So, after two days of the school, we got familiar with high-performance computing in general, different polarization techniques, and yesterday you learned how to efficiently max for molecular dynamic simulations, one of the very powerful methods for studying of molecular systems. And today we are continuing to cover two additional areas that are extensively used in molecular research. These are free energy calculations and docking. So free energy calculations, you might be familiar, is a powerful method, for example, for scoring drug candidates, and in combination also with docking, they give very powerful tools to researchers. So first, this morning, we will discuss free energy calculations in use, automatically, efficiently. For example, Gromax is a backend by using the PMX application that you will learn about. So, it's my pleasure to introduce you to Vitatus Gapsis, who is the main developer of PMX. He will tell you more about the method in general and also the tool itself. And I will find it very useful in your search work. So, Vitas, the floor is yours. Thank you for the introduction, Rassan. And let me share my screen so we can start right away. Now you should see my screen. Let me know if there's something wrong. But yeah, let me then begin the introduction in what we'll cover today overall. So I decided that it would be probably a good start with more broader and more general introduction to the free energies, what kind of methods we have to play free energies from the data and why in general, why they are of interest. So Rassan already remain in applications, but we can also go a little bit into the details why in what particular questions free energies might be useful to calculate. And then in this would be my first part of the talk. And later I would show you in the next main application. So applications we encounter in our everyday work. So I work together with Bert de Groot in the computational biomolecular dynamics group in Max Planck Institute. There we only do computational research and we always ask questions about protein thermostabilities. We ask questions in ligand binding and several examples I'll show you today. And then after the talk, we have also practical hands-on session. So there I would go even further into the details. And then we really can have a look how to set up such calculations and how they can really this BMX and Gromach's combination can do some useful things for you. All right. So let me start right away. So free energy calculations, first, firstly, let's go to basic question. Why care about free energies? Where do we find these questions related to free energies? So the feature that already were mentioned by Rassan, those are affinities. So for example, imagine ligand and protein. So a common question in the drug design field, how strongly does the ligand bind? The binding itself is synonymous to the free energy difference. So if we are able to calculate free energy difference, we actually also already have access to the affinity. In other words, we know how strongly the ligand interacts with the protein. Really for protein-protein binding, it's exactly the same way of, also, stability are free energies. Consider a protein folding example. Protein may prefer to be in its unfolded. So if we tilt the free energy into the direction of the folding state, we then make the protein more thermally stable. This principle, again, is just a matter of calculating energy difference between the unfolded state. Also rates, we often consider rates as a kinetic parameter. But if you express that, yeah, as a relation, rate is a relation to the, has direct relation to the free energy barrier between the potential of mean force, for example, of how iron crosses a channel or water molecule across aquaporin channel, aquaporin protein. It also crosses on some certain geolandscape. And the barrier, then the crossing will be, then the channel will be less conductive. If there is a lower barrier, then it will be more conductive to assess the free energy landscape of this transition. So I hope I convinced that free energy calculations are very many, many different locations. And what methods, what flavors do we have for the free energy calculations? So yeah, this is a very rough sketch. So I don't need to do some attempt at a really fine grained division of the methods. Just my rough interpretation. So let's start from the right side. Those are statistical methods that I separate there in the separate branch. Statistical meaning that we are not using physical principles, or even if we are using them, we are approximating them CV. Those could be ligand based and structure based. So imagine ligand based statistical methods are the ones where we ask if we have now a bunch of ligands, we don't know anything about what they bind to, what they interact to. We simply try to establish a regression between some descriptors, maybe there are a number of atoms, number of hydrogen bonds that the ligand has, number of rotatable bonds and free energy. So these would be very chemo informatics based approaches. Structure based approaches would already take into consider the protein ligand interface and would perform docking into these binding pockets. Hybrid methods, they combine both physical, physics based, for example, sampling approaches as would be Poisson ball and accessible surface. And then regression based on these samples, once it is constructed regression, approximate the free energy differences. Or statistical force field could be used as Rosetta or Foldex, for example, used. They use statistical force fields, but the local sampling in that force field and this already brings them into some hybrid between statistical and first principles based. And first principles based methods, those are the methods that we'll be using today or discussing today and using later. So this is the whole zoos expended there more and those could be free sampling, bias sampling. Then we go into alchemical methods, enhanced sampling, and we will discuss further into those. So let's go branch. So before we just jump in, before we into them methods, how to extract free energy from the calculation. Let's recall a few ways to express free energy. So the first relation, let's have some of the first relation, you probably know it very well, it just relates enthalpy and entropy and the enthalpy minus would be weighted by the temperature free energy. So this is a thermodynamic description of the free energy and another lesson, let's have a look at the how from this point of view, we can express the relate free energy of a state I, we can relate the probability of an article or a system being in that microstate I. And now we'll go over approaches that will utilize, give us insight into these both into both of these relations and we'll express the free energy or extract the free energy difference from the, so let's have a look at the first example and this very simple example, but it already captures or in the mainly intuitive nature of these relations that I just showed. So imagine we have a peptide, which is able to unfold, and we simply monitor the RMSD from the from some reference state, and in the top, we are running at 298 Kelvin at the room temperature we're running it, and we mainly remain below the red light state, sometimes we transition into the unfolded state but very rarely. Now, in the lower panel simulation. Now we are running, sometimes we're still in the folded state, but now almost let's say. percent of the. Higher into the unfolded state, so we keep on unfolding much more often are certain edges. And now the these these these two panels already illustrate very nicely how we can extract the free energy differences, so remember that the three are. Well, they are nothing else, but the probability probabilities probabilities of being in one state, but we simply can count frequency of occurrence of a system in unfolded state versus the folder state and use this relation that I showed. So this kind of definition ratio of the probabilities will come out to relate to the delta G between the two states and we need to look and extract the delta G counting the problem or frequencies and based on the law of large numbers we just relate them to probabilities. Okay, and another feature, we see the effect of the temperature, we change the temperature we change we shift it also the free energy balance to the more to than folder state and we can go back against the happened, so we had our. We all of a sudden increased the entropy contribution, because we just changed the tea to a higher. The gene this case it would be delta G because we consider the two states the difference between the two states we shifted it to another. So you can go even further, so this was just in RMSD space, we can also expand it into the a little bit more into the conformational space density and notice that they were not just folded and folded there were several folded states so that this is just a principle dimensionality that. reduced dimensionality every point now represents a different confirmation of the peptide. And we notice that the basis of different different conformations and all the rest is there unfolded let's call that unfolded, can we extract the free energy differences between those. When let's continue on simply let's count how many points are in each of those, and we can again just let's put the, this is a multi three dimensional. Some coordinate, and we can see those three basins populated nicely, the first one is the deepest, and it has the lowest, the most occupied by that, then there will be the second and the third. And now, if we look at it also, color by the temperature, by which, at which this perform, you can see already that this very nice energy landscape changes substantially the temperature, all of a sudden, where barriers are no longer barriers. The, you could consider it to be much, much, much flatter this. Yeah, so this very simple example already shows us all the particular features of how we energy is an edit sense and how we can control the transitions between barriers, just by changing the temperature. But, say, sometimes will, yeah, we know that using temperature as a trick, we can overcome the transition barriers, we can, therefore, you start constructing some replica chip based replica exchange schemes, and so on. But let's, let's say we are interested at a free energy landscape at a given given temperature. We are faced with a situation where we're where our particle simply can sample just one side of the barrier, the barrier, this barrier, it's, it's quite high and we never cross it so we never observe a transition, but we're interested in learning is the landscape is the free energy landscape and what is the difference between the free energy values on one side of the barrier and on the other. How is this so as time would be umbrella sampling and with an umbrella sampling. And I mean, placing an umbrella potential so what practice, we take this particle, let's, let's imagine now that this particle is actually water molecule. We restrain with a potential at a given position and allow it to sample, sample the landscape as as wishes. And if, if we are, let's say in a completely flat landscape there is no, yeah, no gradient in any direction it will simply sample this ocean as defined by potential. But if there is, if it is on a, yeah, some current free energy landscape, it will slightly tilted. It will have a sample, a little bit preferred direction. Later, we shift them a little bit and do the same. Now, the blue molecule would be in the place of one red molecule. Then we do the same, the same, the same sample across the reaction coordinate. All right. Now, we have sampled them again. As it wishes to sample, we only need to unbiased the and we in principle rely again on our counting. Just count how often the particle prefers to be in one place to another. So the only trick is that we applied also some bias. So we seem, but, but we know how much