for seminars is really going to be about how to interpret your data. And we're going to talk about databases for compound identification. But then we're also going to learn about statistical methods and tomorrow Metaboanalyst to interpret your lists of compounds and concentrations. So for this module, we're going to be talking about the different databases, different types of metabolomic database models. We'll look at some of the NMR and MS spectral databases. We'll also talk about pathway databases. And we'll learn a little bit more about what we call comprehensive metabolomic databases, which are distinct from spectral databases or pathway databases or compound databases. So again, what you did in your lab was basically to get lists. So from spectral lists, the next parts are how do you go from lists to biology, lists to pathways, lists to interpretation. So historically, there have been two solitudes, the same thing, I guess, in Quebec. But you've got chem informatics and bioinformatics. And they've evolved quite differently. And the way you can think about them is that chem informatics has been around since the 1960s. It was developed largely for the pharmaceutical industry and for medicinal and organic chemists. In the 60s, computers and softwares were very expensive and were a way of making a lot of money. And so in fact, American Chemical Society and others have used that model for many, many years and still do. So that most chemical information is not free. Most of it's very expensive. And most of it is very limited to public. It's supported large companies like Bilstein, Sigma, established things, MDLs, another group, CAS. On the other hand, bioinformatics, which came into the fore in the 1990s, was not designed for chemists, but was designed for molecular biologists. And by the 1990s, the web has started to appear. And there are very strong pressures on the US government, in particular, to make everything open access, which they did through an act of Congress that established GenBank. So the result is that in bioinformatics, everything is web-based, open access free. And it's largely funded by federal government agencies from NCBI, Genome Canada, EBI, NIH, and so on. So those two things have sort of made it a little difficult for metabolomics, because metabolomics falls in the chem informatics world, yet it's trying to interface with the world of genomics and proteomics, where everything is free and open access. So that's putting things a little bit in context, and partly explains why metabolomics is lagging a little bit. What do we use databases for? Well, databases, whether they're public or private, are basically a way of information consolidation. But when you build a web base, you can actually link it through a variety of techniques, software, hardware methods. In databases, you can do lots of information retrieval. You can do query matching. Everyone's done that some way or another. Databases also contain lots of reference values, reference data, reference images, reference spectra. And this is how some of the software you guys are just using has been working. So these actually are software tools based on large databases. We also use lots of databases for training and testing things. And this is how a lot of the tools that I've mentioned, predicted tools, have been tested and trained. Primarily in the world of chem informatics and bioinformatics and metabolomics, we use databases a lot, for both similarity searching and prediction. And so structure, sequence, spectral prediction, and for the search. Databases also evolve over time. And in the databases I've built and probably some of you also have been involved in, often they just start as a local consolidation of information you just wanted for yourself or your lab. So it starts off as a hobby database. In fact, GenBank started off as a hobby database. It was kept by a group, I guess in California, where they're just trying to keep a small number of gene sequences related to oncogenes, viral genes. Same thing with protein sequence databases, the 60s. Those are also sort of hobbies. But eventually people start asking for them and they become a little bit more than a hobby and so they start becoming curated and they start becoming a little more sophisticated. They might be relational. And eventually people say, well, I'd like to contribute to the database. And that certainly happened with GenBank and obviously the Protein Data Bank. And no longer is just something that's local to a small community, but it's open to everyone. So open deposition, the result is that many people are doing the same thing at the same time. So it gets a little bit redundant. Often because the database is so large, it also has to become distributed. In the case of GenBank, it's distributed in three different countries, Japan, US and Great Britain. So there's a trend as well. Hobby database is small, limited depth. As you get to larger databases, coverage gets greater, the depth may get greater. And then as you get to an archival open deposition database, it gets very extensive coverage or breadth, but not a lot of depth because you've got everyone who's just racing to put stuff in. There are only a few truly open deposition well-archived databases and most of them have to be run by government agencies. GenBank being one, PDB being another, and Unicrote being a third. Just because the costs and resources are absolutely enormous. As you move up in terms of the scale and scope of databases, also you need to become more standardized. There's greater dependence on being automated and the expectation for querying capabilities get greater and greater. So as I say, it's a challenge maintaining databases. It's easy to start a database. Most databases kind of sit in the middle, curated non-redundant databases and that's where I would say a lot of the metabolomics databases are at. So I mentioned this issue when we saw this picture before about how there's software already that can take an unknown gene sequence and unknown protein mass spectrum and give you information. Give you information about identification, possible concentrations and that. For metabolomics, the longest time there really wasn't that. So over the last 10 years, there's been a real concerted effort to do that. Prior to about 2007, most of the data for metabolomics was in textbooks. Basically it's been lagging behind genomics and proteomics by about 15 to 20 years. And typically because metabolomics covers such a broad area, you can see just in terms of the people here who are looking at all kinds of areas of science. People who are also chemists, people who are software specialists, people who are working in clinical work, drug research, plants, parasites. We've got a whole range of people. And so to try and create a database that meets those needs is quite challenging. The result is that it's led to a bunch of different types of databases that can sometimes are used in metabolomics. So there are the spectral databases, which are crucial. And you guys have seen how these spectral databases are being used in the exercise that you just did. There are compound databases. So these are things that allow you to associate a compound name or like a formula with a compound or a mass that allow you to identify compounds. The pathway databases are allowed to go from the compound list to some biological interpretation. And the comprehensive metabolomic databases combine the spectral databases with the compound databases and the pathway databases with a whole lot of other things, along with descriptions and details. Usually the focus always on a specific organism. But there can also be ones that are fairly comprehensive covering multiple organisms. So I'm gonna talk about each of these databases, but these are the ones that allow you to go from your lists of compounds and concentrations to getting some more information about them. So the NMR spectral databases, there are several that are around. And so we were looking at sort of defined biofluids, but there may be situations where you're looking at very unusual mixtures that are not blood or serum or urine or something like that. So you're having to try and match NMR spectra to something a little more exotic. So these are examples of databases that have reference compound from pure spectra, pure compounds. So one is in Japan, maintained by the Japanese Institute of Standards. It's been around since the 1970s. So it includes not only NMR spectra, so carbon NMR and proton NMR, but also has mass spectra and FTII spectra. You can do a lot of spectral searching with this. However, most of the compounds in this site are not metabolites. There's some, but the vast majority are not. Another one, which is maintained by the Biomeg Res Bank. This is a database that was originally established for protein NMR, but has expanded to metabolomics. And you can go to the metabolomics section there and you can find almost 1,000 spectra, well, actually more than that, spectra for almost 1,000 compounds. There's an average of five or six spectra for each compound. So it translates to something on the order of about eight or five or 6,000 spectra in total. You can search by names and synonyms, inchy keys and symbols, smile strings. Originally it was designed for plants, but it's also extended to include mammalian metabolites. So in that regard, Biomeg Res Bank has some unique compounds that would be of interest in areas of botany, food, and sometimes fungal studies. They've also assigned all of these reference spectra. So another database called NMR ShiftDB, it's version two. This is a community database where people submit proton and carbon NMR spectra, and it covers a lot of compounds. So it's up to 40,000. So it's bigger than the Biomeg Res Bank, it's bigger than the spectra for ACE, and it gets growing. But it's also one that's largely for compounds that are not metabolites. So interesting NMR ShiftDB was started by Chris Steinbeck. Some of you might know Chris who's been involved in leading Kebby. He also has established the database called Metabolites. It's not, as I said, limited to metabolites. There's lots of organic compounds, so I'd say metabolites might represent 1% of the total. It also supports chemical shift prediction. So if you have a compound pasted in, it'll predict the likely chemical shifts, which is very useful. You can search by name and structure. It also stores the files as Jcamp files, which is the NMR standard. It also