So, at the end of the lecture of the lab, my objectives are that you will be able to play with some gene identifiers and gene attributes, that you understand how simple enrichment tools work, and that you get familiarized with the search escape software. So, the starting point of a pathway and network analysis is a gene list. So, it could be like a medium-sized gene list or a very large gene list, and it can come from proteomics or genomics experiments. So what you want to do is to interpret your gene list. You want to know what is interesting about your gene list in an automatic and unbiased way. So, for example, you want to know if your gene list is enriched in some known pathways. So, first, what you are going to do is to find the overlap between your gene list and some gene annotation that are stored in some pathway databases. So, for pathway enrichment analysis, what we use is prior knowledge of the genes that are stored in pathway databases. So, this information is collected regularly, and we need... We say that the pathway are... That we curate the databases. And we visualize the results as a map because it's a more intuitive way to interpret the results. So, now, what do you mean by pathway? So, for biologists, we are more interested by signalling pathway and metabolic pathway. So, signalling pathway that would be like a receptor at the surface of the membrane that is activated, and there is like a downstream signalling cascade. And a metabolic pathway that would be more like a reaction, like more the biosynthesis of vitamin, for example. But we can use... For enrichment tool, we can use also databases that have like a broader content, like drug or disease association, and this can be very useful. And another term that you may hear a lot during this lecture is gene set. So, what is the definition of a gene set? A gene set is a group of genes. So, it's more specific than a pathway. So, a pathway would be... For example, you can have like a big pathway that is divided into sub-pathways, and each sub-pathway could be like a gene set. So, what are the advantages of doing a pathway and network analysis? Well, it saves time compared to the traditional approach. So, the traditional approach would be to read information about new genes one by one. And actually... So, it takes a lot of time to do that, and sometimes it's also biased because you finally you end up looking only on your favorite genes. And pathway and network analysis is also an intuitive way of analyzing the results. So, we all know that within a cell, we have a lot of signalling pathway and metabolic pathway that are interconnected. And when we create a network, it's like drawing this map that we have in our mind. And on this map, we put our data. So, it's much easier to interpret the results. The other advantage is that we can add different layers of information. So, we can overlap different information at the pathway level. For example, you have RNA-seq data, and you have mutation data. And you take your top hits of your RNA-seq data, and the top hits of your mutation data, but they are not the same genes. So, you don't find a direct overlap. It doesn't mean you don't have an overlap at the pathway level. So, maybe these genes belong to the same pathway. So, this is an example. So, you have patient one that has mutation one, patient two has mutation two, and patient three has mutation three. So, you cannot find the same mutations in different samples. So, the mutations are mutually exclusive. So, when you directly compare the patient, there is nothing that is similar. But all these mutations in these genes are belonging to the same pathway, and all these patients are diabetic, for example. So, this is a first example of real data using mutation data and pathway network analysis. So, this is a Nature paper in 2012. So, they had this chromosome 5 deletion that they found in breast cancer samples. So, they look at the genes within this chromosome 5 deletion. They extract the gene list, and they did an enrichment map. So, this network. So, on this network, one circle is a pathway. And what they found is that the chromosome 5 deletion genes are enriched in genes implicated in cell cycle. So, that's the same data, but now it's not what we call an enrichment map. But it's a network created by the Reactome-FI plug-in that you are going to see this afternoon. And now, each circle is a gene. And the different colors are the functional modules associated with this network. Now, another example that we are going to see in the lab practical. So, the same data as we used before with Daniel.  And one major result of the paper is that they found 71 significantly mutated genes. One of them being EGFR, so Epidomal Growth Factor Receptor. And another result, major result of this paper is that some of the mutations were found mutually exclusive. So, now, can we add all these different layers of information in one network? So, all this information were available at separate tables in the paper. Can we add them all in the network? So, this is the network that was created with gene mania. And the layers of information are the frequency of mutations, so the size of the node. The shape is if the mutations are mutually exclusive related to EGFR. The pink color is the pathway that was the most significant pathway in this data set. And we also see the physical interaction and the genetic interactions. So, now, when we add these layers all together, I think we can see that one set of genes can attract our curiosity. So, they seem to cooperate or to work together. So, if I had to work on that project, I would not only focus on EGFR, but probably on these other genes that are related like PIK3R1 or PIK3CA. So, I think it was obvious that these cluster of genes work together. So, a last example how to overlap two layers of information. So, now, this network is a pathway network. So, each circle represents pathways that were enriched in one subtype of the glioblastoma called pronural. And the yellow triangle is the 71 significantly mutated genes. And now we can see where the mutated genes are located in the pathway map. So, we have genes belonging to some pathway that are significantly enriched in this subtype of cancer. So, what are the three elements of pathway network analysis? Then, first, we need our gene list. We need gene attributes that store the functional annotation of the genes that are stored in pathway databases. And we also need enrichment tools that will calculate the overlap between the gene list and the pathway that we are testing. And these tools are also going to tell us if this overlap is significant or not. So, let's talk about the gene list first. So, first, some recommendations between you start a pathway analysis. So, try to get like a clean list. So, as clean as possible. So, try to normalize, to adjust for background, to remove outliers.  So, try to get as much as possible a majority of true positive in your gene list that you are going to use for pathway network analysis. The gene list size matters too. It can also help you to choose the right tool. So, if you have like a very small gene list, then you can use function prediction tool. If you have like a medium size tool, like 50 to 500 genes, you can use simple enrichment tools. If you have like a very large gene list, more than 1000, you want to try to rank your genes by significance. And use tools that use this ranking as input. And make sure that your gene IDs are compatible with the software that you are going to use. So, where do gene list come from? Then they can, from multiple sources and enrichment analysis can work with any of this gene list. So, you have to know that these tools have been developed first for gene expression data. So now, new tools are being developed for more specific case, like if you have methylation data, you can use, for example, the tool rate. Before applying pathway network analysis, it's important that you have a clear idea of the question you want to answer. So, you want to answer one biological question. You make sure that your experimental design to generate your gene list make it possible for you to answer the question is important for you. And then it will help you to choose the right tool and to correctly interpret your results. So, gene identifiers. Gene identifiers are ideally unique, stable names on numbers that represent your genes. They come from multiple databases. And these databases store slightly different kind of information, like gene databases will store gene sequence, protein databases will store protein sequence. So, it means that one gene can have multiple IDs. And also, because these databases are slightly different, they don't completely overlap. So, here is a list of common identifiers for you to know what they can look like. In red are the ones that are recommended.  So, on a daily basis, I use ensemble gene IDs, interest gene IDs and official gene symbol. Gene count is a tool that I like to use. So, gene count is like a, it's a gene database. And when I want to retrieve my interest gene ID very rapidly, I just enter the name of the genes. And I get like a lot of gene identifiers. And if I scroll down, many more information. So, we are going to use this tool in the lab later. So, interest gene ID. This one is my favorite gene identifiers. So, why? Because it's stable. It's a numerical value, so it's very easy to manipulate. And for genes that are, let's say, new or not yet annotated, they already have an interest ID, even if they don't have like an official gene symbol. So, if you use interest gene ID, you don't have to update your annotation all the time. Okay, so we have seen that one gene can have multiple IDs. So, that can be an issue. And you have to be sure that your software uses the identifier that you have on your gene list. So, you may need to convert from one identifier to the other identifiers. And that could be a challenge. So, you have to be careful when you do this conversion. And I think the most frequent mistake is when we use Excel. Because what Excel does is automatically change some gene names into dates. Okay? So, like October 4 is going to change in October 4. And, yeah, I see that every day in my work. So, here on the side. So, at the right side, you have the correct symbol. And on the left side, you have this same gene name that are transformed to date. And if you don't do anything, if you save it again, they will be just numbers. And the problem is that your enrichment tool is not going to recognize these symbols. So, it's just going to ignore that. So, there is a trick. I mean, not a trick, but there is an option in Excel to format your column as text. Okay? So, if you format your column as text first, and then you copy and paste your gene symbol, they should not be changed. But I know it's difficult. So, I think the best advice that I can give you is look at your gene list. If it's not too big, look if you have dates. And if you have dates, then look back at my slide and try to change to the correct name. So, fortunately, we have some tools that have been developed that help us to convert from gene identifiers to the other gene identifiers. And so, one of them is Gconvert that we are going to use in