Two judges in Minnesota's 9th Judicial District have announced their retirements. The Honorable David F. Harrington, who is chambered at Walker and Cass County, and the Honorable David J. Tenich, who is chambered in Brainerd and Crowland County, have both given notice they are planning on retiring. Judge Tenich is a former DFL State Senator who represented District 4 from 1996 through 1999. The Commission on Judicial Selection announced today that the application process to fill those upcoming vacancies is now open. If you enjoyed this segment of Lakeland News, please consider making a tax-deductible contribution to Lakeland PBS.


I have some water with me in this container and some sugar. So what I'll do is I'll take some sugar, I'll pour it, I'll pour it into this container and then start mixing the sugar in the water. So as I try and dissolve it, I wanted to think about when the sugar fully dissolves in the water, what kind of a change will that be? Will that be a physical change or a chemical change? So as you see sugar is completely dissolved, we can't see any more sugar in the container. The turns out this sugar is still sugar and the water is still water. But they have mixed together to form a solution. This change is physical because the sugar molecules have not changed. They have just dissolved evenly in the water. You can separate the sugar from the water if you try and boil this water. All the water will be boiled and all that will be left behind will be sugar. So dissolving sugar in water is really a physical change because you can get back the original sugar by physical means. For example, if you try and boil this solution. Now let's take some more sugar in the spoon and then let's try and heat it. What kind of a change do you think this is? Is this a physical change or a chemical change? Well, turns out burning sugar is an example of a chemical change. When sugar is heated to a high temperature, it breaks down into different substances. For example, carbon dioxide, water vapor and it changes its color and smell. This changes chemical because the sugar molecules have broken down into different molecules and it cannot be reversed by simple physical means like heating, warming or cooling. So once the sugar has been burnt, it is a new substance and cannot be turned back into the original sugar and therefore it's a chemical change.


Hey guys Chris Burch here entrepreneur strategies create DC. We're behind the scenes again in the print shop and Today, I just wanted to quickly go over printing banners What you're gonna need is obviously some form of a printer. We're actually using our smaller machine today. It's a rolling 300 It's a UV printer print bright bright high quality vibrant graphics This is going to be a banner that's actually going in a parade tomorrow So we're printing four of these today, but what you're going to need to get started is you're going to need the printer a Roll of batter material. We're using on this one. We're using a 13 ounce vinyl banner You're going to need a straight razor You're going to need some banner tape and then you're going to need a grommet punch So if you're ready to go behind the scenes with me, let's get started right now And then it'll measure the sheet the the the width of the sheet and then we can take over here Our file is loaded on the computer We have it all set and now what it's going to do is going to rip the job and Then it'll start running This banner. All right, so we have our full-color banner Fully printed Figure that wouldn't bore you guys with the fact of watching the print. It took about Probably about 15 minutes to run this. It's a two foot by six foot banner And then what we're going to have to do is you see you have all this extra white space So we're going to take our straight razor And we're just going to trim a border and if you see here I'm purposely leaving About a one inch border And I'm going to show you why let me trim the other three sides You've got one more cut down this last side and You could use something something like a t-square Just to make it straight but For these purposes is not necessary So now you see we have an even white border all the way around the whole banner And the reason why Is we're gonna flip it over So now we're on the back side And if you've noticed on banners, you might not have noticed there's a usually a seam along the edge So we're going to take as our banner tape. This is just a double-sided Adhesive tape And we're going to lay that down the edge Okay I'm going to trim that off and then you're going to peel The double-sided tape you're going to peel the backing off of the tape So then all you'll be left with is the adhesive and the reason you are going to need this Layer so now you have that adhesive layer and all we're going to do is just fold it over and That creates a seam All right, so now you have for one one you have the durability Now that this banner is going to be double layered around the edge So I'm going to go ahead and tape the all the three sides and then I'll show you how we do the grommet punch All right, so we're all taped up We got seams around all four sides and then with the last step to complete the banner is We're going to put in these little metal grommets And for that we have a manual grommet punch You place the grommet down Put the banner in between And then you can just punch down So now you have a full-color banner We're going to put these grommets all the way around all sides and we're actually because it's the length of this banner We're actually going to put them in the middle as well so they can hang it on this float in this This parade that they're doing tomorrow So as simple as that seems there are multiple steps Do me a favor if this was entertaining it brought value to learning how banners are produced and how we produce them here Do me a favor smash that like button give me a thumbs up Love to hear from you guys post in the comments below and I will see you soon


So I dropped the video on my YouTube channel title, what I would do if I was a 15 year old artist. And this is for all of the young artists out there who feel like they don't know what they need to do. First, upload your music to free streaming sites like I Your Mac, YouTube, and SoundCloud. This will give you practice putting your music out on DSPs and allow you to scratch that itch and put your music out. Next, you want to find communities where you can share your music and get honest feedback. This could be a Discord group, a Reddit group, a Facebook group, whatever that looks like. At this stage, you should be trying to get over the fear of sharing your music with people and being criticized. Next, figure out how to get some money. This is the music business, but things cost from putting your music on Spotify to get and cover art made for your music.  Next, start putting your music on all of the major streaming platforms. Spotify, Apple, Tidal, you know the list. It's around this period that you should also start to consider your brand and what that may look like. Next, you'll want to start to develop your content strategy, all while figuring out everything you can about the music business and music marketing.