includes chemical shift assignments that people have made. Another database maintained by the people who developed Biomeg Res Bank have a bunch of spectra, but also have a whole bunch of literature-derived spectra that they've added to. So it consists of about 20,000 compounds, but only about 2,000 half spectra. So I like the Biomeg Res Bank. It supports searches by names, chemical shifts. You can also do mass spec searches. It has data on chemical formulas, synonyms, species associations, references, and other public links. So those are some examples of spectral databases for NMR. We've also looked at, and I've shown this slide before, but these are MS spectral databases. So the NIST one is one. METLIN is one. We've talked about the goal. So in BASIL, what you're saying is we have incorporated that data in these other databases, or in BASIL, when we pick from library, can you pick these different libraries? Yeah, so in BASIL, actually, you have no library choice. You choose a bio fluid, and it has its own specific library. So those spectral databases I'm talking about are really intended to deal with unknowns that you couldn't, that didn't match anything, say, in BASIL. And so you might have a list of chemical shifts that say, okay, let's hear something way up at 8.9 ppm. It's not fit. What in the libraries have peaks at 8.9 ppm? And you could search through those, and it might suggest a half dozen compounds that have chemical shifts way up at that range. So no, with BASIL, you can't incorporate because it's very strictly controlled by the protocol. And in many cases, the spectra that are collected in these databases were not collected with the same protocol and at the same pH, or they were collected in organic solvents, which can also change the chemical shifts a little bit. So let's say you have your sample BASIL, and because you expect loads of your stuff to come out, you would get something that might be unknown if it's not in that library. That's what you typically do. But as I said, I think the real intent of these libraries is to deal for things that weren't serum or cerebral spinal fluid, which BASIL deals with very well. So if you're looking at, you know, pine sap, or looking at some jellyfish extract.  No. You can only do peak searches. So you have to type in your peaks. You have to type in your chemical shifts that you've manually measured. So BASIL is quite unique. It's the only tool out there, really, that does automatic assignment and quantitation. But it's just limited. But it's just limited. So if BASIL were to work for everything, it would probably take everyone in this room working steadily for five years on every fluid to try and cover that. That's right. BASIL is being modified so that people can create their own libraries and do other fits for it, but it's not really online or those tools aren't really publicly available yet. Next year. So I think, I mean, the point, these are very good points that you've made, but the field is still evolving. Software isn't mature. It's, I mean, you saw even XCMS, which gets, you know, thousands of users and didn't work for everything and still couldn't handle most browsers. And this is because a lot of these things are run internally in their own labs, whereas in the case of GenBank, it's run by a huge government agency with staff of thousands.  So, you know, Gary Shustak is spending, you know, lots of money, probably personal money, just to keep XCMS online working. We spent a lot of money and time trying to make sure that Bazel and HMDB all work for people. So these are sort of again, just our gifts to the community, but they're not as sustainable or easily maintained as, you know, Uniprot or PDB or GenBank. Anyways, getting back to the mass databases, I'll briefly talk about a few of them. The Golem database I mentioned before has a lot of GCMS data. What's nice about it, it has mass spec and retention index data. And it's done this for about 1,450 metabolites. A lot of them are plant metabolites, but many plant metabolites are also human and mammalian and fungal and bacterial metabolites as well. Lots of spectra, all the GCMS data is compatible with NIST format and Amdus software. So it is possible to extract and mine the Golem data and even to search it. As a nice looking website, and if you want to explore, I did encourage you to do so later in this evening. You can do searches with it. Tools, facilities are there. And this is an example of some of the things that you can get from that. Metlin is the database that you guys will have interacted with, especially if you've gone to that site that is in the public share, or if you were successful in getting your data processed. So Metlin has almost a quarter million compounds. And what it's done is, as I said, there's a lot of peptides and then a lot of plants, a lot of drugs, drug metabolites, a lot of exotic toxins, a lot of natural products. So it's very extensive. They have a lot of MSMS spectra. This represents spectra from about 12,000 compounds, of which close to 8,500 are protein, are peptides. So that means there's only about 4,000, what I call organic compounds, or non-peptide metabolites. That's still a very large collection, but it only represents about maybe 5% of the metabolome, at least in terms of the human metabolome. And only what's 1-60th of the number of metabolites that are actually currently listed in Metlin. So it underscores the point is that it's very, very difficult, and will be very difficult over the coming years and decades to actually get authentic mass spectra for all the compounds that we know about. So that's a challenge. And Metlin effort has been going on for 10 years and has had a lot of support from many, many companies and resources, which is one reason why they've been able to collect so many authentic spectra. But most of those spectra, as I said, are peptides. And so that's sort of the very, very low-hanging fruit. And most hits that you get, or most metabolites in mixtures, are not peptides. So with Metlin, if you've got peaks that you've been working with, you can enter a mass, you can choose whether it's positive, negative, or neutral, you can choose the type and adducts. And once you've chosen those things and you can just press submit, and it will list the hits. And you've got information about the compound, the name, and other information. You can do not only a single parent IMS search, we can also do MS, MS searches. So the data can be in MZXML, MZML, or an MZ data format. And so if you submit those, then you can also do MSM search. So that's similar to the CFMID website that I mentioned before. So both Metlin and CFMID support this MSMS search. Metlin searches against its database of measured standards, whereas CFMID searches against both measured standards as well as predicted standards. MassBank is a Japanese effort which is separate from AIST and also from Metlin. It consolidates many, many different spectra from many contributing labs, most from Japan, but a number from Europe and the US. And so it collects spectra not from a single type of spectrometer, but from Qtoss, triple quads, GC, FTICR, iron trap. So it's got a collection of about 15,000 compounds. So in that regard, it's larger than Metlin in terms of the total number of mass spectra compounds. But it's, I think it's smaller than Metlin in terms of the total number of spectra, yeah. So Metlin has 67,000, MassBank has 41,000. But the compound diversity in MassBank is much greater and much more impressive. So in some respects, it's probably a more useful resource than Metlin. So MassBank has a nice interface. You can browse the data, search the stats. You can also do peak searches. So gain allows you to do both paradigm mass and then SMS searches. So those are examples of databases, of spectral databases for mass spectrometry and for NMR. I also mentioned compound databases. So let's say you now identified your compounds. What are they? What do they mean? Alanine, yeah, it's an amino acid, but what's it used for? So there are resources that do have additional information that might be the physical properties of the compounds, additional links, literature, sections, other items. So one database is called KEBI. So the Chemical Entities of Biological Interest. There's about 45,000 compounds right now. Most of the material in KEBI is a derivative of what's already there in the literature or other databases. So they've mined KEG, they've mined database called lipid maps, they've mined drug bank, and they put that stuff into KEBI. So real focus on names and synonyms and ontologies. There's additional information about structure and it's very searchable. PubChem is certainly the database I think all of us have heard of. So millions and millions of compounds and substances. There's limits on how big the compounds can be. It's collected from many, many different vendors and depositors. It has just like KEBI, it has synonyms. It also has chemical property data. It has the structures and pictures, smiles and inchy strings. Some of them have bioactivity. They also have links to NCBI databases and PubMed. And in fact, that's quite useful if you're wanting to do some literature searches and finding out more about how this compound may affect biology or disease or whatever. You can search by name, by formula, by structure, by physical properties. ChemSpider is a European version of PubChem. It's run independently. It has almost as many compounds from a larger number of data sources. It has a lot of calculated properties. They've linked it to Wikipedia articles. They have Spectra for some. They have pharmacology links. They've linked it to mesh headings as well, which I think is now done as well in PubChem. So ChemSpider is trying to differentiate itself from PubChem and has, I think, a lot of really useful resources that most people may not be aware of. How many of you have heard of ChemSpider? One, two, three, four, five, six, seven, eight. Okay, so maybe half. Anyways, just so that you're aware. Ligand Expo. This is the collection of small molecules in the protein data bank. The reason why this is really important is because it actually links chemicals to proteins in a physical way. If it's crystallized, you actually see that compound bound to the active site of this enzyme or transporter. And so it links chemicals and metabolites and drugs to their targets, which is really important biology. You get not just a 2D picture, you actually get the 3D structure of