Now moving forward to announcing who the silver winner is in this category. Let's take a look. Congratulations, MXP Media India Limited for MX Player Queen campaign. Many congratulations to the whole team. Don't forget to celebrate, it's a big win. Moving forward to our next category, which is best use of radio by a brand or a campaign. Let's take a look at who the winners are in this category. Congratulations in best use of radio category. We only have one winner. The silver metal winner here is Frudenberg Gala household product, Private Limited for the brand Frudenberg Gala for the campaign, Why Just Clean Home? Many congratulations to the whole team out there. Ladies and gentlemen, don't forget to send out to your best wishes. Shower some love to our winners by using the hashtag Indian Marketing Awards. Moving forward to our next category, which is the best use of out of home. Let's find out who the winners are in this category. Congratulations here to the team of Madison Media for the campaign, Tata Salt, Walk in Gandhi G's footsteps on 2nd of October. Many congratulations to you. And we also have a gold winner in this category. Let's find out who that winner is. Hi, a big thank you to the jury of IMA and to exchange for media for this award. With every campaign, the endeavor is to do something glutton breaking. And I think with this campaign, the colors marketing team has been able to achieve just that. So once again, thank you so much. Big congratulations to Wacom 18 Media Private Limited, the whole team out there for the campaign, colors Nati Pinky Ki Lumby Love Story. Moving forward to our next category, ladies and gentlemen, which is best use of experiential marketing. Let's find out who the winners are in this category. The silver metal winner in best use of experiential marketing goes to Perfetti Van Mill India Private Limited for the brand Mentos, the campaign Complementos. Moving forward, ladies and gentlemen, in the same category, we have another joint silver metal winner. Let's find out who that is. A big applause for Wacom 18 Media Private Limited for colors Subharam campaign. Many congratulations to both our silver metal winners in the category best use of experiential marketing. Moving forward, which has been the huge impact area for all of us during this unprecedented time. The best use of digital marketing or social media marketing. This is the category I'm sure all of you are looking forward to. We have the finalists and the winners coming up through this audio visual. Let's take a look. Congratulations to the team of Madison Media for the brand Asian Pains of Vido Procek Screen Ka Peherida. Many congratulations to you. In this category, we also have another joint bronze metal winner. Let's find out who that is. Our heartiest congratulations to Wacom 18 Digital Ventures team. For the campaign, would select Wacom 18 Digital Ventures Assur Social Media campaign. Many congratulations to both our bronze metal winners. We also have a silver and gold coming up. Let's find out who takes away the silver metal. Give it up, ladies and gentlemen, for Discovery Communications India team for Into the Wild with Bear Grylls and superstar Rushni Khan campaign. Many congratulations to you. Coming up next is the gold winner in the best use of digital marketing or social media marketing. Let's find out. It's an absolute pressure and honor to be receiving this award on behalf of the team at Minthra. This campaign has been a true labor of love and to be recognized by the stalwarts of the marketing and advertising fraternity is extremely energizing. Thank you very much. Exchange for media and the esteemed jury members. Thanks again. Big, big congratulations to the team of Minthra Designs Private Limited for the brand Asian Pains of Vido Procek Screen Ka Peherida. Moving forward, ladies and gentlemen, to our next category, which is best use of omni-channel marketing. Let's find out the finalists and the winner in this category. Ladies and gentlemen, give it up for the silver metal winner in the category best use of omni-channel marketing to the team of Aditya Birla Health Insurance for the campaign Rakhopura Khayal. In this category, we only have one winner which is Aditya Birla Health Insurance. Moving forward to our next category, best use of integrated marketing campaign. Let's take a look at the audio visual. Congratulations to the silver winner in the category of best use of integrated marketing campaign. And the winner is Titan. Ladies and gentlemen, give it up for the team Titan for the brand Fast Track Reflex Beat for the campaign What You Need. Congratulations. And now moving forward to revealing the name of the gold winner in this category. Let's find out. Many congratulations once again to Minthra Design's private limited team for the campaign Minthra Moments Stiled by Minthra, the gold metal winner in the best use of integrated marketing campaign. Many congratulations to all the winners. And here, ladies and gentlemen, it's time for our next category. And here, ladies and gentlemen, it's time for me to be your lucky charm once again because I'm going to present to you the Carl Nirnek quiz question right now. Do not forget to give out the right answers and email it quickly to the email ID that we tell you. Let's present the question first here. So here is the question three. As per latest IRS, which of the following are in over one crore homes in Maharashtra? A, TV sets or refrigerators? B, two wheelers? C, Carl Nirnek? Or D, all of the above? You have to email your answers to shashmita.niak at exchangeformedia.com. Please check the spellings on your screen for the email ID. Also, it is mandatory to send below details with your answer, your name, designation, organization that you work in, your mobile number, your home or office address and your official email ID. I'm going to repeat the question for all of you one more time. As per latest IRS, which of the following are in over one crore homes in Maharashtra? Is it A, TV sets and refrigerators? B, two wheelers? C, Carl Nirnek? Or D, all of the above? Once again, don't forget to send out your answers to the email ID that is on your screen, shashmita.niak at exchangeformedia.com. And again, let me remind all of you, it is mandatory to send out all your personal details with your answer, otherwise you don't qualify. So don't forget to give out your name, designation, your organization, mobile number, home or office address and official email ID. You stand a chance to win a Samsung tap or an iPhone 12 from Carl Nirnek. So don't forget to participate and send out your answers super quick. I hope all of you are now ready for the big awards coming up next. Our next category, ladies and gentlemen, is branded content category. Let's find out who the winners are in this category. Ladies and gentlemen, give it up for Hangama Digital Media Entertainment Private Limited for the brand Diageo India and Hangama Yara Rangulade Rang. Moving forward to find out who the gold winner in this category is, let's take a look. Hello, and I'd like to say thank you to the entire IMA team for recognizing our work on the Swaxesolo campaign and awarding us in the digital branded content space. We believe that we bring a point of, a counter-culture point of view to every moment. And again, on Valentine, when every other brand was celebrating the love and the mush all around us, we took a counter-culture point of view and celebrated the spirit of singlehood. While I know that this year has been good for us as we won this award, I'm equally confident that with the good work that we are doing, we'll continue to win a lot of awards and a lot of rewards in the marketplace in the year 2021 as well. Thank you very much and wish you all the best. So our big congratulations to the team of PepsiCo India for the brand campaign. Pepsi celebrates the challenger spirit of singles on Valentine's Day. Hashtag Pepsi Swaxesolo. Moving forward, ladies and gentlemen, to our next category, innovative use of technology. Let's find out who the winners are. Congratulations, Street Talk, the team. I'd like to congratulate the whole team out there for their brand campaign, Uttarakhand Tourism. Take the Uttarakhand pause. Many congratulations on winning the bronze medal and we have a silver medal winner in this category. Let's find out who that winner is. Congratulations to the whole team of Flipkart for Flipkart Google OK Flipkart deliver exponential results campaign. Many congratulations to both our winners here. We don't have a gold medal winner in this category. So we're going to move on to our next category, which is marketing on a small budget. How we would all love that, don't we? Let's find out who the finalists and the winners are in marketing on a small budget category. Let's take a look. Congratulations to the whole team of WhatConsult for the brand campaign by George Allianz General Insurance Mental Health Care Kit. Many congratulations to you. In this category, which is marketing on a small budget, we have a gold medal winner as well. Let's find out who takes away the gold. Hi, I'm Neeraj Jain, Lead Creative Strategist at WhatConsult. I would like to thank the jury at IMA for selecting Nikon Instabatch as a winner in marketing under a small budget category. It's true that sometimes and briefs that have almost a low or no budget certainly get the best out of you. And this was the case with Nikon Instabatch too. So a big thanks to the Nikon India brand team for trusting our creative solution and also a special mention to everyone at WhatConsult who was working on this campaign. This one's for everyone. Cheers. Big congratulations once again to WhatConsult team for the campaign Nikon India Instabatch. Ladies and gentlemen, with that, we're moving forward to our next category which is best use of public relations. Let's find out the finalists and the winners. Hi, everyone. It's Willie Balter here from Indigo. I just wanted to say how thrilled we are and thankful that we've won an award at the Indian Marketing Awards for 2020 for the best use of public relations. All the credit should really go to our director of corporate communications, Ms. Leica, who with her team has done a fantastic job in difficult circumstances. So let's hope we can win it again this year. Thank you very much again and best wishes to all of you. Many congratulations to Indigo InterGlobe Aviation Limited for the gold medal winner of the best use of public relations for their campaign, Flying High, Despite Cockpit Battle. Ladies and gentlemen, with that, we come to the end of communications category. But before we move forward, we'd once again like to thank our partners powered by partner Colors and Carl