bias we included because it's simply a harmonic potential. And there is a technique called weighted histogram analysis method allows to remove that potential was from the positions. So we have sampled, let's say, densities, and we recover exactly the same potential as we would as the particle which you've been crossing this range of lines. All right. So this would be an enhanced sampling example. But let's, let's go further. Let's go one step further. So let's, let's look again. The same that we have two states of the system and a large, a large barrier between the two of them. Now we are interested. Again, the barrier is large enough that we wouldn't cross it just spontaneously so we cannot obtain free energy differences just just by pure Sam. We are also, let's, let's formulate the question in the phone way that we are not interested in the height of the barrier. We are just interested in the difference between GA also free energy in the state a and the difference in end state. For example, it could be timing. So the triangle binds to this backman and and becomes a bound ligand bound square. So now we, we could of course do an umbrella sampling, but that would be computationally very expensive and difficult to converge. The question becomes actually somehow learn how to transform this triangle into the square. Is there, is there a technique and this is exactly the alchemical. So let's go now a little bit into the details of that. There are several flavors of the chemical approaches. So here I'm showing I replace the earn now with the, with this ellipse where our question is, can we somehow transform the ellipse triangle and learn what the free energy difference between the two of them is. And there is a way we simply so ellipse is this Hamiltonian. So the potential of the system, which governs the, yeah, the dynamics in state a and potential of the system in state B. All right. Now we can construct a high tone in by simply come back to them with a coupling parameter. So it could be just let's say one my times Hamiltonian a plus lambda Hamiltonian be just a simple linear combination of the two Hamiltonians and control this from the parameter just control it in the simulation. Now, if we very, very, very slowly drag this parameter lambda from one state to another, we can at every point in time we can calculate the partial derivative of the of this combined with respect to lambda. This is the, so the derivative of Hamiltonian with respect to a certain parameter is a force acting along that along that. That coordinate so in this case force along our chemical lambda parameter. And now if we integrate a horse we get work in this case if we do it infinitely slowly. We did the work we don't we don't dissipate anywhere. It becomes the free engine different. And this technique is called thermodynamic integration. All right. So is there another approach and yes there is it. It is a different to consider exactly the same problem it's called free energy perturbation. So for free energy perturbation, we consider system we can simulate the system in state a don't do any perturbation, nothing so far in state B. Okay. So, when we simulated system. We extract the coordinates from the state a. And, and evaluate them with the Hamilton state a. But also we do the. We do the following trick. We take the same. From state a play them also with the Hamiltonian of state B. And now there is a relation the known derived by 20. It's called free energy perturbation that allows to relate the difference between these two Hamiltonians. And their average to the difference, we can also do exactly the same with state B. And it should, it should also come out exactly the same. In state B. Or not evaluate them with the Hamiltonian of state B. So it's native one and the foreign Hamiltonian of state. And their difference in the two Hamiltonians. Exponentially averaged will give us the engine difference. And does it work. Yes, it will. Only friends between the two. Hamiltonians is very small at the order of thermal fluctuations, but usually much larger. To the system where there is no overlap between the two so these two points would be very far from one another but what, but then we can do the trick. Let's simply start putting intermediate states right so we don't directly calculate Hamiltonian and be we divide in the subsection so yeah, XYZ and so on, small, small perturbations, and we do exactly the same math exact. And then we do the simulation intermediate values and collect them into one delta G afterwards. Okay, so this is pretty much a perturbation. And the first conceptual differences between the NTI, they both report on the same free energy difference in the end. But they have different strengths and we FEP is sensitive. In fact, lambda scheduling. Lambda stratification. While TI in principle would be giving a smooth transition, but one is to integrate over over work, work value as you can see, and get the lambda curve and this of course then also FEP would require some equilibration for each and every window and requires one to be always at equilibrium in each and every window and sufficiently converged. So this can be confidential very expensive. But the TI principle relies, or grows standard TI relies on the fact that the transition is always at equilibrium, but this is an an assumption, right. Therefore, there is some certain that the Hamiltonian lag or always introduced work dissipation with this method. So the FEP exponential versus direct calculation for as I already mentioned is actually direct calculation that requires some numerical integration then. And we have more flavor is the equilibrium free energy calculations. Let me also highlight those. So you already saw this of a thermodynamic integration. So we have, we have this way to calculate. In this case, you see, I would double this means that we no longer do this transition from state A to state B, very slowly. So, if we don't do the transition from state A, let's say wild type protein to the mutant protein very slowly. When we make a transition we dissipate work. If we dissipate work well, we call it simply w. So we calculated some work, but it's fine. Now, what do we do with that work. Let's now have a look at a little bit a little bit at that concrete example that I'm sketching here. Let's say we have a wild type protein simulation related very long at equilibrium. We didn't do any magic there. And we made very many transitions from one state to its mutant version. Transitions we calculated this w with exactly this equation that I'm showing you. Now we call values and build a history. This is the blue histogram. Then we do exactly the same but in the opposite direction we simulated the mutant state transitions into the wild type scan. These are the red arrows that we also calculate work values, and we make a red histogram. Now we have problems and they overlap somewhere. And there is a powerful theorem which is derived by crooks. It relates simply probability distribution in the forward direction of work values with the probability distribution in the reverse direction to be different at the intersection point. We can also recover free entry difference this way. Now we dropped the assumption completely require equilibrium durations. Of course the end states these two ellipses like ellipses they need to be an equilibrium but later we don't. So this was the really theoretical part. And let's go now a little bit. Towards the applications. And now you learned a lot about methods that we applied yet, but let's try to bring it to the concrete example. And the example I'm phrasing here is a calculation of thermostability in a protein due to a mutation. So we have the following step, a protein, which can be in two states in this thought experiment. It can be in its folded state and in unfolded state. So we learned that we can extract free energy differences by counting. So let's simply simulate infinitely long and how protein sometimes unfolds, count how many times it is in the folded state, count how many unfolded. Relate these frequencies of the unfolded to the probabilities of the probabilities extract the free energy difference. All right, but this, of course, there is a catch that practically very expensive or feasible because I mentioned that let's do this process infinitely long so it will take a while to reach something right. But let's so let's continue and let's do the next trick. Let's introduce the mutant. So now we have two situations, one wild type situation, and another is a mutant where mutant is folding unfolding folding unfolding and we do exactly the same trick. And now we have systems, which we need to simulate infinitely long. So you may ask, well, just do you made intractable problem twice intractable. All right, true, but now we can that we have also chemical methods, let's do the alchemy now alchemical methods will go in this cycle to draw here, here in the cycle. They will be the horizontal arrows that are connecting the two wild type and mutant in respective folded and unfolded states. So what it means is it means that we are chemically we need to create the Fiancid difference protein in the folded protein and in the unfolded protein, and then from that Fiancid difference. So, in one case, Delta, we recover Delta Delta G, and this Delta Delta G will keep on using this Delta Delta G throughout the practicals and the next examples. This means how much the protein folding Fiancid changes upon mutation. So if Delta G is negative, then we notation made the protein fold. Yeah, strong fold more or the folded state stable that then done full respect to its wild type state. All right, so that's all the magic that we have. Of course, these are more explanations. We still need to build a technical practical framework for that for for running these calculations. So maybe you are already familiar with running standard Fiancid calculations or simulations where you can solve it the system in a box at irons and run the simulation. But now we need to do something a little bit different. We need still to have the same capability of running the simulation, but now our system needs to represent both states, the blue wild type and the red mutated state as in catch at the same time, and we need to have a control between the two states. And for that, so as a dissimulation, but we need a patient for that, we have developed our PMX framework for that. Of which I will talk exactly which which I will discuss a little bit more in detail, but I'll mainly go into the examples showing how we can do what we can do relations. And we'll have a real hands on in the next session on to to where I will show happening to the structures when we do this. When we do this diesel chemical transitions, what happens to the topologist exactly results in and so on and so forth. So we'll now let's have a look what we actually can achieve with the diesel chemistry approaches. So I divided roughly into three categories. Let's have a look at the amino acid mutations, nucleotide mutations and ligand modifications. I'm changing the system in the questions that we ask here and then and this all can be achieved by changing the term. So the protein mutations and ask, so can we predict accurately how mutations change protein thermostabilities. We took the sample protein very well studied by experimental it, it has been all mutated through and through experiment color the mutated in red, and we simply experimentally it has been mutated by and the folding pharyngeal difference. So we can calculate with those alchemical method that I showed before exactly the same pharyngeal differences and compare them to this. So this is a classical way compared to the delta delta G I'll show a few of those plots so let me briefly describe one of the X axis is a G so this pharyngeal difference in the folding pharyngeal difference between pharyngeal upon a mutation as measured experimentally is exactly the same just calculated so no experimental input just