baller lived in another slightly smaller abode in the same neighborhood, as well as an Encino property she once shared with her ex-husband. Candace's newly built home looks like the nicest of them all, spending 6,300 square feet of space, and we'll check out all of the amazing features it packs in, including the pro movie theater and more. In these videos, we don't reveal any addresses, and even though I've done a house tour of my own place, please do not show up at any private residences because it's not safe for anyone. Candace Parker is a professional basketball player for the Chicago Sky of the Woman's NBA, and chosen as the first overall pick in the 2008 WNBA draft by the LA Sparks. She has plenty of awards and accomplishments under her belt, including more than one MVP award, two Olympic gold medals, the 2008 Rookie of the Year award, and much more. Not to mention, she became an analyst for NBA games on TNT and NBA TV back in 2019. At the time of this recording, Candace has scored herself a net worth of over $5 million, considering her NBA salary, endorsement deals, and more.  If you haven't already, be sure to subscribe and ring that bell for notifications because we post brand new videos daily. As always, you can follow me over on Instagram to chat, and now let's get into this video. After Candace ended her marriage with ex-husband Sheldon Williams, the two listed their Encino home they dropped $3.56 million on in 2013. Despite the fact that it was impressive Mediterranean estate, the couple had quite the difficult time unloading this home. They originally listed it in July of 2017 for $4.49 million, with no takers. They put it back on the market in November for a slightly reduced price of $4.39 million. When all was said and done, they finally sold the home in March 2018 for $3.75 million. Located in the highly coveted Encino area south of Ventura Boulevard, the LA Diggs featured a 7,058-square-foot home on a 1.3-acre law and boasted 7 bedrooms and 7.5 bathrooms packed with all the goodies. Originally built in 1964, the estate had been completely remodeled with crown molding, a ton of built-ins, sleek marble floors, and a state-of-the-art sound system. Highlights include a step-down living room and formal dining room, both of which offer plush furnishings, crystal chandeliers, and stone fireplaces. While the spacious chef's kitchen came with Viking appliances, marble quartz countertops, and a stunning island with ample storage space ideal for the basketball star to cook up a storm. Additional kitchen features included an attached breakfast nook and a butler's pantry with plenty of space. Next to this, there was yet another family room to kick back and relax. Probably the most coveted room in the house had to be the master suite, which offered a sitting area, sumptuous fireplace, and French doors that led to a private terrace with stunning canyon views. Additionally, the baller shared a massive walk-in closet that was big enough to pose as an NBA dressing room. Candice's former master bathroom came with its own seating area, a massive marble soaking tub with crystal chandelier hovering above, and a large glass rain shower on black marble. They even outfitted the space with a luxe for a rug, and we can't forget more of those stunning canyon views. That wasn't all for the deluxe features found in both the interior and exterior. In addition, there was a home theater with wet bar where I'm guessing a lot of game footage had been watched, and a huge great room with a view and another fireplace. There were four fireplaces in total. Outside, the property boasted an outdoor barbecue and kitchen alongside the palm-lined pool and spa. Equally impressive was the pool house with sauna, bathroom and wet bar with flat-screen TVs perfect for a team party. Guests looking for a little exercise could do so in the professional-sized sports court, which can be easily lighted at night. Hey, not a bad view either to work on that tennis swing. Well done, Candice. Back in February 2021, Candice parted ways with her first Tarzana home for $2.43 million, but let's take a look. Much like that last spot, again Candice took a small loss on the sale. She initially listed it for $2.8 million in 2020. Considering she paid $2.47 million three and a half years earlier, it wasn't a huge loss for her. Located in a short cul-de-sac in the beautiful suburban foothills south of Tarzana, this estate featured 4,525 square feet and boasted 4 bedrooms and 4.5 bathrooms. While the home was built in 1975, the wood, glass and concrete home had been since updated with first-class finishes. Upon entry, double front doors opened to a double-height dining area with plush furnishings, custom cabinets and sumptuous light fixtures soaring above. The spacious living room sported a pool table, massive marble fireplace, Florida ceiling windows and fleetwood sliders that boasted stunning views. The open concept kitchen offered enormous white marble counters on sleek custom white cabinets, sub-zero and wolf appliances and Butler's pantry with ample storage. Her open kitchen's biggest flex had to be the glass sliders that opened to the resort-style backyard, giving guests the choice of dining indoors or even on the patio. And I think from all this, we can tell that Candice loves a modern and minimalistic look in her homes. You'll see what I mean with the next one, too. Steps away, the home offered a family and great room complete with plush furnishings, concrete fireplace and built-in whip bar perfect for entertaining. The residents hosted a number of guest bedrooms, one of which was a double-height suite with sleeping loft and another with a petite private balcony. Candice's former master bedroom was comprised of a wood-beamed cathedral ceiling, brick fireplace, sitting area and private balcony with a view. The master bathroom came with his or her sinks, a bulky soaking tub and steam shower ideal for soothing, aching post-game muscles. This residence included more bonuses throughout, such as a library office, decked out with custom hand-carved antique wood, colossal shelves displaying all of Parker's trophies and an additional fireplace. We're willing to bet that Candice locked in a few endorsement deals in here. Elsewhere, the residence included a spacious home theater packed with red suede recliners, a games room and a line cellar. Outside, the grounds encompass multiple dining and lounging areas, a small swimming pool with spa, a grilling area and a pergola-covered area. While the features in the backyard may seem a little small and underwhelming, the glass sliders could easily shift the party inside or out. In December 2020, Candice upgraded to an all-new modern farmhouse in Tarzana, which cost her $4 million. Her current home may just be the nicest of them all, built the same year in 2020 and spanning 6,300 square feet on a half-acre lawn. Her home boasts five bedrooms and seven baths, along with both a substantial motor court for six cars plus a three-car attached garage. You know, in case Candice wants to throw a party. The