these molecules as well. And it's searchable like all the other ones through names and formulas and in-cheese. But it's a small database, a few thousand compounds really, but still very useful. There's other databases, less well-known. I've mentioned a couple of them in passing, but for instance, 3D Met has the 3D structure of about 8,000, 9,000 natural metabolites. NAPSAC has about 50,000 plant metabolites. My compound ID, this is a resource developed at the University of Alberta, which is a database of metabolites of metabolites. So I mentioned that before, which are these transformation products, the ones in the liver and the microflora, and this is developed by Liang Li and Guo Guilin. So there's almost 11 million compounds in this database. And when people search against it, they find a lot of hits, especially in mass spectrometry. And then there's another one called lipid maps, which is maintained at UCSD, and it contains about 30,000 different lipids, covering lipids you'll find in plants, animals, microbes, and so on. And it's helped establish a lot of the nomenclature for lipids and the domics. I'll highlight my compound ID a little bit more. As I said, it takes the metabolites of metabolites. It has 76 transformations or fragmentation or modification sets. And so it's taken about 8,000 metabolites from the human metabolome database and transformed them based on their chemistry. And that first transformation generated about 375,000. And then those modified metabolites are then passed through the second phase of transformation, and that generates another 10.5 million, which is why it comes up with almost 11 million metabolites. It's actually a web server, so it's searchable. And so you can submit queries based on the high resolution mass spec of the Paranion mass, and it will come up with a list of possible or potential hits. It also lists all the types of transformations that it supports, and these are examples of some of the output that you can get. So not unlike Mettlin, but in many regards, much more extensive than Mettlin in terms of possible hits. So I mentioned, as I said, that knockout most study, where I said the first two hits that you saw were not the metabolites and Mettlin was wrong, I suspected if you tried searching against my compound ID, you might find something that's more closely matched. Okay, so these are databases, examples of different ones. There's no correct database. Probably detected some bias I have and some probably because of experience in what we've learned over the years. But each can serve their own purpose and each can be quite useful depending on the application you have. And so don't discard any, make sure you can use all of them or at least aware of all of them. So these are potentially useful for, again, identifying compounds, learning a little bit more about the compounds, but what do you want to know? How do you want to deal with them if you want to understand their biology?  The most famous, most widely used is the KEG database. But there's several others that are less widely used or less well-known. How many people have heard of the Psyche databases, Bio Psyche or Meta Psyche? One, two, three, four, five, six, seven. How many people have heard of Reactome? A few more, double hands, sleeping all over. How many have ever heard of SmithDB? That's the people in my lab. Yeah. So anyways, the point about pathway databases is that they're a really rich source of biological information. So they relate the metabolites to the genes, the proteins, diseases, signaling events and processes. So it's not just simply targeting information, it's biology. So the better pathway databases allow and provide tools to support visualization and gene metabolite mapping. A lot of them cover many species. KEG, literally hundreds, Psyche databases, a few dozen Reactome. Initially it was primarily human, but it's expanded again to dozens of other species. So KEG is largely considered the gold standard and it's really a great resource. Its access has become a little more restricted and support is a little lighter because of funding limitations. But I think everyone has probably, or most everyone here has seen a KEG pathway and each pathway is linked to a little metabolite card often. KEG has about 17,000 compounds that are metabolites. It includes a separate database of about 10,000 drugs. Now these drugs are variations. So they're the salt forms of the same drug. So in truth, there's probably only about 16 or 1,700 different drugs but it always depends on how you count them. Very rich in terms of glycan information. It's really unique in KEG and very rich. And with all the different pathways from all the different plants, animals, microbes, other organisms, something like 460 pathways. So very extensive. I mentioned SMIT, Small Molecule Database. I'm bringing this up because not that many people have heard about it. But it was designed in our group, partly because we couldn't find the information we were looking for in KEG and we couldn't find it in Metasite. And there's lots of missing data that seems to be there. So we started filling up and creating our own pathways. And because drawing pathways is difficult, we also developed some software to facilitate that. So right now, SMIT DB has 900 pathways. It has drug pathways, disease pathways, and it has metabolic pathways. These are all for humans. But many of the, or most of the data would translate very nicely to mice and rats, just usually a slightly different name and probably the only pathway that's different between rats, mice and humans is the ascorbic acid pathway. What's different in SMIT DB than, say, KEG and Ractome and Metasite is the fact that it has depictions of cell compartments, of organelles, of organs, protein locations in membranes or other organelles. It also displays a coturnary structure of the proteins. So there's just not a dot as it is in KEG. It's not necessarily a 3D structure, but a depiction of whether it's a dimer, tetramer, and so on. You can map gene, chip, or RNA-seq data, or metabolomic data to these pathways. And you can convert the gene, protein, and chemical lists into pathway or disease diagnosis. So this is what a pathway looks like in SMIT DB. So it's a little richer, I think, than what you might see in KEG. You can see the examples of the organs that are affected. In this case, it's showing, I guess, a phenyl ketone area and a disease pathway. And you can see the metabolites, they're drawn in detail. You can see the signaling processes, positions of where the metabolites and proteins are, cofactors associated with them, different components that are involved in the process and where they sit. Each pathway has a description, a little biography card. Each metabolite is linked to, in this case, the human metabolome database. Each protein is linked to the uniprot. You can map metabolites to the pathway, and so you can color them or highlight them. You can also provide concentration data, as you've measured it, and it'll be displayed in terms of the colors of the metabolites. Then you can also toggle off and on the color. So these are very color-intensive, but you can turn it to a black and white picture so that the colors of the metabolites are more visible. We're also modifying it so you can convert the pathways to look more like keg-like pathways, because people still like the keg wiring diagrams. And so you can flip back and forth between the two. Originally, all of these pathways had to be drawn on PowerPoint slides, and it took hours or even days, and then they had to be image mapped, which took even longer. So what we decided to do is develop some software called PathWiz. It's online now, and it's a web server. So you can generate pathways that look exactly like the SmithDB PathWiz, and it has all kinds of tools that allow you to draw these things fairly quickly. It also allows you to convert all the pathways into Biopax, SBML, and SVGN, and you can save them as SVG or PNG files. It uses a Google Maps-type viewer, so that's sort of a sophisticated JavaScript kind of viewer, and so it allows you to interactively manipulate things. And so it has a pull-down menu, and you can sort of select things. There's a video that I think is about to go on that'll explain how to do it in more detail, but you can add reactions and enzymes and elements, transport. It's sort of drag and drop, and you can adjust things with pivot points. Again, it's like using a standard drawing tool if you've ever used drawing tools for Adobe or Paint. And you can customize things. You can also draw in organs, organelles, cofactors. You can choose your own pictures or specific organs. And once you've lined everything up in the way you like it and adjusted things, then you can just render it, and there's your pathway. And as I say, the pathway can be generated in a colored format or a black and white format or a keg-like format, and that can be saved in all the standard pathway formats. Description just came out a couple of weeks ago and Nikolayk asked its research, and they're certainly welcome to use it if you'd like. So those are the pathway databases, and what I'm gonna now switch into is what I'll call the comprehensive metabolome databases. So these are the ones that combine everything together. So we've talked about spectral databases, compound databases, and pathway databases.  