the lab that is really easy to use. And I would like to mention also Biomart Ensemble. It's not as easy to use, but it has a lot of features. So, you can copy and paste your gene list and retrieve any gene attributes you like, like gene identifiers, but also sequences and orthologs. So, our recommendation is that you need to map everything to interest in ID using a spreadsheet. If you need 100% coverage, then you need to manually create your missing annotations and be careful of Excel auto-conversions. The last tip is that when you have your table of results, try to keep at least two common identifiers of the terms or even more. So, one tool uses one, and the other tool uses another one, and you have all your data ready. So, what have we learned? So, that gene and their products have many identifiers that bioinformatics requires conversion of IDs from one type to the other, but ID mapping services are available, and please use standard and common used identifiers. Okay, so now we are going to start the second part, which are the gene attributes and the pathway database. So, when we use pathway and network analysis, we are more interested by function annotation. That's all the biological process, molecular function, and cell location. These information are stored in different pathway databases, like the gene ontology, keg, or reactor. So, today, this morning, I'm going to describe the gene ontology, and I think you are going to see reactant this afternoon. So, gene ontology is the largest database. It's updated regularly, and it covers many organisms. These many enrichment tools use just GO as the reference databases. So, GO is divided in three major parts. One is cellular component. The second part is molecular function, and the third part is biological process. So, let's say the term plasma membrane, that would be a cellular component. This isomerase activity that transforms the glucose into fructose that would be a molecular function. And cell division that would be a biological process. So, what is helpful to understand is that the GO database is organized as a hierarchy. So, you have more general term at the top of the hierarchy and more specific terms at the bottom of the hierarchy. So, here, you have large gene sets and small specific terms. And what you need to know also is that one term can have one or more parents' terms and one or more child's terms.  So, that's why, because of these relationships between the terms. So, genes are linked or associated with GO terms by traders. These associations are called GO annotations. So, this is an example of manual curation focused on the PERC1 genes. And so, how these GO annotations have been related. A receptor like kinase would be a molecular function associated with the PERC1 gene. Integral membrane protein would be the cellular component associated with these genes. And wound response would be the biological process associated with this gene. So, already three GO terms for one gene. So, there are different ways to get these annotations, not only manual curations. And you can find out looking at these evidence codes. Here, IC, IDEA, TAS, TAS, IEA. And the one I just showed you is probably a TAS, a traceable author statement. So, if you run an enrichment analysis and you have one GO term that is very significant and you want to know more about that term, then you can use tools to get more information. Like quick GO is one of these tools. You can get the child terms and the parent terms and the definition of the term. And a very related tool is AMIGO. This slide is just to remember that we're focusing on the function annotation, but there are many other gene attributes that are available. Like chromosome position, this is association protein properties. And all these other gene attributes can be available from Ansible Biomarkt. Is there differences between quick GO and AMIGO? I couldn't find any differences. And they relate to each other. So, when you're on quick GO, you can access AMIGO. When you're on AMIGO, you can access quick GO. Myself, I couldn't see any differences, but maybe there must be some. Otherwise, why? Why two tools, but I couldn't see any. So, what have we learned? So, gene attributes define functions, characteristics of a gene. Many genes attribute are available in the databases and I've just presented the gene ontology databases, but there are other ones like Kagan and Reactome. So, now we go to the third part, which are the enrichment tools. So, we have our gene list on one side and we have our functional annotation. Now, the enrichment tools, what they are going to do is to find the overlap between the gene list and the pathways that we are testing. And they are going to tell us if this overlap is significant or not. So, many enrichment tools exist. So, they can be classified into three groups. The first one is overall representation analysis. The second one is functional class scoring. And the third one is pathway topology. So, the first one is the most simple tool and this is the one that I'm going to explain later. And it's really good for mid-sized gene lists. So, maybe if you have 100 genes to 500 genes or up to 1,000 genes. One famous tool was David. So, I don't know if you heard about David. So, you can still use David. It's a very good tool, but it's not up to date anymore. So, the pathway database underlying the tool is not up to date. So, that's why we don't recommend David anymore and we recommend a G provider now. And Yuri is the developer of G provider. So, if you have questions during the lab, feel free to ask him. And then the second class of tools is functional class scoring. So, I would suggest that this tool is more for larger list and if you can rank your genes from a significant to less significant one. And one very popular tool is GSEA, Genset Enrichment Analysis. And the third way, the third one is the pathway topology tool. Reactome FY is one example that you are going to see this afternoon. And this one is more complicated. So, it takes into account the relationships between the genes. So, for example, you have a pathway and you have 10 genes that overlap between your gene list and this pathway and 10 genes are activators. They are on the same activating branch of the pathway. This is going to get a better score than another case where you have 10 genes, but 5 are activators and 5 are inhibitors. So, it takes into account the topology of the network in addition to the size of the overlap. So, tips for other represented tools. Again, if you have like a gene list, a small gene list, you can use to like gene mania. If you have like a small, medium sized list, you can use to like gene profiler. If you have more than that, try to order your query. There is an option in gene profiler to order your query. So, you can do that in gene profiler. And if you have like a large gene list, try to order your gene list using GSE or Wiccoxon rank test. So, what is gene set enrichment analysis? So, the first step is to break down the cellular function into gene sets.  So, I'm going to test nucleopor, ribosome, cell cycle and P53 signaling. I'm going to find the overlap between my gene list and these four gene sets. So, how many genes are in common? And then, I'm going to see if this overlap is significant or not. So, here is a very general and conceptual slide that can be applied to many enrichment tools. And that explains the significance of the results calculated. So, let's say I have a gene list that contains 200 genes. And my pathway, let's say apoptosis, contains 100 genes. And the overlap between my gene list and the pathway is 20 genes. So, that's the first result. So, I have 20 genes that overlap. Is this overlap larger than expected by chance? How do I assess that? I can do some randomization. So, there is two ways to do the randomization. First, you can imagine that you can randomize your gene list. So, you replace your 200 genes by random genes. And you do it 2,000 times. So, how many times, if you take these 200 random genes, are you going to have an overlap of 20? You can do it the other way around. You can take your pathway that contains 100 genes and randomize this pathway. You do it 2,000 times. How many times, if I take 100 genes randomly, do I have 20 genes that overlap with my gene list? From this randomization, you are going to get a p-value that tests the significance. So, if you have a low p-value close to zero, it means it's probably not by chance. So, this overlap of 20 cannot occur by random change. So, it's highly significant. So, many of these enrichment tools, like David or G Profiter, what they use is a Fisher's Exact Test. So, let's say you have your gene list that contains four black genes and one red ones.  So, that would be the pathway that you are testing. And 4,500 red genes. So, first, there is like the null hypothesis. List is a random sample from population. Or the alternative hypothesis, you have more black genes than expected in your list. So, the first step of the Fisher's Exact Test is to calculate the null distribution. So, let's say you have 45 red genes in your background population and 500 black genes. What is the probability to obtain five red genes by random change? That the probability is 57%. What is the probability to get four red genes and one black genes? Then the probability is a little bit lower. It would be 35%. Now, this is our case. We have four black genes and one red genes. What is the probability to get that? Well, the probability is very low to get that by chance. So, it's very close to zero. Now, the p-value that you are going to obtain would be the p-value at the cutoff. So, our cutoff is four black genes and one red genes plus the p-value that are less than our cutoff. So, if it's close to zero, it's highly significant. We don't think it can happen by chance only. That's a good question. If we should differentiate up-regulate genes and down-regulate genes when we do enrichment analysis? Yeah, so that's a good question. So, it depends on your input. So, for Fisher's Exact Test, you cannot differentiate. Put your 200 genes. It will not tell you if they are up and down because you don't give that information. So, what you could do if you use these tools is to separate yourself, the up-regulate genes to the down-regulate genes. If you want to see if the up-regulate genes are enriched in some pathway and the down-regulate genes are enriched in some pathway. This is what I usually do. You can use tools like GACA when you ranked all your data from the up-regulated to the down-regulated. And then you have two types of results. You will have like a positive score for the up-regulated genes and like a negative score for the down-regulated genes. So, you can separate the two. No, it's up to you to decide. If you want. Because I think it's easier to interpret if you divide this separately.  