calculation and yeah you see as you see over trend is very, very, there are some outliers there is some deviation, which is below one kilo calorie per mole on average, the gold standard in the field, we are always aiming to reach that standard because we know that below that average error is difficult to force field issues sampling issues and also the experimental error we cannot be exactly exactly on on top of usually, but this limit where that we are able to reach with our methods. You can also change this to protein and I can simply modify thermodynamic cycle a little bit, and all of a sudden, I do exactly the same alchemical mutations but in this case, how, how strongly. I mean I said mutations influence the binding offer. In this case, it's up to to major compatibility complex. That's exposing a path there so I mutated all of these yellow residues and compare them again to experiment. We don't need to the details of this, but it's merely to bring the examples of how these how these collide and again we exactly the same accuracy below one kilo calorie per mole again so yeah let's just look at the left left scatter plot and experiment calculations are very good trend again several outliers we know that they will occur in our calculations and we often alleviate those by sampler or maybe including another force field, but this is a very traditional answer that we expect from. Now again, we keep on pushing this further introducing new questions, and another question could be, could we predict mutation in a protein could be drug resistant. So for that, at first we took HIV proteins as economic. So it's a it's a is usually targeted by drugs in the patients, however, that's what makes HIV so so difficult to treat it mutates, you give a some certain. And then it quickly. There's a few mutations and escapes those drugs are no longer effective, but and it's of course interesting can we rationalize it can we which which I the mutations are making these effect having these effects. So we this time we again thermodynamic cycle where we effect of a mutation so blue versus red, but in this case, not on the folding it before but on the. So, holo versus apple states of the system, and we took all the known or major known. And we used to treat HIV, see that again our experimental versus calculated free energy difference correlates very well, we don't have a perfect. We don't have a perfect. Let's say so we do have a little bit of an offset, but we're very happy because we can distinguish a strong resistance. Low resistance mutation in the drug bindings so we can already have a strong predictive our approaches. We even pushed it one step further, so it is quite intuitive that if the mutations in the active site so the, the previous mutations that we broke. Yeah, if you mutate something in the active site well it will clash with the ligand and it will have a. Direct effect. It's interesting to probe those red residues that I'm marking here that are far from the ligand so they don't have any interaction. And still they can influence the drug binding. So, again, we probed our free energy calculation technique in this. Correlated to the restriction factor which is related to the free energy difference. So to the ligand binding affinity. A very good agree simple we can again discriminate those that will be drug resistant from those that will be less drug resistant. A strength of these molecular dynamic simulations is that they, once we now we have established that we can a accurate predict. We actually have also access to the trajectories and therefore we can understand the mechanisms of what is have the system so I highlighted now the two residues. Well, let's say this is one recipe because it's a homo. We looked closer so why is this mutation, all of a sudden resistant to the drugs in HIV. And it appears that this mutation, it's a little bit to another hydrophilic smaller with residue mutation losing to the alien. What happens there, once we remove losing there. So we replace the alien, all of a sudden, there appears more space in this region of the of the protein. There isn't all of a sudden us in lysine can move closer to us party and form a soul bridge. All right, this happens. All, of course, the loop on which this lysine sits. When it shifts a loop, it destabilizes is indirectly actually transfers the effect of the mutation losing and destabilizes the ligand all of a sudden. The ligand. And the mutation exerts its drug resistant. So, in the end, as a very nice. Well, maybe a nasty mechanism of HIV nicely. It's by the molecular dynamics simulation. All right, we let's move on so I promise to show also something different in terms of what the biomolecules we can treat we can also mutate DNA so we can ask the following we identify how DNA nucleotide mutations would affect DNA protein binding so again I'm just more into the thermodynamic cycle in this case mutating DNA and I'm asking, will the mutation make the binding to the DNA and protein stronger or weaker. And for that we collected very many DNA protein complexes to to thoughts that these are very complex is as well, including the endonucleases and transcription factors, various complexes to look at the correlation. Very similar answer slightly slightly worse and quality than for protein mutate. This could be due to the due to the fact that. Yeah, maybe DNA force fields are not so advanced, not not so often, and therefore not so refined as protein force fields and also experimental data was from very range of sources so. Overall, we still can get a very good predictive accuracy for DNA mutations as well, and we can then further further questions now we have a good message to determine the differences. Can we do something what is often done experimentally so experimentally from, let's say, random binding essays, it is possible to define DNA binding profiles. So what people do they just allow random nucleotides in such an essay bind the random DNA see to the protein and countless from their experimental outcome they count how many times let's say I saw. I didn't in the positive or how many times I was going in a position seven and from these counts, it is possible then to reconstruct such a low thought that tells us what exactly nucleotide preferences are for every position when it's bound to the protein. We can we these established the counts are directly related to the well counts are also probabilities of finding some given position, and this again nothing else but the three energy difference, so we can again. Reestablish this connection between the energy differences which we can calculate the and the and such a probability probability profile. Here I'm showing several such profiles, and the just just by eyeballing I will explain in a second what they are, you will probably see that there are all have the same teach just have small differences between them in particular in particular features they do differ, but they're. The major features are the same, and now the top profile is the computational scan is from our computation all the one to these so for bottom rows are different experiments. Either it is a select six or or within binding my primary experiment. What I'm trying to illustrate here, I also have a quantification of this. Just I find it to see that experiments do have uncertainty, and this uncertainty is makes it a result virtually from the calculation results so in principle, what we compute we it falls well within the of the experimental measurement. All right, and with this we go to the last part for the for the presentation today. So we also can do arguably the most lucrative part, we also can do the ligand modifications, let's say, we are asked. What is the different ligand binding affinity for the red ligand and the blue ligand we construct the following terminal. And now we do the same transition just in this case for the ligand binding, not for them in acidification, not for the nucleotide but for the modification of the. Here I'm waiting with more detailed time cycle we don't need to modify the whole leg and we can just change one particular substitution. Experimentally, often it is difficult because you may need to design a completely different synthesis pathway for those. So it is difficult to expensive also, or maybe not even feasible to quickly get an estimate or so the synthesis of the molecule of the molecules, but we can quickly test that. And yeah, it's been it has a long history because it's such an approach with for the pharmacy industry because it would immediately have consequences to the and the lead optimistic. And the, of course, it has been advertised that this is possible already here I'm showing in 1987 so more than 30 years ago, where people show just on on several molecules that yeah, it works. You can give accurate answers with all these flashy high promises it took it was maybe just a lucky coincidence and and the long time until the field really matured to the to the level where it is now applicable with a routine drug discovery pipeline. 2015 paper where Schrodinger incorporated the the company showed that they do have now access to the routine calculations and they provide them, they established this new state of the art calculation pipeline. And yeah, we wanted to see our PMX and drama can also reach the same accuracies as this Schrodinger corporation, and we made our own application by collecting a large number of pharmaceutical relevant protein ligand complexes and computing simply computing the binding free and we can compare them directly to the, what would we get if we were to use this Schrodinger's expensive commercial see that we get on the right side it's again, as usual, experimental versus calculate the Delta Delta G's for in this case for ligand binding to the proteins and the in terms of the average and certain average unsigned error, we are again in the regime, identical correlation we are again within the uncertainty limits from one between the two methods, you have slightly larger error. And this is mainly because of the methods that we're using to calculate the uncertainty we simply try not to estimate the uncertainties as we run independent repeats from our populations. But otherwise, we are able to reach the same factors as the commercial. And we also can do now, the last two minutes I just want to show that now a talk about mutating something so mutating one amino acid to another mutating nucleotide one to another or changing a small part of the of the ligand, but we can also principle we can do absolute binding operation. Let's say we don't want to mutate just or modify this a part of the ligand we can in principle disappear. Alchemically, just modifying the thermodynamic cycle a little bit and it requires some other computation, some other theoretical and computational considerations, but in principle if we ask the question, can we bind, can we bring this from, from solvent so from water into the binding side of the protein. It is this transition of binding is it's computation challenging but we can do it also all chemically we can disappear the benzene, we can restrain it so place it into the pocket. And then, by removing the restraints and taking their contribution into account a couple system, and we cover exactly the same delta G, but in this case, it's not a relative but an absolute binding. So we, it is also possible. We have probed it in a large large scale scan. So if I just show, we selected from our previous investigation a subset to have many values to play with, we also can recover a very good agreement with experiment there is there are more of the red points that we saw before, but in principle that overall accuracy is very, very comparable so it is also feasible, just the, the, the bottom line that they wanted to bring today. And, yeah, I would like to thank everyone who was involved in studies and tell. Yeah, also other findings, and I'm ready for questions. Thank you because I will encourage everyone to write your