Respirators, nobody likes or wants to wear a respirator. They've been around since the turn of the century. Probably the first respirator was a damp handkerchief tied around your face. And just like the respirators today, their purpose was to clean the area you breathe. There are many different styles, types, sizes and shapes of respirators available today, but this video will focus on the most common use. This is Joe from Silica Dust. We hope it helps you to understand how important your respirator is in protecting your health. Respirators, your last defense. This is Joe. He's just getting started here. We had a few questions about respirators. Let's talk over here. The primary means of controlling silica dust is in engineering controls. This is like ventilation systems, dust collectors. Now, these pieces of equipment, they help keep the dust from getting into the air. But there's also other controls that we need to use, such as limiting the time that you're in a dusty area. Also, good work practice. That's very important in controlling silica dust. EMSHA, that's the Mind Safety Health Administration. They say that while required engineering controls are being stalled, workers also should wear respirators. Under the Mine Act, EMSHA, they are responsible for the health and safety of all America's miners. So they are the ones that regulate how we use respirators on the mining properties. Also, EMSHA says respirators can be used for occasional entry into hazardous areas like confined spaces or in situations where engineering control measures have not been developed. We also use respirators in combination with our engineering and other control measures to provide even more protection than is required under EMSHA regulations. It's currently not possible to measure instantaneously the amount of silica in the air that you breathe. We don't want anyone to even have a remote chance of getting silicosis. It's for that reason that we sometimes use respirators as a backup to our engineering and other control measures in some operations. Well, let me ask you a question. Could I use any respirator to paint my car? No, most of the time we only need protection against dust at this mine. Other hazardous materials like vapors from painting require special filters and canisters approve for that hazard. It's important to know that respirators that filter dust cannot supply oxygen or provide any protection where there's not enough air. They also can't provide protection for any contaminants other than those for which they've been approved. You should always know the limitations of your respirator. Now here at this plant, your primary exposure will be the silica dust. Now most of the dust generated in this plant is from crushing or grinding rock, loading trucks, screening, other material handling. Now the ore that we handle contains silica. Now the other types of mines there or may contain other hazardous materials. And that's why the company sampling program analyzes the ore to understand its potential hazards. You should always know what you're handling. That's right Bill, but our primary concern at this mine is dust that's small enough to get inside your lungs. Dust that is normally considered small enough to enter the breathing system is called respirable dust. We have many processes in industrial sand plants that generate dust particles. These particles cannot be seen by the naked eye. However if there are many of these small particles it may be possible to see them in a beam of bright light. Just remember, just because you can't see the dust, that doesn't mean it's not there. That's why we take dust samples. Now the dust samples determine what parts of the plant, if any, could be a problem. And if they require additional dust control and respirators. First of all we only use respirators approved by NIOSH, the National Institute for Occupational Safety and Health. Come inside I can explain it better in there. NIOSH is the government agency responsible for approving and testing respirators used by workers. The respirator and the filters must be marked as NIOSH approved. The filters must also be marked as approved for particulates if we use them for protection against silica dust. Now we consider a lot of different factors. Not only must the respirator match the hazard, but it must also match the task to be done and the worker too. Some of the primary factors we consider are the nature and characteristics of the hazard or hazards. The environment where the respirator will be worn. The tasks that need to be done. Next, once we know the situation we can choose the type or design of the respirators. For respirators like this one, which filter the air, the two key features are the face piece and the filters. For respirators like this one, which filter the air but also moves air into the face piece, you also have the blower to consider. This type is called a powered air purifying respirator. All respirators can allow contaminated air to flow in if they don't fit you properly. However, you'll be hearing terms negative pressure and positive pressure respirator. These terms describe design characteristics of the respirator. All respirators where your lungs cause the air to move through the filters are negative pressure. If the respirator has a motor or an air supply, it can be either a positive or negative pressure respirator, depending on how it supplies the air and how much air it supplies.  However, if you've not been fitted properly, even a positive pressure respirator can leak inward because the blower can't compensate enough. There are three types of face pieces, quarter masks, half masks and full face pieces. These face piece designs as well as how the air is filtered and provided into the respirator are used to determine the assigned protection factor or APF as we call it. The APF helps determine the maximum dust concentration in which the respirator can be used. A good rule of thumb for face pieces is the more the face that's covered, the better the protection. The filters also have limitations which determine the maximum dust concentration where you can use them. For particulate filters, there are three types, N, P and R, each with three levels of filtering efficiency. 95, 99 and 99.97%, which we round off to 100, so it's called the 100 type. Some particulate filters are affected by oil and lubricant in the environment. The N type filters are not resistant to oils, cutting fluids, glycerin and other lubricants. R filters are resistant to these materials for only one shift. N and R filters must be thrown away after one shift or specified by the manufacturer, unless there has been scientific testing to show they can be used longer. P filters are oil proof and can be worn for a number of shifts until the filters become difficult to breathe through or they get damaged or soiled. Depending on the work situation, you could wear any of the nine different filters for silica. For ground silica operations, NIOSH recommends the use of the 100 series. It's the most efficient for removing small silica particles in the air. We've already completed the selection process for our hazards and we generally use half mask negative pressure respirators with P100 filters. We always use the 100 series here, but you may get an N, P or R series based on the working environment. Well, you talked about the respirator fitting the hazard or the task, but how will I know the respirator will fit me? Well, we'll determine if you're able to wear one by first having our doctor make sure you don't have any physical or medical problems that may interfere with your ability to wear a half mask negative pressure respirator like this one. Sometimes a previous facial injury or an existing lung condition may disqualify you from wearing a negative pressure respirator. However, you may be able to use a powered air purifying respirator. There are so many types of respirators to choose from that generally, if you are physically able to perform the work, you shouldn't have any problem finding a respirator of some type to use. Well, is that all there is to it? Well, Joe, the next step is to make certain that the face mask fits your face so that contaminated air doesn't leak in around the edges. Right. Suppose, Joe, that the results from the doctor are in and the doctors determine that you're able to wear a certain type of respirator. Now we're going to select the individual respirator and make sure that it fits your face. This is called fit testing. Well, why do you do that? Does one respirator fit everyone? No, not everyone's face is shaped the same. So one size or model of a respirator will not fit everyone. Each manufacturer has different styles of face pieces and cartridges, just like car manufacturers. There's lots of styles and shapes. In addition, they all make a number of sizes for each model. We have to find one that fits you best. Although most people are sensitive about anything that touches their face, a properly fitted respirator is also the most comfortable. There are two basic kinds of fit testing, quantitative and qualitative. The type of fit test determines your assigned protection factor, your APF. Quantitative fit testing requires specialized instruments operated by trained individuals to determine how well your respirator fits your face. It actually takes a measurement of conditions inside and outside the respirator and compares them to determine your personal fit factor.  If you can taste or smell the agent, you are not getting a satisfactory fit. Qualitative fit testing is not as sensitive as quantitative fit testing, so you can only get a fit factor up to 10. For us, this means you can only use this respirator in dust concentrations up to 10 times the exposure limit, even if the cartridge would allow a higher concentration. For example, if the exposure limit for the dust is 0.5 milligrams per cubic meter, then you can wear the respirator in areas that have as much as 5 milligrams per cubic meter of dust. If you need more protection than 10, you must have a quantitative fit test. It's also important that fit tests follow a recognized procedure. Even though the qualitative fit test relies on taste and smell, you can't use just anything. There are specific procedures that must be followed in order to ensure you get the right fit. Well, what kind of fit test do you use here? We do qualitative testing, which is the most common type used in our industry. Our potential exposures are well below 10 times the exposure limit.  