So, you see, okay, this is the enriched pathway in my up-regulated genes and in my second list, this is my pathway enriched in my down-regulated genes. But the output of a pathway analysis is really... There is a modification of the pathway, but because we don't know if these genes are activators or inhibitors of the pathway, we cannot conclude too much if the pathway is up-regulated or activated or inhibited. So, you have... You know something is going on with the pathway, but at the second step, after your enrichment analysis, you really need to look at the genes that were in this overlap and look if they are activators and inhibitors. So, the third class of... The third class of tools that I showed you are trying to answer that question, but not the simple tools that I'm going to mention. Sorry, I don't understand how these numbers are... 500 black genes and 45 red genes, and then that's we have 5 red and 45 black. So, that's a second slide. So, the five genes... So, the background population, let's say it's... So, the background population would be your whole genome. So, the whole genome. So, let's say the total number of genes that you have, and the black genes would be the pathway that you are testing. So, just an example. So, let's say you have 4500 genes that are red in your population and only 500 that are black. That would be the cell cycle pathway, for example. And in your gene list, coming from your data, you have four genes that are belonging to this cell cycle pathway, so black genes, and one gene that is coming from other pathways. And now we want to answer the question, is it possible to get four black genes out of five just by random chance? And we know that we have much more red genes than black genes in our population. So, already we can figure out that it should not be bad genes only. If you pick five genes randomly, what is the chance of getting four black genes and one genes if you have this gene in the universe? Very low. So, your p-value calculated but the fissure's exact test is going to be close to zero. No, I just had different numbers here. I had 5 random 45 black, instead of 5 random black. This type of distribution seems to be combined and seems to assume that the gene size is equal to that. So, that's, yeah. The idea is that you're going to do the same thing if you're good. So, say... So, it depends. So, if you are really like a... So, let's say you have your mutation data and everything is ready, you have your gene list and the simple question you want to enter is that are these genes in my gene list belonging to the same pathway? Then, yes, you can use this enrichment tool to give you additional information. So, yeah. Yeah. I'm not sure but I think if you have if your gene list is 10 genes you have 8 black and 2 red, I think the p-value will change. It's the same proportion. Yes, we have 4 numbers that are taking into account the universe size for the total number universe, your gene list size and the number of black and red. So, yes, that would change. What does it make sense to change? Like, is it biologically significant? Yeah, there is pros and cons. I think this is really like a simple method. There is, actually in G-propiola there is also one option I think that's very interesting when you can order the query. I mean, I think Yuri can talk about that later. So, it's the same p-value but if your gene is ranked number one by the significance it's going to be more important than if you are like a gene which is genes number 10. So, I think it will differentiate this kind of of cases. If you like. You said that it does iterations, right? Yeah. Re-samples, it does this over and over and over. In this case, the official exact test calculated the null distribution first based on the equation. But you do them one or the other. You either compute the p-value analytically or you do the re-sampling. So, the re-sampling actually that I showed that, yeah, is more general. Okay? There are different ways to do it. And the official exact test calculates the null distribution, the hypergeometric distribution. This is the way this test works. So, you understand that for the official exact test then the size of the background population change the results. So, most of the case you are going to do like a whole genome experiment. So, your background is going to be all genes in the genome. But in some, yeah, in certain cases you can have like custom chip where you just have, for example, kinases or immune genes. So, in this case the background is going to be smaller and you need to input your background list. So, in general, many of these tools there is an option to upload the background. And that's for that reason. Because the background is taking into consideration to calculate the p-value. What is kind of the bigger question here? What is kind of the signal that we're trying to calculate? Is involved, is activated in this patient or is perturbed in this patient? So, perturbed would be more accurate than activated because, as I said you really need to look at the function of the genes that overlap between your gene list and the pathway that you are testing to see if they are activators or inhibitors. And you may have like a mix of activators and inhibitors. But what it's answering is that you have 100 genes. 100 mutated genes. And maybe, I don't know, 30 of them belong to a cell cycle event. Okay? And that or another pathway that you don't know about. So, it would have been very difficult to know that information just looking at all these genes that you don't know. So, that's the simple question these tools are trying to answer. So, are these many of these genes in your gene list known to be related to each other in any of the databases that you are testing? And even test like a drug database. So, are they to target of the same drug, for example? Are they known to be associated with the same disease? Or are they known to be associated with autophagy? So, that's really the question. So, it's the overlap. Yeah, so it's more that we know the association. So, if we trust the pathway database, we know the association. We know that gene A is in cell cycle. But the Fisher's exact test is trying to give you a p-value to get the confidence. Can we trust these results? Or can we... because we have many false positives in these kind of results. There's what are the most significant to the ones that are less significant. And the way to do it is to as I say to try to randomize. So, by chance only, can we get that by chance only? So, it's really to get the significance out of the results. Not the overlap. Because the overlap, we know. For sure, these 20 genes belong to the cell cycle. But are my data clean? Can we trust my data? Let's say we're carrying over from yesterday's Surab's single nucleotide variants. How much confidence did we have if there was only a few tumor beads that had that mutation? And then now that's become our gene list of mutations. But this is adding a layer of confidence. I'm kind of just struggling with the confusion that I made. I suppose I we found out that yes, this is a significant result. And so these mutated genes are part of the pathway. Yeah, so that's the beginning of everything. Then, yeah, so then I think this is only telling you that given a list of 100 genes any list of 100 genes what is the probability of 10 out of those will be in the cell cycle pathway? And the example is 100 genes but you can have 1000, 2000 genes and you cannot do it manually. So you need to do it automatically using these tools and you need to rank all these results by a p-value for the most significant one to the top. Because sometimes you have many, many, many significant results, many, many pathways that came up significant. So you want to distinguish the more likely true positive from false positive. And then that's the beginning of everything. Then you think, oh, the first pathway, the top pathway is autophagy or cell cycle. So now I need to look at the genes and to understand how it works but then you need to do it manually. Or you need to go back to the wet lab. Sorry, so the data set that we're using we're using from that database as our background what are the mutated genes. In our sample we look at the fish-rex exact test but the p-value that we're deriving is based on the previous data how commonly these genes are mutated in different experiments and then it gives me the test. Is that what you're saying? For example, if I have like 20 apoptosis genes like B-cell 2 and whatever. But then this data sets we'll look at the previous probability based on the previous data that we have and then gives me a p-value even though like if I have 10 genes mutated in apoptotic aspects for example 10 genes so 10 out of like all apoptotic aspects there are like 100 genes and then I'm choosing them from the data but how do I know my data like MDM2 is more important in my data set so it will not answer that question it will not you can add different layers of information when you create the network but for the simple enrichment tool it will not answer that question it's really if you database that you are testing in the pathway database it will just take the pathways okay it will give me just the path how does the p-value come from that so let's say you have so maybe we can take the example of the lab practical so they have this 291 gliomoblastoma samples from different patients and they did this old exam sequencing and they looked to find mutated genes and they looked to find the genes that were drivers they're more likely to be the driver genes and if they end up with 71 genes so these are the 71 significantly mutated genes in glioblastoma so that's their data that's their data that's the omics experiment all the pre-processing has been done and we think that in glioblastoma we have 71 genes that are more likely to play a role in the development of the disease so that's not enrichment yeah that's why just running a muted significantly mutated gene software they ran a muted 6CV basically then they came up with the list of 31 genes that are significantly mutated in that and that that's the data that's not the pathway analysis that's all the pre-processing that you may have seen during the workshop and now you end up with this gene list and you want to interpret this gene list so you can look at the gene one by one and try to understand the function or you can try to do the analysis at once to try to understand the relationship between the genes and the results in this case would be that a few genes are part of the same PI3K pathway and you can do that in 5 minutes instead of looking gene by gene and this is the only information simple tools like this one or G-Profiler will answer for you if you are testing these pathway databases is it the point of having the database to give us a high confidence set of genes that are in pathways already like we don't trust it this sounds like this is a test for membership so the test of significance is high confidence that this is part of this pathway but if our database is already this Q-rated resource that tells us to be able to look up the genes like just count I have a mutated gene X what pathways is it nominated in my database of choice you can do gene by gene if you want so there is two different questions one is to relate one gene with this known function and the other one needs to analyze as a group of genes so now a summary of the different steps of the enrichment analysis so first the overlap is tested with each gene set present in the pathway database so here we show the example with one gene set but actually we can test many