Nirnay Abdo Karoogharome. We will showcase the audio visual and post which will move to the next category, Design. Let's take a look at the audio visuals. And while the audio visuals come on screen, I would like to urge all of you to join us on our online conversation tweet using the hashtag Indian Marketing Awards and give out your best wishes, congratulatory messages, shower some love on your favorite campaigns or brands and give the team some love as they have won the Indian Marketing Awards 2020 right here with us. So ladies and gentlemen, before we move forward to the next category which is Design, we're going to showcase the audio visuals of our partners. To the next category which is Design, we're going to showcase the audio visuals of our partners. We're going to showcase the audio visuals of our partners. We're going to showcase the audio visuals of our partners. The drama's a perfect male. Big Boss. A changeable story. A new show. The new season of Noggin is back again. I will mark the name of every devil in this world. When I make new records, Noggin is the hero of this award. Colors. Colors. Colors. Colors. Colors. Colors. Colors. Colors. Colors. Colors. Colors. Colors. Colors. Colors. Colors. Colors. Colors. Colors. Colors. Colors. Colors. Colors. Colors. Colors. Colors. Colors. Colors. Colors. Colors. Colors. Colors. Colors. Colors. Colors. Colors. Colors. Colors. Colors. Colors. Colors. Here is brand identity or packaging. Let's find out who the winners are in this category. Let's find out the finalists first. Now to announce the winner here, I would once again like to invite Ms. Shakti Salgaonkar, Executive Director, Karnaz to join me on screen. Welcome, Shakti. Hi. The award, the bronze award in this category goes to Kellogg's Granola Brand Launch by Ordinary Brand Strategy and Design. Congratulations, guys. Let's have the audiovisual, please. Once again, our heartiest congratulations to the bronze metal winner, Ordinary Brand Strategy and Design for Kellogg's Granola Brand Launch. Moving forward to announcing or revealing the name of the silver metal winner in this category, let's take a look. Congratulations once again to Perfetti Vanmille, India Private Limited, for their campaign Mentors Complimentors. We don't have a gold winner in this category, but before we move forward, I'm going to quickly take you back to the Karl Nirenek Quiz Contest. Ladies and gentlemen, I hope all of you are ready and set and excited to win Samsung Tab or an iPhone 12. Here coming up is the fourth question of the Karl Nirenek Quiz Contest. Yes. Question four. Every advertiser in Karl Nirenek gets minimum dash impressions. Here are your options. A, 10 crore impressions. B, 8 crore impressions. C, 7.5 crore impressions. Or D, 7 crore impressions. Once again, the details are the same, but please take a note of it. Email your answers to shashmita.niak at exchangeformedia.com. Urge you to please check the spellings on the screen of the email ID. Also, it is mandatory to send below details with your answers, which is your name, designation, organization that you work in, your mobile number, home or office address, and your official email ID. I'm going to repeat the question one more time for all of you. Every advertiser in Karl Nirenek gets minimum dash impressions. Take a guess if you know it correctly. Great. These are your four options. A, 10 crore impressions. B, 8 crore impressions. C, 7.5 crore impressions. Or D, 7 crore impressions. If you know the right answer, quickly type in that email and send it to shashmita.niak at exchangeformedia.com. And don't forget to send your personal details, otherwise you don't qualify for winning this contest. So make sure you send your name, designation, organization name, mobile number, your office or home address, and your official email ID along with your answer in the email. So I hope all of you have started sending out those emails, all the best to all of you. I hope you win away the Samsung tab or an iPhone 12 today. Ladies and gentlemen, with that, we're going to move forward to our awards once again. Our next category, the broad category, is marketing to unique audiences or the regulated environment. In this category, our first segment is not-for-profit sector or corporate social responsibility category. Let's find out the finalists and then the winners in this category. Let's take a look. Congratulations to Briloka Limited Team for the brand campaign Hindwaite Saaf Raho, Safe Raho. We have a joint silver winner in this category. Let's take a look at the winner's name. Congratulations to the motivator team for the brand campaign Himalaya Muskan. And we have a gold medal winner in this category too. Let's find out who takes away the gold. Hello everyone. It's an honor and privilege to accept the Indian marketing awards in the non-profit sector for our Heroes of the Road campaign. This is on behalf of the whole Bridgestone India team. I sincerely thank Exchange for Media for the platform and the recognition. It's certainly a very humbling and proud experience for us. COVID-19 has been a challenging time for all of us. And brands and marketers have a special responsibility towards society, especially in such times. Actually, serving society with superior quality is one of our core motives at Bridgestone. This campaign is very close to our hearts. It not only highlights the unsaid front-line warriors, but also the efforts of our organization towards recognizing these heroes and serving our fleet partners during such difficult times. This has been a full-team effort and a big shout-out to the marketing and CSR teams of Bridgestone India, along with what consult our agency partners for their outstanding work. Thank you again and congrats to all the winners. Cheers. Congratulations to Bridgestone India team for their brand campaign. Bridgestone India salutes our Heroes of the Road. As you can see, our next category is healthcare and pharmaceutical. Let's take a look at the finalists and then the winners. Congratulations to the team of V-Rogue Communications for the campaign Awareness on Schizophrenia Shadows. Many congratulations on winning the silver medal in the category healthcare and pharmaceutical. We're moving forward to our gold medal winner. Let's find out who the winner is. Hi, friends. It's truly a delight to be receiving this at the IMA Awards for the best brand campaign in the healthcare and pharma category. A big thank you to the entire team who's been behind this and on behalf of Team SIPLA, I would like to thank IMA for giving us this recognition. So, cheers to the entire team. Congratulations to the gold medal winner, Shabang, for the campaign SIPLA Breathe Free. Hashtag inhalers hei sehi. Now, ladies and gentlemen, we will announce winners of the two most prestigious categories of IMA, that is Marketing Team of the Year and the Lifetime Achievement Award. Marketing Team of the Year Award is one on the basis of maximum medals won in the 26 categories. And let's find out who the winner is. Can we have the audio visual, please? Hi, Abhi, good evening to the jury members and to the audience watching this particular show. My name is Rohit Kapoor and I'm here to accept and receive the award for the Marketing Team of the Year on behalf of the entire Pafeti team. It's a real honor and we are delighted to accept this award. The team Marketing did some spectacular work last year and I am myself extremely proud of that work. And a big thank you to the jury members for the IMA team, for recognizing and selecting us worthy enough for this particular award.  It was a complete team effort and a collective work which helped us to churn out those beautiful campaigns during the difficult time we were churning out. And I sincerely hope that we continue to maintain this high standard which we have set out for us even next year. But once again, a big thank you to the jury members. Congratulations to the whole team of Pafeti Van Mill India Private Limited. Many congratulations. Ladies and gentlemen, now last but not the least, we're moving towards the Lifetime Achievement Award and this award goes to... It indeed is an honor to receive this prestigious award from the Exchange for Media Group. I regret the inability to be there in person with you and other fellow recipients but would like to commend the efforts put in by the teams and jury to continue with the events even during these times. I believe we at Godridge are privileged to be part of our country's growth journey by being able to serve our nation through our innovative products and solutions. From propelling space missions to providing agri-solutions to protecting the nation from vector-borne diseases or providing sustainable living, the group's various businesses have constantly aligned themselves to India's aspirations. For us, it is most important that besides our strong financial performance and innovative, much-loved products, we remain a good company. At the heart of all of this are our people. We take much pride in fostering an inspiring workplace with an agile and high performance culture. We are deeply committed to recognizing and valuing diversity across our teams. Our work over decades expresses our undying commitment to India and we shall continue with our endeavor to keep on transforming into something new to help bring out the best for our nation. I wish to thank our 750 million-plus consumers in India, our colleagues at Goodridge, the jury and exchange for media team for this recognition. Take care, thank you, and wish you a good year ahead. Thank you so much. Ladies and gentlemen, with that, we would like to give our heartiest congratulations to all the winners of Indian Marketing Awards 2020, powered by Colors and Kaal Nilne. Ladies and gentlemen, I hope that all of you have been sending out your congratulatory messages, best wishes, showering love on our winners. Don't forget to use the hashtag Indian Marketing Awards, but love to be part of that online conversation. With that, I would also like to request all of you to please stay connected to Exchange for Media Group's Facebook official page, because we have a lot of events, a lot of unique IPs coming up. You can be part of them as well. And now I would also like to announce that all those who participated in the Kaal Nilne Quiz Contest, do not forget to keep checking your phone and email because the Kaal Nilne team will get in touch with you shortly after they finalize the winners. So I hope you win that Samsung tab and that iPhone 12. Many congratulations, best wishes to all of you. This is Khyati Kawa, your host signing off. It's been a pleasure to be the host to all of you. Have a good evening, everyone. Thank you very much.