Once you've selected the respirator, try it on. Adjust the straps, wear it for a short time, see how it feels. Next, you would do a series of movements and talking exercises to simulate your movements during the work day. If you don't get a good fit, or if the fit test shows leakage into the respirator, we'd have to try another size or type. So, pick a respirator that you might want to wear. Is there anything I have to do to prepare for a fit test? Well, yes. With a tight-fitting face piece, you must always be clean-shading where it meets your face. That's because facial hair in the sealing surface of the respirator or near valves can cause dangerous leaks. Now, this is true whether the respirator is positive or negative pressure. The only types that can be worn with facial hair are those with hoods or helmets which do not have a sealing surface on the face. This valve keeps the contaminated air out of your face piece when you inhale. Anything that can prevent it from laying flat and functioning properly, like hair, can cause you to be exposed. Well, what if I take the test and there is a leak? Then what? First, we would check the position and tightness of the elastic bands that secure the respirator to your face. These should be reasonably tight and in the correct position on your head. We'd also check to see if the respirator was in the correct position on your face, not too high or too low. If facial hair can interfere with the seal of a respirator to your face, we would check it to make sure you had a good seal. If the respirator still didn't fit or was uncomfortable, the next step would be to select another respirator and then repeat the test. We'd continue this process until we found a mask you could wear and pass the fit test. You would not be allowed to wear a respirator on the job without passing the fit test. By the way, you're scheduled for your actual fit test before you start work tomorrow morning. Joe, to make sure you're getting a good fit out into the plant, you must also do a field check every time you put your respirator on. Now, the type of fit check you do depends on the type of respirator you have. It's a quick but important check you must do every time you wear it. Now, for the type that we use here, you must do a negative and a positive pressure check. You simply inhale with your hand covering the filter. You should feel the mask start to collapse. You can see that right now. The collapse means that the mask is working properly and doesn't leak. The next step is to cover the exhalation valve and blow out slightly to see if you can generate a positive pressure inside the mask. If you detect leakage with either of these tests, adjust the straps or reposition the respirator on your face and repeat it. Well, who takes care of the respirators? Well, Emsha requires that the respirators be selected, used and maintained according to the ANSI Standard for Respiratory Protection, Z88.2, as specified by the standard. ANSI is the American National Standards Institute. This standard requires that we establish a maintenance program which ensures that you have a clean and well maintained respirator. It allows you or somebody assigned by the company to do the respirator maintenance. Whoever does the maintenance must be properly trained and follow the company's program for cleaning and sanitizing, inspection for defects, repair and storage. At this plant, everybody is trained to maintain their own respirator. After you're fit tested, you'll be trained on how to care for work. Well, in general, how do I take care of my respirator? Well, it's important that your respirator be cleaned and sanitized daily to prevent the buildup of germs and contaminants. Also, you need to check each part of the respirator, the face piece, straps, valves, etc., each time before you use it. The flexible rubber parts, such as the face piece, must remain pliable to conform to your face. If any parts are defective, worn or missing, they must be replaced before you use the respirator. Now make sure that replacements are from the same manufacturer. Don't ever substitute parts from another style or manufacturer of respirator. If you have any questions, come see me. How about when I'm not using it? That's a good question, Joe. You should store your respirator in a manner that keeps it from becoming damaged, deformed or dirty. Also, keep it away from excessive heat. The supplied bags or even ziplock bags come in handy for keeping it clean. It's a good idea to mark the storage bag so you can tell yours from someone else's. Also, check the manufacturer's instructions before marking on your respirator. Before you ever use your respirator, you'll attend training sessions. This is when you're first hired and every year after that. It'll teach you everything you need to know about your respirator. Now they'll cover in greater detail subjects that we talked about today. The hazards, proper respirator selection, respiratory protection program, medical surveillance, the limitations of your respirator, fit testing, maintenance, cleaning, storage, proper use. They'll go over the company's respiratory protection evaluation program. Well, we'll keep records of your fit testing and training, which we recommend be done annually. Some key points to remember, you must always check your respirator every time you use it. It's your health that's protecting. Wear it correctly. The respirator straps must not be worn over your hard hat. You must change your filters according to the type of filters you're using. Remember, all filters are changed whenever it becomes difficult to breathe through or if they become damaged. And in our filters must be changed after every shift unless the manufacturer specifies differently. If you have any questions, ask. Well, I certainly have learned a lot about respirators. You guys really know your stuff. But how do you remember it all? Well, you can always refer to the company's respirator policy, which is in your health and safety manual. I have another way also. I use the letters in the word respirator. Responsibility is everyone's responsibility to ensure respirators are used safely. Environment. Be aware of the hazards in your environment. Selection. Select the proper respirator for the hazard and the job. Proper fit test. Only wear a respirator if you have passed a proper fit test. Inspection. Inspect your respirator before ever used for warm or damaged parts. Respect the hazard. Silica Dust is a serious respiratory hazard. Adjust. Adjust your respirator properly to maintain a good fit. Training. Take training seriously. It can save your life. Organize programs. A written program consistent with ANSI must be in effect. Respirator. Use it. Wear it. Get your life event. Respirators. Nobody wants or likes to use a respirator. They have existed since the change of century. Probably the first respirator has been a wet cloth tied around your face. And as today's respirators, their purpose was to clean the air you breathe. Today there are many styles, types, sizes and different forms of respirators available. But this video will focus on the most common use, protection against silice dust. We hope it helps you understand how important your respirator is in the protection of your health. Respirators. Your last defense. Hey George, you have a minute. I present you to Joe. He just started and he has some questions about the respirators. Let's talk here. Now, the main way to control the silice dust is through engineering controls. This is like the ventilation systems and the dust collectors. Now, these teams help the dust not enter the air. But there are also other controls that we must use such as limiting the time that remains in a dusty area. Also, the good practice of work is very important in the control of the silice dust. The EMSHA, the Ministry of Health and Security in the Minas, says that while required, engineering controls will be installed. Workers must also use respirators. According to the Minas law, the EMSHA responsible for health and security of all American miners. Thus, they are the only ones who regulate how we use the respirators in the Minas. The EMSHA says that respirators can be used for an occasional entry in dangerous areas with closed spaces with situations where engineering control measures have not been developed. We also use respirators in combination with engineering measures and other controls to provide more protection from the requirer. Currently, it is not possible to instantly measure the amount of silice in the air you breathe. We do not want anyone to have the remote possibility of contracting silicosis. That is why we sometimes use respirators as a support to our engineering measures and other controls in some operations. Well, let me ask you a question. Can I use a respirator to paint my car? No. Most of the time we only need protection against dust. Other dangerous materials such as the paint vapors require special filters and cans approved for that danger. It is also important to know that the respirators that filter dust do not supply oxygen or protect some when there is enough air, nor can they protect against other contaminants for those who are not approved. You must always know the imitations of your respirator. Here in this plant, your main exposure will be the silice dust. Well, most of the dust generated in this plant comes from the mollusk or the wetland of the rock, the trucks of cargo, debris from the crib, or other material manipulation. Now, the mena that we manipulate contains silice. In other types of mena, the mena can contain other dangerous materials. And that is why the Ministry of the Cooperation analyzes the mena to understand its potential dangers. You must always know what you are manipulating. That is correct, Bill, but our main concern in this mena is the dust capable of entering your lungs. The dust that enters your respiratory system is called a breathable dust. We have many processes in the industrial sand plants that generate dust particles. These particles cannot be seen simply. However, if there are many of them, it may be possible to see them in a ray of bright light. Remember that it is only because the dust cannot be seen if it does not mean it is not there. That is why we take dust samples. Now, the dust samples determine which parts of the plant, if there is any, can be a problem and additional dust and respirator controls are required. First of all, we only use approved respirators by the NIO, the National Health Institute of Copational Security. Come in, you will be able to explain better. The NIO is the government body responsible for approving the respirators used by workers. The respirator and the filters must be approved by the NIO. The filters must also be approved for particles, and we use them for protection against silice dust. We take into account many different factors. The respirator should not only be suitable for danger, but it must also be appropriate for the task to be carried out and also for the worker. Some of the fundamental factors that we consider are nature and the characteristics of the danger or the dangers, the environment in which the respirator is going to be used and the tasks that must be carried out. Then, once we know the situation, we can choose the type or design of the respirators. The two fundamental characteristics for respirators like this, that filter the air, are the face part and the filters. For respirators like this one that filter the air, but also move the air in the face part, you must also consider the compressor. This type is called a purifying air purifier. All respirators can let air flow contaminated if they are not placed correctly. However, you will hear terms like positive and negative pressure respirators. These terms describe the characteristics of design. All the respirators in which the lungs make the air move through the filters, are negative pressure. If the respirator has a motor or an air supply, it can be positive or negative pressure, depending on how the air enters and supplies itself. In a positive pressure respirator, the contaminated air does not flow into the face, when it is inhaled, because it has so much air from the compressor's face that it only flows out. However, if you have not placed it correctly, even a positive pressure respirator can filter into it, because the compressor does not compensate enough. There are three types of face parts, masks of one quarter of a face, half masks and full face parts. The designs of these parts, as well as the way in which the air is filtered and supplied, are used to determine the signal protection factor or APF. APF helps to determine