many gene sets more than 3,000 gene sets so now we need to order the results by the enrichment p-value for the most significant gene set you want the lowest p-value and then because we have tested so many genes we need to adjust for the multiple hypothesis testing and this is what we call the FDR and we usually use the Ben-Germany-Horberg method so the FDR corrects for multiple hypothesis testing so if you have like FDR of 15% it means you have like 15% change that your result is a false positive so 85% change that is a true positive and why statisticians want to do that they say that because we are testing so many pathways at the same time we are increasing the chance of making one decision so they think that the p-value are overestimated so like p-value of 00001 estimated we use the Ben-Germany-Horberg method to correct that and maybe now our adjusted p-value is going to be 00001 so a typical output of an enrichment analysis is a table that contains at least the pathway or gene set names that we are testing the number of overlapping genes the number of genes in the original pathway the p-value that test the significance of this overlap and the adjusted p-value and the typical output is a long table with many many gene sets row by row and it's usually very difficult to interpret and it's difficult to interpret because the gene set they contain genes that are redundant so they are not completely independent this gene set so maybe your gene set 1 that is very significant has 50% of genes in common with your gene set that is number 10 so that's why we use network visualization so we are going to our last part of the lecture that is the intradiction to network visualization using cytoscape so cytoscape is an open source software for visualizing complex networks and there are lots of apps that are available so networks, the advantage of networks is that it can represent relationships between the data so you can discover new relationships and also you can visualize multiple data types together so two important words are edges and edges so nodes are the circle and edges are the lines that connect the nodes and so if you do pathway enrichment analysis you have two types of network one that I will call gene-gene network which is actually more like a protein-protein network but one circle would represent a gene and the line would represent the association between these two genes and the other kind of network would be a pathway network where one node represents a pathway and the line between two nodes represents the genes that are overlapping between these two pathways so one another important information is the layout so the network layout so if we don't apply any layout it will look like a hellball so the most common layout in site escape network is the force directed layout in the force directed layout the nodes are repelling each other but if they are connected by edges the edges pull the nodes together like springs so you have two forces repelling and pulling so from it was a bit difficult to understand so I could see it as videos so if you are interested you can do at the workshop go to these links and see the video of the force directed layout and here are like a few snapshots so that would be step one it's a little like a hellball and you see that some nodes are overlapping but then we are applying the force directed layout and the nodes are going to repell each other and you see that the distance between the nodes are increasing but this one for example here they are highly connected by edges so they kind of form a cluster then we go to step three and we see now that it's spread out and here I would say this is the end result so there is no more overlapping between the nodes so for us it's better to visualize but still the nodes are highly interconnected then they form these clusters so that's what we call the force directed layout so before we start the lab an introduction to cytoscape so when you open cytoscape you have three different panels on your left you have the control panel at the bottom you have the table panel and at the right you have the result panel and so you can save your session so you have when you have done your network you can save your session to reload it later you can save the network image as any format as you want pdf, png also another feature is to be able to navigate for the network so you have your network on this window you go in the control panel you click on network and you will see like a blue square and you can move this blue square to navigate for the network so force directed layout is the most popular layout but you can try different ones like the sickle layout you can play with colors and you can play with shapes and you can end up with a beautiful network so what have we learned it's useful for seeing relationships in large datasets that they are useful for integrating several datasets and types together it's important to understand what the nodes and edges mean automatic layout is required to visualize the networks and visual attributes enable multiple types of data to be shown at once so it's useful to see the relationships between the genes and there are many applications available for site escape more than 200 so you can go to the site escape app store and click on the icons to see a description of the applications so they have different categories so apps involved in pathway analysis some involved in gene expression analysis or literature mining or pathway comparison so depending on what you want to do maybe there is an app that is available for you and it's a software that is in constant development it's an active community with more than 5000 downloads per month Slicer from Gary Quaid, Lincoln, Daniel myself and we are ready to start the lab practical


Everyone, Myself, Darshan Pandit, Assistant Professor, Department of Computer Science and Engineering from Walton Institute of Technology, Solapur. Today we are going to discuss about basic drawing function in computer graphics. So here at the end of this session student will be able to draw different graphical object using C functions. So in this lecture we are going to see how to draw polygon, bar, pie slice, sector. So here function to draw polygon, so the polygon is any two dimensional shape formed with straight lines, rectangle, quadrilateral, pentagon and hexagon are examples of polygon. So here with a given set of points we need to draw the polygon. So in this way we need to declare an array which includes all set of points. So int x1, y1 will give you one point, int x2, y2 will give you one another point, int x3, y3 will give you a third point and int again x1, y1. So in polygon one thing you have to remember starting point and the end point should be same. So that will form a closed object. So here we are using these functions drawPoly which has parameter int number of points int array name. So number of points is nothing but so in above array how many points we have used. So we need to specify that number of points and name of array that is point that we have to specify here in array name. So just we will see the example. So here we need to include the set of files that is graphics.h, conio.h after that we need to declare graphics driver and graphics mode and here we are declaring array in which we are specifying number of points that to be used to draw the polygon. So here we have used 500, 200, 400, 600, 200 and again 1500. So this is the, so here first and last point are same. So after that we are initializing the graphics driver and graphics mode and we are specifying the path of BGI. So after that drawPoly function in which we are specifying number of points. So here number of points are 4, 1, 2nd, 3rd and 4th. So these are the 4 points and name of array is point. So that we have declared in drawPoly function. So after that gets here and close graph. So this will give you output in this way. So by using drawPoly function the polygon will be drawn. So here we are having another way to draw polygon with a given set of points. So that is instead of using drawPoly we may use fillPoly function that draws the polygon and fills the polygon. The parameters are same, we need to specify point array that is array after that. In array we need to declare set of points so that we have declared after that instead of using drawPoly we have to use fillPoly functions. And parameters are same like drawPoly that is int number of points and int array name. Just we will see the example. So here we have used fillPoly instead of drawPoly. So this fillPoly function will fill the polygon and also it will draw the polygon with a given points 320, 150, 440, 340, 230, 340, 320 and 140, 150. So this is first and last point are same. So here in this way you will get the polygon which is a solid filled polygon. So how many points are required to draw polygon? So you can answer, so how many points we need to pass in a drawPoly function or how many points required to draw a polygon? So here n plus 1 points are required for n number of vertices of polygon. So the polygon should have at least three sides and angle and if polygon suppose if polygon is having three vertices then four points are required to draw a polygon as start and end points are same that is start and end point are repeated. So the next function is how to draw bar? So bar is used for drawing bar charts. So here we need to pass x1, y1, so here bar int left, int top, int right and int bottom. So here left is nothing but x1, top is y1, right is x2, bottom is y2. So here you can see, so this is x1, y1 and here x2, y2. So let us see example. So here bar, so in this we have passed x1, y1 and x2, y2 that is 50, 100, 200, 400. Again second bar it will be drawn using this coordinate 250, 100, 400, 300. So third bar will be drawn using this coordinate 450, 100, 600, 200. So in this way you will get the output. So three different bars you will get based on the specified coordinates. So next function is to draw a pie slice. So this will be helpful when you are drawing pie charts. So it requires a parameter int x, int y, int st angle that is int starting angle, int end angle, int radius. So here you are having x and y that is from where you need to draw the pie slice. So here you are having starting angle, this is the end angle and this is the radius. So when you specify this parameter pie slice will be drawn. So let us see the example. So here we have drawn pie slice with x200, y200. Starting angle is 90, end angle is 0 degree with the radius 100. And another pie slice with coordinate x coordinate 400, y coordinate 300, starting angle 90, end angle 360 and 100. So this will give you the output in this way. So this is 90 and this is 0 degree and here starting angle is 90 and end angle is 360. So function to draw sector. So in sector same like we need to pass int x, int y, int starting angle, int end angle and with this we need to pass int x radius, int y radius. So this will draw a sector and it will fill the sector from center. So an example we have drawn sector with the point 300, 300 and starting angle is 180, end angle is 0, x radius is 100, y radius is 200. So this will draw a field sector in this way. So these are the references used to prepare this PPT. Thank you.