modern farmhouse getaway comes with an open concept floor plan that includes a living room, fully furnished to Candice's liking, built-in fireplace and oversized windows giving tons of light. Throughout the home, there's also wide-plane cardwood floors and a pretty vibrant color scheme with shades of white, gray and warm acorn. A center hallway on the main floor leads past a black and gold office up to the massive grey room. This upscale room has a formal dining area with Web Bar, family room with yet another fireplace and backlit built-ins, as well as a courtyard you can access through glass doors. Here you'll find the courtyard has its very own outdoor fireplace too. There is even space to accommodate a large event and some chefs to prep food. There's a casual breakfast bar, sunny breakfast snook and access to the yard via walls of glass. The mansion is complete with five private bedrooms each with their own ensuite bath. Candice's upstairs master suite is the best of all with massive space, a marble fireplace, a showroom closet that's bigger than a shop and a spa-style bathroom with both a soaking tub and one of the biggest showers I've ever seen with multiple shower heads, no less. Candice kept her movie theater game strong in her new digs, however this one is definitely the showstopper of this house tour. The soundproof movie theater features 10 leather recliners, its very own TV lounge and a games room with a Web Bar. Not a bad spot to kick back and relax. Candice can also enjoy the work hard play hard lifestyle in her lush backyard, complete with saltwater pool and spa with Baja Shelf, barbecue area, kitchen and fire pit. Additional features include an adjoining pool house with Web Bar and, of course, her very own half basketball court to work on that jump room. Alright, so I think I'll bring this house tour to an end here. We got to take a look at WNBA's superstar Candice Parker's Homes in Encino and Tarzana. After checking out the three impressive homes did you have a favorite? What about the features? I liked her newest house the best, that's no-brainer. It's a bit more modern than my taste but there were so many cool features it's hard not to love. From the backyard to the movie theater there was really something for everyone here. Not to mention Candice can feel like a queen in that master suite. Be sure to let me know what you liked or didn't like about Candice's homes down in the comments. Thanks for tuning in. Don't forget to follow me on Instagram and I will see you all in the next video. Bye!


alogue with local government, I have done more than any other minister in this government to cultivate a strong and positive relationship with local government. However, when we get to the point that we are putting money into a settlement and seeing teacher numbers go down not up, then I think that the Government is entitled to call time on those particular arrangements. When Mr Gray was asked by Mr Crawford what money would the Labour Party put in to try to protect teacher numbers that are apparently committed to protecting, the answer was absolutely none whatsoever. I painfully regret the fact that I have not been able to get to a mutual agreement with the Convention of Scottish Local Authorities on the issue of teacher numbers, but if I am to respond positively to what is clearly a majority opinion in Parliament that teacher numbers should be protected because this Government believes that, the Labour opposition apparently believes that we should maintain teacher numbers, I am bringing the constructive and positive action to actually tackle this issue here to Parliament and that is what we will put to local government in Scotland. The final point that I want to make is about the investment that we are making in our young people and in the wider investment in the educational environment in Scotland. We set out our determination to tackle educational inequality and we have made the first clear statement today of £20 million of further investment in doing that. Today it looks as if the Labour Party is going to vote against £20 million to tackle educational inequality in Scotland and that is a matter of profound regret. It will also vote against £526 million being allocated to the college budget in Scotland on the basis that that is not nearly enough, while £526 million is more than any Labour administration in Scotland ever delivered for the college sector in our country. The minister is in his final minute. The setting of budgets is a difficult and challenging task for any administration and in the financial context in which we are operating, in which Mr Fraser, our budget has reduced in real terms by 10 per cent since the Conservative Government came to office, we managed within that context to deliver public services in an effective way for the people of Scotland. However, what the Labour Party has demonstrated today is that it is unfit to come to Parliament and deliver a coherent argument as to how to handle the budget. Apparently it was all to be about health, then it was to be about local government, then it was to be about colleges and actually it did not amount to enough money for any of them in terms of the proposals. This is a budget that is strong, a budget focused on the needs of the people of Scotland and I encourage Parliament to support it. That concludes the debate on the budget of Scotland number four, Bill. The next item of business is consideration of business motion number 12230, in the name of Dolfitt's Patrick. On behalf of the parliamentary bureau sitting out of business programme, any member wishes to speak against motion should press a request speak button now and I call on Dolfitt's Patrick to move motion number 12230. Formally moved. Thank you. No member has asked to speak against the motion therefore I now put the question to the chamber. The question is that motion number 12230, in the name of Dolfitt's Patrick, be agreed to. Are we all agreed? The motion is therefore agreed to. The next item of business is consideration of a parliamentary bureau motion. I have asked Dolfitt's Patrick to move motion number 12231 on sub-committee membership. Moved. The question is motion will put decision time to which we now come. There are two questions to be put as a result of today's business. The first question is that motion number 12226, in the name of John Swinney, on the budget Scotland number four, Bill, be agreed to. Are we all agreed? The Parliament is not agreed. We move to vote. Members should cast their votes now. The result of the vote on motion number 12226, in the name of John Swinney, is as follows. Yes, 64. No, 53. There were three abstentions. The motion is therefore agreed to. The budget Scotland number four, Bill, is agreed to. The next question is that motion number 12231, in the name of Dolfitt's Patrick, on sub-committee membership, be agreed to. Are we all agreed? The motion is therefore agreed to. That concludes the decision time. We now move to members' business. Members should leave the chamber, should do so quickly and quietly.