Hello students, we have talked about many technologies both label based and label free platforms and what you may appreciate that omic tools often generate huge data as you have seen in the case of microarrays, SPR and NGS platforms. So data could be obtained from different equipments sometime when we are doing measurement using microarrays, then you are measuring the intensities. When you are looking at a mass spectrometry type of platforms, then you are looking at area under curve. So different instrument give different ways of measurement, but finally what we get, we get huge data set and then what we have to try to do further from that how best we can make sense of this biological data set, how much it is relevant to address the primary question which we are interested to address. In the previous lectures, you were exposed to the basics of statistical analysis and the importance of having clear understanding of the statistics that are to be implemented to get the meaningful insight and interpretation of your data. The data obtained from these high throughput omic technology platforms mostly comprised of huge range of values that need to be scaled down to perform the analysis. Also the big data that comes with lot of variability, sometime it is day to day experimental variability, sometime instrumentation error, batch effects, of course manual errors will be there as well. So therefore, there is a need to look at how best to normalize that data and how to remove or minimize the technical artifacts to obtain the logically relevant and meaningful data set. So we need many preprocessing steps which includes data scaling, normalization before we actually proceed for the statistical test. In today's lecture at hands on session, I have two of my research scholars Deepthrub Biswas and Arjit Mukherjee who will walk you through different steps which are required for the analyzing big data sets. They will further elaborate on different aspects of statistical analysis and data visualization and plotting. I must would like to mention that there are many very nice open access platforms and softwares available now and good idea for all of you students and participants to explore many of these software tools to make best sense of your data, not only how to best analyze the data but also how to interpret data and finally how to effectively present the data. And this is where even data visualization becomes very crucial. The data generated from these techniques comes in the form of big tables but big tables does not convey the biological sense of what we want to address. Therefore, you have to convert all these findings into the graphs and the visual way of displays. So therefore, we thought you should be learning some of these softwares and tools which can help you to best utilize and visualize your data sets. So, in today's session, we will have the basics of secondary data analysis and we will walk you through different steps involved in preprocessing of the data, basic statistical analysis, data visualization and plotting. So, let us start today's lecture. So, let us take an example that we have we have designed an experiment where there are 13 controls and 15 disease samples. So, after running all these samples individually, we got 28 files of which 13 are control file and 15 are disease file. So now, we need to analyze these files and need to know how the controls are different from disease. So, the first thing we should do is we need to take these files and do a primary analysis. So, after primary data analysis, what you will get total number of files which we took an example of 28. When we are talking about all 28 files, there is number of things that come into play like technical variation, experimental variation, batch effect and all this kind of thing. That means, we need to apply some strategy or to normalize the data. So, next part of the data analysis is the secondary analysis where we first transform the data that can be locked to transformation and try to understand whether the data is following a Gaussian distribution or normal distribution or not. Followed by if data set is skewed, then we go for sample wise normalization and if needed, we do data filtering and followed by different statistical test. So, after the analysis of the 28 files, what we get this kind of distribution and with the help of the normalization transformation and scaling, we need to get the data like this so that we can proceed for defined statistical test and statistical plot. And finally, after the secondary analysis, we try to link our significant protein list with biological pathways, protein-protein interaction and different pathway enrichment networks to understand the biology behind the disease. So, now, let us welcome Arijit Mukherjee, a junior research fellow in proteomics lab who will be giving a detailed information of secondary analysis. You are not aware of the overall overview of the complete data analysis workflow. So, in data analysis, we go through three steps mainly primary analysis, secondary analysis and then to the tertiary analysis. So, what you get after the primary analysis is the list of masses with their respective abundances or intensity. Now, from these respective abundances or intensities of a list of proteins. So, what we have in thousands of proteins, we need to make valid inferences based on some statistical test. And this statistical test and other forms of transformation scaling normalization, these steps comes under the secondary analysis steps. So, let me take you through the steps in secondary analysis. What we get after the raw intensity values, we do missing value imputation. So, missing value is a common feature in a omics dataset. This may occur due to some technical limitations or due to some manual handling. So, we will be talking in detail about this point about missing value imputation in terms of KNN that is KN nearest neighbor main replacement median replacement. Next step is to do log transformation and scaling. So, we do generally log 2 or log 10 based transformation and scaling and centering in order to be able to compare different kinds of distribution because different samples may take up different kinds of distribution after log transformation and scaling. And the next step we can perform is the normalization. Normalization comes under median absolute deviation normalization or quantile normalization. There are many other kinds of normalization that depends on your data structure that we can perform, but here we will focus on these two kinds of normalization specifically. And next comes the statistical analysis steps. Statistical analysis is performed to test a hypothesis basically. So, depending on your experimental question that you want to know the significantly dysregulated genes or proteins or mRNAs that is present in disease sample as compared controls. So, we need to perform T test or ANOVA test or regression analysis. These things will be covered in this lecture and finally, comes the data visualization and plotting. In data visualization and plotting we can draw hit maps or cluster maps box plotting volcano plots etcetera. So, this is the overview of the lecture that is going through the secondary analysis that we are going to talk about. Now, coming to the first step after you get your raw spectra if you look at the mass spectra abundances it comes in the terms of e to the power e to the power 9 e to the power 10 and so on. So, from these numbers it is quite difficult to play around this and to make valid assumptions. And as you can see in the figure on the left side that with the 4108 elements that have been plotted on the in a frequency distribution curve this is quite skewed and this kind of skewed distribution we cannot make any statistical inference based on this distribution. So, what we do we perform some log transformations generally we take a log base 2 or log base 10 and transform the data on the right hand side of the figure you can see the log transform data distribution. It is not perfectly normal distribution, but it is assuming to be a normal distribution. So, this log transformation brings the data range to a workable range that is 15 to 40 in this example. So, these numbers are quite easy to make inferences with this kind of data. So, coming to the importance of log transformation that as you can see in this figure that the skewed distribution is transformed to normal distribution or becomes less skewed and this allows you to interpret patterns in the data and always we can perform some inferential statistical test that is t-test or ANOVA etcetera. In as you know in t-test the basic controlling assumption is that your data distribution should be normal distribution. So, this allows to help us to perform such inferential statistical tests and in omics studies we generally perform log 2 or log 10 transformation that depends on the objective of your experiment. In biology generally we do log 2 transformation because we are interested which genes or proteins are in the full change of twice as compared to the normal or dysregulated or downregulated at a full change of two values. In that case it depends the scaling whether you take base 2 or base 10 that depends completely on the objective of your experiments. Now, next coming to the topic of scaling and centering if we take the log transform distribution it looks like a normal distribution, but when let us suppose we are taking control and disease samples the control samples will have a different mean if we log transform that values and the disease samples will have a different mean if we log transform the values, but these two distribution however it is normal we cannot compare the two means when the two means are very different. We need to scale and center so that the distribution takes up the similar mean or median values and they are distributed around those mean or median values and this is the purpose of scaling and centering that we can compare different samples since all samples become normally distributed with mean of 0 and this is performed in terms of z score. The z score as you can see in the formula is the x minus mu upon sigma x is the individual values of the variable, mu is the sample mean and sigma stands for the standard deviation. So, this z score reflects distance from the mean in terms of standard deviation. Assuming a normal distribution the z score should range in minus 3 to plus 3 because in normal distribution 67 percent of the population lies within the mean plus minus 1 standard deviation and 95 percent population lies within the range of mean plus minus 2 standard deviation and 99.9 percent that is almost all your data should lie within 3 standard deviation plus or minus of your mean or median. So, this is a generally very convenient method of scaling and centering that you can perform comparison of two or more different distributions in terms of z score. So, what are the other methods available for scaling and centering? In z score we assume that your data distribution is completely normal distribution or Gaussian distribution, but it is not always the case in real example or in real life examples we can see that the distribution may not be perfectly normal and in that case assuming that to be a normal distribution is not the right way to go on. So, in that case we can take up other scaling methods such as we can scale our data from a 0 to 1 scales. This is particularly useful when variables come from possibly different distributions, but it preserves the shape of its distribution while making them easily comparable on the same scale. So, here if we call the value as a score you can see that x minus minimum upon maximum. So, each of your observation is subtracted with the minimum value of all observations and then divided by the maximum value of all observations. This brings your data to the range of 0 to 1 and no other data will lie above that range or below that range. So, in this way we can compare different distribution in terms of scaling and centering. Now, next our point is the missing values as we have already told that missing values are a common feature in omics data. Now, let us think about what could be the source of missing values. The first reason could be that peptide is present in some samples and absent in some samples that is it is always generally a case that if you take 10 control samples or 10 disease samples or cancer samples out of 10 only you could get in 6 or 7 samples that particular protein and the rest of the 4 samples if this protein is not present and this can happen this can happen due to various reasons. The reasons could include the peptide may be present in below detection limit in the sample or the peptide is detected, but abundance is reported as NA not applicable due to technical limitations. If in data analysis step missing values can be imputed and there are many other methods for imputation of missing values and missing value imputation should ideally rely on the reason behind getting a missing value. Now, next we talk about the categories of missing value the categories of missing value is based on two properties missing completely at random that is m car abundance dependent missing values missing completely at random values occur due to some technical glitches in instrumentation such as pore ionization other peptides competing for charges etcetera and abundance dependent missing values occur due to peptides below detection limit or even they may not be present in the sample or it may be the case the detector got saturated and fails to record the abundance. So, these are the main two types of categories of missing values. Now, coming to the imputation of missing values here we will be talking about imputation using mean or median or lowest values. So, the mean can be used for each protein across the samples in a particular control or disease group we can use the mean to replace the missing values there or we can use the median values to replace the missing values. However, this method is