I'm Shannon Mustapher, an award-winning mixologist, educator, and author. Like a confession booth, the bar is a place for all walks of life to come in and share their stories. So today, I'll be sitting down with innovative creators to turn their journeys in the classic Grand Marnier cocktails with a unique twist on a tradition. Yo, Otis, what's good, man? Shannon, how are you? What's up? It's been a minute. Good to see you. Good to see you. What's about your music and, you know, what brought you here? Yeah, well, I'm Otis Cain. I am a writer, producer, artist, play instruments, a mix. I do the whole thing. I've been doing this for a very, very long time. Now, tell me a little bit about your background and your heritage. My dad's side of the family, full Mexican. My mom's side is Jamaican. I'm very shaped by American culture, but I do have, you know, that blood running through my veins.  Like, what kind of music do you do? I would say that my music is very funk, soul, pop, R&B. Obviously, one of my biggest influences, Stevie, Sam Cooke, you know, those guys. Love songs are a big part of the R&B genres. How do you draw inspiration from that to create like a new twist on it for the listener today? Love songs are very important to me and making love songs in today's world is pushing the culture forward because black artists aren't usually shown in that light. You know, the music that is promoted the most heavily isn't promoting black love and I think what's really important about what I'm doing is being the voice for that. I'm curious. Is there like a particular moment or a story when you had that light bulb moment? You're like, it's time to start building Otis. I was kind of burnt out on creating careers for artists when I knew I wanted to create one for myself. You know, I've always wanted to use my voice. It was August 2020 when I released my first song and I wasn't doing sessions with other artists. So I had already made the decision to not produce but now there was no turning back. You got started in your own direction. When did you start to see it as paying off? Like when did you start to see signs that you were succeeding in making it as an artist? One thing that made me feel like this is going where it needs to go is I started to get a lot of playlists on different streaming platforms and that is kind of the first step for an independent artist because that's the hardest thing to achieve in this industry. From a kind of big picture point of view, what does success look like for Otis Kane? As long as I'm happy doing what I'm doing, that's success. The message is about this resonant with me. I sent this to my ex. I sent this to my wife. I played this at my wedding. I love this song. I love the message. So all these things are reinforcement that what I'm doing is touching people and bringing light to people. Now onto the drink and what you're into. My style, as you know, is rooted in classic cocktails with a little bit of a Caribbean and a tropical flair. I'm an old school Southern girl, but I like to explore global flavors as well. I think I have something in mind for you. Okay. So just sit tight. Okay. I'm going to put it together for you and then we'll see what you think. Amazing. I'm excited. Before we go into the drink, I want to sample you on the star. This is Grand Marnier, a blend of cognac and orange liqueur. Wonderful way to elevate any classic cocktail. Have a taste. Now the thing I find really wonderful about this. Oh, you're saying it all. So I definitely taste the vanilla. I taste the hint of orange. I know it's orange liqueur, but just it's hidden in there. It's not overwhelming. I love it. It's basically going to amplify all the other flavors that I want to add to this spin on the Margarita that I'm calling the floor to have makeup. Let me stop talking and start pouring here. So I'm going to make one for myself as well. I'm going to start off with fresh lime juice. Now next, I made a custom syrup for you based on the dried hibiscus. I'm going to add some tequila. Tequila, yeah. Yeah, I'm using a Blanco. It's not really light, clean, just like the classic. Now next we got the star, Grand Marnier. I'm going to get a little more citrus here and I would say this is kind of the thing that's going to pump up the volume on all the other elements here. So this might love song to you. Oh. And to the Margarita. Yes. For the Jamaica. I hope that this drink is like a fitting way to celebrate your transition into making your own path as an artist. Let's try it. Cheers. Cheers. Wow. That is excellent. Can we call it the Grand Otis Cane? I like Grand Otis Cane. I know this cane. Grand Otis Cane. It has a musical ring to it. All right. You came to the right place. Thank you, Shannon. Cheers. Cheers.


This paper presents a new type of bionic gliding robotic dolphin which has been designed to mimic the movement of real dolphins. The robotic dolphin features a yaw joint which allows it to move in different directions while also allowing for more flexibility in its movements. A dynamic model was then created using the Newton Euler method to analyze the propulsion of the robotic dolphin in both dolphin-like swimming and gliding motions. Several simulations and experiments were conducted to test the robotic dolphin's capabilities in both modes. This article is offered by Yang Zhang, Zhen Cheng Wu, Zhen Wang, and others.


Everybody force and Jake here at being a personal for Christy. It is a 2023 leaf ultimate hockey Good luck Chris See you Vaughn All right, here we go Mount Rushmore quad relic.  That's pretty sick 7 to 25 How Lindsay Sauchek and Lindstrom an auto of Barry Melrose cool is the rule one of three on that one Barry Melrose auto and triple autograph one of 12 Mario Lemieux Eric Lindros Mark Messier Big time triple right there Congrats Chris really good box there get those cards out here. We appreciate the order


Globalization is rebooting itself in all sorts of ways. This is a world in which a lot of the global south is acquiring agency. Many developing countries want broader representation of their interests. What do we do to make sure that the technology that we're developing will spread to the 30% that is disconnected already globally, outside of the digital landscape? We need to change because of climate. We need change because of digitalization, but we need that transition from here to there to be fair for everybody. The institutions of the 20th century, we need to be retrofitted to be made more agile and more responsive to manage the technological age. The entire system is to enable increased connectivity. That's where the world is going. We need to innovate faster. And to do that, we need to move to a different paradigm. What's important about trial and error is about experimenting. Interdisciplinary is vital to tackle today's big challenges. We cannot work in isolation. We have to make the right choices, whether it comes to finance, whether it comes to innovation and technology, whether it comes to governance. The nature of the task at hand is a man. We need all hands on deck.


I think we'll start this out with the demo. We want to know what changes you're going to have. I'm going to drive a bit loud. You're going to find the function full of her. We might be wrong. We're buddy. We're driving. Trying to figure out the best way to explain about how reinforcement kind of gets going, right? And what makes something reinforcing into itself. And there's a lot to this, folks. We could talk for a while about how to establish a reinforcer. We can get into classical conditioning. We have primary reinforcement, secondary reinforcement. That's not where I want to go with this. Sorry, it's just a little warm. I'm getting distracted. I really want to get into the concept of motivation because motivation is really what makes a reinforcer valuable, right? Because we know about these things called motivating operations. So we take the concept of motivation and behavior analysis and break it into something observable. So as we break it into something observable Sorry, as we break it into something observable we run into the concept of motivating operations. And motivating operations are really events, stimuli in your world, events that happen to you that make certain other stimuli reinforcing or not reinforcing for that matter. So we can focus on establishing operations and we can focus on abolishing operations. But you know what? I, it's, folks, I'm sorry, it's 100 degrees out here. In fact, my wrist tells me it's 100, well, not quite. Close to 100 degrees out here. Don't look too closely. You'll find out why. It's the hair. Anyway, I'm done. I have had enough of the videos. I'm done. I'm out of here. See ya. Hey! What? What do you want? You gotta finish the lecture. I have water. On a day like tonight, you think water's going to be reinforcing. Is that what you think? Yeah. Because you're right! You know why? It's hot! And we've established a motivating operation that water is reinforcing to a person that's hot and thirsty and hasn't drank in anything in a while. Do you understand now why reinforcers are based on motivation? Because if you establish the contingencies correctly, if you establish the environmental stimuli appropriately and you, engage in some, he's not even giving me the water yet. If you engage in appropriate motivation. You contingency wielding... Anyway, so just because he's a behavior analyst doesn't mean he can reinforce... It does mean he can reinforce me, folks. I need to calm down because he's probably going to start worrying about my magnitude before too long. Anyway, the point is by running in temperatures like this it probably genuinely establishes water as a motivating operation or the water becomes, you know, the thirsty, the experience of hot and dry and cold and I'm even out of words because I need the water so bad. So it would probably really reinforce me to continue on with my lecture if you gave me a little bit of water because I think my behavior is getting a little weak. Thank you, sir. Much appreciated for hanging on to the...  Much appreciated for hanging on to the water while I give my pseudo life for behavior based lectures and trying to demonstrate what motivating operations are all about. So keep that in mind with anything.  You probably established a motivating operation. Oh! My gosh! Hear that? You probably created a establishing operation. You established the effectiveness of a reinforcer by depriving of it or putting somebody like me with hair like that in temperatures and sun like this and in making me go run probably establishes the value of this stimulus as a reinforcer. So reinforcement then going to happen without some motivation, folks. Motivation is stimuli. Exposure to stimuli. Repeated exposure or deprivation or whatever. There's all sorts of different things to it. You have to watch lectures on that too. So hopefully you don't have any questions because I can't answer you right now. But probably will if you write something in the comments.