So there's this misconception out there that there's no good men out there and all men are commitment phobic. And I'm just getting tired and tired and tired of hearing that rhetoric from women.  Everybody is gonna ghost you and disappear. And I wanna say that that's bunk. I mean, it is bunk.  So I can understand why many of you feel like there's no good men out there that all men are commitment phobic. I get it. And it is important to address this. I don't wanna minimize this. And even though we're gonna be talking and knowing that about the secret thoughts a guy has when he's into you, the reason why I'm sharing this content today is because there are really good men out there that genuinely want to be in a relationship. The challenge is sifting through the weeds to find the, I was gonna say flower, but flower represents more of a feminine type of thing. So you're sifting through the weeds to find the tree, to find that strong sturdy tree out there. And part of the problem is you ladies yourselves because as much as men are delusional and I'm gonna say this, men are rather delusional. And what I mean by delusional, human beings are delusional in the fact that they overrate themselves on what they can actually attract into their lives. They overrate themselves. In other words, typically men want that, I'm not saying all men, but it is traditionally, this is the case, men want that young, hard, hot body and women want that financially successful, gorgeous, attractive man. And I know many of you say, no, I don't want that. No, I don't want that. But the reality is is I highly doubt many of you would date men who live in their basement with their parents or with a parent, okay? And you're probably thinking, well, how could a 45, 50 year old man believe me that happens frequently more often than not, especially after the pandemic and a lot of the financial distress we've dealt with in the last 20 to 30 years. So I'm here to say that we're coming back to the fact that yes, we are experiencing a lot of dysfunctionality within human beings themselves. Ladies, you are just as equally as dysfunctional as men. And just because you may have a propensity to want commitment, maybe more so than men, pointing the finger at men is not going to help you attract those men who genuinely want to be in a healthy, happy relationship. And if you follow my work, and I'm gonna go into a little bit more detail today about something I've talked about. But if you've been following my work, you know about the three types of people who are actively dating today. And I wanna show you this chart. And you can see right up here, it says users, spenders and growers. And roughly 20, by the way, this is not a fact. This is merely an opinion. Roughly about 20% are users, 20% are growers and builders. Most everyone's a spender. Now users are those people in it for short-term game. Those are the guys who love bomb. Those are the players. Those are the women who are gold diggers. Those are women who are entitled. Actually gold diggers entitled might be in it for a longer game, but they only care about themselves. Those are the users. And the growers and builders, they seek long-term commitment. They're emotionally grown up and with good relationship skills, and they have their act together. And the challenge with the spenders, and as you can see, the arrows are going either direction. The spenders, they seek connection. They seek companionship. They seek coupling and or sex, but they have no real direction. They're uncertain. They're fearful. Usually they have a dysfunctional life, and that's what I wanna talk about today. What I mean by dysfunctional life is their emotional maturity, their level of emotional maturity, and they oftentimes lack relationship, good relationship skills. And by the way, this isn't just men. This is women as well.  And the delusion I was talking about earlier is everybody thinks they're the grower and the builder. Everybody thinks they're good at relationship skills. And do you know why I know this? Why is it that 50% of divorces end up in, or excuse me, marriages end up in divorce. Second marriage is that 65%, and third marriage is it's 75%. There's gotta be a reason why this happens. Usually, and I don't care what age a person is, it's because there's a level of dysfunctionality. Now you might be thinking, well, Jonathan, this is just too much work. I can't deal with this. Well, it starts by doing the inner work. It starts by healing yourself. If you're not familiar with the book, I talk about this frequently, the Hoffman process, the Hoffman process. By the way, there's a link to get all the books I recommend. The Hoffman process is a deep dive into healing childhood wounds and traumas, and adult traumas that cause the negative patterns and beliefs in your life that cause you to choose the wrong person. And I don't care how much you think you have your act together, everybody needs a level of healing. In fact, and this is why it's important to also seek out men who are doing the work. By the way, I will tell you this. I have a client who I've worked with for gosh, going on four years now. They're finally, excuse me, three years. They're finally moving in together. And this was a man she met in his mid 50s when she was mid 50s, he's mid 50s. By the way, both 55 when they met, I think they're only a few months apart. So this whole thing, men want younger. I mean, you know, they're the same age. He had been going to a therapist for years, not because he needed anything special. He wanted somebody to talk to on a regular basis. There are men out there that do the work. There are men out there that want significant relationship. There are men out there that want to be with you. The real trick is learning how to find them. That's where my coaching comes in. You know, my area of expertise as a coach is teach you how to ask the questions based on your personality to determine if a person is right for you. By the way, there's a link below to schedule a call with me because that's how I can help you as a coach. And I can also help you if you're with the wrong guy right now. Okay, so most importantly, coming back to those spenders, the big, the reason why I want to spend a minute or two extra on the spenders is the spenders are the men who want companions, or people who want companionship, connection, and sex. But maybe they don't have their act together. Maybe they're going through a difficult divorce. Maybe they were going through a breakup. Maybe they've got health issues. Maybe they've got a contentious ex. Maybe they've got issues with their children. There's a level of drama going on in their life. And when someone has drama in their life, the ground underneath them doesn't feel solid to actually lean into a healthy, happy relationship. So this is all predicated on your ability to choose better. But some of you might be going, Jonathan, it's so hard to meet people. Listen, folks, I hate to break it to you. But as a matter of fact, I talked to a woman today. I go, how easily do you meet men?  I go, really? When was the last time you had a first date? She goes, a year ago. I'm like, you said you meet men easily, but it's been a year since you've had a first date. Well, yeah, I have to go outside of the house to meet people.   There's no doubt about it. But if you're not on the dating apps, then it requires to get out of the house to be seen by single eligible people. And you can, this is where the delusion is.   And by the way, I'm talking about men and women alike. By the way, guys are just as delusional in what amazes me here in the United States. We have roughly a 55 million, there's 55 million people who are single. Think about that, 55 million people are single.  