the maximum concentration of dust in which the respirator can be used. A good general rule for face parts is, the more covered the face, the better the protection will be. The filters used also have limitations, which determine the maximum concentration of dust in which they can be used. There are three types of particle filters, N, P and R. Each one with three levels of filter efficiency, 95, 99 and 99.97%, which we round to 100, so that it is called the 100 type. Some particle filters are affected by the oil and lubricants in the environment. N-type filters are not resistant to oil, fluid of cut, glycerin and other lubricants. The R filters are resistant to said materials only during a turn, the N and R filters can be thrown after a turn or according to the manufacturer's specification, except that there has been a scientific evidence that they can be used for longer. The P filters are an oil test and can be used during a number of turns until breathing results are difficult or are damaged or dirty. Depending on the situation of work, you can use any of the nine different filters for silice. To operate on land with silice, Nios recommends the use of the 100 series because it is the most effective. We have already finished the selection process for our dangers and we generally use negative pressure respirators of half a mask with P100 filters. We always use the 100 series here, but you could use a N, P or R series according to your working environment. Well, have you talked about the respirator that is suitable for the danger or the task? But how if the respirator is suitable for me? Well, let's determine if you can use one, ensuring that you do not have physical or medical problems that can interfere with your ability to use a negative pressure respirator like this one. Sometimes a facial wound or a pulmonary condition can disqualify you to use a negative pressure respirator. However, you could use a purifying air respirator if necessary. There are so many types of respirators to choose from that you should not have any problem in finding an adequate respirator. Well, that's all. Well, Joe, the next step is to make sure that the mask is suitable for your face so that the contaminated air does not filter around the edges. Correct, let's suppose now, Joe, that we receive the results of the doctor and he has determined that you are able to use it. Now let's choose the respirator and make sure that it is suitable for your face. This is called adjustment test. Well, why do you do that? The respirators don't look good to anyone? No, the face of everyone has the same shape so that the respirator does not adjust to anyone. Each manufacturer has different styles of face parts and cartridges like car manufacturers. There are many styles and shapes. In addition, everyone makes a number of sizes for each model. Although the majority of people are sensitive to anything that touches their face, an adequate respirator is also more comfortable. There are two types of basic and qualitative adjustment tests. The type of adjustment test determines your assigned protection factor or your APF. The quantitative adjustment test requires specialized instruments operated by qualified individuals to determine how well your respirator adjusts to your face. In fact, it takes a measure of the condition inside and out of the respirator and compares them to determine your personal adjustment factor. The qualitative adjustment test is already in the ability to taste or smell a test agent like banana oil, saccharin or irritating smoke that is roasted out of your respirator. If you can taste or smell the agent, it does not adjust satisfactorily. The qualitative adjustment test is not as sensitive as the quantitative one, so it can only achieve an adjustment factor of up to 10. For us, this means that you can only use this respirator in powder concentrations of up to 10 times the exposure limit. Although the cartridge allows a greater concentration. For example, if the exposure limit for the powder is 0.5 mg per cubic meter, then you can use the respirator in areas that have 0.5 mg per cubic meter of powder. If you need more protection than 10, you must perform a quantitative adjustment test. It is also important that the adjustment tests are still a recognized procedure. Although the qualitative adjustment test is already in taste and smell, you cannot use anything. There are specific procedures that must be followed in order to ensure that you will obtain the appropriate respirator. Well, what kind of equipment and adjustment do we use here? We do qualitative tests that are the most common in our industry. Our potential expositions are below 10 times the exposure limit. Now, well, before we do the adjustment test, you will have to choose a respirator that you think is the best for your face. Once you have selected the respirator, try it, adjust it, use it for a while and look how it feels. Then you will make a series of exercises to simulate your movements during a day of work. If it does not adjust well or if the adjustment test shows the filtration in the respirator, we would have to try another one, so choose a respirator that you want to use. Is there something that it must do to prepare for an adjustment test? Yes. With an adjusted face piece, you must always be affected in the area that the mask touches your face. This is because the facial veil of the respirator close to the valves can cause dangerous filtrations. This is how it is whether the respirator has positive or negative pressure. The only types that can be used with an official veil are those with hoods or helmets that do not have a sealed surface on the face. This valve maintains the air contaminated outside your face when you inhale it. Anything that makes it not flat and working correctly like the veil can expose you. But what happens if there is still a test and there is a filtration? First we will verify the elasticity of the elastic bands that make sure the respirator touches. The same must be done without the correct position. We will also verify if the respirator is too high or too low. And because the facial veil can interfere with the seal, we will verify to make sure that it has a good seal. If the respirator still does not adjust correctly or if it is uncomfortable, we will have to select another respirator and repeat the test. We will continue this process until we find a mask that I can use correctly. It will not allow you to use a respirator without having passed the adjustment test. Whether it is the step, you have programmed your adjustment test for tomorrow morning before starting to work. I, to make sure that you receive an adequate respirator on the plant, you must also make a field verification whenever you place the respirator. Now, the type of adjustment verification depends on the type of respirator you have. It is a quick but important verification that you must do every time you use it. Now, for the type that we use here, you must make a positive and negative pressure verification. Simply inhale it with your hands covering the filters. You should feel that the mask starts to collapse. You can see it right now. The collapse means that the mask is working correctly and does not leak. The next step is to cover the valve of exhalation and blow gently to see if you can generate a positive pressure inside the mask. If you detect a leak with any of these tests, adjust the straps or reposition the respirator on your face and repeat it. Well, in general, how do I take care of my respirator? The MCA requires that the respirators be selected, used and maintained according to the standard ANSI for respiratory protection. CE88.2 as specified in the standard. The ANSI is the United States Institute of National Standards. This standard requires that we establish a training program that ensures that you have your respirator clean and well maintained. It allows you to have someone assigned by the company to do the maintenance of the respirator. Whoever wants to do the maintenance should be properly trained with respect to cleaning and disinfection, disinfection of defects, repair and maintenance. In this plant they are all able to maintain their own respirator. After you have passed the adjustment test you will be able to take care of a respirator. Well, in general, how do I take care of my respirator? Well, it is important that your respirator is cleaned and disinfected daily to prevent it from forming germs and contaminants. You also need to verify each part of the respirator, the face part, the straps, the valves, etc. The flexible rubber parts like the face part should be suitable to adapt to your face. If there are defective parts they should be replaced before using the respirator. Now make sure that the answers are from the manufacturer. Never replace parts from another manufacturer. If you have any questions please let me know. This is a good question, Joe. You must keep your respirator in a way that does not damage it. Also keep it away from excessive heat. Provided bags or even ziploc bags are useful to store it. It is a good idea to mark the bag where you keep it, so you can distinguish the yours from the others. Also check the manufacturer's instructions before marking your respirator. If you do not have a respirator for the first time, you will have to go through training. This is done when you are hired and then every year you will be taught everything you need to know about your respirator. Now we will cover in more detail the topics we have talked about today. The dangers, the choice of the respirator, the appropriate, the respiratory protection program, medical supervision, the limitations of your respirator, adjustment tests, maintenance, cleaning, storage and correct use. They will review the evaluation program of respiratory protection of the company. We will save the precedents of your adjustment test and training that we recommend that it be carried out annually. Some key points to remember you must verify your respirator every time you use it. It is protecting your health. Use it correctly. The respiratory currents should not be placed on top of your helmet. You must change the filters in appropriate form. Remember, the filters must be changed once it is difficult to breathe through them or if they are damaged. The N and R filters must be changed after each turn unless the manufacturer specifies it in another way. If you have any questions, ask. Well, I really have learned a lot about respirators. You know the subject, but how do you remember everything? Well, you can always submit to the company's respirator policy that is in your health and security manual. I also have another way. I use the letters of the word respirator. R for responsibility. It is the responsibility of everyone to make sure that the respirators are used safely. E for environment. Be aware of the dangers in your environment or environment. S for selection. Select the right respirator for the danger and work. P for adjustment. Use a respirator only if you have a corresponding adjustment. E for inspection. Inspection your respirator before each use to see if it has damaged or damaged parts. R for respecting the danger. The powder of silice is a serious respiratory danger. A for adjustment. Adjust your respirator as it corresponds so that it is properly adhered. D for development and training. Take the training seriously. You can save your life. Or for the organization of a program. Be aware of the dangers in your environment or environment. R for respirator. Use it. Put it on. It is your last chance.