So last time we spoke about function behavior and structure. We discussed that functionalities are realized by various structures that interact with each other. These interactions are known as behaviors. Yes, we also looked at the function behavior and structure elements of a mood-based music player. But you also mentioned that we need to integrate the function behavior and structure. Why do we need to do that? Yes, that's an important question. So learners, what will happen if you do not integrate function behavior and structure? Pause the video, write your answers in the notebook and then continue. If you do not integrate function behavior and structure, the design would be many solutions and incoherent. For example, mood-based music player, if you had only focused on the functionality of mood detection, we would have created many solutions for achieving this functionality. This is not decidable. Okay, so we will have an incoherent solution if we do not integrate the function structure and behavior. But what are the benefits of integrating them? Creating a conceptual design, it is useful to integrate function structure and behavior for the following reasons. It helps us create an integrated view of the design solution. We also can ensure that the solution fulfills all the requirements. This unified view can be used to communicate within the team and the various stakeholders. It's also a useful technique to avoid getting stuck while creating a design. Okay, so now I have these individual fb and s. Now, how do I go about integrating the function structure and behavior? Is to start from known solution. For example, we know that there are cameras that can deduct mood automatically. These could be the structures in the design solution. But we also need to evaluate if these structures are applicable in the current problem and adaptive. In the next step, we can start thinking in terms of end user as well as a system to generate behaviors. For example, a user would place the camera to automatically start playing songs. The system could fetch song list based on a particular mood. And so on and so forth. In the next step, we map all the structures that work together when a behavior happens. In this way, we identify structures, behaviors and map them. The next step would be to identify how the structures and behaviors satisfy a particular requirement. So now we have identified structures, behaviors and functions and also mapped them. Okay, so this looks something like a graph, right? Yes, it is called the function behavior structure FPS graph. The nodes are the FPS. Links connect the FPS. Just like a graph, you could start from any of the nodes F, B or S and move on to the next as you identify and connect them. So learners, here is a question for you. Can you think of combinations of FPS graph and the kinds of graph that can be created? Pause the video, write your answers in the notebook and continue. On the screen, there are combinations of the FPS graph. In this graph, the behaviors are mapped to the structures and then the function B, S, F. In another graph, functions are broken down and mapped to structures which are combined to different end user behaviors. F, S, B. So there can be many types and kinds of FPS graph. Oh, okay. So there are many possible FPS graphs for a given problem. But how will I know if the graph that I have created is good enough? That is a good question. There is a checklist of things to keep in mind to self-evaluate a FPS graph. Okay, that sounds interesting. You know, there is a learning environment where you could do all this. Create a FPS graph and self-evaluate it for different software design problems. Really? Well, that sounds like a very cool solution. Yes, Tink and Link is a learning environment where the learners are taken through the steps of creating the FPS graph as well as evaluating it. In this video, we saw the importance of integrating F, B and S while creating a design. We also saw that one of the ways to integrate them is to create a FPS graph. Yes, in an FPS graph, the nodes, they form the function structure and behavior and these nodes are connected via links. We also saw that there are many combinations and kinds of FPS graph possible for a software design problem. A checklist is available to self-evaluate the FPS graph. In the next LED, we will explore how to use Tink and Link to create and self-evaluate a FPS graph for various software design problems.


Thank you for staying with us. You're still watching the breakfast on plus DB Africa and now it's time for a hot topic. But for us less just browse through the papers a little bit. So most of the papers this morning had the, well, sad incident that happened in Ibadun, which is the explosion. So on the garden, we have outrage as Ibadun explosion exposes illegal mining week regulations on the punch. It says residents blame maliant illegal miners as federal government probes blasts. And on the daily independent, it says federal government committee to investigate breach of explosives control law. And finally, the nation says Ibadun explosion three dead 77 injured 58 houses damaged. Well, we have other other topics here or other headlines here. So if you want to catch up, you can, you know, check out this newspapers. But now let's move over to our hot topic. And our hot topic this morning is reps iterate commitments to pass and build on state police. Now, one of the members said community policing is actually one of the very smartest way to police your people. Someone sees something, say something, know something, and they need to speak at the right time. But joining us to talk about all of this and how state policing can actually help us is comrade Mark Adebayo. He is a spokesperson. Good morning, Mr. Mark. Thank you for joining us. Yeah. Good morning. Thank you for helping me. Thank you so much. Yes. So I mean, we've seen we've seen what happened in Ibadun, sadly on Tuesday night, and we see how the explosion have, you know, claimed some lives, which as of right now, the toll is about three and about 77 or even more people were injured. And we're talking about state policing. What is the benefit of state policing? And how can we even start to use that in our system? Does that work? Because the reps right now are talking about the commitment to pass and build on state policing. As of right now, everything comes from a central place. But the the distribution or the division of labor, as we like to say, for the state. Now, do you think it's a great way? And how can we start to go about this? Yeah, well, first and foremost, my heart goes to the families of the 15th of the unfortunate blood that clean the ice. I'm destroying the properties in Ibadun. Yeah. My heart goes to them. I pray for this introduction to the public recovery. Yeah.  It's the same thing. The issue of the blast and the issue of security, the issue of state policing is something, you know, all of them are one and the same issue. If there was no failure of intelligence, if we had effective intelligence operations, people cannot gather explosives in the domestic area of a city like Ibadun. And it would be left there and then up to this disaster happened. On the issue of state police, state policing, community policing and reservoir. As good as it may seem on paper, I was part of the people maybe over about two decades ago that we started this, you know, focusing for state policing. But one factor has become, has been a constant denominator in all of these agitations for state policing. As good as it may seem, the major one is the misuse to which the state police can be put by the state governors that are notorious for their dictatorial tendencies, for their tyrannical tendencies and the disruptive, you know, orientation to reality, you see. If you get, let me give you an example. Look at the state-dependent electoral commissions. How well have the governors been managing that? How independent have those state-dependent electoral commissions have been? How fair are free and credible are the relations being conducted by those state-independent electoral commissions? You see that in most states, it is the party in power in most states that will claim to have won all the seats in all the local governments of the state. Is that party so popular? Is that government so popular to the extent that it will clear all the seats, all the council offices in those states? No. The question is that they simply just write results. The same way they are mismanaging the state-independent electoral commission is the same way they are going to be mismanaging the state police. Their opponents will no longer be able to breathe. They will not be even during elections that are conducted by the Independent National Electoral Commission. They will lead the state police on their opponents. They will use the state police to disrupt the campaign. In the major fair, we have really common denominator in the role of this argument, in the role of this agitation and focusing on state police has been the misuse to win the state governors to put the state police as wonderful as need food as this might be, because of the obvious and clear failure of the federal police in policing and securing our lives. We might say, okay, let's have the state police, but there is the fear of the misuse to which these state police elements could be put by the state governors, because by the time you now give control and command and authority of the state police to the governors, we don't know what will become of that. It may later become counterproductive. That is the fear, except there is a robust legislation that will make it impossible for the governor. I don't see how that is going to happen, because they are going to be answering to the state governors and if they are answering to the state governors as their commander achieved, they will do the bidding of the state governors. Look at the way during elections, state governors are employing dogs to do their dirty work. So they will not need that, again, that they have official, uniformed people they are going to use to do whatever they do. That is the major fear. Just a moment. That is the major fear that the Indians have been expressing about this state police. Mr. Adebayo, just a moment. Are we not focusing on the negative at this moment, because no matter how we see it, even now, the state governors, yes, they have some control on the police that is federal. They are the ones who buy the cars. They are the ones who give them all other things that they use to function in the state. So if they want to use the police against their opponents, they can still use it. So if neighborhood watch, for instance, is working in Lagos and then Amotecun is also working in the Southwest, don't you think that's a semblance of what the state police could have been? Because I personally haven't seen when these people have been unleashed on the opponents of whoever is in power. I'm just saying. Yeah, yeah, yeah. Thank you. Look, yeah, the state governors are mostly, you know, responsible nowadays for the maintenance of the so-called federal police. You are right about that. But they have limited control. They have limited control. They don't have for limited control of the state police. The constitution does not give them even the, because you're basically clear, before the governor can talk to the commissioner of police in the state, he needs to talk to the president. The president will talk to the IG, the IG, you know, that kind of crazy arrangement that he said. Of course, you know that they have