So keg qualifies sort of as a comprehensive metabolome database. Eco-psych is another one that's quite comprehensive, very good on E. coli, metabolites, human psych. So to be a comprehensive metabolome database has to have at least 1,000 metabolites. Most of them are organism-specific. They have to be continuously updated, and they have to contain a combination either of chemical data and pathway data, chemical, spectral, and biological data, chemical, pathway, or spectral data, or everything. So some of them are different combinations. So keg and eco-psych are chemical and pathway data sets. Human metabolome database would be chemical, pathway, spectral, biological data. And metabolites would be chemical, spectral, and biological data. So metabolites is making an effort to become the GenBank for metabolomics. So it's maintained out of the EBI in Chris Steinbeck's group. How many of you have heard of metabolites? Two, three, four, five, six, seven. How many have deposited data into metabolites? Okay. So at least you've heard a few. Now, today, everyone will have heard about it. The point is that it is intended for upload, and in fact, there's gonna be likely requirement for many of the journals that anything you publish in the field of metabolomics will have to be uploaded either to metabolites or to a metabolomics workbench at UCSD. So it's gonna be essential for publication. And the idea is that you're gonna be able to upload the experimental data, the spectra, compound lists, if you've got the biological metadata about the process and sample collection. Right now, it's the fastest growing database at the EBI, at least in terms of amount of data being uploaded and rate, but it's also the smallest database at the EBI. Anyways, it supports a variety of search options. It's linked to Kevi, and as I say, if you wanna get spectral data, this is a great place to grab some. And it's compliant with the metabolomics standard initiative, that's MSI. So it's important that people know about this, but it's also important that you guys start thinking about depositing your data there. If we don't, as a community, A, these databases will disappear, but it'll be increasingly challenging for metabolomics to be viewed as a legitimate science. So metabolites is at the EBI. The other major resource is actually the University of Alberta for metabolomics databases. My lab or my group has developed, I guess more than a dozen different databases. You'll hear about the human metabolome database, the drug bank, the yeast metabolome database. We've worked on phenyl explorer, which is a plant compound database, E. coli metabolome database, the food database, cow metabolism database, the toxic exposome database, SMIT DB database, and then serum blood urine CSF metabolome databases. So a lot of these databases grew out of a project that started in 2005, which is called the human metabolome project. How many people have heard of the human metabolome project? Okay, that's better than it used to be. Anyways, it was sort of the project that no one had ever heard of. It was sort of the equivalent of the human genome project, but it was intended to essentially identify and quantify metabolites in the human body. And a lot of that was to try and get material that had already been collected because people have been studying humans for centuries, but a lot of it was stuck in journals and papers. And what we were tasked to do was not only try and grab that data, but also measured ourselves, confirm it, validate, extend it, and then make all that data freely accessible. And that's what we've been doing and have done. We also developed a lot of tools and technologies, software, analytical techniques, protocols that helped improve metabolome coverage and throughput. The interesting thing is that when we started with the project in 2004, the literature review that we did at the time suggested there was about 690 metabolites in the human body. So that was what was listed in KEG, that was listed in the human psych and cross-checking, and they were pretty much convinced they'd covered everything. So in 2006, we were pretty excited when we actually had more than 690 identified or compiled almost 2,200 metabolites. We actually had a press conference about it. And it got coverage in nature and science and everything else. But later on, we discovered there was lots of missing metabolites. And so by 2009, we were up to about 6,400. By 2013, it's 37,000. Today, it's about 42,000 compounds. But really what we think is probably likely well over 100,000 metabolites that are detectable in the human body. So it's grown pretty rapidly. And in fact, whereas we used to depict the human metabolome as being much smaller than the genome, it's now inverted. And the metabolome is much, much larger in terms of diversity than the human genome. We've seen this picture already, although it's a little distorted for whatever reason. But again, a whole range of different metabolomes from exogenous to endogenous. And so these are housed in different resources, drug bank for drugs, T3DB for pollutants and toxins, HMDB for most everything, and then food DB restricted largely to food metabolites. So the HMDB, as I said, about 42,000 metabolites, we've been able to identify 170 that are really unique microbial metabolites. In reality, our microbes probably produce about 2,000 metabolites in our bodies, but 95% of them overlap with other metabolites that are already in our bodies, so we can't tell whether they're microbial origin or not. It has a lot of data on abnormal and normal concentrations, lots and lots of disease links, lots of spectral data that we've collected, some of which also has been mined from other resources. Because it links metabolites to genes and proteins, you can do sequence searches. Because it has spectral data, you can do spectral searches. And because there's a lot of data, it has a lot of browsing tools. It has pathways and pathway search tools, structure searching tools, you can browse by different biofluids, very extensive text searching and also full data downloads. So these are just some screenshots of what's inside the HMDB, shots of the spectral viewing tools, browsing tools from metabolites, details, pathways, data fields, there's more than a hundred different data fields. Variety of spectral searching tools, not as good really as what you'll find in CFMID. But we're working on improving those and making them a little more extensive and robust. So you can do MS as well as tandem MS searching. You can also do spectral searching by NMR, typing in peaks. You can draw a structure in there with a Java applet and it'll look for similar structures or identical structures and list their level of similarity. You can also look through different biofluids and look at all the different diseases and abnormal and normal concentrations. You can look through different biofluids and also look at the different concentrations. And it was really designed for clinical applications. What's actually much more popular than HMDB is the drug bank database. And the reason why it's much more popular is because it's the first database to link drugs to their targets. So in fact, it's used a lot in drug discovery and drug development now. So it's a small database, there's only about 15 or 1600 compounds in it, at least of drugs. There's a lot of drugs under development that are also listed in the database. But it has a lot more biology and this is because people have studied drugs to death. So it has a lot of stuff about absorption, distribution, metabolism, toxicity, mechanism of action, pharmacokinetics. It has a lot of drug metabolites, a lot of transport data, lots and lots of data on drug-drug-drug food interactions. But it's also allows you to do all the sequence and spectral searches that HMDB has. So it too has a lot of views, a lot of images, it's formatted very similar to what drug bank, which HMDB is formatted. You can look through drugs by categories, you can draw structures, you can search drug targets by the sequence, and then you can extract data through the data extractor, which is essentially a very sophisticated MySQL query system which allows you to put together queries without knowing SQL. Toxic exposure, and this is the one, as they say, which is trying to address this emerging area of environmental metabolomics or the exposome. 92% of all deaths in North America can be attributed to exposures of either small molecules or microbes. That's, I think, quite striking because most of us are inculcated with the idea that we die from genetic failures or something like that. But really, when you look at it, and this is not new, it's just that no one's really talked about it, but there's many papers that have highlighted this, and this is what epidemiologists always bring up. And so being aware of these compounds that ultimately kill us is, I think, useful to know, and also useful to try and understand so we're not exposed to them. And it's not like Agent Orange or Sinai that's doing it. In fact, many of these compounds that are in the toxic exposome database are produced in your body. These are things like uremic toxins and oncometabolites that have rather significant roles in development of disease. So this database structured very much like Drug Bank and has a lot of sane type of information, but obviously not everything in it is about drugs. But the way that toxicologists speak is very much the way that drug pharmacists and pharmacologists speak. Foods, we eat other metabolomes, and so trying to capture that information is something we've been working on for a long time. 10 very years here has been working on it longer than he wishes to, but it's probably still gonna be worked on for the next few months. So beta release is out there. People can look at it. It's still being added to, but it's one that covers a lot of information about what we eat, including some of the food additives that are there, and information about flavor, aroma, color, as well as effects on human health. So there's lots of data on humans, but obviously people study model organisms. And so we've been trying to extend this to other model organisms like yeast. So there's a yeast metabolome database. It's also relevant because yeast is used in a lot of food productions, whether it's wine or beer. So it was quite interesting to explore that. We enjoyed trying lots of different wines and the excuse of measuring the yeast metabolome, but anyways, it's fairly extensive and covers a lot. Another one is obviously E. coli, so that's another model organism. And in this case, it's been studied much more extensively than yeast, and we're actually doing another update on it in this next couple of months. So this database, when it's done by the end of summer, actually will have pathways for every single metabolite. So that represents hundreds, if not thousands of pathways that are being prepared and drawn. So right now we have 125, but as I say, this will have pathways for every single metabolite, which will be the first organism to actually have that done. So you can look at different databases and you can compare them based on what they have. And some are more extensive than others. Some have, were developed for specific needs or niches. This is not intended to be exclusive or covering all databases, but it's useful to look at what's there and to be aware of what's there. So I think we're coming up to the end of our time. I don't know if people had any questions with respect to what I've covered so far in terms of databases or how to help interpret your data.