has some pitfalls because this method may not truly reflect the biological variations since we are substituting the multiple entities with the same values. So, it is not a true reflection of biological variations. Now, in this slide you can look at the example of missing value imputation using mean, median or lowest observed values. As you can see in the matrix here that we have genes on the column and column side and samples that is divided into controls and desist on the row side. So, here are four control and four disease samples you can see this kind of values are lot of there are a lot of missing values for gene A this is missing for disease 3 and disease 4 sample gene V for disease missing for control 4 sample and so on. So, this kind of missing values always interferes with further data analysis and based on the distribution and based on some logic we can impute the missing values. So, that we get a complete matrix for further analysis as you can see the on the right hand side there is replacement values by means that is the rate ones are replaced by the value of the mean. If you see at the disease category on the right hand side the 35 and 36 are the values for disease 1 and disease 2 sample, but the disease 3 and disease 4 samples are missing. These values can be imputed with the mean that is the first two disease values the mean value is 35.5 and we can impute with the mean values. So, this is a method of imputation for using mean values. Again if you look at the replacement by medium that is highlighted in blue color that you can see for gene E that the control samples are having a median of 23 in 23 25 and 22. These are the three ranges of the control 3 4 control samples. So, we can impute the missing value as by median using 23 for the gene E in control 3 sample. So, this is another method of this is the method of median based missing value imputation and the last one green highlights the replacement by lowest transit value. So, it assumes that the peptide is below the detection limit in the sample that is why we are encouraged to use this kind of missing value imputation by the lowest observed value and this is shown in green. In green color as you can see for the control samples the lowest observed value is 21 in all the for all the genes. So, we can ideally substitute the 21 values in all the missing places with this kind of missing value imputation procedure. So, this is you got a general summary of how the missing value what the missing value is and how can they be imputed. Now, coming to a another missing value imputation approach that is KNN that stands for k nearest neighbor and KNN uses an unsupervised algorithm to impute the missing values. Now if you look at the graph that the star shaped point now we need to classify we do not know what which category it is going to be. So, we need to classify this star star shaped point whether it goes to the circular type or goes to the rectangular type. Based on this the KNN algorithm what it does it computes the distance between all the unknown points of all the observations and from the all the observation it takes the nearest k that is any positive integer it should be any positive integer and here in this example we are showing that it is k equal to 3. That means, it is taking the three shortest distance of the circular type and the rectangular type and from these distances the lowest distance it have the sum of lowest distance it will be categorized as the particular category of the unknown variable. In this case you can see that the it this star point might go to the rectangular side. So, this algorithm works like this and based on this if it is a rectangular type or a circular type it may be assigned a value based on the total distribution value of that particular category and this is the underlying assumption of KNN based missing value imputation. Now, the next topic comes to the normalization. Normalization is a pre-processing step in omics dataset in order to compare different samples to make valid inferences. Normalization is performed to remove the technical artifacts in experiments. Technical artifacts may come from changes in column, may come from manual handling or may come from changes in day to day temperature also. So, these kind of technical artifacts are different from the true biological artifacts. So, the whole aim of the normalization process is to remove these technical artifacts and without disturbing the biological variations. So, from these technical or biological variations we need to classify whether this dataset could be used for competition and to remove these technical artifacts we use various normalization methods such as quantile normalization and median mad normalization. Median mad means median absolute deviation normalization. Now, here in this slide we have shown you an example of quantile normalization and quantile normalization is a technique for making two or more distributions identical in statistical properties. So, let us take a case of 4 genes with 3 cases. The values are filled arbitrarily and here I am going to take you through the steps in quantile normalization step by step. So, from the raw data the first task is to rank the values from lower to higher. As you can see in the ranking of the values of lower to higher in case 1 the value 5 is given a rank 4 that is it is taking a rank from lower to higher order. The next step is to sort the data according to the rank. So, we will rank the we will sort the values according to the rank that is 1 2 3 4 there are 4 genes here and after that we would replace the values by means of that particular rank. So, in terms of if you see at the raw data and the final replaced by mean data that the rank remains the same the order of genes A B C D remains the same, but the values are changed and they are having the similar identical statistical properties. So, this is the underlying assumption of quantile normalization. However, there are some pitfalls in quantile normalization that extreme values could be masked. Hence, we may lose potential biomarkers because biomarkers are the proteins of interest that is having dysregulated expression level that is in the normal distribution curve they might be in the tail region. So, quantile normalization may mask your potential biomarkers from the analysis and so, it should be utilized or implemented with proper care and quantile normalization is generally used for microarray data analysis. Next comes the median absolute deviation normalization what we call in abbreviation as mad normalization. So, the mad normalization is preferred when the distribution of the data is not normal and it is particularly useful in that case. This score is particularly applicable when we assume that the data distribution is quite Gaussian and normally distributed. So, mad normalization is similar to z-score scaling and the only difference is that it takes the median of absolute deviations so that they are not affected by extreme values and this is particularly useful for non-normal distributions. The median absolute deviation as you can see in the formula is the median of the absolute deviations. Now, let us talk about the data visualization and plotting techniques. If you look at any omics paper in proteomics or genomics these kinds of plots are very common that is heat maps, cluster maps, box plotting, volcano plots, PCA plots so on. So, let me take you through the through the glimpse of these old data visualization and plotting techniques. Let us talk first about the heat maps or cluster maps. Heat map is an useful tool to visualize the patterns in large scale biological data. So, what is basically is that there you can actually visualize thousands of genes and their pattern across numerous samples. As you can see in the figure that the rows are the list of proteins and on the column there are list of samples and you can see for a particular protein if you go through but go through the row you can see the pattern of this particular protein across different samples. And this this allows us to get a get a visualize an overview of all the patterns in the large scale biological data. And this curly scale that ranges from minus 4 to plus 4 this comes from the zis code scaling. So, after you lock transform your data you do the zis code scaling and you can plot this as a heat map. The blue ones are representing the lower expressing values and the red ones are expressing the highly expressed proteins. So, this is a very useful technique in omics studies that you can make patterns out of this whole large scale biological data. Next we talk about the Volcano plots. Volcano plots are a quick and easy way to visualize the genes or proteins that are statistically significant. In Volcano plots what we plot on the x axis there is locked to fold change values and on the y axis there is negative logarithm of p values. So, after you perform your t-taste or ANOVA you end up with some p values for a particular protein and you can also determine the fold changes in comparison to the control samples for the disease samples. So, this disease samples and control samples ratio is called the fold change in terms of expression values. So, if we plot the locked to of fold change and the negative logarithm of the p values we can know what are the points that are very much statistically significant in terms of p values and they are significant in terms of fold change. As you can see in the figure the red dots are above the fold change values. So, these are having significantly significant p values and the green dots are on the lower hand side are having a locked to fold change in the range of minus 1.5 to minus 2 that is these proteins are quite down regulated as compared to the reference samples and on the right hand side the green dots denotes these proteins are up regulated as compared to the reference samples. So, this is a quick and easy way to visualize your data and which proteins are of your interest in terms of biological questions. Now, coming to the box plots box plots are easy way to visualize the spread of your data points for a particular gene or protein. So, when you have narrowed down to your proteins or genes of interest this box plot can be an easy way to quickly compare the distribution of the abundance values across the samples. As you can see in the example on the right hand side that we have taken the thymocene beta 4 like protein 3. This protein we have plotted the fold change values across different types of samples and this is compared in grade 2 samples grade 3 samples and grade 4 samples. You can see the data distribution of this protein in terms of fold change values across those samples and this is generally plotted in terms of box plot. And in box plot you can see that there is a media line in the middle of the box and this represents the median value of expression of that particular protein and after that you have on the upper hand side you have third quantity and on the lower hand side you have first quantity of the expression values. So, these are the parameters of box plots that is used to actually visualize the spread of the whole data. Now, let us talk about PCA or principal component analysis. Principal component analysis is a dimensionality reduction technique to interpret the clustering of similar type or grades of samples. In PCA generally what you see is a two dimensional plot. The x axis represents the principal component 1 and the y axis represents the principal component 2. So, the principal component 1 represents the maximum variation of your sample. So, this is the line where you can group or cluster your data and see how they are clustering. As you can see there are different colors of grouping of individual samples into different clusters. That means, they are quite different from the other samples. This PCA plot is another easy way to visualize the grouping and clustering in your samples for making further biological inferences. I hope today's session was informative and you go to learn many new tools which you should start playing with it. There are a lot of data sets available in the public repositories. Of course, you should ask even our lab to provide you some more raw data set which you can play with you can try to analyze yourself. And with these sessions I hope you are getting good understanding of different steps involved in data processing and analysis. Of course, each instrument platform whether we talk about microarrays or surface plasmon resonance or next generation sequencer or mass spectrometer. The initial raw data processing is very unique for each type of instrument platform. But once you have obtained the data in the excel sheet format, then a lot of things are pretty much common because you want to make biological sense out of these raw numbers and these big data tables. And what you are looking at, what was your background? What is the noise? What is the actual signal? So, many things for example, the processing steps remains very similar irrespective of which technology platform we are using. Of course, there can be uniqueness looking at the constraints of each technology as well. But finally, what you want? You want to make a biological sense of the data and you want to plot the data in more meaningful way. The different ways of visualization of data and tools available which can effectively represent your data are very crucial. So, some of these tools I hope what you learn today would have given you much better understanding that how effectively you can start looking at your data. And in the upcoming lecture, we will continue further on this and show you more software and analysis which can now try to build from your protein and gene list, how to we can make the networks and pathways and make better biological sense looking at the complexity of the problem which you are working on, how to make sense of that information by targeting different pathways and interaction networks. We will continue more of the session in the next lecture. Till then, thank you.