found a way to bypass that and the state governors and the state CPs are working together. We are, they want to work together because it's not in all cases that the police, you know, as a matter of fact, the police, the CP is more powerful than the state governor when it comes to security management, because it is not acceptable to the governor. So, in the situation whereby we have had a situation in this country, when former governor Rotibe Amici, the former minister, was governor in the first state, we had a situation where a CP told the governor, you cannot pass this place. Told the CP, a CP told the governor, block the road and say, you cannot pass this place. So, the governor said, my state came down, just as I said, go back. So, the governor had to go back into his vehicle and go back. We have had a situation whereby police kidnapped the city governor in this state, in Alhambra state, was kidnapped by the police, you know, locked inside the toilet in this country. So, when it comes to security matters, when it comes to in the state, the CP has more power, has more authority than the governor, because he can disobey his orders. This is my concern. This is my concern, Mr. Adebayo. The concern is that the way you're talking that the state governors do not have power over the police, you seem to be exonerating them from all the heinous acts that have been happening in their states and they are the chief security officers. Are you saying this is just a name and they have nothing whatsoever that they could have done to make security better in their states? The governor could do a lot to improve security in their state. And because of the frustration that the issue of Amantecum has come up, the issue of Neighborhood Watch, like you have said, I give them that. We have not seen a situation whereby Amantecum or Neighborhood Watch or the rest of that have been reported to Mahandu citizens. We have not had such cases. And even those such cases happen probably to be very far and in between. So now the governors, you know, people make a mistake. They think that people do not think, let's talk constitution, let's talk law. There's no place in the in the 1999 constitution of the Federal Republic of Nigeria, that says the governor is a chief security officer of the state. There's no where, there's no place, there's no segment, there's no section or chapter of the Nigerian constitution. That said, the governor is a chief security officer of the state. It is not written there in black and white. It is not there. It is just a nomenclature given to the governors by people who do not know that there's nothing like that in the constitution of the Federal Republic of Nigeria. Yes, he receives the security vote every month. He receives a huge security vote that has no everything and the rest of that. Now we, if it's not as if I'm throwing a video idea that maybe that I'm not saying it's okay, let's see how the people watch us, they are not and they are not as armed as the Nigerian police that we are talking about. If it is a dissent, advancing to the state governors, that they have no authority and control on command over the commissioners of police in the state. The only thing that they have is that they can inject a CP posted to the state, although it is at the discretion of the president of the IGP to move that CP. It may just ignore the governor. But where you have a good working relationship together, the police and the state governors have collaborated excellently where to manage security. But we still have all these lapses. I'm just wondering how much effective this state police would be. They would be effective in the sense that they know the terrain of their state. They know the community, the notion of the commissioners and the rest of that. What is important is that we must make sure if we are able to guarantee that they would not be a source of headache to the public, to the citizens, that they would not be, you know, escorting the citizens, the disturbing people that are going above their local duties.  The way we have been seeing it, it would be excellent. I believe that they would do a good job because they know their community is very, very well. All these things we do better, but we must ensure that no injustice is meted out to any citizen of the Federal Republic of Nigeria. We are never, this week, of it. It is very, very important for us. But as of right now, that still happens sometimes. But as of right now, that still happens sometimes. There are people who are driving, minding their business on the highway, and they are being stopped by this policeman to check their phones, check their boots, you know, they even do stop and search. So if you're talking about the security of people and injustice, it still happens even though the police is not being controlled by the state. So does it mean we will scrap it? Yes, exactly. In the US, 49 states, you know, have state police. The Hawaii, you know, has a shared division. Don't you think this is a good way for us? Because at the end of the day, we're looking for better ideas to ensure that we have good security. You're seeing insecurity, you know, in the North, you're seeing insecurity everywhere. Even on the highway, if someone is traveling, even for instance, if I'm traveling to the East, right now, there's insecurity. Even if I'm just minding my business somewhere, there's insecurity. And now there's even this issue of the explosion that happened in Ibadu. So if we don't have state police, because most times, it's always going to come from, you know, the federal level. How do they know the terrain? How do they, you know, have that trust with the people that are there? Because sometimes, you know, it's even people from the North that are being posted to the South, which is great as well. But we're talking about state police and we're talking about, you know, things becoming better. But if your your issue is the fact that there's injustice and, you know, it would be twisted, you know, they would be twisted and turned to the governor since the governor is now the one who's, you know, in charge of them. Then if that's it, then I'm sorry, I don't think that's valid because it still happens So what is your response to that? That happens now by the field and police. There's always the truth in fact that we shall lie to our government and state police. But maybe you don't know what I'm trying to say. What I'm saying is that I'm saying that is a good idea, you know, is a good idea. The issue of terrain, they understand the terrain, they understand the news and carries of their communities and things of that. Maybe they'll be able to, I mean, they'll be able to locate where criminals, high doubts can be and they'll be able to take action about that. It would be wonderful. Every of that element of the entire government of this state police argument is very okay. But the trepidation, the concern is that the governors might misuse the authority over these police. Comrade Femfalano, SCN, you know, about 24 hours or 48 hours ago also expressed the same concern that the state governor, that the state governor was alleged to have elucidated a work back as part in the state and up to today, nothing was out of that. The issue is that there was no state police and there's not been state police by then. There is this fear, there is this fear that the governors might just misuse this state police. So I guess that's fear of dealing with this. But that does not mean, follow me please, that does not mean that we should throw it away, maybe with the bad ones. We might, it is now the use of the citizens to stand up on our feet and call out any governor that misuses this, because already the chief has learned, we need to, we can't stop it again, we just have to go ahead about it. But the governors must have listened to and made sure that they don't misuse this police personnel to harass their opponents, to intimidate the citizens. The federal police wants to have an evidence that you have been harassed, but we see what they have been doing now, but they know that the young lady who was harassed in Augustine, whose food was smashed down by one inspector like that. And that inspector has been arrested, he's on the road, he's on the road trying. If the same thing, the same discipline can be brought into the body of the state police, we wonder for all of them. You see, the state police, we lose its usefulness entirely if it is unleashed on the citizens in a way that is not beneficial for security and our common interests. That's what I'm saying. We need the state police, because sometimes, hopefully, maybe this one can just bring different attitudes into the whole policing issue in Nigeria. Maybe now, the issue of the America that you are referring to, that is the different climate entirely. No comparison with that kind of climate, that kind of advanced democracies. That kind of advanced democracy. We shouldn't have to be baking, working to be better. Yes, absolutely, we have to be police, but the governors, the governors in those places are not missing those personnel. We are entirely different, we are not the same, the attitude of our leaders, of our rulers, of our governors and everything, from the way we conduct our affairs here, is totally different from what they are doing that please. We should not even compare ourselves to that lady. Yes, we are showing a sample of what they are doing with their own police. Can we replace them in Nigeria? That is the big question. We can try. Okay, sorry, Mr. Adebayo. Well, let's say if you want to learn a language, the best way to learn a language is by speaking it. So if we don't begin, we would never know the challenges that we need to surmount or not. So we do hope that we can begin something and see how it goes and be tweaking it a little bit as we go along. But now, the concern is the coalition of United Political Parties. What's your stand on the insecurity that's happening generally in Nigeria? You see, it is regrettable, it is dampening, it is actually very tragic that we have reached the level of this collision with Ghana. It is horrific. You wonder that we have almost 10 you know, intelligence agencies in this country. We have the DIA, we have the FIB, federal intelligence and intervention group. We have the state security services. We have the directorate of briefing intelligence. We have the national intelligence agency and so on and so on and so forth. And because if failure of intelligence is a guarantee or a security, if you do not get your intelligence gardener right, the society will pay for it. Failure of intelligence is what made America pay for that. Failure of intelligence was what led to it's paying for October 7th. So, but in this country today, you wonder what all these intelligence agencies are doing. Right before the analysis, Boko Haram was gathering, was mobilizing, was recruiting, was bringing in arms into the country in Borno State. They were training their personnel right under the analysis, they were training their personnel, they got arms, they got weapons and the only terror on this country and we are simply with it today and we are here to. So, the intelligence agencies seem to me to be completely useless. I mean nothing. If they could train in America to go and, you know, extraordinarily additionally from Kaya to Nigeria, how is it so difficult for them to be able to pinpoint the gathering, the only way to guarantee me is about the terrorist and bandit that's gathering somewhere and then move me, you know. How do you explain that bandits