Welcome to the reading from Trichy Assaram, today I am going to present a case regarding scapula winging, a book of case series. Myself, Bhagadalavek, Nammasartha 1st year, author to R. Chandru, not 3, V. Kavukovalan, our guide, Professor K. Venkateshallam, in his three chiefs, Department of Authorities. Introduction, scapula winging is a finding seen with clinical examination of the shoulder. The shoulder blade effectually sticks out rather than lying flat along the back. This outward and upward motion causes the scapula to resemble a person. Trauma is usually the inciting cause, although a variety of etiologies can cause injury to the associated nerves, including viral illness, allergic reactions, toxic exposure, and surgical complications, medications, and pre-existing or developmental medical conditions. Another injury results in subsequent weakness and paralysis. Causes of scapula winging. The condition of scapula winging or scapula protection occurs because of injury to one of the three medial stabilizing muscles that include the serratus anterior, trapezius, and rhombus. Causes of scapula winging. The function of these three muscles is to keep the scapula stabilized medially against a posterior chest wall, with injury to these muscles causing the scapula to lift from the chest wall. Based upon which muscle is involved, the scapula can deviate either medially or laterally due to the unopposed contraction of the remaining functioning muscles. Medial winging occurs from injury to the serratus anterior, and lateral occurs from trapezius or rhombus. Medial stabilizers are scapula. In the diagram, we can see the trapezius, rhombus, and the serratus anterior muscles, which are the primary medial stabilizer muscles of the scapula. Here, the anatomy of variations of the lateral lines can be seen. Case illustration 1 of trapezius. An 18-year-old was involved in a motor vehicle accident 8 weeks prior. He reports pain and weakness with deformity of the scapula. MRI was performed as the muscle tear or other injury. The A.P. frontal X-ray is non-contributory. MR imaging. A coronal fat suppressed free-to-weighted sequence demonstrates diffuse abnormal and homogenous hypointome signal within the right trapezius muscle without atrophy, which can be seen with the red arrow. Diagnosis. Isolated trapezius muscular denogation from injury to the spinal axary nerve. It is a 11th cranial nerve, clinically representing the scapula winging. Case illustration 2 of serratus anterior. The serratus anterior is critical for allowing the shoulder to take and move forward, increasing upward reach. This movement is exaggerated with activities such as punching a bag or swimming crawl and butterfly strokes. Conditions affecting the nerve and producing winging include blood trauma caused by pressure from crutches, sharp trauma from the causes, such as shoulder surgery, most commonly repetitive action trauma from sports activities. Other non-parametric causes include infections such as influenza, tumor, toxin, autoimmune, and inflammatory conditions of nerves, such as personage-turner syndrome. A long thoracic nerve in the real is its changes in the muscle, and this nerve is particularly susceptible given its superficial course along the side of the chest hole. Condensation. The axial STIR remains through the right thorax of a 22-year-old weight. Weakness and decreased mobility demonstrates increased signal within the serratus anterior to present with the arrow, positioned between the right scapula and posterior lateral chest hole. Case illustration 3.  The serratus anterior muscle between the second rib and scapula demonstrate enhancement. Case illustration 4. Colonial oblique T1 weighted image centered over the scapula in a 23-year-old with needle scapula pain for five years. There is a loss of muscle bulk and intervening fat intensity within the serratus muscle belly represented with this arrow. Case illustration 5. Serratal oblique STIR image parallel to the scapula rashes and middle to the scapula shows hyperintensity throughout the trapezius muscle. Represent with the arrow, with the maintained bulk suggesting acute denomination. The muscle lies immediately superficial to the rhombidinosis. Case illustration 6. 15-year-old main presence is trapezius pain with C34 denomination on EMI. Coronal STIR image showing asymmetrical diffuse left trapezius muscle edema. Corresponding to denomination injury of the spinal accessory nerve. Case illustration 7. The rhombidinosis lies deep to the trapezius and both the major and minor components pass inferior lateral from the vertebrae to the middle scapula margin. Denomination of this muscle is caused by injury to the dorsal scapula nerve which is often thought to occur as the nerve passes through the scapula. This is a rare injury with the incitinous cause, usually sports activities. Damage causes the swinging of the scapula and results in difficulty with throwing and overhead movement. American short denomination changes both acute and chronic. Continuation. Accelative and weighted image through the upper thoracic level reveals month atrophy and fat infiltration of the right rhombidinosis represent with the arrow and compared to the contralateral side. Case illustration 8. Alastrofibroma or scapula winging simulator differential diagrams. Accelative and weighted image demonstrates the mass lesion between the chest wall and more superficial serratus anterior at the level of the inferior angle of the scapula containing internal elements of lower signal and increased signal corresponding to fibrous and fatty elements of an alastrofibroma. Case illustration 9. Scapular osteocondroma or scapula winging simulator differential diagrams. Scapular osteocondroma is a rare finding resulting in scapula winging. These benign humans, contiguous with the bone cortex, can arise from the internal margin of the scapula, producing mass effect within the constricted scapula thoracic space. Continued, surgical detu weighted image in a 16-year old female demolishates a large one excrements contiguous with inferior ventral scapula cortex represented with an arrow, projecting intricately to about the chest wall. Notice the dissonant scapula thoracic bursa with the thick wall pseudo-capsular concession with bursaries. Case illustration 10. Venus malformation. Exile XT IR, trillium in the top, and exile P1 weighted at the bottom images revealing a mixed signal Venus malformation situated anterior to the rhombide major and medial to the serratus anterior. There was no scapula winging and this was discovered incidentally. Case illustration 11. Dissecting intramuscular ganglion cyst. Exile T1 weighted sequential images represented on the top, demonstrated dissecting intramuscular ganglion cyst within the trapezius emanating from the AC joint and coarsen medial. Serital and coronal fat suppressed T2 weighted images represented in the bottom reveal the intramuscular position in 60 characters of the lesion. Case illustration 11. Serratus antidive lipoma.  Case illustration 13. Plexiform neurophoropoma deep to left trapezius. Fat suppressed sexual post contrast T1 weighted and exile T2 weighted images reveal a large infiltrating enhancing mass represented with an arrow deep to the left trapezius muscle remodeling the left P6 lip extending from the left mural foramen consistent with the plexiform neurophoropoma. Case illustration 11. Intramuscular mass in right trapezius.  The differential included tumor and sarcoma with the pathological diagnosis of proliferative myositis. We can in the diagram we can notice the preserved low signal of hyperindex striations. Case illustration 15. Muscular strain in inferior trapezius.  Scab-lovinging treatment. Needle scab-lovinging is usually treated conservatively with observation for at least 6 months taller for nerve healing. During this time, physical therapy for muscle strengthening and tapping or bracing is performed. This is a long process and recovery can usually take up to a year or longer for resolution but is successful in great majority of patients. Surgical options for failure of conservative treatment include a split pectoralis major transfer whereby the sternum head of the pectoralis major it transferred to the inferior border of the scapula. Later scapula winging from the injury to the axary nerve often does not respond to conservative treatment. Here we can see the diagram of the lateral winging and medial winging and its treatment. Continued surgical considerations for failed conservative treatment include exploration of the injury site with muralizes often more favorable results compared to end-on-end switchering and now report. Most patients post-recovery from surgery continue to have a wing deformity. The injury is isolated to the trapezius muscle a special procedure called Eden-Lang muscle transfer can be performed whereby portion of the levator scapulae rhombite major and rhombite minor are diverted to the sites of terapia attachment. The lateral scapula winging from the injury to the rhombite muscle scapula nerve is usually treated conservatively with a cervical spine stabilization and physical therapy. Stiller injections and massaging have been noted to occasionally help. Suggesting this condition involves two facial lateral wings, the first connecting the lower medial border of the scapula to the spinal muscle and the second spanning from inferior angle of the scapula to the latissimus cluster. Conclusion, the scapula winging is a dysmorphic abnormality usually caused by injury to one of the innovative nerves of the posterior medial muscle stabilizers of the scapula. This condition causes pain and produces significant complications of the movement, thereby making activities of daily living difficult. Weakness is a commonality amongst all patients with scapula winging. The diagnosis of scapula wing is usually made cleanly, but inciting cause is often not obvious. Continued, electromiographic studies can help localize the muscle involvement produced in the scapula winging. Signistic imaging is not always necessary but high resolution imaging such as MRI can assist in identifying muscle involvement and allow localization of affected area where nerve impingement or damage occurs. Treatment is usually conservative and focuses on strengthening the surrounding non-affected muscles while providing pain relief. Sometimes, surgeries perform from refracted cases that do not respond to less invasive treatments. Here are the references which I have been using. Thank you.