Okay. Welcome everyone. So today, great comments. So today, I'm just going to be talking about differential analysis and how to handle that for your newly called gypsy peaks. Ignore this taxi. Sorry. Moving on. So some of the learning objectives I will try to cover today are what are the kinds of questions you can ask to better inform your analysis as you're looking at your gypsy data.  One we've touched on, but I will go in more in depth is doing comparing it via genomic coordinates. So that's to identify overlaps. And elements that are not overlapping between your files. So to do so, we are going to use a bed tools. We will also include the context of coverage and amplitude. We will have to use a couple tools for that, but the main, the main statistical driver for that would be HR. And then we'll cover some, we will cover some ways we can handle triplicates. The main one I'm going to be talking about is defined. The pen is showing, right? Yeah. Awesome. Thank you. The main one I will be going over is defined in our package. And then I want to take some time to highlight other considerations when we are analyzing our chip seek just because sometimes it is not as straightforward as we would like. And then I want to highlight some additional resources for you guys. We have another tool called deep tools just for QC or generating plots and some existing chip seek pipelines out there that you guys can leverage. Okay, so, um, day one, we aligned our reads called our peaks.  So, what can we do with that? In terms of analysis, there are two features that we get are two analysis pathways we can do. And if you recall that your enriched regions are really just a set of coordinates. You can find those coordinates if they you can find whether your set of coordinates overlap. And you can also find whether your set of coordinates do not overlap so they would be considered unique to each condition. Now, the context of whether of how to interpret overlap and your unique peaks vastly differed according to your model or experiment type. What I mean by that is let's go into the first example. So, say your example for example a your first set of files are a solution for condition a, and your second set of files is K for mono methylation for condition for conditioning. When we perform a overlap and find that we, what we are essentially getting is a bunch of regions that are enriched with both 27 a solution and K for mono methylation. So, what does that mean? Well, what in, if you recall the, the study that Martin highlighted where they found regulatory elements that had certain histone marks, 27 a solution and K for mono methylation often co occur in region and distilled regulatory elements that we called active enhancers. And so active enhancers then go on to affect promoters, increasing transcription, etc, etc. For unique peaks, what does that mean to have a solution on its own that doesn't coincide with K for mono methylation. Well, going back to that same paper, 27 a solution can co occur with other marks at different regulatory elements. So, for example, 27 a solution can occur at active promoters. So this, so in those promoters, they would also share with K for trimethylation. They can also be bivalent promoters. So they could be at a location at a promoter where they share it with 27 trimethylation. What does it mean for K for mono methylation to be on its own. Well, we've seen examples where K for would be a primed enhancer, and then following some change, it becomes active. Let's take a look at another example. What if we are comparing say 27 a solution across to cancer types. If we find shared peaks that would indicate that they perhaps share common regulatory elements such as shared enhancers or shared activity at promoters. So that suggests that they have common mechanism or regulatory pathways regarding unique peaks. For example, that could mean disease specific 27 a solution. And to cover one more model, say you had a cell line that you then induce and uninduced for, I don't know say like a transcription factor, for example. And if you were looking at 27 acylation shared peaks shared peaks between your induced and your induced state would mean that your 27 acylation mechanics are independent of the treatment. But then if you find 27 acylation unique peaks. That would mean that's your one set of acylation was gained following the induction of your transcription factor, and then a set of 27 acylation peaks was followed was lost following the induction of your transcription factor. So really just to summarize all of that. The types of analysis we can do is to identify overlaps and unique peaks, but the interpretation of your overlap and unique peaks will depend on your type of experiments. So just keep that in mind as you are asking questions and trying to formulate a, a, a, a analysis around your experiments. So, how do we identify identify genomic overlaps. You will find in a lot of papers they have a figure such as this, where it is a Venn diagram showing how many regions are exclusive to one condition how many regions are exclusive to another condition, and how many regions are shared. How do we get to this point. Well, recall, recall that your enriched regions are in bed format and really what that is is a set of genomic coordinates for each feature between your two files. So we can find the common alley or the overlap between your two files by using a tool called a bed tools. I'll go into more detail, but that tools later. So to identify the commonalities what we can do is just use the command bed tools intersect, we would list our first file by doing dash a file a and doing dash be file be that will give us the the intersect so the left outer joint. So that would be the 61,000 and 147,000. If we were to do the same, but to switch our file a with file be so the command would become bed tools intersect, and we want to report the number of elements in file be that intersect. Oh, sorry, that's sorry. Let me try that again bed tools intersect dash a and file be that would just give us the overlap in the middle here. Likewise, if we were to do the identify the file be that overlaps file a that would return us the intersect so the 147,000. Adding dash be will alter the command. So instead of giving us returning sorry, instead of returning us the intersect of file a and file be instead it will give us elements unique to file a that are not found in file be so in this particular example, bed tools intersect dash be dash a file a dash be file be this will give us the 61,000. Now the neat thing is file be can be any can be a variety of types of files. It can be a set of. It can be a bed file so containing a set of coordinates. It can also be a BAM file. It can also be a VCF. So what that will give you is bed tools intersect that file a and sorry, file a and file be if file be will say a BAM that will give you all the regions that contained reads from your BAM, or if you were turning VCF effort, sorry, if your file be was a VCF that will give you all the regions that had a that intersected with a VCF. So one problem when trying to generate this kind of an analysis that I would like to highlight is that your intersects are not always one to one. Let's take this example down here where this is your file a this is your file be and each of these blocks are a elements from each of the files. So we can see here that in file a this particular this particular block intersect multiple in file be this will affect how you count because do you report this number relative to a or do you report this number relative to be so one method to solve this issue is to merge your intersect together and by merging what what that means is you take the left most coordinate and then your right most coordinate and report this whole intersect or this whole section as a single element. And how do we do that. We can do that through the additional features of bed tools. So bed tools is actually a really great tool that I hope that everyone will become more accustomed to using as you are exploring and learning how to perform gypsy analysis just because it is so rich in features. Being a tool that can work on a bed file and a bed file can be really a set of German genomic coordinates for anything and bed tools offers a lot of functionality. The one featured I mentioned was merge. So, given your set of bed, bed files where you have various elements. And you supply a file, it will then merge those elements into a sync merge elements that are overlapping into a single one. If you provided a certain distance that that can also merge your elements into a single one as well. It will also merge bookended elements into a single one. So, very practical and useful. Another function I would like to highlight is called map. So say if you had your file a so this could be your enriched regions, and your file being this could be, I don't know, your regions of regions of methylation, for example. What map does is it takes the overlap from the elements in be that intersect with a and then can summarize the values that accompany each of those elements, for example, you can and use that to calculate the mean methylation that co occur with a. So this allows you to combine your analysis with other epigenetics such as by self fight. And one of the last features I would like to highlight for bed tools is the closest function. So given file a given file be it will just report the closest element in proximity to your file a. It hasn't been extremely useful for me. Say, for example, you find an enhancer. You have, but the problem is, it's hard to contextualize what your enhancer is doing. One common assumption is that your enhancer is operating on a nearby CSS. So using something like closest can then help you identify the closest CSS. And just to highlight the interoperability of bed tools is available command line, which we will primarily be using. You can download it through conda. It has a wrapper and are Python, and it's also available in a Docker. And as I noted before, it's not just limited to your peaks as bed files could be any feature, including plus or minus two KB of your CSS, it could be exons, it could be methylated regions, it could be tabs. So you can do a lot to manipulate to manipulate your files. It's also good for speedy and analysis. So one issue I would like to highlight when you're just doing a genomic overlaps is that they make very simplistic assumptions. For example, here I have K for trimethylation for two conditions. The highlight I want to show is that in one condition, I have two peaks here and then in this in the opposing condition, I'll have only one peak. The highlight indicates a peak that is supposedly unique to my above condition. So if we check the amplitude in the tracks, that is not the case. So this would be a false positive of following a binary comparison. If I were to do the same and look at it in this scenario where, sure, this peak and this peak for the 27 oscillation for these two conditions. So they are technically both present, but if we compare the height and the amplitude of the tracks, one is vastly higher than this one than the other. I think this is also another very obvious example. So the question then becomes, how do we account for difference in coverage difference in amplitude. To do so, you'll often find analysis such as this where your X and your Y axis will be a signal or coverage or amplitude from each of the opposing conditions. So where each of these dots will be a peak from a unified peak call set. So that means we would take your peaks from file peaks from the this example this wild type, and then peaks from your mutant. Read them together, sort and merge them and to get a unified peak set. We then have to assign, because that's just a bed file, we then want to convert that into a bed graph file where we want the fourth column to be coverage and pile up information for for the X and the Y to inform our X and our Y values. So to do that, we will use bed tools, utilizing the feature of intersect and counting the number of reads that intersect each element in our unified peak call set. To do that, we basically have a data frame that we can then read into our, once we read into our we can apply a, we can find differential peaks through full change. We can supply it to other statistical software that can apply normalization for library depth, for example. Sorry, to account for library depth, for example, such as a common, sorry, commonly used ones such as HR and DC to that will give us a combination of full change and AP value.  If we were to just use bed tools and only annotated the regions that were exclusive based on based on genomic coordinates, what we essentially get word would just be the regions on the Y and the X axis. So we would be missing out on the context of all all this in between and along the roughly along the diagonal. So to summarize how this workflow would look like. We start off with our peaks, we start off with our bands, we read our peaks into bed tools and then we merge it to get a unified peak sets. We then supplied this peak set with our bands into bed tools where we would enumerate each peak with the number of reads that can be found there. This produces a data frame, we can read that data frame into our and use statistical analysis package such as such as edge art to calculate that.  