Welcome to Longmont Voices and Vision, a project of Longmont public media. In the midst of the darkest period in our lives, when we're bombarded 24 hours a day with news of the coronavirus and the human and economic carnage it's causing in our society, we're challenged to cope with our fears and anxieties, we're remaining hopeful about what lies on the other side of this crisis. This project presents an opportunity for Longmont residents to share with others how they're adjusting to new realities of social distancing and the kind of future they hope to experience on the other side of the crisis. I'm Tim Waters, host of these conversations in a Longmont public media volunteer. In this series I'll be asking Longmont residents, many of them your friends and neighbors, three questions. What are you doing to get through this crisis? Even though we cannot be together right now, how are we staying connected to friends and families? And what's the future you are hoping to see and experience on the other side of this crisis? I hope you'll stay with this series and enjoy listening to your friends and neighbors and learn from them how they're getting through and what they're looking forward to in a new reality on the other side. Vito Montone, thank you for lending your voice and your vision to this Longmont Voices and Vision project.  Yes, hi, I'm W. Vito Montone, I go by Vito. I moved to Longmont about two years ago now, or is it three? I don't know, I'm losing track. I run a strategic agency that does work for people around the country. And I am the new president of a board of the Longmont Council for the Arts. Well, three questions you know I'm going to ask right now, given where we are in this, in this experience, right, with this pandemic. The first question is just being curious about how are you getting through this current situation? Well, you know, it's, I don't know if other people can relate to it, but for probably, I don't know how many years of being self-employed, we've run a virtual company. So for us to be home more than normal is normal, you know, we're normally here all the time. And I guess a year or so ago, a year ago, we were blessed where my son, an adult son, chose to move to Longmont and then we chose as a family to buy a house together. So our core family is together. That's also normal for us. And what we've been doing, frankly, is re-experiencing all the Marvel movies in chronological order. So not the way they were released, but the actual storylines and it's unbelievable what, you know, what Stan Lee created and what he was able to keep together. So every night, we watch a movie, we're halfway, you know, so that's what we do and we look forward to that every night. So in a moment in history when physical separation and social distancing are now expected, but so unprecedented for us, what are you doing to stay connected to that? You've already answered part of the question in terms of staying connected to nuclear flame, but the rest of your family and friends, how are you staying connected? Well, we've used Google Meet and we use, of course, we're part of the wave of using Zoom, which is fascinating that one of my divisions of my son's company actually supports Zoom, so they're pulling their hair out with all the new demand. And of course FaceTime and Chats, because we're normally a virtual group, we just stay connected that way anyway. And on a practical basis, we're, you know, Amazon Prime members, we get a lot of our staples that way, except for toilet paper or fears. And even our protein, we get it from a farming system run by the individual farmers called Moink and we have all our protein delivered.  And everyone's, at least in our neighborhood, which is not far from yours, we're very, everyone seems to be pretty respectful of the distancing, you know, just opposite sides of the sidewalk or whatnot. So, you know, I don't, you know, because of people that might be having hardships, I hate to say this, but it's kind of normal for us. Yeah. Well, my third question, as you know, going into this, whatever was normal before this pandemic and social distancing and the isolation, coming out the other side, whatever the new normal is likely going to be different. Life is likely not going to be the same. So given that, the third question is what's your preferred future? What do you want to move to or toward on the other side of this? Well, that's obviously the most thought provoking question. And certainly, it seems that we're being taught a massive lesson that we're a bunch of human guests on a planet that ought to be working together and being cooperative, and being prepared for listening to experts as to what we should be being prepared about. I think it's the most disconcerting condition that we watched, you know, a very large economy, pretty much knock it out of the park on fighting the pandemic. And we still don't seem to be really listening. And we did a variety of things a few years ago that disabled our ability to respond. And I think breaking down those walls that are silly and politically biased, you know, partisan, when we're supposed to be taken care of each other, I just, it's almost stupefying to me. Like, we're supposed to be intelligent and powerful and smart.  You know, it's really fascinating. So I'd like to see a more related intellectually caring world of each other moving forward, even in our small town. We're part of the system, right? So. You know, thank you so much for contributing to this project.  And we'll see you on the other side of this pandemic. Absolutely. You too. Stay safe. Tallis, thank you so much for your contributions to the Longmont Voices and Vision project. We've started all these interviews by asking those involved to talk about themselves.  Yeah, thank you so much. I appreciate the invitation. I'm Tallis. I am the host of the savvy entrepreneur on Longmont public media, as well as on my own YouTube channel. I have been an entrepreneurship since 2004 when I started my first company. I have since been involved with multiple companies and multiple different industries from financial technologies to aggregating apps to market and efficiency plays to pharmaceuticals, including a chemotherapy drug that involved with a laptop surgical tool, other medical devices, as well as aerospace and graphene nanostructure technology. I also taught at the University of Colorado. I was lucky enough to get the opportunity to teach at the age of 30. So I was one of the younger faculty members there. I've also written a book called the Savvy Entrepreneurs Business Handbook. And yeah, I am happy to be with you today. Thanks. First of three questions. We're in a moment of kind of unprecedented in any of our lives in terms of what we're going through. How are you getting through this current situation? I think I'm doing pretty well. I've been focusing on making content. I've been speaking with a lot of professionals regarding this COVID, whether it's accountants or other business professionals talking about what they've done in order to make this hard times or people who've been displaced because, you know, financially displaced because of the situation. Yeah, stay at home. I'm lucky I have a backyard. I have an awesome wife that I can hang out with. I just got three ducklings. So I plan around them just built them a duck house and actually, before we jumped on this call, I was setting up to build my raised planner beds that I've been meaning to put together. There you go. We're in a moment in time where we are kind of the physical isolation and social distancing is unprecedented in our lives. How are you staying connected to family and friends in this time of social isolation? You know, that's a great question. It's one of the topics that I spoke with a PhD psychologist about and if you, not to do a shameless plug, but if you want to check out my content about this, I've got great tips and tricks from PhD talking about how social distancing and isolation can impact the psyche. So, yeah, a very fascinating topic. I have been staying connected with my in-laws and my direct family members through video conferencing and through telephone calls. I have had friends, a couple that are being impacted by this. A couple of them are depressed and one of them was severely depressed. And so it has definitely taken tolls on people who are close to me. And luckily, you know, with technology, you can communicate readily. I can't imagine what it was like in 1919 flu epidemic when people had no corn peening and you have to write a letter and then hope that the mill carrier didn't die. And then, you know, it's quite a different time. So I think we're in a much better place than people have been in the past. The third of my three questions really is about the future. And it's safe to assume that whatever the new normal is on the other side of this pandemic, life will be different than it was on the front side of this pandemic. So for you, assuming we can influence or help shape some of what that new normal is, what's your preferred future? What would you like to see and experience on the other side of this? Another very good question. I would like to see people understanding the value of social programs. I think that people should realize after this that universal health care is something that we actually need. For cases like this, people are going to realize that a universal basic income is something that isn't just a pie in the sky idea, but it's something that can actually be implemented and could be very beneficial in times like these. I think that, you know, property values for commercial real estate are going to drop, making it a lot easier for businesses to get off this space if they need to. And I also think that a lot of the growing pains that have prevented us from working at home more are going to be overcome after this. People are going to realize how awesome it is to just rule out a bed, make breakfast, grab a shower and get to work within 30 minutes rather than spending an hour commuting. I've been working from home for a very long time. Luckily, I'm an entrepreneur. I have the flexibility to do that. Many members of my team are international and across the US. So I've been doing this for a very long time and I think kind of a best kept secret kind of thing that a lot of people are going to wake up to. One of my colleagues is a professor at MIT and he was saying that MIT is not going to even bring in new freshmen until next January. So a full year without class and they've been trying to implement at home learning with good success considering their top two university with a ton of resources. But I do see education changing along with working in the fact that a lot of it will be done at home. I do acknowledge that there are people who maybe won't thrive in that type of learning environment as well as working environment as others might. But I do feel as though this is a more efficient use of labor when possible to have them just work at home. The other great thing is Longmont has Next Light Internet, right? So I'm here with Gigabyte Speeds and I think our picture looks really good today. Well, yours probably looks better than mine. Talis, thank you so much for your willingness to participate in this project.  