The signal oil program the whistler That whistle is your signal for the signal oil program The whistler and the whistler and I know many things for I walk by night I know many strange tales hidden in the hearts of men and women who have stepped into the shadows Yes, I know the nameless terrors of which they dare not speak Yes friends, it's time for the signal oil program the whistler Rated by independent research the most popular West Coast program in radio history In gasoline, you know it takes extra quality to go farther and Signal is the famous go farther gasoline So look for the signal circle sign in yellow and black that identifies signal service stations from Canada to Mexico And now the whistler strange story The blank wall It was five o'clock and the crowd in the locker room had slipped into the usual routine of easy chatter About wives and food and the thousand things that happen every day in a bank as busy as the Crane national On one side the men on the cleanup crew were getting into striped overalls with a night's work ahead of them On the other the guards and messengers were exchanging official Navy blue bank uniforms for street clothes To Walter Mason though. It was a day of days He didn't say much as he stood in front of the locker at the end of the row Half hoping the crowd would forget it and let him go his way in peace Yet wanting at the same time to stand up on a bench in the center of the room and shouted at the top of his lungs As it turned out, however, he didn't have to make a decision Charlie Bryson another messenger made it for it. What's the matter Charlie? I just happened to think excuse me Joe All right everybody hold a minute. What do you I almost forgot this is Walters big night Oh Charlie for the love of my don't be bashful. They don't care about that much They don't want to be nice to walk from now on fellas. He's liable to be the boss one of these days I want you to know we think it's great. Well, thanks Charlie Out at the crane place on Long Island soup and fish Yes, you know they say there are two sure roads to success one of us to be smart the other is to marry the boss's daughter Yeah, but this isn't just almost we're gonna keep our eye on you water 25 years as a messenger isn't it well not quite I'll bet they give you a great big shiny desk less than a month after your daughter marries young crane You know Walt it's hard to believe Mrs. Crane will take this land down What do you mean? Oh, you know Donald's picture in the paper with his society gal and that one Naturally mama expected him to marry one of them. Yes, but Donald had different ideas. Oh, I know that only in a pinch I still bet mama told young crane what to do wait now you've got the kid all wrong He's you know, I'm very well Walter. Sure. He's clicking right here at the bank, isn't he working himself up from the bottom? Just like his father What's wrong with that? Well, Donald crane isn't exactly working himself up now wait a minute My daughter Kathy is going to marry Donald crane I'm happy about it. Naturally, but not because he's part of that Long Island crowd I'm happy because both Kathy and I think he's as fine a man as his father was they don't come any better than that That's all So long fellas Yeah, Charlie Good luck tonight. Thanks You know Charlie This is going to be the greatest moment in my life And you meant it Walter It's the most wonderful thing that ever happened a party at the crane home on Long Island Announcing Kathy's engagement to the son of Alfred Crane And that's the missing part of the picture, Walter It's five years now since mr. Crane died and you meant it when you told him He was the finest man you ever knew That's why everything has to be perfect right down to the carnation in the lapel of your rented dinner jacket And that's why even though it costs six dollars You ride out there in a taxi instead of the pre-war coupe in the garage at home Yes, Walter Everything has to be right My name is Mason beg your pardon. I said my name is Mason. I'm Miss Mason's father Would you mind taking my hat and coat Oh, of course. Let me help you. Thank you. I'm sorry sir. It was quite a shock. I I Thought I recognized you. It's nothing. It happens all the time. I'll tell Mrs. Crane. You're here. Thank you That was strange wasn't it, Walter?  Oh, it's nothing Kathy. There's nothing to be afraid of darling What do you mean forget you work at the bank daddy? These people are really very nice. Oh, it's not that You're worried about something I can always tell why it's just that I don't think it's right for you to go Walsing off with an old fogey like me while Don stands around with his hands in his pocket. Do you mind mr. Mason? Oh Donald. I'll speak of the devil. I'm just like my father can't stand competition. You see Kevin I'm only kidding matter of fact mother'd like to talk to you mr. Mason Of course, and it's about time you two got to know each other You know, I I want you to be as close to her as you were to dad. I hope so Donald Where is she in the library? Probably tapping her foot and by the way, whatever she says don't take it too seriously Please sit down mr. Mason. Thank you, Mrs. Grain and sharp. Yes, ma'am Would you bring us some coffee, please? Of course ma'am. I want you to know mr. Mason How much I admire your daughter? She's a charming girl. I'm very proud of Kathy. You have a right to be Donald worships her of course. I'm sure they'll be very happy. I Yeah, I Don't know quite how to say this mr. Mason I'm afraid I'll have to put it a little bluntly. What's that Mrs. Grain? I'm not entirely convinced. It's going to work out Donald and I have discussed it many times But of course he's a stubborn as his father. What are you talking about? It's hard to reason with young people at a time like this. I Opposed the marriage from the very first, but Donald is going ahead in spite of me, but I don't know what you let me finish I Was forced to look at this thing rather coldly I'd prefer to stop it now rather than see Donald do something he'd regret but wait, Mrs. Grain you don't Mason There's no point in disguising facts You must know Donald's decision came as a shock to me when I realized that thing was serious I took it upon myself to investigate your background quite thoroughly you What I hired a private investigator. He's been working for two weeks and he found out What nothing? Otherwise, I never would have gone through with this announcement party tonight. However Until the wedding, it's not too late and I think it's only fair to warn you that there are still some questions unanswered My investigator seems to have run into a blank wall You could find nothing of your whereabouts or Occupation prior to the time you came to work for my husband in 1922 and what's all this got to do with Kathy and Donald? everything people in my position of responsibility Can't be too careful mr. Mason. Oh, but they can mrs. Crain. I wish your husband were alive I wish he could have sat in that chair over there and listen to you talk about Right of you to investigate me What difference does it make where I was in 1922 or 1916 or any other time for that matter? You've taken a great deal up on yourself mrs.  Mason Sharp I didn't mean to intrude man, but your water doesn't tell it. Thank you sharp Poor mr. Mason a cup of coffee You will excuse me mr. Mason certainly Why didn't you tell her the truth? Billy Billy Billy Ford sing sing 1916 to 22 Charge manslaughter and don't tell me I'm wrong. I was there sharp Roy sharp It hit me between the eyes when you walked in tonight, but I wasn't sure until a minute ago There's a reason for the blank wall the gum shoe ran into isn't there? I I didn't try to fool anybody I told mr. Crain. He helped me. He's dead and I'm afraid the old lady hasn't got his understanding hard You know Billy as of two hours ago. I started believing in miracles again. What do you mean? I'm in a spot. I need ten grand the worst way wait a minute Roy We're gonna get it for me ten thousand dollars. I can't you ten By tomorrow night at ten Crescent bar and 58 and third. I'll be there about wait a minute Coming ma'am shop wait a minute shop. I can't the old lady's calling Just remember to meet me tomorrow night Billy or some of those questions of hers won't be unanswered any longer With the prologue of the blank wall the signal oil company brings you another strange story By the whistler There were so many comments about the information. I gave in last week's announcement. I'm going to repeat it tonight As I stated since the spring marks the 25th anniversary of the founding of signal oil company I looked up some facts which I thought would surprise you as much as they did me and here they are 25 years ago here in Southern, California Regular gasoline sold for 23 cents per gallon without any taxes On the other hand in 1940 Regular gasoline sold for only 18 cents per gallon and that price included taxes of four and one half cents per gallon Which had gradually been added? So the price of the gasoline itself without taxes had actually gone down about one half All right now. What about today's prices? Since 1940 items in the cost of living have climbed 53% Compared with an increase of only 13% in the retail price of gasoline get that since 1940 a 53% increase in the cost of living Compared with only 13% increase in the cost of gasoline And I might add the gallon of signal that you buy today is a vastly superior product the finest gasoline ever offered to motorists without extra cost So when you drive into your signal gasoline station and say fill her up Remember, you're not only getting the tops in gasoline quality You're also getting one of the most remarkable values that your dollars still buy today Now back to the Whistler Well, Walter after 25 years the blank wall that separated Billy Ford ex-convict From Walter Mason respectable citizen has given way It came at a terrible moment, didn't it Walter and you know that one word from sharp will put an end to the marriage of your daughter Kathy and Donald Crane You make an excuse to Mrs. Crane leave the party early arrive home before midnight and Spend six hours sitting in an easy chair in the living room thinking The morning light is gray on the windows when you rise Wearily start to put on your working clothes. There's no other way is there Walter Kathy's happiness comes first now Sharp must have his money. Good morning Walter. Uh, not so good. Mr. Cromwell. I'm not feeling very well Too much party last night. I'm not sure but I Was wondering mr. Cromwell if I felt like going home this afternoon. Would it certainly Walter I understand You should have found this morning. It wasn't necessary to come in. Well, I I thought I'd be all right. Thanks, mr. Cromwell Your savings account sure give you the balance in a minute Let's see Melford manning Mason.  