questions in the chat. And we can take it from there. So, have a question from either how much computational spend time are needed for PMX of a protein complex. For instance, I guess the question is if you have complex and you want to calculate the absolute binding energy interpretation of the question. Okay, let me give a bit of an overview so now these calculations that I showed all of using non equilibrium free energy calculation protocol. So, the timings that we would need for that are approximately say, in orders of up to 100 nanoseconds per PNG value in total. So, irrespectively of the of the question that whether it's protein complex or protein ligand complex. In total, it would take, let's say 100 nanoseconds or seconds to get the things that I reported. Now, it of results in a different user time needed, because the larger systems if you're interested in protein protein complexes will have many atoms you will. You will have a lower lower simply lower nanoseconds per day output for that. Yeah, but but that's probably another question. Is there a big difference because if you're in a small legal compared to this much larger molecule from a complex that it will introduce additional. Yeah, that's also a good point. Of course, the perturbation size also matters. We always play an perturbation that are not as small as possible because then we simply retain the system as close to equilibrium as converge quickly. Of course, for the absolute free energy so I only touched very, very lightly on that, but those would be much more to converge them one needs a 10 times longer something than them to converge relative changes that's why relative changes are so they are easy to. And the perturbation sizes of. Yeah, I don't know up to up to atoms should be should be easy to punish. Thanks. So there's a question from Gregor, who's asking whether accuracy compared with MGB SA or MMP BSA. Yeah, certainly these these methods of chemical math are more accurate rely on this assumptions that the that the approximate methods. Also, yeah, we don't have a problem of incorporating or combining add adding with the anthropocontribution.  So it's already cheating is involved and then when you add now a little bit from another method and this Frankenstein of course can be fit into a good agreement. But it's by no means robust and rigorous. For question. You see difference in performance of the various force fields that you have used in the. Yeah, that's a very good. We do see difference with, let's say, probably amber for the force fields and charm family for our performing best in our experience. Maybe PLS is. This is a little bit less accurate, but we are always using the non commercial PLS version. Yeah, publicly available. And what the interesting thing that we've noticed that the force errors usually cancel outs force fields do make errors. But they are often in different directions or in different parts of the mall than they reduce one another. Mainly, I could explain this by the fact that different force fields molecular mechanic for currently have very simply simplified forms. So there is only fit with such with such high highly restrained functional forms. And, for example, amber is trying to the optimally the electrostatic potential surface representation of small molecules. At the same time charm is not trying that and to optimize the the interaction with water for small molecules and coordination with the water. These two may not be necessarily compatible in one single force field but if you were to simulate to obtain else with two different force fields, have a much more accurate representation of what's overall happening in the system if you're if you're simply able to find that console both of them. It's difficult to say how to find this consensus, but for free energies, it's very trivial in principle, just a value so you get one value from amber. Another value delta G from charm, and we noticed that these two force fields are very well compact simply calculate average of those. This will be always at least as accurate as the more accurate force field alone. Thank you. We have another question regarding outliers. So in the examples that you're showing there were outliers, but do you that people validate or benchmark results for every application today? Yeah, certainly. It's always very good to very good to have some experiment data on the system or a similar system to compare to before starting to make predictions. So there could be, of course, with either with the force field or with a particular setup. For example, just from the personal experience, always an issue if there are additional factors involved, simply the accuracy drops significantly. But if you have a handful of data to compare, it's always good practice to establish the relation. So what will you expect from your prediction? Sorry, I was muted. Thank you. So I don't see any more questions in the chat. So now it's 1030. So I suggest we take now the half an hour break before we continue with the practical demonstration at 11 o'clock. So thanks everyone. We'll have a break and we'll come back again in 30 minutes.


Cardio is overrated for cutting and dieting. Cardio is overrated for getting lean. Good morning. Good morning. Come to story time around the fire. We're talking about bulking.  I got six sets of two. Bulking baby, that's what we're doing out here. You just don't want to get shredded. What I want to do and what I'm doing is different, you know? Sometimes you got to do what you got to do, whether you want to do it or you don't want to do it. Anyone ever tell you that? Yeah, you tell me that all the time. My mom used to yell at me all the time. Sometimes, Mikey, we got to do things we don't want to do. That's what you say to us.  Dan, Seabass, and Kyle are hanging off of a cliff. Who are you saving first? Oh, who are you saving? I feel like I'm saving Seabass. Well, first of all, I probably can't pull any of them that regardless of. Yeah, you can't. You think so? You got to look 430.  Okay, what if Mike's hanging? Oh, if it's just, is it just you? No. Do as I say, not as I do. So with bulking, right, there's things and tips and tricks that I do that we're going to go into in this video. Like everything else we say, as strict as your goals are, as strict as your plan and your action and execution need to be. If you're trying to be Mr. Olympia, or you're trying to be the greatest power lifter that ever walked the planet, you should probably have a stricter guidelines and plan than I have. If you just want to build some muscle, some strength and look pretty good when you decide to cut back down, which we're doing in January, stay tuned for transformation challenge. Not really a challenge, but documentation. Then you can get away with a couple other things. I don't know exactly one, two, three tips, but something easily I do. The team killed it on our launch this week. Seabass and Aview are here grinding. So I bought cookies for the team and I ate a cookie where when I'm dieting, I don't eat no cookies. One simple trick that I do like to do is obviously when we're dieting or cutting, I'm tracking every single thing I eat when I'm bulking or it's off season or I'm not as worried about my body weight. I will just track my protein. So right now I'm making sure I get 200 grams of protein a day and then I eat kind of whatever else I want. Trying to get some veggies in, trying to get some water in, and there's some basic rules I still follow. Like I try not to drink my calories. Maybe I'll do a cappuccino or a latte or something small, but I'm not chugging Gatorades full of sugar. So anything that's crazy calorie dense, even like the peanut butters and the guacamole's and stuff like that, I still avoid when I'm bulking just because I know how easily it is to go from 3,000 calories a day up to like 6,000. Little changes I make and I enjoy switching from like chickens and turkeys, 99% ground turkey. Now I'm eating a little bit more beef. I want to taste a little bit better. It's got a little more nutrient dense, but it does have more calories. So I eat a lot of 90, 10 beef, but I still, I still meal prep. I'm still like this week.  Like a tip necessarily, but I guess like a suggestion or something that we should think about is depending on how drastically you've dieted, right? If we've gone from 30% body fat to 25% body fat, this may not apply to you. No offense. You may have some energy stores still left in you and you may still be feeling good. But if you've gotten, you know, below 15 or something or like as you guys saw the last transformation I did, I dropped like 65 pounds, which is a significant amount. And I got fairly lean.  I'm not JPG.  Just a dude who lifts weights here hanging out with y'all. But I got fairly lean and I had to diet pretty drastically based on my lifestyle, genetics, et cetera. So my energy was low going towards those last like two months of training and my training didn't like suffer a ton. Leverages got weird. So strength wasn't necessarily there. I traveled a bit. So that affected it. But my general, how hard I could push on all my lifts went down. All right. I have less calories in less energy in less energy. I have to put out. So when you bulk, you may not just automatically hit a hundred pound deadlift squat PR. But I do know and think you'll be able to push your volume and strength on your accessories a little bit more me and see Basser even just talk about that. You know, a lot of times as a strength athlete or even a bodybuilder or aesthetic trainer, anyone that goes to the gym, we get acclimated to the same weight. So we use the same thing week after week.  And we just found a weight that's comfortable, but challenging. We just stay there.  It's time to get a little bit better. And so how do we build more muscle? How do we build more strength? Start chipping that thing up. I use the hammer strength chest press one of my favorite machines and I'll add fives, 10s, quarters, when I can, I'll do another extra set at my heaviest load where typically I'll do a top set and I'll back down one or two sets lighter weight. If I'm bulking or right now I'm feeling better. I'll jack that up. We're just talking about I do belt squats. And Joe, shout out to Joe standing, my coach right now has it prescribed it like RPE eight. And so with a heavy band, I'll do two or three plates. But now that I'm eating a little bit more, I'm a little more acclimated. I'm a middle of the training cycle. I feel a little bit better. I'll throw on three plates in a quarter and you got to start to chip yourself and push yourself. And that's kind of the benefit of the gym. And that's why the gym is our microphone. And that's why I'm here talking to y'all is because when we start to realize how we can push ourselves, how we can succeed, how we can build a goal or have a goal, build a plan and then execute that plan and push ourselves day in and day out showing up. That's where the magic happens in bulking. Sometimes it's easier to see and feel because we can add weight where when you're cutting sometimes we can go a month and because we see ourselves in the mirror every day, we don't feel that we're making progress, but we can visually see and feel progress as we're lifting and bulking. And so that's another reason I'm a strength training advocate is because I don't care if your goals are strength or if your goals are aesthetics or health or I don't really care. When you focus on strength, regardless of all this, you can continually see the steps it takes to get better and push yourself through progressively overloading. Cardio is overrated for cutting and dieting. Cardio is overrated for cutting and dieting. Cardio is overrated for getting lean and Cardio is underrated for building muscle and a bulk. Now, I think Cardio having a baseline of it will help our work capacity. We can dig into GPP and just general fitness later, but having a base of healthy heart, healthy cardiovascular system will benefit you in the long run of life.  