Moving on, I would like to talk about triplicates. So one strategy we can use is called a diff bind. So diff bind is a wrapper essentially where after we get our so assume each of these is a experiment and each of these lines are a track and each of these bars is a pile up for each of your replicas. So we then perform a peak call with our same max to we get enrichment and enriched regions. So in the red, what we will feed that all into diff bind and what diff bind will do is similar as before, it will merge all our peaks across all our three replicas. And so you just get one set of unified peaks. It will also do the count for us where it will count per replicate the number of reads in each peak. And then it will apply normalization, and then perform a statistical test to give us the in the statistically valid and full change appropriate full change will start a calculator Q value for us and then we can filter by a full change in Q value similar as we did before. Some of the cons for using diff bind, for example, is we need replicas so more than three. One problem is your merging can go poorly, if your peak call wasn't great to begin with. And so this makes it hard, or at least your certain, sometimes your analysis with roadblocks doesn't go according to plan, go according to plan so it might not perform that well. But if you have three replicas, this is a great package to use. If you don't like the default overlap function, it has other overlap functions that you can use to make it a bit more specific. And you can, as a rapper, it also has access to both edge R and DC to so you can use those to find your significantly enriched regions, according to your choosing. Regarding edge R and DC to I've mentioned them quite a bit so I just want to give a refresher to those who are not as familiar with it. I'm not a statistician so I might butcher some of these explanations but bear with me. So both are soft, both are software very commonly used in RNA seek. They make the assumption that different differential events are rare. So they follow a bind negative binomial distribution. Some of where they can differ is in their normally, sorry, normalization methods, in which edge R uses something called trimmed mean of M values. It looks at each, each sample, and then tries to in assuming the log ratio is one given that nothing should be too different. And then calculates the weighted mean of a log ratio per, I believe it was each combination, and then scales it accordingly, so that it is as close to one as possible. For DC to do it does something similar where it looks at each of your samples. And within each of your samples also looks at each gene and tries to find where that gene is relative to all the other samples in a term in terms of a geometric mean. And it finds the median of that and then scales each length accordingly to that median ratio. Also, in terms of the type of statistical tests that edge R does and DC to does edge R has two functions. If you are just comparing two groups, you get your age. Edge R performs an exact test. As of late, if you are performing an analysis on multiple groups. Edge R uses a can use a generalized linear model. And then for DC to it just uses a wall test. So, again, just summarizing things. For your experiment, you can ask yourself, do you have triplicates. No, we can then go into the previously described pipeline where we take our peaks, we merged them into a unified peak set. We then use a tool, use a tool to look at the coverage for each of our bam files in that unified peak set. We then create a data frame, supply that into edge R. And then we do a, we can then filter for significant and reads, sorry, peaks of a search in full change defined is nice because it basically does all that for you and you can call it in R. Edmund sorry to interrupt Monica has her hand up. Just before you continue. Um, like I use HR for for transcriptomics bodies like my, my preference. Is there any reason why you prefer HR here also for for chip seek or either way like DC to will work similarly, or why do you prefer HR here. In this particular scenario, I'm just using HR because it works better for our for the, sorry, for the tutorial. Yeah, for the tutorial that we're about to go through. But really, you can switch that between a DC to or HR. I do believe DC to only plays nicely if you have triplicates. Or at least more than. Sorry, 2 or more per group when you're doing your comparison. So if you only have 2 samples that you're comparing, you might want to use HR. Okay, that makes sense. Yeah. Before we move on, I think there's a question in the slack that Jose is answered, but he also says he wants your opinion on from Desmond, a bit of a theoretical question, but I recall Max to assumes the chip seek data follows a Poisson distribution, both both edge are and De seek to assume the data adhered to a negative binomial distribution. Wouldn't this lead to a bias in the downstream data analysis or does the NB models each package use is generalized to the Poisson when working with chip seek, or is this not even an issue. I think my take on that is the 2 assumptions being made for. Max to chip seek calling is going to be a different assumption opposed to. Opposed to what edge are and to assume, because again, that's. Sorry, go ahead want to interject what I answered admin, but this is how I see it is is that the assumption is being applied on different targets. So what Max to is analyzing this reads what edge are and the seeker analyzing our genomic features, whether they're peaks or something else. So the distribution is. It can be different distributions and still not be contradictory, right? The peaks can have 1 distribution, but the underlying reads can have a different distribution. I will say also that the, the negative binomial distribution is like due to the next generation sequencing data per se. So that's why it's used now. Well, maybe just to interject. I mean, the negative binomial distribution works for for expression for gene transcript. Abundances. But, you know, does that model apply in the context of chip seek distributions. You know, I think that's. I haven't seen any evidence that that's the case. I think DC two is, I mean DC two is designed for doing differential analysis across large groups of samples, where you have, you know, many, many measurements, 10s or hundreds of measurements. You know, assumes a mean and uses a negative binomial to provide a significance for the differential expression chip seek is a different beast because you almost never have that number of samples and so, although the new version of DC two of course allows you to do a lower number of replicates. I'm always very cautious and using DC to for differential analysis and prefer to use diff bind or or deep tools to do to do any differential analysis that depends on amplitude. I mean, this is a big discussion and maybe maybe we want to work work through it but but even thinking about what amplitude means in the context of chip seek data. What does amplitude mean amplitude is a reflection of occupancy of that mark within the population of cells that you're measuring. It's not a measure of strength in the sense that we think about it in gene expression amplitude and gene expression means that that gene is expressed more amplitude and chip seek means that more of the cells within your population have that market that particular position, and we have no understanding of what the minimum and maximum is. You know, taking into account amplitude I think you have to be very, very cognizant of the, you know, of the biases in the data and the challenges there. Well, you know, well, well undertaking such an analysis. And I mean I think the first step is to do a binary analysis with, you know, FDR cut off peaks that you use that have been generated using a Poisson distribution that would be my first approach. And I would be very cautious and using amplitude, but that's just my opinion and obviously this is an area of of ongoing research and development. But, you know, you really need to think about what amplitude means in chip seek which is different from transcript. Anyway, I'll be quiet and please. No, definitely appreciate the input from everyone. Thank you. Okay, moving on. So another method to utilize your triplicates if you don't want to call peaks of on them individually and analyze them individually. What you can do. So again, each of these tracks would indicate a separate replicate and each colored bar would indicate the reads corresponding reads there. And then you would just combine your three bands into a single one. And then you can perform the same binary analysis as we have highlighted previously some pros and cons for this particular approach. This particular approach allows you to use the same strategies that we highlighted in the binary comparison. One issue is assumes that there is tight correlation between your replicates. So for example in this taught in this crude drawing, you can see that one replicate is vastly different from the other two. So it may represent what you are showing. Or it may represent some of the results. So let's start coming back to the problem with Broadmarks that I briefly talked about. So when you're working with Broadmarks, the merging result can be get messy. For example, if you are looking at triplicates here. If we were to merge that you essentially get this extremely large feature and honest and it's hard to say because maybe you do want something as big because 27 and me three can look that large occasionally. But because of Max two and how it performs a broad peak calling thing, it doesn't always have the. What's the best word to call it the best resolution for boundaries. So some additional ideas I would like to highlight when trying to wrangle your Broadmarks is that, like I mentioned. Broad mode works, though it can, it can have for boundary resolution for your broad peaks. So if you are just trying to identify a differential. Broadmark, sorry differential mark what you can do is instead of peaks generate a set of overlapping windows using, for example, bed tools. This will essentially bend the genome, and then you can apply the same approach where you just do a recount per each of your enriched regions. You can do a recap for each of your windows and then apply apply a apply a statistical test to find enriched regions and use those. There are other tools that are that also try to address this broad peak calling problem. For example, we would be cicer to there are additionally certain are packages. This one is called seesaw that tries to use the overlapping genomic window approach to help find differential broad peaks. So, so either items I would like to highlight as you are trying to perform your gypsy analysis is keep in mind what keep in mind of the big picture what are you after are because what we really want is to identify a set of regions to gain insight into a cells population and epigenetic state. Even if you find, say, a one a different even if you find a differential for say, a solution. Based on what we learned from the end code paper that usually co occurs with other with other measurements, for example, if we found, for example, for the differential. We can learn a lot more by looking at, for example, k for mono methylation to identify if it is an active enhancer. If it is an active enhancer, you would usually see that chromatin accessibility is also increasing at the location to allow for recruitment of say transcription factor. Speaking of transcription factor, you might see increases of that. Additionally, if it is a newly formed 27. Sorry, if it is a newly formed enhancer, does it have any consequence downstream to the distally regulated gene that is that is acting upon. Do you see increase in transcription, for example. Another idea to also keep in mind is ultimately we want to link our epigenetic state change to some sort of functional genomic feature. So say, for example, you called your K for me three peaks, you found your differential K for me three peaks, and then you intersected them with your promoters and essentially what you have are a set of promoters with differential K for me. Three events. That is one method, but perhaps a quicker way to get that result is if we just looked at our genomic features directly. We did a count found and found a, sorry, did account for K for me three and and applied as the, sorry, applied the workflow of counting how many K for me three reads are there and applying a full change and a significant special to just gain to just get all K for me three promoters. So