You've got exactly thirty two hundred and six dollars and eighteen cents Thirty two hundred oh Thanks, Jim. Thanks a lot Charlie I I don't know how to ask you this exactly. What is it? Well, you're some kind of trouble Charlie How much have you gotten the bank? Huh? My savings. Yes, I I know it isn't a nice thing to ask It's no secret, but it's no fortune either About two thousand bucks two thousand and I know I couldn't ask you for all of it What I should say you couldn't the old lady to ring my neck. How much could you let me have Charlie? You know, I'd pay it back. Oh Sure gosh, Walter. You're my best friend. I'd do anything for you only this is all right forget it But Walt there must be something I know no no there isn't Charlie. Just don't mention this to anyone. Please don't Well, what do you think I Don't know car shows care. All right. I bought it from the original owner Motors had a complete over. Yeah. Yeah. Yeah. Yeah. Yeah, I know Well, I guess it's worth the five bills only five hundred dollars for a car is over pal.  I know you're the one person who'll give me a fair price Mr. Mason a fair price How can I put a price on anything like your colleagues? That's very nice of you Arnold But it's been taking up too much of my time lately and I've decided to quit. I cannot believe that you You'll drop out of the society completely No, no, what is happening? What if I've used it a long time to sort it on and I've made up my mind How much what is the collection worth? If I may suggest Mr. Mason I would prefer to to give you the money in the form of a loan That way I could keep your stamps here for a while all together in case you change your mind You're a very unusual man. Oh His business Mr. Mason business. I I just don't want to lose a good customer. I know better than that All right, mr. Businessmen How much of a loan can you make on it? $1,000 would that hill a thousand for a little collection like Arnold it's a tragedy how few of us ever know our real friends You added up Walter after leaving Arnold's tiny shop add up every cent you were able to raise It comes to a little over 6,000 not what sharp asked for but you have to agree have to be satisfied because There's no one else you can turn to That night at 10 o'clock you enter the Crescent bar He's already there waiting an empty cocktail glass on the table in front of Hello, Billy, please shop. All right Sit down Walter. He doesn't time to pay for the drinks. Well, yeah, I don't care for anything Well, it's all right to let's get to it bring the money when you I want to talk to you about it first He's talking about it. I just want to see it. I uh, I can only raise 6,200 I Asked for 10. I thought I made it clear the best I can do sharp you don't know how much trouble it was I don't care about that. I Told you it had to be 10,000 I meant it. I Have my troubles too Billy. Oh, there's nothing more I can do. That's why you're wrong. I Don't understand. Ah, maybe I shouldn't make it so easy for you, but we can get that money another way If you're suggesting I'm not suggesting anything Billy. I'm telling you what you're gonna do I've I've looked into your job at the bank. What about my job? It includes some interesting responsibilities For an ex-con the people I work for have faith in me if that's what you mean I'm surprised the bonding company slipped up so badly. I'm not covered by your bond. Oh Alfred Crane was alive when I got out. He was decent enough to give me a chance bless him Billy bless him It gives you another chance now. I don't think so. I do I Understand you'll be carrying a batch of negotiable securities tomorrow morning. What of it? You're taking them to the Lincoln Investment Company at 1030, right? I know what you're thinking Roy.  You've got it all figured out, haven't you? I've got you figured out That's why I'm so sure you'll cooperate tomorrow When you take that morning stroll It doesn't seem real does it Walter you wonder home in a day and there's another sleepless night While you tell yourself over and over that this thing can't be happening after all these years of peace The next morning you arrive late at the bank still days shaky wondering what will happen when you set out for the investment company and Roy Sharp walks up to you on the street But you don't have to wonder long water. Mr.  Oh sure. Mr. Cromwell. I I'm feeling better Charlie Bryson figured you needed a little rest. He took over for you took over those securities for Lincoln Investment He's going to deliver them. What? Oh, but he doesn't have to do that. I'm perfectly able. Mr. Cromwell I'll go tell Charlie and Suddenly you realize Walter how impossible the whole situation is The phony hold up the inevitable questions and then sharp coming back to you with another proposition and another You're certain of one thing if sharp were to see your friend Charlie carrying that satchel in your place He'd never dare go through with a robbery Instead he'd go straight to mrs. Crane with a whole story And that doesn't matter anymore does it Walter because you've made a decision an important one Hello, Kathy. I have something to tell you dear and there isn't much time Sit down here right next to me. What is it dad? What's happened? Kathy you remember that young man I used to tell you about Billy Ford the one who went to prison. That's right He was a nice kid Kathy had some tough breaks got into trouble did five years in sing-sing I remember dad you thought a lot of him. He died a long time ago. Didn't he? I thought he did What do you mean Kathy? I'm Billy Ford.  Mrs. Crane's butler Roy sharp. I see Where are you going dad believe me this doesn't make any difference between us, but I want to be honest with Don now While he has a chance to change his mind. I'm sorry Kathy. Please darling. It's all right. I'll go with you No, no, I'd rather face them alone The whistler will return in just a moment with the strange ending to tonight's story Meantime here's a thought for you drivers to consider If you had to wear heavy winter clothes right on through spring, you wouldn't feel very peppy. Would you well? That's just how your car feels about wearing tired old winter motor oil and gear lube now that springs here Yes, if you want to put spring into your driving It's high time you were treating your car to a spring changeover at your signal service station First step is a motor drain and a refill with signal premium motor oil The new type lubricant that guarantees you a sweeter running motor by keeping motor six times cleaner and reducing cylinder wear one third Next step is fresh signal gear lube for transmission and differential and a signal double-check lubrication for the chassis And at the same time your signal dealer will be glad to check those other points that need attention every five or ten thousand miles Such as air cleaner oil filter or front wheel bearings Man a car just can't help feeling peppier after a spring tonic like this So give your car a break see your neighborhood signal dealer this week for a signal spring changeover And now back to the whistler So she's gone now Walter and you sit alone in the living room exhausted numb Filled with a black kind of despair that blots out everything else Yes, Kathy's gone to tell the man. She loves her father was a criminal an ex-convict There was no other way The terrible coincidence that placed you and Roy sharp at the same party ended all hope Yes at that moment the blank wall was breached and Billy Ford and Walter Mason became one and the same man You find yourself staring at the black butt of your 38 revolver lying on the living room table The gun you would have carried this morning if Charlie Bryson hadn't taken your place At one point you start to reach for it and Decide that's never the answer You aren't conscious of time But it's past noon when you decide it wasn't fair to let Kathy face them alone and an hour later You walk into the library of the crane home on Long Island As you expected Mrs. Crane is terribly upset They'll pounce on this like a flock of vultures the papers the tabloids Just a minute, maybe I'm sorry Kathy. It wasn't fair of me. Wait a minute dad. What what's the matter Donald? Show him the paper sure here. You are mr. Mason Attempted robbery broad daylight. It's the butler here dad He was killed this morning in a gunfight on Broad Street. He tried to rob one of the messengers from the bank sharp killed Gunfight wait a minute Charlie Bryson. What about Charlie Charlie the messenger. He was the one who I'll get it Roy's shop Then he didn't know mr.  Oh, thanks Yes, Charlie. Are you all right? All right?  Oh about that money if you still need it Brother, I've got it. No Charlie. Thanks, but everything's all right now Let that whistle be your signal for the signal oil program the whistler each Monday at 9 Brought to you by the signal oil company marketers of signal gasoline and motor oil and fine quality automotive accessories Signal has asked me to remind you To get the most driving pleasure drive at sensible speeds be courteous and obey traffic regulations It may save a life possibly your own Featured in tonight's story where Charles seal and Jack Petruzzi The whistler was produced by George W. Allen with music by Wilbur hatch and was transmitted to our troops overseas by the armed forces radio service This is Marvin Miller speaking for the signal oil company to CBS the Columbia broadcasting system