And if you no longer bench press, we can't grow big titties. And that's what we're trying to do here. So while you're bulking, I suggest just like everything else, five to 15 minute warm up, 10 to 30 minute cool down, call it cardio, call it GPP, call it a recovery walk. I don't care what you call it, just do it. And same thing goes when cutting. Let's manage our calories better.  I get it. I'm there. But when we're dieting, nutrition, when we're bulking, base level of fitness, because then also when we flip it around, we're probably going to cut eventually and we may need to up our cardio to supplement our dieting. And then you're going to be so out of shape that it won't benefit us by the time you get in shape, your cuts going to be over. Hopefully that makes sense. Do a little bit of walking, do a little bit of fitness, be a human, take care of your heart, take care of your vastness.  The truth is our metabolism is way overrated. So the variance in what each of us need to live every day, the genetic difference is so small. The bigger difference is how much you move or not. And like me, I don't move a lot. I train. I try to walk a little bit. And otherwise I'm working at a desk all day literally sitting down all day long, where if you're a nurse or you work in a warehouse or any job, really, you're going to end up walking more.  All right, you're going to end up walking. And although it seems little, it can add up really quickly and you can burn a lot more calories. So those people, and then the other part of that is the mental aspect, which we could call genetic or nurture. It doesn't really matter where some people don't realize how many calories they eat. And a lot of people, either for too much or too little, some people like, dude, I eat so much. I can't gain weight. You're not eating that much. So tracking while you're bulking seems kind of silly for a lot of us, but for a lot of those hard gainers, which I hate that term because it's literally false. You're just not eating enough. So for those folks, ignore tip number one and start slamming peanut butter and avocado, et cetera. For you to track your food and get an idea of how much you need daily is going to be key for a bolt. Certain of us, right? There's always those people that don't track, you know, I'm making this up at 60% of us are going to gain weight when we don't track. 20% of us are going to lose weight. We don't track and 10 to 20% are going to stay the exact same. If we don't track, you have to know where you sit in that kind of spectrum, mentally, physically, et cetera. And if you're dedicated enough, if you want to build muscle, if you want to build strength, you're going to have to track while you're gaining weight.  And for someone like that, you may have to meal prep and get more meals in, you know, the six meals a day has nothing to do with metabolism has nothing to do with digestion. You're not going to be more efficient. That's just some old school bro science. There is a little bit of truth to dosing out your protein evenly through the day that will help build muscle, but it's miniscule. I wouldn't over exaggerate it. But if you have trouble gaining weight, you may need to eat five, six, seven meals a day, smaller, medium, larger meals to get in those times.  So we'll catch that in the vlog for sure coming up. Appreciate you guys, man. 3SB.co for all the new fits. My favorite zip up hoodie. Telling you cozy. I'm telling you cozy. And I know I made it so it sounds like I'm biased and I am, but it's honestly the best because I'm picky about what I wear. I'm picky about what I share with you all. If you all followed for the last 10 years, you can count in one hand the amount of things I recommend and sell to you. So if you want to support, you want to be part of the crew driven by community culture, 3SB.co, good company. Southern Mike, we're out of here. New videos every single Monday and Thursday. Catch you soon.


Hello, everyone, and welcome to Playground AI. This may be the best free image generator on the internet. And I don't say that lightly. And when I say free, it's basically 1,000 images part of me per day, which is technically free. But if you go over that, you just create a second account. So whatever. Let me show you how to do this. Let me show you a little bit about it. If you look at this is the original screen, PlaygroundAI.com links in the description. Just look at the quality of this. This is generated using this software. And it is wild. It is browser-based, so you don't have to download anything or go anywhere. You just go right to the website and off we go. OK, here we go. Let's start it off. Let's click on Create and let's get the ball rolling. This will be a very brief overview of the product if you want to log in and try it out yourself. I recommend it because there's lots here and it's hard to go over everything all at once. So first off, the big thing is prompting. This is basically, like I said, an AI image generator. And it relies on your prompt. Now, you can type in anything you want and it will do the best it can to render it. That said, it does a pretty good job most of the time. Every once in a while it gets it catastrophically incorrect and those are hilarious, so in a lot of ways it's worth it. So keep that in mind. I'm just going to type in here like I did. French Bulldog on a beach, Blue Lake, Mountains in the Background, something simple like that. And then I'm going to click Generate. And then while it generates here, you'll see here it counts down how long it takes. I'm going to talk a little bit more about some other options while it does that. Oh, look at that. I did that quickly and I like it. That is great. The left eye here is a little bit too dark on the left side of his face, but that's okay. So right there, we've got a great image. And remember when I said it was free, it's free. Just click on this little button here, download it. Click on what you downloaded. You got the image right there. It's a little guy, but that's okay. If you want to make the images bigger, let's, well, let's just get right into it here. First off, let's go to the model. There are three models you can use. Two of them are technically free. Dolly too, I believe used to be free, but now they charge you. So you could stare clear of it. Stable diffusion 1.5 gives you the most options, but stable diffusion 2.1 is also quite good, although it just gives you a lot fewer things you can do with it. If you want to create different sizes, I created a nice simple square, but if I wanted a bigger one, and why not? It's always better when it's bigger. Why not? Just click on that. If you want it wider or narrower, there are a few different options here. 768 by 512, then 512, 768, et cetera. Okay, good. Prompt guidance is basically like seeding and styling. Like how much does the software have to stick to your prompt? How much creative leeway are you giving it? Higher values will make your image closer to your prompt. Lower values will give the software or the program a little bit of freedom to do what it wants. And then quality and details really quickly here. It defaults to 50, and that's a pretty good one. If you get something you really, really like, you can go all the way up to 150, but that said, you can only run at 150 for a few ones before it starts asking you for money, or alternatively you just drop it back down to 50, and you stay free. Okay, the last thing I wanna show you on the right side here is this right here. Very, very important. Instead of the number of images setting to one, set it to four. I mean, if you're gonna do something, do it four times, it's better to have four bites at the cherry than one. And it's the same cost, nothing. So there you go. Okay, the right side is out of the way. Let's look at the left side here. So we add in our stuff here. Now here's the other thing you can keep in mind. You can remove things from the image. For example, in mid-journey, if you type in, make a character and it comes out with glasses on, sometimes you don't want it to wear glasses. You don't want it glasses free. So I would just type in here glasses, so that it's being removed from the image. In this case, I don't need to, but something to keep in mind. And then the big one is filter. This is a little bit of a misnomer. I don't think it's quite what it means if you think about Instagram filters and stuff like that. This is actually more like style. This is more like, you know, if you were to make something in the style of using mid-journey, for example, this is kind of that thing. So if you want to make it cartoony, if you want to make it woolly, which is like emigrammy or something like that and the other program, I just learned about it the other day. You want it to make it anime, et cetera, et cetera. You want it to be an oil painting, cinematic. You know, it's all in here and you can try these out. They're quite good. In this case, I'm going to go with, I don't know, what we just went with nothing, but if I wanted to make it cinematic, we could do that and then I can click on regenerate and see what happens. We're going to get four cracks at the cherry here because look, we set the number of images to four and now we're regenerating with the same prompt, but we're using the cinematic filters. So let's just want to show you the quality of the filters and boy, these are pretty good. Yeah, you know what, not too bad. I don't know why they added in a second bulldog, but so be it. Look at this one. This one's very, very nice. We're going to click on this one. And again, it's free. Just click on download, but there are other options. If you like this one, but maybe you want some variations, add in some variations. I clicked on the plus sign. Variations are coming. If you want to edit this or make alternative actions to it, just click on this button. For example, there's edit, there's make it private. I don't know why you do that, but go ahead. There's in painting where you can sort of paint around it and kind of make some changes to it. Face restoration. If you love the image, you can upscale it for X. There's stuff there. You can click on the edit button here. Looks pretty cool, but maybe I want to mask out something. Maybe his eyes should be a different color. I wouldn't do this, but I want to show you the techniques. So this is where it kind of combines with like Photoshop skills or Photoshop. And then watch. You can change the size of the brush. So I'm going to make it a very small brush. And I'm just going to kind of dot over his eyeballs right there. And if you zoom in, you'll see here that we've kind of just put some red on his eyes. And now I'm going to add another instruction. So I'm going to make his eyes pink, something like that, okay? And then click generate. And because we masked it, it's going to try and make that area pink. So I've got this good looking bulldog. And hopefully he comes up with pink eyes and it did not do a very good job. So I apologize. But again, what can you do? We could try again. We can make it bigger. So we'll make it like this area just to see if we can get this to pink. And I'll be in make it pink and click generate. And again, I just did this very, very quickly. I just want to show you what this program is capable of. And hopefully it does it. And let's see if it wins me an award. And it does. He's got a pink forehead. How awesome is that? Okay, so there you go. I'm going to cancel out of this because I don't want to pink pupper. But I wanted you to see it. Now here are the four generations of the variants that we created while we were sort of chilling here. And I like this one. He's got a pretty wild-looking face on him. Whatever. All good, right? You want to download it? Click download. Remember, all those other actions. You can in-paint it. You can do these changes to it. Get a thousand of these a day. Amazing software. Let me know what you think. Thanks for watching.