And if you're gonna make the effort, then you better be prepared. That's why I recommend reading books like my book called What the Heck Is Self Love Anyway? A Journey of Personal Development, Self-Alfit Spiritual Work, by the way, a link below to get my book. This is a book to help prepare you so you can be better at picking people, but also because you're aligned in your sovereignty, your self-worth, your self-esteem and your self-confidence. Okay, here's the bottom line. You put yourself out there and you've got a date. Yay, you've got a date. And he asked you out on a second date and he asked you out on a third date. And now you might be wondering, what is he thinking? And how can I tell if he genuinely wants a relationship with me? Well, let's talk about those seven secret thoughts a man has when he's into you. And these are things men don't share. But I can tell you, I have a dear friend who's, listen, this past weekend I was speaking to him, he's my age, he's in his mid-50s and he was sharing with me, he would consider getting remarried. So this delusion that men don't want it, they do want it when they're with the right person. And we were talking about, I was asking him a few questions, what he thought of in the beginning. And he even shared some of the things that I'm sharing today, but this is what I've outlined in previous videos and I'm going to share this again.  People who are not into you or lose interest stop looking you directly in the eye, directly in the eye. You kind of do this. In fact, I once went on a first date with a woman and I swear to God, this is the way where she was talking to me. So she's right there, but we'll pretend this is me and this is her. She was like, hi, Jonathan, how are you doing? I hope you're having a good day. Did you have a good day? I hope you had a good day. I mean, literally she gave me no eye contact.  And by the way, she talked to the person next to her, direct eye contact. So when a guy gives you direct eye contact, that's a good sign. Number two, either wants to be friends with you on social media or follow you on social media. And I know this is a bit corny, but guys, at least these are for the guys who are into social media. Not everybody's into social media. I'm into social media. So this one is one of my own that when I like someone, I wanna follow them on social media. I wanna look at their past posts and get a sense of their lifestyle. So that's a good sign a guy might be into you. Number three, this is probably the most important one. He wants to be your boyfriend. He thinks about it ahead before he asks. He's intentional. That's what's going on in his head. He goes, I met someone special and I want her to be my girlfriend. Men want to stake a claim with the people that they like. So men definitely, and I'm gonna talk about this in the next one, they're territorial. So when they like someone, they want you to be their girlfriend. They wanna be your boyfriend. The problem is, again, those dysfunctional men who don't have the ground underneath them solid, they want your companionship, they want your connection, they want your sex, but they don't want anything solid. This is, by the way, the easiest way to avoid the spenders and spenders mean they just spend time with you.   It's just a matter of degrees. So the fourth thing, and I talked about this a second ago, he gets a little bit territorial. He gets a little jealous if you're talking to guys. I get that way.  I mean, I'm certainly, my philosophy is when I remember I dated this drop dead gorgeous woman and lots of men would talk to her. And my feeling was, well, I'm the one going home to having sex with her. So I felt good about myself, but I still was feeling a little jealous. It's very natural because as I said earlier, men can be territorial. Number five, he's thinking, can she fit into my life and vice versa? And this starts on the very first date. This is why a lot of guys do love bomb. It's because they're futurizing because they're trying to determine if you could fit into their life. So that's not an uncommon thing. The love bombing is just the overboard version of it and it's usually driven by the sexual piece. So that's how you differentiate love bombing because love bombing is lust and limerence.  But guys like me, we'll futurize. I call it futurizing. We try it on for size. We're trying to figure out, can she fit into my life? And we may say it out loud. We may say it out loud. Number six, he'll make an effort for you.   Listen, even an alpha guy who really likes you, he will make consistent, consistent effort. Because he genuinely cares about you. He's into you. And number seven, this is one from my own playbook. So when I really like someone, so when I really like someone, I mean, this is what's going on. I gotta clean my place. I gotta clean my place. And I have to make sure the toilet seat is down. Let me repeat that. I've gotta clean my place and make, that's, I mean, when I like someone, this place is spick and span. Because when I'm living on my own, I'm not necessarily perfectly clean. But when I have someone I'm into, I keep the place clean. And I always make sure that the toilet seat is, excuse me, down versus I said up, but I'm gonna say down. That's a sign and he's thinking about it. That's a true sign he's into you when that toilet seat is down. And by the way, I've made a mistake and once the two in the morning girl I was in a relationship with fell in. Oh, I felt so terrible. I could hear her. Cause I got up in the middle of the night to go take a leak and just totally Jonesed. Okay. So these are the seven secret thoughts a guy thinks about when he's into you, you may not know this out loud, but his actions would certainly demonstrate this. And as I said before, while there are a lot of users and time wasters out there, there are a lot of good men. There are a lot of good men. And this is why I want you to be intentional in the dating process. And before that penis ever gets to go inside the vagina, read chapter one of the book, eight dates, read it together. So you can read this book so you can get the mechanics to a healthy, happy relationship.  By all the books I recommend because the reality is, as ladies, you all think a lot of you think you're so good at this, you're not, you're terrible at it. And then you wonder what's the definition of insanity? Doing the same things over and over again, expecting different results. This is why I scream at the top of my lungs to recommend a better way to do it. So my hope is you're finding value. Did this resonate with you? If you did hit that like button, please share this video. Please subscribe to my channel. I'd really appreciate it. Check out the links below to all the different offers I have. All right, we're gonna do a short Q&A right now. So if you have a question for me, write the word question and post the question thereafter or purchase a Super Sticker Super Chat. All the monies from the Super Sticker Super Chat goes to a scholarship fund in the name of my son, Connor Asley. That's my son who passed away. That's a picture of him there and there. I love this one. And that's him right there with his brother. I love that one. And all the monies, there's a little dollar sign there. Or you can hit the super thanks and all the monies go to a scholarship fund to defray the cost of personal development. Also donate to the Hoffman process inside seminars just to name a few. So we've already given away a couple thousand dollars from all the Super Stickers. So we're time to take questions. And again, if you're listening to the audio portion of the replay, you won't be able to see any of this. Okay, I saw a question come in. Nicole writes, question. A man should know they want to marry you within four months of dating, right?  