I'm jumping on the bandwagon and I'm checking out the Creality LD002R resin printer. My name's Jim and this is The Edge of Tech. So like I said, we're going to check out the Creality LD002R resin printer today. Creality did send this printer to me for a video. They didn't ask for anything in return, so that's why I'm doing the video now. So this video is coming out a little bit later than some of the other people have already done their reviews, but I wanted to do wait a little bit and get some time with the printer before I put it out. So one of the cool things is today we're actually going to go straight into the prints and what we got off the build plate and then towards the end of the video we're going to do the unboxing and you can see how easy this thing was to set up. It's a really cool machine. It has a 2k screen. It's five and a half inches. It pretty much shares the screen with the Mars and the Photon I believe. So if you're looking to replace them I believe that screen will fit in any of these printers. So we already know that screen is out there readily available and if you ever have an issue with your Creality screen because it is a consumable it's really cheap to get your hands on. But I think we should jump into the prints. Let's do it. Future Jim here. I got my test prints done and I'm going to show you what they look like a little bit closer now. So this was the test print that was on the card. I really think it came out very nice. It looks really good. There is some bumps back here because I did not clean up the supports. I just pulled them off to cure this and then show you. But I mean everything looks really nice. The details in the face are really good. I think overall this was a very good print for the first print on this printer. Next I printed an Aria Dragon. I did this with no supports and it looks absolutely gorgeous. This is with the Anycubic Grave resin and it looks super good. It's really hard to show but that you can see the fangs here. You can actually see all the little details there. This one came out just absolutely gorgeous. I really love the Aria Dragon and I really love the detail you can get with the resin. Next I did one of my Maker Coins and if you follow me on Twitter you realize that I accidentally dumped some green in with the gray. So that gave us this weird kind of green gray color but it's not too bad. I really like how the Maker Coins came out. It looks really awesome in there and it's really smooth. I should have probably cleaned up in here a little better. So even the little glasses all the way around came out and that's really awesome. Same thing with the Patreon Maker Coins. This is a special Maker Coins for my patrons that I just came up with and it really turned out awesome as well. Something I need to work on is cleaning these up a little better especially in between here. So I'm gonna get a little toothbrush and work on that but that's not the fault of the printer. That's my thing but the detail is awesome. Like I said the glasses all the way around they came out really well. When I was getting this off the bed I actually cracked it right there so that's my fault not the printer's fault. I just chiseled too hard and cracked it myself. That's not the printer's fault that's totally my fault. Otherwise this thing came out super good. I'm really proud of this Maker Coins. Special shout out to Tyler West for hooking me up and you know I really like this printer. This is the four test prints I did for this video. I really like how they turned out. As you can see they really turned out great. I stuck with the default settings in the slicer and that's how I got these three. This one was the test print on the card. For my first prints with this printer and not knowing the printer it really did a great job. I'm really happy with this and the link will be in the description for the ARIA. This one again was the test that came on the card and these two are my special coins. I'm not gonna release the files for those yet but maybe someday I will do that. But ARIA Dragon's link will be in the description below and I hope you guys like the prints. Now let's build this printer. Alright so I got this thing unboxed and it came very well packaged. It was surrounded in Styrofoam and as you can see there's Styrofoam actually inside of here as well. We'll get to that in one second. It did come with an extra FEP which is great. It came with a power cord and then it came with a bag of stuff right here. In the bag came gloves, some masks, a couple of resin filters, two different spatulas, a paint brush, the standard Creality Allen wrench kit with the screwdriver, a USB thumb drive here and the manual. So I mentioned that the printer came packed in Styrofoam so let's check that out. I'm gonna take the cover off here and set it next to it and it looks like the whole head here is completely packed in Styrofoam. Inside of here is the plate so we'll take that out and set that to the side. We see the plate that came with it here, the vat, the lid and the printer. So for the instructions I plug the printer in here but now we'll power it on. We're going to go to tool manual and we are gonna raise the bed enough to put the plate on it. It's very greased in here from the factory which I like a lot so we will take and we'll put the plate on it here and we'll tighten that down. Now what we need to do is level the platform. So let's take off the vat we'll untwist our thumb screws here. Once your thumb screws are loose you can remove the vat like this and set that aside. So I noticed that under here there is a film we need to remove so grab a hold of the yellow pull tab and and pull that out like that. Now what it tells us to do is go to tool manual home and that will bring the bed down. It does say that it has been leveled from the factory and we're gonna find out. It did come all the way down but I don't know how to tell if this is level because I don't know if it really is leveled from the factory. We need to loosen these four screws here and I did that already. Now we're gonna hit the home button and it's gonna come back down. So as you can see it pressed down a little bit and the plate is staying on the bed. So the next thing we need to do is hold this down really good and then tighten all four of these bolts and once you get that done your bed should be leveled.  I'm gonna take a microfiber and just wipe the screen down here so there's no dust and I'm gonna wipe the bottom of our vat here and I'm gonna wipe the inside because there is some stuff on the inside and we don't want that to be inside of our prints. Now screw your vat back to the body here. Now that your vat is screwed back to the body we need to load some resin. One thing I really like and I hope that you can see is that in here there's some steps and the bottom one's a hundred and forty milliliters then 170, then 225, then 285 and you can gauge how much resin you put in off of the steps in here. I think that's really cool. So I'm gonna take my anycubic gray resin because I did not get any resin with this printer. It is mixed up. I have my gloves on and I'm gonna put in resin. As you can see I put about 170 milliliters according to the chart in the vat. Now I take the USB stick that came with the printer and pop it in here and I really like that the USB port is on the side in the front. Some of them are hard to get to in the back and I don't like that but this is nice it's right in the front. I'm gonna do the test print that came on here and to do that we know that my USB stick is in. We're gonna go to print. It looks like there's a couple different options here so I'm gonna hit play and it should start to print now. All right well we've seen the test prints we've seen the build and it was super easy to put together. It came very well packed and it does print really well. Overall I really like the Creality LD002R resin printer. It performs like it says it's going to and I really like that it's really simple to use. Creality you guys did a great job with this one. Well I hope you guys enjoyed the video I hope you learned something and as always keep printing. Hey everybody I hope you guys enjoyed the video. Don't forget we do a live show every Monday night right here on the edge of tech at 6 p.m. so come join us for that and we're gonna start putting out videos on Thursdays so check out that. If you like the video give me that thumbs up. If you haven't subscribed yet please do that. I'm trying to get to 10,000 subscribers and I need your help so hit that subscribe button down below and if you want to be notified next time we do our live show and we go live Mondays at 6 p.m. or we put out a great video on Thursdays hit that bell and YouTube will notify you. Thank you guys for watching. Check out that video.