Thank you for being here. I don't have slides, so I'll just speak to the points. It's a joint paper. I'm writing with Dr. Leanda Kanbigi who also works with the Center for Migration Studies here at the University of Ghana. So we are looking at the challenges of stakeholders in the return and reintegration of Ghanaian returnes from Libya. So a bit of introduction, then history of Ghanaian migration to Libya, conceptual framework methods, findings, and then the conclusion. Return migration has been in the Ghanaian development discourse since the 1990s when the country regained economic, social, and political stability. There are various programs to encourage the return of Ghanaians abroad and friends of Ghana for development purposes. Some of these programs include the Emancipation Day celebrations in the 1990s. We had the Homecoming Summit in 2001, and then the establishment of the Non-Residents Ghanaian Secretariat in 2004. And then there was the recent Aspera Homecoming Business event that happened earlier this year. Many of these programs are targeted at returnes from the global north. And this is in spite of the fact that the majority of Ghanaian migration are to other African countries. The emphasis on returnes from the global north is perhaps because of the amount of remittances that comes from the global north. The Bank of Ghana recorded that between 2004 and 2008 remittances from the global north, so Europe, US, Canada, accounted for between 90 and 97% of total remittances received by the country. We argue that because the emphasis of return migration is on returnes from the global north, the states and other non-state stakeholders were ill-prepared for the sudden return, and therefore reintegration of Ghanaian migrants from an African country in this instance, Libya. We interrogate the role of the states and these institutions in the return preparedness and reintegration of these Ghanaian migrants. Before I delve into our conceptual framework and then find in just a brief history about Ghanaian migration to Libya, Satya Spiga, 2005, notes that Libyan oil exploration and production, together with its mines and farms, served as pool factors for most of the immigration to the country from other Sub-Saharan African countries. Ghanaian migration to Libya was also boosted by bilateral agreements between Ghana and Libya to send teachers to teach English in the country. So 200 teachers were sent in the mid-1980s. This subsequently served as a pool factor for both skilled and unskilled migrants to move to Libya. Libya subsequently transformed from a destination country in its own right to a transit country for many low-income Ghanaians. They have notes that, though the Libyan 2006, he notes that though the Libyan government relaxed its immigration rules because of its pan-Africanist role in Africa for some time, irregular migration was on the increase. So the Libyan authorities intensified their immigration crackdown on irregular migration. So thus, prior to the political crisis in 2011, they were already dealing with irregular migration. So that between 2000 and 2012, 12,201 Ghanaians were deported from Libya. Klaj 2017 also argues that the political unrest and agitation coincided with these immigration policies already. So the Ghana Immigration Service reported that in 2014, for example, 1,415 Ghanaians were deported. So this makes Libya the country that deports Ghanaian migrants the most. So many of these migrants, when the political crisis unfolded, they were put in between the walls with both sides of both Gaddafi forces and anti-Gaddafi forces, accusing them of siding with either forces in the conflict. So many of these migrants who occupy very low socioeconomic positions in Libya were caught up in this. And there was a need to evacuate them. So then we look at the role of the states in doing this. In understanding the role the states played, the states and other non-state stakeholders played in the return on the integration of Ghanaian migrants from Libya, we draw on Casarino's 2004 conceptual framework, where he questions the adequacy of the main economic theories of migration, so the new classical theories and the new economics of labor migration theory. In understanding migration and return decisions beyond the economic factors. So he argues that decisions by individuals are likely perceived as income maximizers without taking into account the contextual factors within which individuals make their decisions. So he argues for a broader framework, structural framework, in understanding individual decisions that migrants make with regards to return. In this paper, we are not so much concerned with individual decision-making processes by the migrants. These have been explored by other scholars. So we have Candeligui 2017, Clyde 2017, and Mensa 2016. They have looked at individual decision-making processes of return needs. We want to move this theory forward by focusing on the structural forces at play, particularly when we consider that irregular of individuals were caught up in a situation where their plans for return were thrown into disarray. What role did the states and other stakeholders play to facilitate their return and subsequent reintegration? Research on return migration and development is mixed with some scholars arguing that it needs two developments. So Waba, for example, is of this opinion. And then others are more skeptical of this opinion that return migration can foster development. So we have Nimbex Orson and others 2012. So we are arguing that when people return because of a crisis situation and they are put in a situation where their plans have gone awry, if the structures are put in place, it could facilitate their reintegration. Of course, this is by no means saying that migrants do not exercise or returnes do not exercise agencies. And as we see later on in the conclusions, because many of them were sort of unhappy with the return and reintegration processes that were put in place by the states, many of them took their decision to re-emigrate back to Libya, even though the conflict situation was occurring. So then by way of methods, we used many qualitative research methods, in-depth interviews and focus group discussions among 32 participants from six categories of actors. So we spoke to return migrants, family members, civil society organizations, community leaders, intergovernmental organizations. So the data was collected both in Accra and in the Bronga Haful region, which is Accra because state institutions, many of them have their head offices here, then intergovernmental organizations as well. And then we decided on Bronga Haful because of the 19,000 Ghanians who were deported from Libya, nearly half or almost more than half returned to the Bronga Haful region. Yeah. Okay. So then let me quickly go over the findings. So we look at the various state agencies and the road they played to facilitate the return of these migrants. So for example, with regards to the Ghanian state and its agencies, one of the things they were tasked with was to evacuate Ghanians from Libya. But Ghanian state or Ghanian government have a policy and institutional framework for the evacuation of people in crisis situations. So this meant that Ghanian migrants in Libya were among the last to be evacuated. So as a former diplomat at the Ghana Embassy in Libya indicated, the mission therefore, the Ghana Embassy in Libya had to rely heavily on international organizations to provide much needed evacuation terms, of course, providing airplanes, et cetera, and healthcare during the evacuation. The other thing, the embassy, the Ghana Embassy in Libya also faced had to do with what we are calling trust deficit. So the Ghanians in Libya didn't trust their embassy and the vice versa. So many of them therefore contested claims that the embassy did all they could to help evacuate them. So as a 52 year old returning put it, the embassy's officials are useless. They don't help anybody. The officials in Libya do not help at all. The other thing the states tried to do was to provide migrants or returnees with a source of livelihood. And they could have done this by speaking to employers and businesses to take on these people with regards to the skills they have acquired in Libya. But what they did instead was to provide financial assistance for these people who had returned to Accra to return back to their villages and towns. So they provided an equivalent of 25 cities, to 30 cities, which is roughly five, six to seven euros, and for them