Stay safe. Yeah, thanks Tim. Good job on the project. I commend you for such a great endeavor. Thanks. Take care. Karen Dyke, thank you so much for lending your voice and your vision for this Longmont Voices and Vision Project. A good way to start these interviews has been to learn a little bit about who's being interviewed. So tell us about you. Okay, excuse me. I'm Karen and I'm a retired nurse. I worked primarily, excuse me, I worked primarily in nursing administration, ICU, ER type directors. Excuse me, I have to start coughing. This is new. This isn't the virus. It's important to point out to everyone. Yeah, yeah. You don't need a mask around you, but anyway. So I'm a retired nurse. I moved back to Colorado and back to Longmont when I retired. I have four grandchildren here, two sons and their wives and my four grandchildren. So what I do now is I play bridge and we figured out how to play bridge on Zoom and a website, which is good. And I'm also an activist and I work on environmental issues, primarily about fracking because of the air pollution with that. And then also climate change. And so I keep really busy with those things and I like to garden. So I'm glad if I'm stuck at home that I'm at least able to get outside and look at the tulips that are blooming and such. You know, I'm going to ask you three questions. The first is in this period that's unprecedented for any of us, probably unprecedented in human history. How are you getting through the current situation? You know, I have my jigsaw puzzle back here. I'm doing jigsaw puzzles. You know, I'm kind of, I've always been that news person always reading the newspaper and such and and have the news on to anything exciting happening or disastrous happening. And I found I've had to back away from that. I do like to read but I find that I have difficulty holding my attention to books. It's hard to get into that. So jigsaw puzzles have been good too. I can watch something mindless on Netflix or Hulu. My kids don't want me to go out anywhere. So I've been very few places since late March. I haven't been anywhere where I've been out of the car. I went to the bank and at the ATM and such. My youngest son brings me groceries a couple times a week. So, you know, it's at some point isolating, but we figured out the zoom is a good way to connect we did. I did a book club this morning with my book club and and most of the people are older than I am and all but one managed to get on to the site so that was really good. So, yeah, just, you know, I did walk down a golden ponds a couple times, but I see the people walking I live real really close to there I can walk to there. I see the people walking look very sad and it makes me sad so I'm better off in my garden I think then out walking I do just go to walk to mailbox here and such so I'm getting through it. So, you know, he is isolating that I've gotten kind of used to that and I talk on the phone a lot. I really don't enjoy talking on the phone that much, but I've talked on the phone a lot now. You may have already answered my second question, and that is how you stay connected to family and friends. Anything else to add to that? Probably the other thing is staying connected with my activism work, because, you know, we know that when we get through this, we have a crisis worse than this pandemic facing us which is climate chaos. And we've already watched bits and pieces of that we saw Australia on fire, who we've seen in the past huge fires in California in our own forests and so, you know, that that that is still out there pressuring us. So, I continue with meetings and, you know, emails and online working on that and I do work with a sustainable resilient Longmont we do our meetings online now we have our renewable energy committee, which is the one that pushed for the ready for 100. So we've been working very diligently with PRPA is changing their, they're working on on their process for what their goals are for the next 20 years and, and unfortunately they've put natural gas in there. That doesn't work very well for me and I think most people who live in Longmont we voted strongly we didn't like natural gas so why we've moved to natural gas when we know how polluting and damaging it is to the atmosphere makes no sense so continuing to work with several different groups and along the activism line, trying to keep our, our, our path open. We're probably going to do. I know we're doing an Earth Day thing here this week here's just the 50th anniversary of Earth Day and we can't celebrate like to. But we'll be doing a webinar for Earth Day that has some children, youth, they're not, they're not kids their teenagers, a panel with them and then we'll be doing a webinar in May. About renewable energy, just to keep that going forward so I stay very busy with activism. Well, much of what you just described I suspect he's at the third question and that is. It's safe to assume that on the other side of this pandemic, whatever the new normal is will life will be different than it was before this pandemic. We have the opportunity to shape some of that, whatever the new normal is so for you what's the preferred future, what would you like, what are you drawn toward what you what would you like to see an experience on the other side of this. Yeah, I think, you know, a number of things I, as I look at the future I think we, we have to take what we've learned about this pandemic and build a better future off of that. We've seen that our federal government hasn't worked. At least in my mind, they've kind of stood in the way we've had like city states doing the work themselves we've had the states trying to do all of this and it's, it's very mismanaged and I say that from a healthcare background. We need, we need strong leadership from the federal government to prepare us for what's coming to help us define a path that isn't based on profit but it's based on the needs of the earth. And we need, we, we are going to have to deal not only with the US but we need strong leadership here so that we can around the globe work on how we defeat climate change. We haven't done a good job of that with this pandemic. Other countries have done a much better job than the US have and it kind of hurts my heart. My age, you know, I've always had this thing that the US is great and you know we're very innovative and creative and we can do anything. We have really, this has really been bungled and that kind of hurts my heart and how we haven't been the leaders, you know, we, we just haven't been out there being creative and and as a result, thousands of people died that probably didn't need to and and one of the ladies that died last night, I know, and that's very discouraging to me that, that she had to die. And yeah, that's just very discouraging and we could have, we could have done better, you know, South Korea and some other countries did a much better job than we did of flattening the curve and keeping it from spreading. Karen Dyke. Thank you for your activism. Yeah. Thank you as well for your contributions to this project.  And when we can all come back out of our homes. We'll see you outside. Okay, great. Thanks.


sometimes you have to create your own closure in life. You have to curate and tailor in a bespoke way what looks like closure for you. It may not be what you first anticipated it would look like because what you anticipated and your preferred version would have involved someone's cooperation. The best closures are like that I think but many closures in life cannot be that way because the person that you need to give you that perfect closure either can't because they've passed away or they're not capable or able to because of some physical disability or something in the aging process or you can't find them and they're not contactable or you can but they don't want to be participating in it which is my father's case and so you can't rely on their cooperation to give you the kind of closure that you really wish you could have.


It's a moment to step back and really rethink and we set up a new world order after World War II. We're now in a different world than we were then. There is an opportunity to put those funds straight into jobs that will accelerate the pace of the decarbonization. So we actually just had a report from the International Renewable Energy Agency about the trillions, literally, that are there that are possible in investing in renewable energy, which is cheaper in most places than fossil fuels, specifically in the electricity sector, but also moving into the transportation sector. So we have a chance to both put people back to work and to chart a different future in a short time frame. Dealing with climate change isn't a luxury. Dealing with climate change is a life-and-death issue and it has massive economic consequences also for people, for farmers who are suffering droughts in their countries, for particularly people in harm's way of intense weather events. And those are things that are happening right now.