No.  By the way, I just was talking to a therapist who said that during COVID, a lot of men to save money have been shacking up with the women or having women shack up with them. I'm using the word shacking up but move in together. And they're finding out that they're in dysfunctional relationships. She said 70% of her practice is men. Now, I'd say within four months he knows if you're a viable candidate it begins to start to, if he wants to get married only if he wants to get remarried. If he doesn't want to get remarried that may take a lot longer. So for those men who are growers and builders yeah, we would know within, I would say six months to a year. Six months to a year is a good timeline for a guy to know is this woman long-term commitment material or not? So, but it's rarely is it ever four months? And if it is, there are probably guys who have only been married, never been married before. All right. Denise says, let's do a singles retreat. I'm in all in favor of that. All right, let's go swimming. Do you have a question for me? All right, looks like we have a bashful group. Oh, Dale writes, okay, question. Aren't some of the most dysfunctional peeps some of the, wait, let me read a bit. Aren't some of the most dysfunctional peeps some of the most well-adjusted? This is dysfunctional society, right? Well, here's the thing. Spent, okay, remember I said spend the dysfunctional people, the spenders, okay? Typically they don't, the ones who aren't able to be in a fully committed relationship are usually when the ground underneath them isn't solid. Now, if the ground is underneath them isn't solid then you have to determine how emotionally mature is this person to be in relationship? And if you're not familiar with my emotional maturity relationship skills chart, I want you to see this is not a fact, it's an opinion. Roughly 20% of the population has clinical issues. I'm talking about borderline. I'm talking about narcissism. I'm talking about drug abuse or addictions and whatnot. Okay, and while I say 20% are healthy I'm being rather generous. Most humans have dysfunctional relationship skills. This is human beings, not women or men. This isn't men, this is human beings. This is why, it's funny, I talked about this recently. I saw a commercial where these three young women are talking and they're asking about this woman's date she had and she goes, oh my God, the guy checked all the boxes. He was tall, dark, handsome, good job, good relationship with his parents, blah, blah, blah. And she says, why aren't you interstituting again? She goes, when I asked him about therapy, he goes, what's that? I thought that was kind of a funny commercial. But ultimately, the point she's trying to make is if you want to grow as a human being then it's important to invest in personal development, self-help or spiritual work to be able to build the roots to trust most likely for a long-term loving relationship. So anyway, coming back to your question, yes, could they be well adjusted? Most often in midlife, they've got some level of chaos, some level of chaos, an angry ex-spouse, issues going on at work, health issues, maybe child issues, there's usually something going on. That's their escape clause because they can have all the sex with you and go, oh, I'm dealing with an ex, I gotta take a break from this relationship. Be careful of those exit clauses, Dale. Hope that helps. One day, I don't know how to pronounce your name, question. Dating a guy for four months, he's great in every way but recently freaked out when he was referred to me as boyfriend, is that a red flag?  Folks, if you're not familiar with my relatively new dating vow, I want you to understand something, folks. There's an old saying, women are the gatekeepers of sex and men are the gatekeepers of commitment. Calling each other boyfriend and girlfriend is a level of commitment. So I have something called the dating vow and it goes like this, you both recite this to each other, you both recite this to each other and it goes like this. I agree to explore the process of getting to know you with the intent to declare something serious in the next three to six months, you each say this. I agree to be monogamous sexually while we have regular sex together. I agree to not actively seek to meet others while we're in the dating process including taking my dating profile down and also calling each other boyfriend and girlfriend. I agree to speak up if this isn't working for me versus pulling back, ghosting or disappearing. And lastly, I agree to invest regular time together in the process of getting to know you which looks like calling each other boyfriend and girlfriend. We see each other doing social activities, hobbies, mutual interest, spending time with family and friends. Now ladies, 90% of guys will bail on this but let me tell you, they're usually the users or the spenders that are bailing or the emotionally dysfunctional people or the ones who have clinical issues. So does it really matter that you lost the wrong guy? Don't you wanna set some standard for yourself? I'm gonna share with you.  Now if you're not having sex, then it's not a red flag. Sounds like you guys have a casual relationship or just hanging, sounds like you're just friends. You're both in the friend zone if you're not having sex but that's just my opinion on it. I hope that helps. Jennifer says, do you think age difference matter in dating and marriages? Sometimes yes, sometimes no. I know I have a friend who's 22 years younger than his wife. She's 72, he's fit, no. Yeah, she's 72, he's 54. So maybe 18 year different, excuse me. I know I have a friend who's in his late fifties and he's in a relationship about to marry a woman who's in her half his age. So no, I think anything is possible. Hope that helps. All right, Kimberly writes, what kind of date should be an ideal type of date for you? What do you do on your first dates? My typical first, if you're asking me personally, my typical first date is go out for drinks. Go out for drinks, that's usually it. I like to go for a walk sometimes but I love where I live. There's a beautiful marina and having drinks there is a great way to connect. Sometimes it's a coffee date. Sometimes I don't like to do anything fancy until we've felt like we've gone out a couple of times. So that's usually my MO. Thank you for asking, Kimberly. Michelle says, how do you see through the love bombing and sincerity? How do you see through it? Versus, if it's, well, first off, love bombing is just typically a person's drive to get you in their pants. It's, listen, a well-adjusted person doesn't, might get excited but they don't go over the top and ultimately you have to learn to trust your intuition. Your intuition is speaking to you frequently. If it feels off, it usually is. The problem is many women are just as narcissistic as men and you're needing validation. I need you to love me so I can feel good about myself. I need you to love me so I can feel good about myself. I need you to love me so I can feel good about myself and I'm getting all this love bombing that's so wonderful because I'm getting all this special attention, I feel so special all of a sudden. And then he's gone. When you feel good about yourself, you don't need love bombing and you'll actually be detested by love bombing. That's just my, that's my perception anyway. Take it for what it's worth. All right, thank you, Michelle. Have you ever read John Bradshaw in The Family? No, I have not, thank you, Dale. Suzanne writes, why is a guy that's so agreeable and wants a long-term relationship, respectful wants to jump start into me. He doesn't appeal sexually to me. Am I superficially distrustful or too fussy? Yes. You know, I can't speak for you. I can't speak for this. It's, I do believe our egoic side. Well, you know why? I'm gonna tell you why. He doesn't represent something familiar to us. By the way, most humans choose dysfunctional relationship because they're trying to heal a childhood wound from their parents. So I highly recommend reading this book, Suzanne, Getting the Love You Want by Harvelle Hendricks and Helen Hunt. So I want you to specifically pay attention to the Amago. So pages, I want you to read pages 33 through 42, page 68, page 153 and 156. If you read this book, there's a link below, you'll understand why you're not choosing a great guy. It's because you most likely need a dysfunctional man because you haven't healed within yourself most likely. Read this book, change your life. Everybody read this book, change your life. All right, this is gonna be our last question for the evening. Oops, let's go here. Merita says, Jonathan, I find sexual innuendos during first dates off-putting and not classy. The word porn should not be part of a first date conversation. Am I overreacting? What's your take? No, you're not overreacting. I can respect that that doesn't feel good for you.  