that will give us the same result essentially. And also, consider the computational cost. As Martin highlighted, ventals and genomic overlap is sort of the first step is sort of the first way to start exploring your data. It's also one of the fastest ways. But as we've highlighted, there are some caveats to that. Edge are defined. All of these are packages in our are isn't known for speed, and it can have trouble wrangling large data sets, if you have a lot of them. So you might have to come up with. Sorry, you may have to come up with workarounds such as doing the counts, the overload counts yourself, and then supplying that into our to perform her analysis. Another item I would like to highlight is called the when you're doing analysis for say a taxi and also surprisingly applicable for a chip seek is the idea of irreducibility discovery rate. So the So the idea here is asking the question of if I have two technical replicates or two biological replicates or even pseudo replicates. So I, for example, I subset my library down to 90% and compare how often am I able to call that same read, or sorry, how often am I able to call that same peak. So in the about in the diagram a and diagram B. The idea is that yours, regardless of regardless of depth in your two technical replicates biological replicates or pseudo replicates, your signal, your relative signal should always be the same, or be similar or along this diagonal. And so your, your, sorry, your let's say you sub sampled your peak, and then the signal jumped from originally here. Sorry. Yeah, it deviated off the diagonal of this which suggests that it is subjective to have a link and that means it's not highly reproducible. Similarly, if we were to look at it based on ranking. You would expect your one of your top piece to be consistently our, you would expect your peak to be ranked the top regardless of whether it was a whether your library is at the normal depth, or if it is 90% if it vastly changes. It can, if it vastly changes that might suggest that that peak isn't particularly reproducible. So this is an example of a good library that has a high reproducibility, a example of a poor of a, sorry, of a example with low reproducibility is that, as you can see, after you sub sample your peak, the signal vastly changes and isn't doesn't scale accordingly. And even in terms of rank, if your peak, for example, was ranked with waypoint but then it after you sub sample it falls to the wayside all the way to the bottom, it might suggest that I get that this peak isn't particularly reproducible. And I would like to highlight this particular set of software to use for a tax seek is because, again, your attack seek doesn't have background. So applying this IDR threshold could be another way to get to have a more high confidence set of Pete. So, a few more items I want to highlight deep tools is a awesome tool with a lot of feet, rich features. Like Max two, it can generate big waves, where you can then use those big waves to produce figures. One figure I would like to highlight, you might see very often is the meta plot, and this heat map, the idea of this heat map is each row would be your set in of your set of peaks. And the heat here would just indicate the normalized coverage at those peaks. So TSS and TES would just be the start and end of your peak. And then you can see that it extends a little outwards from those regions. Using this, you can demonstrate the peaks that have low strength, or sorry, low signal, the peaks that have high signal. You can use this to compare across various marks. So the idea that would be, let's look at this one, for example, you can see that this one would be. If you were to look at this particular replicate, for example, this replicate would be showing no. Sorry, would be showing no signal at these peaks. Well, the red replicate would be showing a lot of signal at these high peaks. So this particular, oh, sorry, I also forgot to mention this is the meta plot. This is just a summary of all the information here just to denote the trend. This particular analysis is also useful for doing QC. For example, if you're, if you supplied instead of peaks you supplied region, promoter regions at say the TSS, we typically expect to see enrichment at the TSS for K4 ME3. If that deviates that might suggest either a reflection of biology, or something went wrong such as a sample mix up, for example, and etc etc. Deep tools is also good because you could use that to do a correlation plot. This will give you a denogram and then you can see the correlation between. Sorry, so this is a heat map you would see you would see the correlation across between all your marks. So this is useful in when comparing a set of narrow marks, they tend to be more closely associated. Broad marks tend to be off by themselves. So this is good for doing again QC to see expected relationships. I do want to warn that if you are incorporating multiple sample types. The biological difference differences tend to overwhelm the sorry, the biological differences can tend to overwhelm the histo marks in certain cases. Additionally, the assuming that you understand chip seek, and you know what you understand chip seek you know the steps you know what analysis to do.  So various consortia have already designed a chip seek. So you can analyze them in the same way that these consortia have the plus benefit of doing that is you can then use the outputs and compare it to the outputs that the consortia has generated. And then they will both be general, they would be both be analyzed in a uniform manner. I'm just highlighting the chip seek pipeline developed by encode that Martin and I believe Guillaume is part of the taxi pipeline. You should see a lot of familiar elements such as you have the reads you align your reads you.  Another set of pipelines I would like to highlight if encode wasn't your flavor and of course also has a set of pipelines. Each for a chip seek and another one for a taxi. Again, you would see a lot of this familiar elements fast QC to just take a look at your files, BWA to perform alignments, but card to mark dup filtering and somewhere along here is Max two to call your enrichment. Okay, so in conclusion, consider the types in kinds of questions that you can ask when you are comparing comparing your peaks across different kinds of experiments. One of the first analyses you can do is just use bed tools to compare overlaps and find to find overlaps and unique peaks. The general methodology for utilizing amplitude when comparing your peaks is to have a unified peak set. Get contextualize that with recounts have a signal data frame and then perform some sort of statistical analysis to get your differential peaks. We also briefly talked about ways we can handle triplicates. For example, you can find your differential peaks, you can use bed tools for the genomic intersect, you can feed your signal data frame to edge are or define for example. And I hope I also was able to highlight some of the considerations and complexities that you may encounter when you are working with peaks. So I think this is a good part where I can ask any questions.


Welcome to Toffey TV, this is the Everton News Daily. Jarrah Branthwaite has given an interview about how he started the season at Everton. He did reference his lone spell at PSV, where he spent last season, come of age over in Holland. He said, looking back, it was the best thing I could have done. I grew as a person, I'm more confident on and off the field. And we've seen that since he's come back in to the football club in the summer. He's forced his way into Shondite to start in 11 and set up a good partnership alongside James Tarkovsky. On Tarkovsky, he said, Tark is a real leader and talks you through the game. As a young centre back, having experience next to you helps massively. And Branthwaite has been impressive in this short period of time since he got into the first team this season. So much so that he's obviously attracted interest from Manchester United and Manchester City according to reports today. Everton are keen to tie him down through a new long-term contract, so hopefully they can get that one sorted soon. Everton are amongst a number of clubs tracking out if he had Jota. The former Celtic player was left out of the squad for the season. He went over there for £25 million. It raised a few eyebrows when he did go over there. But there's been some kind of break down between the two, as he had still in negotiations to release him from his contract so he can return to Europe. If that's the case, then the former Celtic player in Portuguese under 21 striker would be free according to reports, massively sure. On that one, that's what they're saying, and Everton are one of the clubs tracking him apparently. Obviously did really well for Celtic and got a good reputation West Ham. Also looking at that as well, so that's one probably to keep an eye on at the moment. Former Everton manager Frank Lampard is the favourite to take over at Rangers. Rangers repart a company with Michael Beale on Monday. I've identified Lampard apparently as the favourite. Lampard, who left Everton at the end of January, also had an 11-game interim spellers manager at Chelsea for the end of the season. So he could be retaining into football at a good big or good job for him as well in Glasgow. And finally, Everton's links to kit manufacturer Castore continue. The Blues contract with Hummel ends at the end of the season and news broke the other day that Everton are set to replace them with Castore. They are at the centre of a lot of negative things at the moment shall we say. We've seen obviously the issues with Aston Villa's kit, which the kit absorbs all of the moisture rain sweat, whatever. Villa simply aren't happy with it and they want to break their contract with them or end their contract with them. Newcastle have already announced that they're going to end their contract with them this season as well. And Genoa, who are part of triple sevens group also breaking their contract with or got rid of them as well. So it is a little bit of a one that's got a lot of people talking about whether Everton should be going with this manufacturer. But the positive way to look at it is surely Castore will learn from this and by the time they come to do Everton's kit, if that is indeed the case, they will have got past these issues and can have a smooth transition with Everton's kit. We'll see. We will see. That is it for the news day. Give the video a thumbs up, subscribe. If you haven't and if you want to become a Trophy TV Premier member, the link is in the description. QR codes on the screen now. See you later.


Hello and welcome to this presentation of the STM32U5 embedded operational amplifier. It covers the features of this IP which is widely used for conditioning analog signals. The STM32U5 integrates two op-amp modules which are very similar to the STM32L5. These are general purpose analog amplifiers which reduce the need of an external standalone op-amp. As these op-amps can be configured in standalone mode with all terminals available for the user, it's possible to use them as a voltage follower, non-inverting and inverting amplifiers, as well as analog filters such as low or high pass filters. They can also act as a pre-amplifier for the ADC input. The table shows performance parameters for the STM32U5s and STM32L5s op-amps. Compared to the STM32L5, the analog supply voltage range of the STM32U5 is extended from 1.6 to 3.6V. The gain bandwidth is also extended to 2 MHz in normal mode and 0.5 MHz in low power mode. Note that the op-amp module present in the STM32U5 supports a high speed mode that is activated by setting a control bit. The slow rate characteristics are improved in the STM32U5's op-amp module. By using an internal feedback resistor, the op-amp can also be used as a programmable gain amplifier or PGA. When enabled, it can be in calibration mode, all input and output of the op-amp are then disconnected or in functional mode. To facilitate PCB design, a SPICE simulation module will be provided upon demand. This table shows the internal and external connections of the op-amp inputs and outputs. Each op-amp module has two inputs, inverting and non-inverting, and one output. The non-inverting inputs can be connected to either a GPIO pad or internally to a DAC output. The inverting input is used for feedback when PGA mode is selected or can be connected to a GPIO pad. The output is either connected to a GPIO pad or to an ADC input. By connecting the op-amp to its dedicated inputs, it can be used as a transimpedance amplifier. A transimpedance amplifier is a current to voltage converter. In addition to this presentation, you can refer to the following presentations for the peripherals connected to the STM32U5's operational amplifiers. Analog to Digital Converter or ADC and Digital to Analog Converter or DAC, General Purpose Input Output or GPIO.