Hey there friends, thanks for checking in today. We're looking at a Henry homesteader Autoloading 9 millimeter carbine made by Henry. They are known for lever action guns This is the first auto loading rifle that they have in production. It's a very nice carbine with a 16.37 inch barrel American walnut the craftsmanship with Henry rifles are amazing and their shotguns I have a lever action axe and I'm still impressed with it very impressed with this homesteader now This is available in three separate configurations this one has Glock mags available. I had to switch out the mag adapter with the Glock mags So that'll fit Glock mags, but it comes with two Proprietary Henry mags a five and a ten rounder and that model goes for an MSRP of 928 this goes for 959 So it has the Henry mag only the Glock mag adapter and another model with an adapter for mmp9 Magazines and sig p3 20 mags So that's how this rifle comes, but the American Walnut is absolutely beautiful now It's a bit heavy right under seven pounds but what it offers is Quite a bit a ton of fun. I'm going to get it over to the table so we can take a look at it I have a lot around through it already. I just got it I fired it quite a bit very happy with the craftsmanship The accuracy of it being that it is heavy You hardly feel any of the pushback a little bit of pushback that nine millimeter offers But a nice all-around package. Let's take a closer look. We'll start with the stock Very nice American Walnut stock With a rubber recoil pad goes a little over a half inch has a bit of a spongy feel to it a rear sling mount right there The checkering on the grip of the stock is very well done. We see that same checkering on the forend there Very nice with a sling stud up on the forend I mentioned it has Glock mags or it is suitable for Glock mags All you have to do is punch out these three roll pins and Take out the old mag the Henry mag and put in a Glock mag It does have the magazine release right there, and I currently have a Glock 19 mag in there the charger is ambidextrous It's right there Charge the rifle all you have to do is pull it out and move it to the other side If you want to charge it from the left side of the rifle the ghost ring site The rear ghost ring site is right there. It is adjustable No need to adjust it for me right out of the case. It worked fine, and then the front is a blade One of the things that is super cool about this is this lever here. It is also ambidextrous We see it on both sides. All you have to do is pull the bolt back Push that upward and that holds the bolt back. You could run a boar sling through it to clean it The barrel is threaded. There's a thread cap there You can run a can on there if you wish and there's a closer look at the front sight Very nice package, and I like the adaptability of putting the Glock mags or P320 M&P 9 or just use the Henry mags right out of the case take a look at the top strap of The rifle the American walnut I Think makes the rifle heavier, but it is just beautiful the craftsmanship of this check out the safety thumb safety on the Tang of the rifle, but the craftsmanship is second to none Really nice setup Really happy with it. You know I've taken quite a few shots now what I've noticed is That the the sights you put that ghost ring you put that blade right in the middle of the ghost ring and you're pretty dead nuts Each time I shot it it is drilled and tapped to put a rail on there in case you want a There's a motorcycle riding by here have been the trails, but you can put a rail on there put an optic I don't know. I kind of Kind of want to keep it the way it is. You know, it's a it's a Henry which in my estimation is Fine out of the case, but there's a closer look at it beautiful rifle very accurate No pushback at all with the 9 millimeter Henry homesteader. I told you the MSRP is in the mid to upper 900s However, I did a quick search and you can find this anywhere between 800 to the MSRP prices, but if you get a chance check it out It's a beautiful rifle very easy smooth shooting rifle made by Henry the craftsmanship is amazing The accuracy is really nice And I like having a rifle within an up without an optic for once because it takes me back to that old time Shooting if you like videos like this, please subscribe and share. I always appreciate thumbs up button Thanks for watching and you guys be safe


Hi there, my friend and friends. When you first start learning HTML, it can seem really straightforward. You have a paragraph. Oh, you just need a little p tag for that. Then you learn that there's different types of headings and that's not so bad. And then you have lists and there's actually two different types of lists. And then there's the items inside of that. And then there's the strong and the M and there's links and there's images. And the list just seems to keep growing. And you wouldn't be wrong because there's actually 118 different elements in the current HTML living standard. But don't worry, you definitely do not need to learn all of them, especially not right now. I'm assuming that you're very early on, you're just starting to learn HTML and maybe sprinkling a little bit of CSS in there. And if that's where you're at right now, in my opinion, there's only really 20, maybe 25 tags that you actually need to learn and remember sort of what their purpose is, especially early on in your journey. And to help simplify things a little bit, I also like to group these offs and we have less than 10 in every single one of the groups. And by doing that, it makes it a little bit easier. So we have a meta group of tags. I have a layout group of tags and what I like to call the content related tags. Now, I like grouping them off because I find by giving them sort of a more specific purpose, it sort of just makes life a little bit easier. Like I'm putting in content or I'm making a layout. I should be looking for one of these ones. And these are completely made up by me. These aren't like official categories or anything like that. I just again think it helps simplify things, especially when we're learning. And one thing I just want to say before we jump into these different categories is this video is really intended for people who have at least started writing a little bit of HTML and CSS. Because we're going to be going through an overview of tags. If you're looking to get into web development, you haven't even written one line of HTML yet. This video might seem like a little bit much. And I'm going to link to a playlist just down below this, which is my complete beginner series on getting started with HTML and CSS that really starts from like absolute zero of how like what an HTML tag or element even is and the syntax behind it. And it starts very, very simple. But if you have started playing around with it a little bit and you want to know the things that you really need to focus on, especially in the very early stages, because there's a lot more than what we're going to cover in this video. But these are the ones that you need just when you're starting off and then you're in the right place and we're going to start off with that meta category that I just talked about. Now in this meta group, there's actually a meta tag that's going to come later. You don't even have to worry about that one for now. So hopefully that's not too confusing, but in this grouping of meta tags, the first one there is your doc type.  And this is just telling the browser that it's an HTML document that it's seeing, and it's also actually telling it that this is an HTML five document. This is important because we've had previous versions of HTML that were actually a little bit different in the rules and the syntax that had to be followed. So it's just saying, you know, I'm using the most current version of HTML. Just so you know, and it just helps the browser know what it's looking at. Then of course we have the HTML tag itself, which opens at the start and then ends all the way at the bottom because that's where all of the HTML is going. And I know it can seem a little bit silly that like we just told the browser it's looking at an HTML document and then here's where it starts and here's where it ends, but when we're dealing with computers, we usually have to spell every single thing out. And that's exactly what we're doing. We're giving it a file type, doc type at the top. And then we're saying, and that here is where that HTML content actually lives. Our next up, we have two very important tags, which are the head and the body. And the head is really where like the meta type things live. It's the things that the person doesn't see, but just like those other ones we've looked at, which is why I sort of group all of these together and the whole purpose of the head is to put information in there. That's like extra information. And it's a little bit like when you take a photo with whether it's your phone or a DSR or any digital camera, it's going to store a whole bunch of extra information about that photo from the location to the time of day to a whole bunch of other stuff, the ISO you have and all those types of things. I'm not much of a photographer, but all of that information is stored in the photo. But when you look at the picture, you don't see that. This is what's in your head. It's extra information you're providing to the page, but it's not actually things that are showing up on the page itself. So one of the important things we have in there is our title and the title will show up in the tab and it also shows up in search results. So this is a very important thing to have in there because this information is available, but it's not actually part of the content of the page itself. And you just include whatever the title of your page is. That's nothing too complicated. And then after that we have sort of a grouping, which is either going to be a style tag where you can put your CSS or a link to an external style sheet. So if you're very early on, you haven't maybe touched CSS yet or you've just started looking at CSS, you might have seen the style tag. And if you've been doing this for a little bit longer and you're just watching this as a bit of a refresher, then you've definitely seen the link tag, which is linking off to an external style sheet. If you've just been doing like the inline CSS where you're going on the individual tags and applying CSS rules, you're very soon going to find your style tag in a future tutorial. So just know and get used to putting that there or the link to your external style sheet, which is generally how we do things. Though a lot of the time with early content, we use a style element at the top to put the CSS in. I've already mentioned the body, but this is where we finally get to like put content on the page. And these next two categories, which are the content related ones and the layout related elements, they are all going to be included here. And I've just realized I've been going back and forth. Sometimes I'm saying tags, sometimes I'm saying elements. The tag is we have an opening and closing tag, of course. Right. So the P for our paragraph, which we're going to talk about in a second, the opening one and the closing one. And then the content goes in there. So if we look from everything from the beginning to the end is our element and a lot of learning material you do, whether you're watching my channel, you're reading blogs, you probably see them used a little bit interchangeably. They tend to mean the same thing unless people are being very specific about an opening or closing tag, but sticking with that P tag, that's one of the essential content related tags that I do like to talk about. It's usually the first one that people learn, or they learn the heading tags first, the H1 through the H6. And I sort of group these as like, if you know one of them, you know all of them. So you have a heading one, a heading two, a heading three. And so for both of these, it's we're sort of telling the browser what the content is. I have some text here. What is that text? Is it a regular paragraph or is it a heading? Now, one thing that's really important about the headings is while they come with default styles on them, where when you do an H2, it gets a certain size. It gets a bold font on it and stuff like that. You don't actually want to be using these for styling purposes, because it's actually creating the document outline for your document or for your page. So it's a lot like a table of contents. And if you look at like a textbook or something like that, where you have a table of contents, where you have like a main section, and then there's subsections within there. And then there might be even subsections within the sections. That's a lot of what the headings are doing. Or it's exactly what the headings are doing where the H1 is your page itself, the title for the whole thing. You should only have one H1 per page. Then the H2s, they're the next level down. So this is sort of creating like the sections of content in the document outline. Then you might have an