So I wouldn't do it with someone I wasn't gonna go along with it. But for the most part, if that's not your personality and they say that, probably a red flag means ask more questions. And a red flag might simply mean you want to slow down the process and ask yourself, really ladies, I want you to ask yourself after a first date, is this person worth seeing again? That's simply the only question you ask yourself. Is this person, let me reframe that. Do I want to see this person again? And then go from there. And then the next date, do I want to see this person again? And the next date, do I want to see this person again? Make it very simple on yourself. So I'm gonna share a prayer with you so we can invite love into our lives so we can invite that man who has these seven secret thoughts and that prayer is simply this. God, universe, spirit. I invite love into my life. I invite healing into my life. I invite joy, abundance and happiness in my life. And I invite in a partner and I am gonna allow that partner in my life, somewhere where we have amazing chemistry together where we can't rip each other's clothes off. And we have a great level of communication where we're with each other and we can go banter for hours and hours on end. And our lifestyles are compatible with one another. And we have mutual emotional maturity with one another and we will build the deep roots of trust doing social activities, hobbies, mutual interests, spending time with family and friends, traveling together, teamwork, building skills both in our personal professional life that leads to us either moving into getting married together. I invite this into my life, God, universe, spirit. And that's my invitation for all of you to invite that into your life. Are you in agreement with that? Give me an amen. Listen, we're gonna do a short live stream today, a short Q and A. Please purchase a super sticker, super chat before you wrap up just to show me that this mattered to you. This resonated with you. Also, you can purchase a super thanks if you're watching the replay. All right, we're gonna wrap up today as I always do.  I'm gonna reach into the camera and give you a hug of love if that's okay. I'm asking you to turn to someone, a pet, a teddy bear pillow and give Iter them a hug of love because hugs are a great source of love. And let's face it, we could all use more love in our lives. I wanna thank Michelle and Susan and Christie and Joan and ABD and Kimberly and Warrior Banners and Grace and Maria and Marita, Laura, Nicole, Hailey, Lucy and Marie, Laura, Mr. C. Everyone, thank you so much. Wishing you a fab evening. Bye now. Hey, other.


A brief essay on human culture was written by Fabled Teflink. You can find it on the scp wiki at https://scp-wiki.wiki.com, forward slash a-brief-sa-on-human-culture. You'll find a link in the description below. I often walk down the halls of my site and hear various people discussing how familiar some of the anomalies in our care are creatures and beasts that spark the imagination, reminding people of stories they heard growing up. It's true, honestly, and not insignificant number of anomalies in the foundation care are creatures and concepts that are familiar to the general public, as they have a place in human legends and folklore. The question that normally follows is why? Why is the public allowed to effectively know that these creatures exist? These are anomalies, things that go against the natural order of the world, and by all means, should be hidden away and never heard from again. The foundation has the means to effectively wipe all knowledge of these entities away from the general public subconscious, so why doesn't this happen? Webster's defines culture as the customs, arts, social institutions, and achievements of a particular nation of people or other social group. Culture, though, is who we are as a people, and it's unique to every single person. It's informed by your lived experiences, where you come from, how you were raised, everything you've gone through in life helps to shape your personal culture and identity, and in turn, your culture and identity shapes that of others around you. Culture is how we as humans create a unified understanding of how we interact with one another, and how the world interacts with us. It creates our beliefs, our customs, our understanding, and so on. So what does this have to do with anomalies? Well, once again, if you look through our files, you'll see familiar names and descriptions, creatures iconic to various cultures from around the world, from the South American Yate Vio to Chinese dragons. Truly, nearly every mythical beast either is real or was real at some point. Our stories come from a place of truth, so why does the general public get to keep these stories? Culture is a multifaceted thing. Each of these creatures plays a role in our culture. It's well known that humans have been telling stories since they first learned how to communicate with each other. Basing these tales on the world around them, the crackling thunder was a furious god, and the beast with red eyes was something to be feared. These stories created understanding, and then they became tales of morality and lessons to internalize. To answer the question I keep hearing, why don't we just eliminate them from the public's memory? The answer is simple.  We would lose the stories that shaped us as people, and help us understand the world around us. Our culture, our identity would be fundamentally changed. These stories have their place in inspiring awe and fear in us, and as a result there's no way to remove them without fundamentally changing what it means to be human. Besides, isn't it nice for the world to feel a little bit more magical? Dr. Farron Karoway. Thank you very much for watching. If you enjoyed the video, hit the subscribe button. You can probably hear in my voice that it's a little bit rough, or maybe you can't. I don't know, but I'm having a little bit of a cough this week, but I think it's getting better, and I think next week I should be able to get back to making videos on the screen again. I actually kind of enjoy doing some of these readings every once in a while. Anytime I can find an essay or a first person perspective tale, it's fun to get it out of the way. I like doing these. But if you'd like to support the channel even further, head on over to patreon.com forward slash deesmerian and pledge at any level like everybody here on the screen already has, including MC Kejmal, who has pledged it $50. It's nice to know that I'm not alone out here, and I will see you all again on Tuesday.