As part of efforts to strengthen the health sector's capacity in responding to emergencies and enhancing health care delivery, the Project Implementation Unit through the Health System Strengthening Project, HSSP, recently handed over vehicles, pharmaceuticals, medical supplies, and laboratory consumables and reagents in a small ceremony. The Ministry of Health was handed over two panel vans, supply of pharmaceuticals, infrared thermometers, and universal transport kits. The St. Jude Hospital received one panel van and one double carb pickup van. The Ezra Long Lab of the OKU Hospital accepted receipts of laboratory consumables and reagents. Program Coordinator of the Health System Strengthening Project and the OECS Regional Health Project, Jeanette Jean-Louis Euse, says it is a hope that the donation will make the beneficiary departments and hospitals more efficient and also boost the health sector. Today, we are delivering to the Ministry of Health Wellness and Elderly Affairs, to the St. Jude Hospital and to the OKU Hospital, specifically the Ezra Long Laboratory, much-needed resources which should assist them all in improving the delivery of care at the various institutions. Our focus at the Project Implementation Unit is not only on the provision of resources, but we are concerned about the expected impact which the use of these resources can have on the improvement of the delivery of care. We all know how the lack of resources can affect health care performance. Acting Permanent Secretary in the Ministry of Health, Jenny Daniel, expressed gratitude to the HSSP for this donation and vowed their commitment to utilize these resources in the best way possible. We also recognize the significance of the World Bank Group, which continues to finance all major components that lead to the strengthening of our health care system in preparation for the gradual implementation of universal health coverage. I would like to thank the Project Implementation Team of the Ministry who have worked adversely over months towards the procurement of the components donated here today. The Ministry pledges and commits to the purposeful and strategic utilization of all components in our move to strengthen our health care system. Minister for Health, the Honourable Moses Jabatis, welcomed the support provided under HSSP and says this signifies a move in the direction of achieving universal health coverage in St. Klosho. The donations this morning, while they may be physical, vehicles, pharmaceuticals, reagents, while these may be physical, they all form very important parts of this whole milieu, if you wish, and this whole puzzle, which we have to put together to achieve universal health or universal health care in St. Lucia. I want to thank you for your commitment and your dedication, and I want to say to the staff of the St. Jude Hospital, staff of the OKEU Hospital, and also the other departments of the Ministry, those professionals will be benefiting from the use of the vehicles that we care about you, we're thinking about you. Everything is not right yet, and it may take a while. But I want you to be not comforted, but I want you to know that we are working towards this goal. The Health System Strengthening Project, HSSP, invested up to 2.2 million EC dollars through Component 4 of the project, that is, Contingent Emergency Response Component CIRC. Reporting from the Project Implementation Unit within the Ministry of Health, Wellness and Elderly Affairs, I am Fennel Neptune.


Good this is Thanks for coming forward So everything I begin with it's always like this I always put you know Listeners advisory unclear language, you know, I'm not speaking about It's just that some some of them Blockchain or whatever and You know, I was like the big jokes about You know para advisory, you know, there's a kitty by vote at autumn months will tell you why you know stuff like this Yeah, so TT docs you was wondering the you know blockchain is always about Blockchain is always about Payments FinTech financial centers trade finance and stuff like this why Shipping is like I always got a lot of people and tell a lot of people Why is like why shipping why logistics, you know, it's supposed to be like Bitcoin it's here, right? In fact wrong a bit of my background is I used to work in this company I believe you still you guys see it about it. It's called Mark's and Spencer So I was with there to understand the business process There was going in round maybe oh nine oh eight. They almost go in the midst of bankruptcy and they come out of plan B and stuff like this and basically They didn't couldn't figure out their supply chain and Stuff like this and IBM failed them tremendously.  Don't be shy. Yeah. Yeah. Yeah some more some more Okay, what about it here him? You're still regular people. Yeah, okay Anyone at the back have bitcoins? No, oh Yeah, that's one Come a look for me. I'm gonna show you this amazing hardware. Anyone know what is this? Yeah, yeah, this is amazing right so Anyone doesn't know so I don't about half of them doesn't know what is Bitcoin or blockchain on So it's actually not You see, there's a lot of KAM uncle Sam and the Singapore uncle Sam is going to come down to you and say Oh, we're gonna do a token sale. That's like Where is your anonymity? Where's the KYC? Where's the AML? So if you did it successfully, that's fine, but sooner later uncle Sam is coming after you Singapore one and the FCC to watch out for that So I don't need to read this. It wasn't on control by any central bank So there is something like digital assets. So this is the white paper abstractions you can find it out and figure out Basically is About a mining stuff like this, but we're not doing mining Our stuff is on a private blockchain. So there's no mining. It's just consensus and If anyone want to know about smart contracts That's amazing guy over here Paul. This is helping me as well on the smart contracts if you know about Anyone knows about project who been he's the guy. Yeah, sorry. I don't mean to put it on the table, but yeah You're cool so Yeah, this is what I explained about blockchain Blockchain.  You need to understand two core things. I mean, do you remember two core things? One thing is cryptography. If you understand two five six hash simple enough, bam, you boom the second one is Decentralized distributed leisure If you don't know what is that is just Google it up. It's amazing is you there's no single point of failure and Forgive me ever wrong. Okay. So this part has been done. It's immutable So it's a fully secure and stuff like this Blockchain what it can is really soft thing about something which is email or Music if I send you a copy of it, you have no problem with it but if I give you something a Currency or assets or digital assets for example Then it will be problem. How did he then fight this is real and I'll make sure I don't have this copy with you. Okay, so Blockchain it really helps to solve this problem, but there's a lot of dispute in in outside and I believe everyone have some of them you raised up the hand of the Block chain. No, no, sorry the Bitcoin who holds a Bitcoin Do you know there's a civil war outside? Tomorrow and right now. Yeah, but it changes is tomorrow 1st of August if you don't know about it Read about it. All right. That is a total civil war When I was in Hong Kong And if you read my profile on the scaling Bitcoin I met this amazing guy called who owns BTCC his name is Bobby When I met him he Was like, yeah, hi, I'm BTCC. I'm Bobby. I was like, okay I was like then when I go to back to the website and look at his website. It's like BTCC able to mine 1000 Bitcoin per day. I was like, whoo You must be something Then The next guy I'm gonna meet is a young kid Who built a POS system and this POS system is like, I'm so cool I'm gonna show you how to pay by Bitcoin. It was like what? Bitcoin It was like scam must be a scam But when he showed me right in front of the face of the debt POS misses the IoT device built in front.  I was like, this is so awesome right So Let's go a bit laughter down here What are the common problems in the shipping and logistic industries? I Don't need to explain this you can read it up. Take a screenshot of it. Take a photo and You can this things are yeah, common problem when a shipper fades Things like I gotta need to buy my insurance exporter need to talk with the communicators and importers is like, hey It's a ship right on time and my freight forwarders gonna do my job So a bit of conclusion if you can't see it on the back and Shipping process most of the common problems is the ship missing documents Documents not informations that which is not exactly right and most of the shipping documents Manual process and it's physical form and Stuff like this What things can go wrong? I was like, you know, if you guys Anyone in the shipping industry logistic, we love to talk to you come and look for us Basically what things just go wrong is all the classification of information and the goose and stuff like this Basically what we try to do is we can't try to consolidate Everyone on board or the cloud. Yeah, it's like, oh, it must be another eye cloud But no no no no way this is just the beginning of the first race It's like 10x having the credit cards and stays, you know, it's so cool. I can pay by credit cards using a bit coin So what we're trying to do is something like ebio leading so we put them on the cloud and digitize them and If you go remember the back in the days you call the someone else. It's like, hello.  Nah, I think email right now. Yeah, but the fax machine is very simple I just fax you you get a copy already and I have another copy already So this is what I'm trying to do But in future we're gonna give you that away from the fax machine so we're coming to on board with everyone and everyone on board on the cloud right This is the scale that How tedious the process of the shipping is you're just like I'm going to go to ship out one place and Imagine you guys are just buying for things from Taobao or eBay or you know any commerce platform You was thinking that hell I'm just going to wait and buy and this that's it, right?  