and terrorists will gather like 500 in number and move over 100 or 200 kilometers to go and keep people in Borno State, to go and keep people in New York State, to go and keep people in the animal state. All the time they are mobilizing, all the time they are gathering, all these intelligence agencies are not aware. What are you paying them? Billions of naira for every year. Every year, these guys collect the mongos amount of money every year for mongos food every year. What are they doing? So, it was specifically telling us the failure of security and that is the problem we are having. We have some machine that I'm computing that they don't know, they are only on, they don't know what to do about this security. That is why it is bourgioni, almost on a daily basis and it is you and I that are the regular Nigerians and the masses that are bearing the brunt to travel from now. Again, it seems that labels in the expressway doesn't seem to be, let me say the group has got the kidnappers and the bandits are back there. But thankfully, if not for the amotecum introduction in the south-west and that is why we just need to give the police a try. Even for the amotecum outfit, only those who have become of the south-west by now. So, we have the police, we have the army, we have the air force, we have the navy, we have what you call them now, civil defense and rest of that. Yet, insecurity is decreasing every day. How do you explain that? The seat of power, the federal capital territory are bourgia. We are the president in the side, we are the chief of army staff, we are the IGVs living, we are the chief of air staff, chief of navastar, LSDC, everybody, DSS, NIA, that is their headquarters. If a bourgia indeed cannot protect a bourgia, it is a very dangerous signal. Tending towards the failure for this country got to be. It is scary because nobody can, it is indivisible. That within a bourgia, we are left here because of all these circumstances. We are the president in the sides that we are unable to protect a bourgia. You see, on the daily basis, I had a security analysis saying that minimum of 12 to 13 cars are totally in a bourgia either snatched or removed via the apart in a bourgia or the daily basis. And people are kidnapped anywhere in a bourgia. Even on the day, you know, the issue of one child is more prominent and deadlier in a bourgia than any other place in the country today. So, the issue of one child, the police were on top of this before. I mean, sometimes, three years ago, one year ago, you see that the two policemen, and I don't know what has gone wrong with the game in a bourgia. So, if what is happening in a bourgia tells you what is happening, what is happening in a bourgia, if they are unable to protect a bourgia, how do you expect them to protect a bourgia in a bourgia, told them in a bourgia? Over 900,000 square meters, how are they able to protect the rest of the territory of Nigeria? We are unable to protect the police in Nigeria, capital, where all the power in the country reside and work. That is an issue. Hopefully, the security agencies will just sit down and do something about this thing. It is not a thing, it is so esoteric that it is impossible to secure this country with all these security agencies, the multi-layer, multi-level security agencies that we are after. That's why I'm wondering what the state police will be able to do. Federal capital territory, you are federal police there, why are they able to secure that federal capital territory? How come? So, is that not telling us something? And one thing that I'm sure about is that even Nigeria Army wishes to be effective, they will be, because we have one of the best trade mission personnel over the world. It is that knowledge globally that the Nigeria Army personnel have one of the best trade in the world. What you have to say, what is happening on this station? We have this call on the issue of corruption before. Look at the Mungos Corruption Association that has been, you know, concerning how corruption has been fouling the Kuku Aram students in Eastwap, because money is meant for armed and religion. So, we have to wrap up quickly, but I just want you to just touch on this. So now, I like the fact that you said we can try the state police because you just have to do a lot of things to see which won't work. Now, in trying the state police, do you think the powers should be limited? So, they're still stapled, because your concern right now is going to be harassment of citizens, injustice by the state governor, since he will be the one in charge. So, do you think the powers, we can have the state police, but their powers can be limited to ensure that this is not happening? Because at the end of the day, in as much as we're trying to battle insecurity, we still want the lives of Nigerians to still be secured and they're not being harassed by other people? Well, constitutionally and legally, their powers are not limited, because there are things they cannot go into. There are things that the federal police, only the federal police can handle. You know, their powers will be constitutionally limited, that's what I'm expecting to see. One of the things, one of the best things we can do is that to encourage the state governors to partner robustly and effectively with the civil rights organizations, you know, understand, they need to partner to ensure that this issue of harassment of citizens does not happen. You know, you need to partner, there must be a content in the curriculum that allows for the mental human rights chain of the citizens that they know how to handle to deal with the general public. Because it will be attacked with the general public. So they need to be effectively trained in inter-tribal relationships and for them to understand the fact that they are primarily attempting to secure the citizens and not to harass them and not to extort them. And then to put in place, you know, there will always be, but no matter how best or how well you train them, there will also be violence, there must be safety, you know, instruments put in place, there must be mechanisms put in place for them to know that there is a system of reward and punishment for the good and the bad. So, and I believe that's good. And more importantly also is the fact that before they start this state policing, they must ensure that they will take care of these policemen and women, they must make sure they are playing their surrounding well, they must make sure that they are in the state that will be responsible for their kids, their uniform, their car, their boots, their bed. All right. So, the situation will probably be corrected up for you. You know, one of the reasons why these people are very, very oppressive and aggressive against the citizens, the fact that they are not going to take care of, they are not going to take care of. And we all, we need to take that out very seriously. Yes sir. I want to say, it comes to me that there are police people. All right. This is where we have to wrap it up, sir. Sadly, we have to wrap it up here. So, there was a very important mention. Yes sir. Thank you so much. Thank you for joining us. Thank you for sharing your valuable contribution. We appreciate you. Thank you so much. All the way. Thank you so much for being here, thank you very much. Thank you. Thank you. All right. You've been speaking to Comrade Mark Adebayer, he's a spokesperson of the Coalition of United Political Parties, CUPP, and we'll be talking about the fact that the reps have reiterated the commitment to passing the bill on state police. We'll go on a short break and when we return we'll be looking at our next hot topic, but let's look at the weather first, see you soon.


Well it was a blessing for me to be able to go to Olivia as the first time coming out of the country to really to take the gospel and preach the gospel out there to unknown people and different race, different culture, very poor and it was just so humbling. I remember the incidents which we were going to downtown to the main Plaza to open and preach over there and I was very excited but we were had to take a bus it was like a 40-minute drive to downtown and remember this is a bus you going through hills I mean it was kind of interesting scary going through cliff there was no barriers in the in the side of the road and there was a lot of people coming inside the bus they were picking up people that were having that direction and it was not a very large bus but it was packed they packed people inside of buses like Sardin so our leader Charlie decided that we should go ahead and preach inside the bus what a best time to do it were a lot of people and so he started for like maybe 20-25 minutes and people were very listening to what was said and then after he got done I had the opportunity to start preaching in the in the bus and it's kind of interesting how these people so simple and poor but really what a hard to listen and when they feel that they are being pierced by the word of God the expression that they do is that they bow their heads and they don't even look at you and when I was opening and preaching inside the bus I was drawing drawing the attention that we have an issue in which that we are strapped this is an analogy that our brother Mark used in one of his message that we are strapped with this bomb that is called death and we don't know and this bomb that was dropped everybody is strapped with it it doesn't respect the young the child and my emphasis was like if you die today when you got into a lot of you into heaven so I really started to preach the the coming of Christ and not only the coming of Christ that is so imminent that we don't know we're gonna die so what are you gonna do in the day because the Bible calls it says very clear that it's appointed man died in the judgment so what are you gonna do in that day of judgment when you're gonna stand before God your eyes will close when you die and open your eyes before the throne of God and what you're gonna do and say so it was very impact I was an impact by the humility of the people people wanted to hear no one stood up and say you too loud screaming while you're it was just people listening to the preaching so that was very interesting and we got to the Plaza I really think I didn't preach in a plaza because it was so totally different atmosphere it was a lot of argument and debates going on in the Plaza but one of the brothers started to preach there at the Plaza and just seconds the crowd just drew next to him and started asking questions and a lot of stuff broke out and it was kind of scary but thank God that God was with us and God planned this trip and I know that God moved in a mighty way so it was a privilege to be part of this


How late it was this you spin me what round baby right round like a floppy disk Okay, so I've got my five and a quarter floppy disks not a good singer good engineer Got my five and a quarter drive wired up to my feather and I'm now running the mass storage demo For Tina USB, which means that this is actually showing up as a 360 k Double-density drive. I had to do we factor a little bit here so you can tell it Hey for the MFM decoding you're expecting 360, but here's the good news It just shows up as a drive and if you're like, hey, I want to know what's in this read me dot text It's a mouse driver for you know an old IBM PC, but it shows up just like a disk drive This is a good example of why you can't just use a USB floppy disk because there's no such thing as a 360 k IBM USB floppy drive, but now there is