H3. So the H3 would be a separate section within that H2 that was just before it. And then you might have several of those. And then you go to another H2 and that's sort of pulling, okay, we have another separate section here. So the way that you use your H elements, the H1 through H6, it's really to create the document outline and treat it as if it's creating a table of contents, because that's basically what it's doing behind the scenes. And then, of course, we have our lists and there's two different types of lists. There's actually another type of list that we're not going to talk about today, but there's the two ones that you see the most often, which is the OL and UL, and that's for ordered list and on ordered list. And then whenever you have these lists, first we're saying I have a list and I have to say every item that you have in there. So then we have our list items, which is our li tags. So again, it's you put a UL, then you put all your li's for each one of the different bullet points or the numbered lists that you might be running there with an ordered list. Is each one of your li's that are in there. And then at the bottom, you can close that off and keep going with some other content. The next two that I like to look at that are sort of treat them similarly to one another is the strong and M tags, first of which is strong, which actually stands for strong importance, which by default will make your text bold. And then the EM, the M, as you'll often hear it is your emphasis tags. You're putting emphasis on that text. And these both have semantic meaning to them, just like our H1 through H6 help define the document structure. There's semantics and there's actual meaning behind when we use the strong or the M tag. And that comes to like how the text is read. Because obviously a big difference between I love you and I love you, right? And if some, depending on how someone says that. And so that's if you have the love in the M tags there, you get the more, the more serious one, maybe there, you're putting extra emphasis on that text. And it's not just about the visual of it being italic, but some people will be using screen readers that read a page to them. And it will, this helps inform the screener how it should be reading the text aloud. It's giving that extra information to the browser as well. Now up until now, most of them, I think are pretty straightforward, though strong and M, maybe sort of like it's strong importance, but you get there. Then you have the A tag and an A is a link and that can seem a little bit weird. But what it actually is, is the anchor tag and you can see it as it's anchoring that text that you have to another location. So when I click on it, it's going to bring me to that. So it's like tethering it or anchoring you to another spot. And if ever you have trouble remembering it, that's just what it's that's what it is. And it's nice and short, which is good because we tend to put a lot of links on our pages and there's two different ways we can use links. We can link to another page or we can link to within the same page. And if you want to link within the same page, you would link to the ID of the element. And then when you click on it, it will bring you to that part of the page. And of course, if you're linking off to another page, you just put the URL for that page and you can be brought over there. Next up is another one that you probably end up using a lot, which is our image tag and that one's pretty straightforward. I am G. It's a little easier to write than writing out image and you just tell it where the image is and you have an image. So that one's nice and easy and usually pretty easy to remember as well. And the last one that I want to look at that's more content related is the span tag and span is similar to your strong and your M. But the difference with it is it's it doesn't come with any default styling and it has no meaning to it. So you might be going, well, what's the point of it then? But the point isn't to take the default styling and it's nice that it doesn't come with any meaning. And the idea is that I can select a piece of text and I can style it. So if I have a heading and I actually need like say it's a two word heading and you want the color for each word to be different for a design that you're creating, well, then you would use a span tag to be able to select one of those two words and change the color of it. And those are the essential for me, the content related ones where we're really working with content. We're telling the browser what the content is like, this is a paragraph, this is a heading, this is a list and things like that. This is a link and image and all of that. So like for me, those are very content related. I like grouping them like that. The next group that we're going to be getting to is more of the leot types of things. We're still telling the browser and giving it information about what each piece is, but it's a lot more leot related and grouping related, I guess you could say. But I just want to let you know like right now we're going through all this. It might be a lot of information. If you want a cheat sheet that has all of this information in it, I have made one. There's a link down in the description below where you can get this. I list all of these tags out with little descriptions of what they are and include a couple of links on some of them that might need a little bit deeper of an explanation. But with that out of the way, let's get to it. And the first one I want to talk about is the main and the main element or the main tag open and close. It's a little bit like the H1 where we should only have one per page. And the idea with the main is this is the main content of the page. And you might be saying, well, Kevin, the entire page is my main content. That's why I have a page. And if that's the case, just throw one in there because every page should have one main. And if everything is your main content, then just main at the top, main at the bottom and your content goes in there. No problem. But you might have other things on your page as well. One of those things being the header. And a lot of the time we have a header at the top, then we have the main and then the other one that fits after that is the footer down at the bottom, the header, which often is at the top of the page. And most websites these days will include your logo and your navigation because that's not really part of the main content of your page. That's like the before your main content of the page, right? Because it's always there. It's not like if I go to like a pricing page, the navigation is not really related to that pricing. It's related to everything on this website that I'm on, but it's not about the pricing itself. The main content is the pricing. So I have this extra information that's coming before I get to the main stuff. Unlike the main, you can actually have multiple headers. We'll talk more about that in a second, though. The other one that I just mentioned though is the footer. And the footer is similar to the header where it's something that's at the bottom that's not related directly to the main content but is usually related to the entire page you're on. So often this actually has like a different navigation in there on a lot of pages now. You may have your copyright information there. There could be like extra other information that you find down in the footer and it's usually repeated on every page though it doesn't necessarily have to be. And in both the header and the footer, I said that you often find navigations and that's where you would have a nav tag. And our nav tag is used to say this is a navigation. Importantly, though, it should be like an important navigation. So it's not every group of links that should necessarily be a nav. If you're just throwing like here's like a couple of pages you might like. That's not really a navigation. That's just a group of links. But if you're doing it as a navigation, so like this is how you should get around my site. Here's the primary, all the primary pages that you might be interested in. Then you can use a nav and list out those pages that you might be going to. There you have your nav and then in the nav you'd have a list of links that you'd be using to list out all the different places that people could get to. And next up is two that are a little bit more people get a little bit confused between the two of them, which is the article and the section. And an article is something that should be a standalone piece of content within a page. And that means like if I took it out of that website and I brought it somewhere else and placed it there, could someone make sense of what's going on in this piece of content. And so this could be a recipe, right? Like say I'm on a recipe site that has all these different recipes. Every page might be a different recipe. But could I take that individual recipe off and go like I could I could take hopefully your recipe from your page, bring it over to my website that's a web development and put a recipe there. It might be weird, but it would make total sense. So that's the idea with an article is the standalone piece of content that can stand on its own. And the other one is the section. And the section is more about like sectioning off content. So here is, you know, you might have a homepage that has like your main area at the top that's talking about your product, then you have like a little about us section, then you have a pricing section. And it's one page that's broken up into different pieces. And those pieces are sort of related to one another, but you're you're separating the different sections. It could even be within an article, you could have several different sections within a larger article. So they don't really stand alone. They're related to the other content that's around them, but it's different sections that you're sort of organizing your content by. You can use your section element. I have one more element that's probably the one you're going to use the most. But before we get to it, I just want to circle back to the idea of the header and the footer, where as I said, you might have a header at the top of your page and a footer at the bottom of your page. But you can also have headers and footers in other places too. And one of the common areas you will see them is used within articles because you can open an article and then your article might have a header where it's sort of the title of the article and it's introducing the article. Then you have the main content of the article. So the article itself, don't put the main tag in there. You have main tags on the outside probably, but it's the article itself. And then you might have a footer down at the bottom. You do not need to have a header and footer within an article. You might only have one or the other and not both. And if you're not too sure about it, don't stress too much about it. Header at the top of your page, main inside your main, you might have an article or a few articles and then a footer down at the bottom. And the very last thing that we're going to talk about is the div, which is your division, which is used all over the place. And divs is we will call them is something that we use to divide our content up into ways to organize it. And the only real reason you'd be using it is when you're going to be using CSS to create a layout, because this is how we sort of group different things. So when we need two things to go next to each other, we sort of have to wrap them up in a little box to put them next to each other. And in general, I'm going to encourage you to go and use one of those other more layout related things before you use a div. So maybe an article makes more sense or maybe a section makes more sense. You want to use something that actually has meaning to it. But if, you know, I just need to create these boxes because I need to organize content in a very specific way when I'm making my layout, then the div is completely fine. And that's what we use it for. And we use them all over the place. And you'll definitely see them used a lot. But if we can use the correct semantic tag, we want to use the correct one. But you might say like, no, this really isn't a section. I have a section of content. I just need three columns in there or whatever it is, then just put each one of those in a div. There's no problem at all. So once again, that cheat sheet is linked down below if you'd like to get that. And if you really are just learning HTML and you're very early on in your journey, you might also really enjoy this video that's right here where I look at five important HTML concepts, specifically for beginners that are just getting started with it. And with that, I would really like to thank my enablers of awesome TT LLD Andrew James and Rico Michael Simon Tim and Johnny as well as all my other patrons over on Patreon for their monthly support. And of course, until next time, don't forget to make your corner the internet just a little bit more awesome.