to return to their home countries without taking into account the fact that many of these people perhaps lived in Accra or had other social networks in urban areas that they would want to stay with. So of course the amount was woefully inadequate to facilitate their return to their hometowns and villages. And then also another thing they tried to do or they tried to do was to host these migrants when they returned. And so there was no facility to immediately house these returnees. Instead they were hosted at a military stadium, Elwak, and thus expose them to elements of the weather and it made their registration processes and immigration processes very, very difficult. And so it's also hampered the performance of their duties including providing therapy, psychosocial and post-traumatic stress, and therapy for psychosocial and post-traumatic stress disorders. Of course this also exposed the officials of the national disaster management organizations, NADMO, who were in charge of these two elements of the weather as well. And we are unable to do their job properly. So with regards to the role of governmental organizations, like I said when I was discussing the role of the states, they helped with the evacuation of Ghanians from Libya. But their job was made difficult because of the lack of data and then financial constraints. Lack of data in the sense that it was difficult to estimate the number of Ghanians who were going to return. So therefore to provide the needed planes and other modes of transportation to bring them to Ghana. So without knowing the scale of mass return, it was difficult to envisage the number of people returning and the sort of support they might require. So the magnitude of this challenge is captured in what some of the staff of the International Organization for Migration Ghana had to say. So they said when we were first putting together those contingency plans to assist these Ghanian nationals to return, I think the Ministry of Foreign Affairs came up with a figure close to 5,000. Then they made our scale is up to 8,000. Then they said that to be on the safe side, let's expect up to 12,000. That would be the maximum. Now, as you know, close to 19,000 did arrive. Not to talk about those who did not arrive. Those who went to other countries. So the issue of data and statistics, we need to really look at. It was a major, major challenge. The other thing, intergovernmental organizations like the IMO and UNDP tried to provide was training programs. So to train these returnees with regards to making a livelihood for themselves. So the International Organization for Migration, for example, has returned packages for voluntary returnees where they undergo training, et cetera. But they didn't have something for return migrants who were forced to return. But they made provisions, so then they provided some form of training for these people on a temporary basis in order to contribute on a temporary basis to help them cater for themselves. And the return packages they gave, the training they gave to these people did not include cash with the assumption that they did not want to enable them to find their return to Libya. And the assumption was that we're going to do this once they were provided with cash. So they focused on training, providing training with regards to livelihoods, hairdressing, farming, et cetera. And then business startup kits as well. So the role of civil society organizations and community leaders. Many of the returnees we spoke to were happy with the role of civil society organizations and they were happy with the role of the community leaders. So for example, the director of Doma FM station, which is a local radio station in the Bronga Haful region, used the medium of radio to establish contact with many of these Ghanaian migrants who were trapped in the political crisis in Libya. So they could establish contact with their families. So they got into contact with their families and they found out how they were doing, et cetera. They also used the medium of radio to get in contact with their members of parliament and other government officials by phoning in into live broadcast at the peak of the crisis and making direct appeals for evacuation. Their harrowing stories helped governize public opinion in favor of government action to charter aircraft, to extract them from Libya. So with regards to the role of another civil society organization, we had this school in transit. Together with IOM, they helped to provide the training programs for these migrants, but it tended to be ad hoc and on a limited scale. So for example, out of the 900 returnees who returned to Incoranza, only 50 benefited from the distribution of the startup kits. So though laudable in its intentions, the school in transit were also accused of favoritism because it also gave supplies to other poor people in the community and this agitated and upset many of the returnees because resources were scarce. So we include a role of the family in our discussions of stakeholders because family members are both beneficiaries of remittances and barriers of the burden of hosting forced returnees. Unplanned returns from countries in crisis are promptly curtailed sources of remittance income and sometimes exact expectations of unfordable reciprocal benevolence from family members. So adverse effects recorded in this study on families with regards to the return, with regards to the unplanned return of guardians from Libya included loss of social standing and respectability, being saddled with caring for mentally and emotionally distressed returnees, sharing limited family resources with people who have returned as failures because their plans didn't go through. And then also, so some of these frustrations were summed up in what some family members said. So for example, Adjoa mentioned that we started everything from scratch. I mean, two of us, no support from the church, no support from the community, no support from governments, no support from the assembly, no support from anywhere. Now the major challenge is that he's unemployed. Aside from farming, he does not do anything. I would like him to go back to Libya. Everything shows that he's not a happy, a happy man. So because many of these returnees were unhappy with the way they were handled, with regards to state and non-state stakeholders, many of them returned to Libya at least once during the conflict situation. So out of the 11 migrants, we spoke to six returned to Libya during the heights of the crisis. So quickly by way of conclusion, this paper concludes that even though trapped Ghanaian migrants were eventually evacuated, there was to a large extent institutional failure involving both Ghanaian state agencies, intergovernmental organizations, and local non-state actors who were ill-prepared to assist migrants on their first return. That's frustrating their reintegration into various communities in Ghana. The coalition of credible migration data and real-time access to migrant stock, as well as migrants' flow figures are critical to international responses to crisis situations where migrants are implicated. In addition, the evacuation, repatriation, and reintegration of returned migrants are shown to be collaborative tasks that are needed to be executed by diversity coders at different spatial levels. This study therefore highlights the need for a clear national policy on evacuation of migrants implicated in crisis situations. Such a policy needs to be costed and budgeted for with clearly delineated mandate assigned to relevant stakeholders and adequate training provided to facilitate quick and efficient response in these situations. Also, in order to enhance the efficiency of the National Disaster Management Organization, the organization needs to coordinate the management of disaster incidents. Moreover, a well-resourced local disaster management organization with access to funding sources beyond government intervention would greatly improve its operational capabilities. In line with this initiative of NADMUN to establish a disaster management fund should be strongly supported. Also, the construction of a purposeable reception center comprising among others a reception unit, a psychosocial orientation unit, and a temporary camp and offices for medics in order to facilitate the process of screening, profiling, and record-taking of returnees from countries in crisis in a more humane manner is critical.  I'll leave it here for now and then we can follow up with the discussions later. Thank you.