Ladies and gentlemen, it is my very great pleasure to welcome you all to the University of Auckland Aotearoa Māori Business Leaders Awards. An event featuring more glitter than a Marae driveway made up of Smash Power Shell. Tonight's main purpose is to celebrate Māori excellence and success in business. We do so in order that it may serve as an inspiration to others and we have an exceptional group of Māori leaders to honour tonight. Being Māori is our secret sauce. It's our superpower and I feel incredibly lucky to be Māori every day. Through courage, through conviction, through perseverance, through sacrifice, we can shape the world to be absolutely whatever we want it to be. So I say be bold, be tenacious, be selfless and continue to be driven.


For the CircuitPython Parsec today, I wanted to show you how you can use a step size inside of a range when you're looping through things in order to change the increments you use in the loop. Often when we use the 4I in range and then some number, the default is to increment one value at a time. This is a really common way to sweep a value up. Say you're bringing up the PWM value of an LED, you can loop through it with 4I in range and then a number. But you can also use 4I in range, start value, stop value, and size value. What that does is allow you to say, here's where I'm going to start, here's where I'm going to stop, so that's the range. Then the actual size of the steps, so it'd be like plus I equals I plus 2, I equals I plus 2, I equals I plus 2, so it keeps incrementing up by 2s or 3s or 4s or whatever you need. I have some numbers printing out when I press a button on the circuit playground express. When I press the B button, you'll see it's going to increase that step value, so now I'm increasing by 2 every time through. Now I'm going by 3, so 0, 3, 6, 9, 12, and so on. Now I'll go up by 4s. The way to do this is right here in the main loop. When I press the button, one of the things that's happening here is for I in range, start value, I have that set to 0, stop value, I have that set to 41, and the step value or size of the steps is 1 to begin with, but every time I press this button, I'm increasing the number there, so I'm going through different values. I can also decrease that using the other button. That is how you can use the range command with start, stop, and step in order to change how you loop through a set of values. That is your circuit python parsec.


Good morning, everyone, Basel Chapman here on the 30th of September, last day of the month. You can see the candle on the right for the Dow, a leg seat to the downside. You can see the arch formation taking out the left side, low of Jude in the weekly chart. And we're holding the low of four days ago, so far, 28, 9, 5, 8. In the Dow, Dow's up 23, attempting some kind of a little H pattern U-turn. We'll see if that can happen. The histogram of the MACD has started to improve. That's the first sign. The stochastic is now over 10%. That's good. S&P. S&P at this moment is trading up 15. Now, that's a really good sign. It didn't take out yesterday's low, and it's moving higher. But if by Tuesday of this week, we can see a move into the 30, it's a 36.56 up 16, into the 36.90s, especially the 3700s. That'll be the first time that we're seeing the overboard signals where the technicals start to improve with price. And that's going to be important. Otherwise, taking out the lows of yesterday would be very, very negative. The QQQ, nice move up 2.19 at 274. It's got a lot of work to do. The technicals are improving very slowly, but 278 is the big resistance to take out next week. IWM, the Russell 2000, much better move actually. It's a 2.4 up, up 1.43% at 163.39. Let's see what happens next week. Looking at the gold. Gold right now is trading up 12. This is a very good move in gold. The technicals have improved, but the 9 is still far from closing over the 14. If you're looking at the dollar, the dollar actually is up 19 ticks. It's just sitting on the 14-period moving average at a peak D. And that just says, yes, there's a really good chance of the dollars and have a little bit of a digestive phase right here. If you're looking at the TLT, that is bonds down again. It's down 21 ticks and one or three yields moving up again. And if you're looking at Crude Oil, I didn't do much of that in my Tiger Technicians Hour, an hour at this past hour when I did my show. But it's down 75 cents. Crude Oil is not yet really moving to the upside. So the commodities in general have been kind of weak. So we're looking at a very important two days coming up today. Finish today, see what happens with the chart. But how, if there is a strong move, it has to be start early next week. By Tuesday, Wednesday, we need to move much higher. Taking out the lows that we made this week would be very negative. Have a wonderful weekend. Check out my opening call. And stay tuned. You've got a Steve Rhodes recording show earlier today. It'll be coming up. Great program today. See you on Monday. And I hope those of you in Florida are really surviving well. And we are fortunate. Have a great week.