A magistrate court sitting in Yola has denied Honorable Lawal Uba a former governorship candidate of Africa Action Congress Bill for allegedly insulting Governor Amadou Umaru Fentiri. The politician has been remanded for over 30 days at the Correctional Center after being accused of sharing videos in which he accused the governor of betraying the PDP presidential candidate, former Vice President Atikua Buboka and others who helped him win election. Magistrate Aminu Abutkadir Ma'ilha dismissed bail applications by counsel to the defendant while a similar application at the High Court was dismissed. At a resumed hearing today, the judge adjourned the case till Thursday to allow the prosecution to prepare for presentation of two compact discs containing some evidence. The man has been in detention for almost more than one month and he is a gobernatorial candidate. He was the gobernatorial candidate of AAC in 2019. He contested with the incumbent governor. So why should he be subjected to this kind of physical mental torture? Just because of these simple remarks he made. Because it's a case with political undertone. Because what they are alleging that he did was that they are alleging the allegation is that he made some defamatory remarks about the governor and what are they classifying as defamatory remarks is they are alleging that he said that the people that supported the governor, the incumbent governor, to get the seat where there is occupying and that the governor has let them down and that the governor has equally disappointed his political godfather in the presence of Atikua Buboka, the presidential candidate of the PDP. So the question is that those comments, if you look at them microscopically, are they defamatory? The answer is no, they are not defamatory. Those are comments anybody can make about a politician, about a political officer. The message I have for the governor is that first and foremost, he should know he's a leader and a leader is there to take any responsibility and is there to bear in mind on whatever situation comes his way. Because he lead people and among these people there are different people who display different characters and you are one to lead them. So I think you should have that mind of accommodating all sorts of things. You are there to sort out if something of that nature happened when people have thrown to you for you to sort it out. But now as they say did and they relate it that based on what you have ordered to the governor that the government is interested in the case or is the government that is going is moving with the case. To me I am not happy about it because what I know about a leader is anything that you should manage.


Welcome to film my run and Little Hampton for the beach five-miler. The Little Hampton beach five-miler part of the Wesse Six Fun Run League So I'm here running with Worthing Harriers again 10 minutes to start of the race We've got quite a big crowd of Harriers running today. We've got a few fast runners today Olly's running Shaun's running Quick quick runners As you can see there are a lot of people here and it starts with a complete mele round the field One kilometer in, three kilometers in can't keep yourself in the wind Four K in halfway now on the way back Wind behind us That's five K now six K in now. She's got nothing in my legs That's seven K last kilometer Less than a kilometer. Oh that was horrible. I just had nothing in my legs So I think the gun's the ultra On Sunday Definitely taking out of my legs there Shaun, how'd you do? Fourth again. Fourth again? You all right Olly? Yeah, yeah, there's only this little bit left


The title of this video is not Skinner's Self-Management, however you might be able to interpret it as such if you pay close attention. There are two terms in this video, one is the controlled response, one is the controlling response. The controlled response is the response that you want to control through the controlling response. This is what self-management is all about. You emit a response, controlling response, in order to control another response, a controlled response. If I was trying to quit smoking, I want to reduce that, because I have a behavioral excess, and there's another term we should define later. I have a behavioral excess, we need to get rid of it, so we need to decrease smoking, so I'm smoking. I need to engage in a controlling response to decrease this controlled response. So what could I do? I could break my cigarette. That's good! Or I could hit myself on the head. Ah! That hurt! I should put my cigarettes down. Or I should just not go by them. The controlling response is the response that you need to learn in order to control the behavior that is the problem. There's a little trick here, though.  And there's a whole bunch of problems with this, and self-management is very, very challenging. But we need to know what the two core components that Skinner talked about are with regard to self-management, which was the controlling response, one that you emit to control a particular behavior. You control the problem behavior. That's what we're after. So it sounds like there's a lack of control. Pretty much.