This is Witchbase news for Friday the 27th of March 2020 I'm CMDR Buur In this week's news Frontier break their silence on fleet carriers We have new details on carrier costs how they work and when you can get your hands on one If you enjoy this video consider subscribing and also click the little bell icon to make sure you don't miss any of our future videos Frontier finally broke their silence on elite much delayed fleet carrier system last night with the announcement of some new details on the behemoth ships a date for the live stream reveal as well as incoming beta tests before the carriers finally go live in June I'll say that again Frontier have confirmed that fleet carriers are arriving in the live game in June You'll be able to test them out before then however with two beta tests the first of which starts on April the 7th for PC players with a second beta test for PCs and consoles in May the much speculated purchase price for the vessels has also been revealed and it's an eye-watering five billion credits Frontier clearly designing these colossal ships to be a significant investment for their owners The carriers come with 16 landing pads of various sizes and will require a new fuel tritium to power their frame shift drives which are capable of jumping the Titanic ships up to 500 light years those jumps can also happen whenever the player chooses but there is apparently a build-up and call-down period between the jumps as part of the announcement across all Frontier's social media platforms they also stated that since the initial carrier reveal last year they have revisited the design of the vessels cookie cutter loadouts and instead opted for a more flexible customization design as with all the other ships in Elite Dangerous they're saying that this design rethink will allow players to make their carrier unique and allow for player-to-player interaction the like of which Elite Dangerous has never seen before the post from Frontier then goes on to say that players will be able to manage their carriers finances by setting tariffs and adjusting the buying and selling prices for commodities traded at the carrier's marketplace it's safe to assume from this statement then that your carrier appears to be designed to make money for you via the commodity market given the use of their phrase and i'm quoting directly here player-to-player interaction like you've never seen before in Elite Dangerous unquote you could speculate that carriers will see the start of a true player-to-player trading system and player-driven economy an email announcement from Frontier later on Friday stated and i'm quoting again fleet carriers provide new opportunities for expeditions and long-range travel the ability to reload refuel and repair for conflicts and campaigns as well as tools and storage for mining operations far from home that last line is of particular interest tools and storage for mining operations the direct implication here as we've speculated before on this channel is that carriers will essentially introduce player storage space meaning you can finally swap to a smaller ship while hanging on to your cargo which will be stored for later use a press release appears to have gone out to gaming news outlets as well that includes one extra detail not mentioned in Frontier's other communiques quote carrier owners will be able to set tariffs on all goods traded on board to support weekly upkeep costs if owners consistently fall behind on their payments the fleet carriers might ultimately be decommissioned and sold for parts unquote perhaps understandably the words weekly upkeep costs and decommissioned did cause a degree of anxiety in the community but as we've stated above and this press release from Frontier seems to support the idea the carriers do appear to be able to make money for their owner meaning in theory at least they could be self-sustaining with enough traffic visiting them in a direct response on twitter to help calm the upkeep and decommissioning anxiety Frontier did respond with the following quote you need to incur on huge in capital letters debt for that to happen and our devs know that you'd want to go on vacation from time to time unquote that statement does seem to imply that you'll need to essentially ignore your carrier and indeed the game utterly for a significant period before decommissioning becomes a consideration but without more clarification on running costs for these vessels it's hard to know for sure any developer worth their salt will know that there is a balancing act between any potential grind to maintain the upkeep of something like a carrier versus the reward for owning one by the same token to make the carrier worth having and feel valuable you'll need to put more than two tomatoes and a bag of cats into it every week given the hints that we've seen in the last 24 hours on what carriers will be bringing it'll be particularly fascinating to see how organizations like the fuel rats operation Ida the East India company and the AXI use them given that Frontier have revisited the carrier design and changed how loadouts are going to work and that they are now finally actually giving details on cost fuel upkeep etc coupled with a clear roadmap on when we can get to try these things out before they actually launch in June it does feel like they're listening to the community and reacting accordingly the beta test will be a fantastic opportunity for the player base to grapple with carriers try everything out and then give constructive feedback on what they like and what they don't like and I for one fully intend to avail myself of that opportunity we're getting further details in a live stream on April the second and then the beta for carriers on pc goes live just five days later if you've enjoyed this video consider subscribing to the channel and be sure to click the little bell icon to make sure you don't miss any of our future videos and if you want to help support the work of this channel you'll also find links below to patreon that's it for now thanks very much for watching we'll be back later this week with more videos until then 07 commanders follow the greens on the way out and do keep clear of the toast rack we very much look forward to seeing you next time


Welcome to the original gangsters podcast. I'm your host Scott Bernstein quick kid edition. We're going out to New York and Talking about the West Side Genevieve's crime family court news Carmine Pizza Polito Acting capo of the Genovese's little Italy crew also has territory other parts of Manhattan and Queens cop the plea from his Racketeering sports gambling extortion case this past week Sentencing guidelines for this case are pretty Moderate and I would say that this is a great deal for him. He's gonna do anywhere between 10 months in three years Probably end up doing about I would guess 18 months on a case where he was threatening to kill people. They got him on Recordings telling someone that he was gonna break their face and another debtor He was telling he was gonna put him under a bridge Carmine Polito is a serious individual he is a big-time earner and He's known as a big-time tough guy and as we know in this world that can be a Powerful combination and lead you up the ladder in any crime family Carmine He's a favorite of You know a lot of the leadership of the Genovese crime family and has been going back 30 or so years he beat a murder rap Back in the 2000s after being convicted of it. It got tossed on appeal beat it at trial. No seven This is a guy that Back in the day before he got his button Or right when he got his button It's interesting to note that he he had hundreds of thousands of dollars of gambling debt, but you had a number of Very influential figures in the Genovese crime family fighting over his services and there were a bunch of sit-downs multiple crews trying to claim him and For a guy that had a lot of debt it shows you how valuable he was and He's very well liked by barry Barney below mo the godfather of the Westside the boss of bosses. I always laugh when I say that I'm sorry boss of bosses I'm joking because those people give me grief for for referring to him as the boss boss We know he's not officially the boss of bosses, but most powerful Don in New York City He apparently has a soft spot in his heart for Polito. He's been a golden boy For a while, but it looks like he's gonna do go do some prison time But really good deal that he cut a shout out to Jerry Capacia gangland news who broke the story of the global plea Case came down in late summer early fall of 2022 took down I think six or seven Gambino or sorry six or seven Genovese Polito was at the top of the indictment as a number one defendant So a global plea means that everybody cops and you know the the prosecutors can clear the case and Everybody can just kind of do what they have to do for punishment and go on with their lives and not get this thing into trial or These huge back legal back and forth. So everybody copped the plea most of these Codependents are probably to do a year or less I'm guessing polito is gonna do a little bit over a year. You could do three But even if you do three you're only doing eight, you know, 18 months until you hit the halfway house, so Only other thing to point out and this was in Jerry's column on Thursday that I guess the hearing that Was held in a Brooklyn federal courthouse last week to come to these pleas got a little dramatic a little explosive one of Polito's Codependence friend of his Mark Fuhrer. I believe his name is a former car dealership owner He eventually copped the plea but he was like screaming at the court hearing then throwing around Accusations that he is that he was framed his attorney claims. He was just a gambler just a big gambler had nothing to do with booking bets or collecting bets and then it should be noted that a polito since his bus in 22 and getting bonded out has been according to federal prosecutors still You know working every day in the Genovese crime family handling business as usual conducting calls and meetings with Other members of organized crime within the Genovese crime family that he's not supposed to be specifically. There was a lot of communication with Anthony Rominello couple out of Queens who's now in prison on his own extortion case, so Just keep an eye out on Carmine pizza You know, he's acting capo for Johnny Hollywood handles everything day-to-day In Little Italy and parts of Queens other parts of Manhattan, but he'll be doing a little bit time in college probably about a year year and a half, but For what he was looking at this this turns out to be a great deal All breaking news out of all the five families in New York come to the og come to games to report I'm Scott Bernstein og pod out


And I practice this move too because it makes you really use your feet. But is this not the move you have your horse doing when it goes across the pen? Say the cow is out there. What do you want your horse to do? Is it come across the cow like this? That's the exercise. That's why we exercise that part too because it's more important. It's really important how you use your feet. So you see those markers out there? They're stepped off. This is where you guys heard some of you guys, I want to be able to, I don't ride my horse to the end. He's cheating me on the left side too much. You're not working your pen even. So if he's cheating you on the left side, is it because he's too long on the right? So what I'm going to do right now and what I do every day, just like what I do riding two-year-olds, the first thing I teach a horse or a rider has got to be rate. Rate is how fast is it going? And that's a feel. That is feel. Rate. Rate is just like when I train a two-year-old. The first thing you do when you're a teacher horse.


everyone and welcome. My name is Melissa Armel and I am the Stock Swoosh. I get this question often people ask me again how much time they need to spend trading golden gaps. Golden gaps is a strategy that I invented and created a very long time ago. Now I started trading in 2008 and it took me about three years to figure out my system. So I've been trading now for gosh since 2008. That's absolutely crazy for 14 years. And so I've had the Stock Swoosh for about 10 years. So it's very interesting because I teach people and a lot of times I teach people that have been trading for longer than I'm even alive. But the fact is that they don't know what I know, which is about gaps. I train gaps in a very unique way. And when I started trading, I decided that I wanted to do gaps. It's interesting because even 14 years ago compared to even now, there's not much out there about gaps. It's actually good information. Most of the information when you Google or look for stuff about gaps is wrong or bad. And you know that if you've been trying or attempting to trade gaps for a while. The reason I like gaps though is because again, you can trade them quick and fast in the morning. So people ask me how much time do you have to devote to this? Listen, the class itself is a two day course that's on Saturday and Sunday and a weekend. But after that, once you are starting to do it, once the time investment for you is Monday, Tuesday, Wednesday, Thursday, Friday, you only have to focus on the first half hour out of the day. Sometimes we're done in 10 minutes, five minutes. We were done super different fast today on a Friday, which was great. That's really what I wanted. We got a quick trade. We're out. We're just looking for a move, a five minute, six minutes, 10 minutes, whatever it is. The whole idea is to chunk it out. So I don't spend a lot of time actively day trading. I prefer to spend as little time as possible so I can have the rest of my day to myself. And then I could do TV or whatever I want to do in New York City, which is where I live. So it's very interesting. Again, people think they have to spend all this time from 930 to four o'clock eastern time trading. That is not true. You have to find stocks that move so you can get that move in those couple of minute periods. But that's what I'm looking for. And again, that's what I took the time to develop my system to try to figure out what stock is going to be a big move and what is going to have a fast move.  If you'd like more information on the Golden Gap course that I teach, email me on Melissa at thestockswish.com. Have a great weekend, everyone.