doing research. So our first speaker is Professor Hyun. Please welcome him. Okay, good morning, everyone. So, thanks for introduction. And actually, our chair, I mean, phrase everything I want to say at the beginning. So last week, we had some school basically introducing the basic concept of the statistical physics, but there's no, basically, there's no, I mean, lecture on the life science component. There was nothing about the life science component. So maybe next two days, we can actually hear something about the life science and application of the statistical physics in the life science. And maybe this is the first example. So molecular chaperone, what is a molecular chaperone? Okay, so molecular chaperones are cellular machinery that actually consume large amount of ATP to help protein or RNA to fold into their functionally-competent state, native state. And actually, the history of the molecular chaperone is actually very long, and it feels kind of dominated by the experimentalists, the biochemists or molecular biologists who are very well familiar with the equilibrium chemical thermodynamics. And the, oh, sorry. If you look at the chaperone for protein and RNA, it looks very different, but there are some underlying principles for their action can be described as some unified framework. And I want to, maybe I can actually emphasize the non-equilibrium nature of a chaperone-assisted folding in this talk. Okay. So let me start with the protein and protein folding and protein chaperone. Protein folding is basically defined by the, so three-dimensional sequence, one-dimensional sequence, which includes a three-dimensional, native structure of protein. And for a given sequence, well, if, as long as the sequence is already designed, evolutionarily, it actually defines a unique native state. And the folding passway is kind of smooth, folding landscape is smooth, smooth such that as long as you are sitting in some right condition, let's say low temperature, then you can get to the native states very easily via smooth landscape. Okay. Hydrophobic patches are in the native states, hydrophobic patches in the sequestered, sequestered in the interior of the structure, such that there's a kind of structures that are very compact and that there are low hydrophobic patches exposed to the exterior of the structure, that, so that you don't need to worry about some sort of aggregation kind of stuff. Okay. Or however, not all the protein can be folded nicely. I mean, at least 30% of the entire protein has some problem of the spontaneous folding. It doesn't actually fold into the right structure. So in that case, you can actually imagine the very long in the landscape like this. So in this kind of very long in the landscape, only a fraction of the population can fold into the native state and then rest over them to trap in the kinetic trap. So one minus five over them trapped in the computing base of attraction. So you can actually think about some folding passway in many different parallel passways. So in this kind of scenario, folding kinetic can be described as this kind of kinetic scheme. And the five, which are called the partition factor is defined by the two kinetic rate, cholesterol rate to the native and the disordered state. So this process going directly from the disordered state to the native state is very slow because of a large free energy barrier that actually doesn't occurs on a biological level on the time scale. So essentially, folding can occur in this direct passway, but this fraction is very small. And then most of them are trapped in the disordered state and trapped in such a long time that you don't see the transition from here to here. So then the problem is, this is a kinetic expression for the entire folding as long as the KS is kind of not too small then you can actually discover the native states, the kinetic rate to the native state can be described by this kind of a kinetic rate. However, when the KS is very, very small then on a biological level on the time scale only you can actually see the, only fraction of the population is being folded into the native states, okay? So we have the problem with this hydrophobic patches. So in the cellular condition, when this kind of disordered state is like a dominant, then you need to worry about the aggregation of this disordered, among the disordered proteins. So that caused a lot of problems for the cellular function. So cells that are equipped with this kind of protein chaparone, one of the best studies of protein chaparone is grow here. And it looks like some sort of like a cylinder, better like a structure. It actually have a cavity interior of the structure. And then we actually involve this misfolded, it recognize the misfolded structure, hydrophobic patches of the misfolded structure and then it endures it inside the cavity and then spit it out. So, and you need to, I wanna emphasize that this machine actually consume a lot of energy from the ATP and they actually undergoes large conformational change. This large conformational change is coupled with this, coupled to be the protein and that exert a lot of mechanical force to the proteins. So this trapped misfolded protein trapped inside this cavity or maybe it doesn't need to be trapped inside the cavity but whenever this misfolded states or protein in terms of the chaparone, it can feed into the some first, exerted by the chaparone in action and then it unfolded. So what happens in terms of, so when this cycle can be actually, when this cycle is repeated, then a lot of misfolded protein is unfolded into the states, okay? So what happens in terms of the landscape, this is the landscape and the chaparone actually capture the misfolded protein and then unfolded into the HLA, reset the structure into the folding states. So in a sense it actually performs some sort of error correction, okay? So when this process is repeated, iteratively repeated, then what happens is that it actually anneals the system into the native states, native population. So when protein, the chaparone, recognize the misfolded protein and unfolds and unfolds and unfolds and after an iteration, this is a population of unfolded states and then population of native states actually even like this equation, okay? As time goes on, then this is the population of native states actually in the cell environment, population of native states increase like exponentially and the time scale of this exponential increase dictated by this tau zero, which is a cycle of one cycle of chaparone in action divided by the fraction of this partition coefficient of the spontaneous folding, okay? So typically this gloria, this tau zero, the one cycle is about two seconds and for protein like a viscose, this five can be as small as 1%. So the total amount of time needed for folding the entire population into the native states is about the 200 seconds. This is the estimate you can actually make. So shifting our gear to the RNA folding, the story is about the same, but slightly different. I mean, so the story is the same in a sense that the RNA is defined by the four different sequence, G, U, C, A, G, C, A, U and then it actually first collapsed into some secondary structure and then under some proper condition like a magnification ion, it actually is assembled into the three dimensional structure. In this particular RNA is actually, I'm showing the group on internal ribosine, it actually displayed the catalytic activity on the self-possessing catalytic activity. However, this folding, even though we're folding effectively, so folding is very low, is the five only 0.088, so only 8% of the population can fold the native states or spontaneous read, actually read some help from the RNA-SHA pool. Okay? So it's also equipped with this kind of another machinery, this RNA-SHA pool, is the name is actually site 19, it's just one of the best studies of RNA-SHA pool, it was discovered in the early 2000 and it actually showed helicase activity, it online the surface exposes the helices. So whenever some helices is exposed to the surface, they actually, it actually online, it recognize this kind of helice surface exposes the helices, and online this helices into the single-stranded RNA. Okay? This is what happens. So site 19 can recognize, but because of the exposes helices, it's also in the native states. So not only this is native state, but also these foreign states have this kind of exposes helices. So what these people found in their studies that when you add the site 19, it actually unfolded the native states, native RNA as well as misforded RNA. So, and also they found that regardless of the initial population, native and misforded RNA, which is a steady state of value, which is not equal to one. So in the case of Gruyere, if you actually add the protein chaperone, I mean you add the chaperone, entire population of the protein get to the native states after a long time. But in this case, if you wait for a long time, it actually some value, which is not actually zero. And