I cook it or whatsoever Just imagine the amount of companies controlling your daily products is all here Just imagine Jack Daniels here, right? Who like to eat Oreos is over here Unilever and store on coke and stuff like this Yeah, this is the whole thing that is one of the largest main capital market exists in the world and 85% of your products are shit by containers Well Yes, it's like you're fine, but If you're going to put in your file in the manual process and the papers and stuff like this There is no trust because I can just simply Temper the documents and says hey, you know if I'm going to import the stuff from Somewhere else and Singapore GST over $400 after the pay tax, right? So let's do tricky a bit. Let's put the documents and says This stuff is actually worth $1,000, but I'm going to put the documents and say say Just 399.99 so I can save That bloody GST That's like Yeah, but you can save it once twice not a problem But think of companies with the biggest scale Auditors before it's going to come to you and it's like yeah, what's going so wrong? Are you trying to do something else? There's a test okay, and nothing is to tease so I was like Why your company is TT dogs as like yeah trust and transparency We're trying to create transparency Trust and transparency in the platform We're trying to build this on a consortium blockchain We don't see Competitors if we see competitors, it's like we are scared or come we don't want to scare the competitors We're like 10x and we see we see other competitors. They're doing they're validating us. They're developing here We are not the first mover We will found that over the market. We're definitely not the first mover and We found that there's a lot of people using blockchain and blockchain, but they really don't understand the key values of blockchain key values of blockchain is only two things as I said again cryptography and decentralized Discipline ledger so we're trying to bring them on board and We're doing a private blockchain if any of the competitors are doing a private blockchain, we love them to work with them so We're trying we're not doing a lot of people a lot of my friends down here. I think Prakash over here. I say hey Dude, are you trying to do ICO? It's like no no no no no no no Make something useful first. Yeah, the whole key is is there anyone using it? it's not the token sale or whatever and We can understand the value of what is the token is As I always will consult Professor Paul here I don't put you in but it's the best guys to look for smart contracts if you already want to know about smart contracts this guy So what we try to do is a validation We're trying to we haven't do a white paper yet But we're soon going to do a draft of white paper, but I don't know when I can guarantee you in that but it's we're trying to do is validation between imported and exporter to do a validation and Agreement agreement will be in the future. We'll come like It's on this is on a smart contracts level already Agreement is mostly more something like you agree on something I burger and something and let's do automation vending machine and it was executed the calling the rules and Utilities is this blockchain We want to create something unique to the supply chain Logistics and I would like to thank Glenn for coming as well. It's amazing guy for the supply chain so we can consult on and Just which is the record and the compliance Which is on the dead that you have that rule to set? Let me tell you what is smart contracts about. I know some of them you're bored already. It's like Yeah, I know what is much contracts. Yeah, okay So as I said fax machine PDF invoice, I don't I think you guys you receive it every day during your work You definitely have a copy of a copy, but it's not going to work on this contract. It's just a copy of the contract So what it's going to do is when you execute on the smart contracts If you have a digital assets, it can be a property of land or the house or whatever that you agree on You have 100 shares and so on so stuff like this. Yeah Hmm, you're wondering This is this is by tech crunch I don't need to explain this anyone knows R3 networks you should look for them and This is there's amazing that Goldman Sachs is there's a potential market value of that 37 B outside Not male or this be USD 100% of them do work with Blockchain on the banks. I believe you heard the news as oh, yeah, OCBC is transferring some money to To other branches using blockchain. It's like all right and Does bang really understand blockchain? I'm gonna take you later But what we try to do is TD dogs. We are very very early stage company. We once again, it's a very early stage company We're still going through that validation process We are validating your partners and we will enroll for programs and stuff like this. Smartport challenge is one of them and the quantity of transparency and stuff like this and which is Perfect for perfect fit for this shipping in the logistic field Competitors If you don't tell you about the competitors use like you must be kidding me No, we really did a lot of homework. We went through months of Finding out competitors even talk to them as I think as customers. It's like, hey, I want to use the service Are you using real as a blockchain? It's like, yeah, there's a list of competitors this over here one of the one of the rated ones as SQL chain and Cargo chain and stuff like this. We even pitched to Merce We went to Merce and just pitch to them and then no, it's like. Oh Yeah, doing this with IBM is like We should go to IBM and then we did talk to IBM and they love us Sorry That's banged really really ex really really familiar with blockchain. This is give you the answer by Bloomberg by PwC and this is the whole chart of the journey that we're trying trying to Validate over the blockchain But right now in the current stage, I can tell you honestly if you go to pitch it at anyone else Not a lot of people know blockchain and it's like they only want to know three things to call three things. Can you do cheaper faster and Anything better than my competitors or in the market does and Right Anyone knows what's this Anyone has seen this Can I just give a shout? Yeah, okay, so one thing is SpaceX create something which is a reusable rocket. So if you build a rocket, it was gonna shoot up once It's going to waste a million dollars. So we're trying to do something a reusable blockchain Which means that in the future Everyone is using blockchain. I'm just trying to advocate that until the day I die because I believe in that and If you really really believe in the old eyes If you don't come and look for me, I might give you a bit of Bitcoin part of it. Okay, and So to to use done that we is is a long long way, you know when SpaceX created this is I'm gonna you reuse go rocket. I'm gonna send people on Mars and people just tell you Crazy, are you really saying you're Mars and then you look at that and what we're trying to do and The shipping and the logistic industry has not been changed ever since I left my job and Right now I came back. I would like to thank Singapore government. It's like good Lord They create this amazing holy church and For me to say here in to talk about blockchain. Otherwise, nobody understand them We love to talk to you and please come to our website and give us a light or whatever And we love people to advocate about blockchain. So We love to come to you to me after this session. We have community discussion and I think basically thank you. That's all I love to shoot out any questions or any doubts Sorry a bit long, but come on. Don't be shy. Yeah, go ahead. It's not only about tokens currency, I think it's more about using supply Blockchain to change to do something good to change what we are doing That is something I really like so you only talk about the logistics and shipping and that is part of supply chain So my question is what other area in supply chain do you think can blockchain can help to transform our trade trade finance You look at this our trade is amazing When you look at our trade is trade finance It would you have engaged a smart contracts and automation everything You agree a buyer and exporter and seller and Buyer agree on that everything will become like a vending machine. So We we ourselves we see that potential down there But we are now we and we are nobody so we are very early stage. We're trying to create In near future. I don't know when but we're still looking at that in near future that we are trying to create tokens for that shipping but in order to convince because our business model is purely B2B and You asked any companies down here. Hey, please buy my token. They'll tell you.  Yeah, so they'll rather give you fiat currency and that so once we the option rate kick in You see that people are using it there where is the ICO and everything goes come in So right now is we don't see we need to substance ourselves So we learn do a digital view of leading first to something to ourself and slowly we're just going to step it up and Next few years. I hope that we can become like flexplot does Yeah That's Jeffrey. Jeffrey knows about flexplot. Yeah, any other questions? Yeah, go ahead Hi, you see you mentioned that I bang down transaction in the blockchain Yes, you could you share a little bit more details on how was the transaction being done? Is it to crypto or other type of currency? Banks right now days where I know that they're using private blockchain They they don't do public blockchain because of the gas price and the mining fees That's one thing. So they transfer different branch in different countries within their own networks If you look at historical records RBS Royal Bank of Scotland Royal Bank of Canada RBC sorry Royal Bank of Canada is the first First banks that using blockchain technology to do a transfer which should each other and for the shipping and logistics It's about last year it was Barclays that using the R3 networks to ship a physical goods From London to all the way to South Africa and It's a real trade It's a real physical containers with around probably about 1000 bitcoins of the goods and For Singapore wise OCBC on last year on the fintech festival they have brookly man says that hey Are we have transferred? money physically from a digital assets from Singapore Malaysia Hong Kong then back to Malaysia then back to Singapore then back to Hong Kong again back to Singapore So is the they using? Crypto tokens on on their own digital leisure. Yep Any other questions does it answer the question sir, okay So just imagine that next time you can just go to Taobao and use your Bitcoin and I just went to Tanjong Baga and says that there's a shop that said Bitcoin as well Yeah, so any other yeah, my question is can't you guys start off like flat flex port? instead of waiting two years to become like flex port Flex port is very interesting You can ask Jeffrey flex plot we haven't pitched Jeffrey before so I'm sorry about it. Okay, but Is flex port is only the barriers from pot-to-pot it doesn't really cover the whole container journey right, so flex port only find your your Something like your aircraft you when you buy a plane ticket you need to know your Nick's of kin is on the aircraft and Is it your next skin is safe on the aircraft? So they will go down to the this platform of flex port and you find it's like a little dashboard and It goes up to one place to another from time to time so There's a little two down there. Yeah Any other questions does that equals your questions? Yeah, sorry about it Right, okay. Thank you. Thank you so much, JC