then another point is that if you add more chaperone, native states, population actually decrease. This is what they found, which is kind of weird, but I'll get to this later. I'll try to explain, discuss this observation later. So, in terms of this kind of log in the landscape, what other chaperone does is the following. It actually recognize misforded states and then reset the system into non-forded states, intermediate states. And not only that, it actually also recognize the native states and then reset the native states to the initial condition, but different extents. So it recognize misforded states better, but the kappa is actually, so therefore kappa is smaller than one, is the number is between zero and one. So the iterative value in the kinds of, I write down the equation has to be modified to some other form. This is the equation which I derived from the earlier, but other than chaperone, you have to write down, you have to need to include the effect of kappa in the equation. We need some modification. So we need to generalize this iterative value mechanism by other than chaperone into this way. So let's assume there is some sequence of RNA. It actually fold into the native and misforded states. Chaperone recognize this misforded RNA and then reset the system into non-forded states and then give another chance to fold. Then this population is divided into the folding state and the misforded state again. This is what defines the lawyer, right? But not only that, the chaperone actually recognize the native states as well and then unfold. It actually recognize the expositiveness and then unfold it. So that even the native states are given another chance to unfold. So a fraction of kappa of them are misforded and the one minus kappa of them are left folded. So this is the population of being, not being affected by the chaperone and this population will be given another chance to fold and unfold. So therefore, at the second stage of this iteration, what happens is the population, this is the first stage of the spontaneous folding. This misforded the population and second stage, second iteration, this is the population of misforded states, right? M2. And this top population is actually folded states. So if you repeat this process many, many times, then you can write down this kind of recursion relation and then you can actually solve the equation. So this one coming from this pathway and this one coming from this pathway and then you have to add these two. So you can think about some sort of this kind of logic. Then finally you can get a native population after any iteration of this kind of process, right? So the native is can be obtained by solving this recursion relation and this is an expression, okay? So which is a function of kappa and an iteration and the partition factor phi, okay? So if you plot this partition, that native yield as a function of iteration time or time, then this is the curve which apparently parameterized with respect to kappa. When kappa equals zero, the case is exactly the same as the gray year. So every population, all the population reaches the native states when kappa equals zero. However, when kappa increase, the steady state population is not one but some number which is more than one, okay? So and the steady state population is you can actually obtain by putting this n equal to infinity. So this is the expression, okay? And kappa equals zero, this is the expression for, it's the same expression as gray year. And when kappa equals one, which means that the native states and this for these states is recorded as equally by the Arlene Chaperon, then there's nothing happens, right? I mean, because the Chaperon acting on the native states as well as the, this for these states equally likely. So therefore there's no effect by the Chaperon. So the native yields just remain as phi. So nothing happens when kappa equals one, okay? So, but what is the meaning of this kappa? To get a better idea, we can actually map this entire landscape picture into three state model. Like if you see it, there's a, I mean, air going down and air going up and there was some transition from this for the state native states, potential transition between these two states. Therefore you can actually map this one into the three state kinetic model, reversible kinetic model like this. And this is just a very prototypical example in the non-equivalent statistical dynamics. So you can solve everything. So this magenta arrow is due to the Chaperon action, which increase with the, so because of it burning ATP, the rate of this Chaperon action, it can be increased. So it actually function of ATP. So if you add more and more ATP then this rate increase in a Michaelian type, hyperbolically increase like this, okay? And this three state kinetic model can be solved analytically and you can actually get the expression for the current flowing in this cycle. So this is an expression for current. So in balance of the kinetic rate, actually produce some sort of a non-vanishing current like this. If we kill the other, there's no current in this cycle. I mean, all the I and M and N species will be accumulated via the Boltzmann rate. However, if there's any imbalance, I mean, K, I, M, M, N, and K, N, I and something like that. So I mean, for all the rate and reverse rate, if there's any imbalance, then there will be current. So that's the signature of a non-equilibrium. And also you can actually get the steady state population of the native states or misworded states and even I states. So these are the expression, okay? So, so two, I mean, so these are the, so the expression for this native rate as steady states, as I mentioned, can be obtained by putting this N equal infinity, this the steady state expression for the native states. And you can now compare this expression and this expression in a certain condition. So this entire full expression can be actually compared with this expression if you think about, in our previous scenario, I said that this not, this, I assume that this rate is very small, okay? And then, so what happens is the following, okay? So I mean, if you actually put this expression into this steady state expression, native yield is expressed by this three kinetic rate and the kappa, right? And however, in this expression, if you actually assume this rate is very small, then what you get is the following expression, okay? So you can compare this kappa and this, this kappa can be expressed by this two rate, the ratio of this two rate. This two rate is nothing but a ratio between these two arrow from unfolding rate from native state, unfolding rate of the, before the state by the chaperone, okay? The ratio between the two, okay? So in the absence of a chaperone, KMI and the KMI is actually zero. In this case, you can actually erase these two arrow, the reverse arrow, and then what happens is that, you know, this RNA or protein actually get into these two states, L and L. And then, as long as you can wait for a long time, then these two species will be equilibrated. So this is just a hypothetical situation. So if you wait for a long time, then, in the absence of chaperone, they'll actually get to the equilibrium distribution, okay?  So steady state expression for native states is nothing but equilibrium distribution. It's equilibrium probability for the native states, which is to take about stability difference between the misforded and native states, right? So, in fact, this is the conventional view. I mean, what is the aim of a molecular chaperone? In conventional view, people consider that the biochemists typically consider the molecular chaperone consume ATP-free energy to establish native areas about the potential distribution. This was the conventional view, but the IR view is a little bit different. Molecular chaperone actually consume a lot less and less of ATP-free energy to establish a non-equivalent steady state distribution dictated by this kind of rate, a magnetic rate constant, okay? So therefore, this equilibrium distribution and the steady state distribution should be different, but the confusion was that for the case of Gloria, which has to be studied for the last 30 years, this is a bit hard to actually dissolve this distribution, this distribution, because for the case of Gloria, kappa equals zero. So, when kappa equals zero, as you can see, this steady state distribution for the native state is just a steady state, even the native state is just one, right? And for the case of many proteins, the stability difference between native and misforded states is very large. So, equilibrium distribution is also close to one. So, there's no way to actually distinguish whether this mechanism is due to the equilibrium or non-equilibrium, but at least for the RNA case, this is kind of obvious. I mean, as I showed you, RNA, native state is not, the probability is not wise, it's just a non-zero value, right? So, apparently, we actually try to compute how many ATP has to be consumed to actually, to fold misforded states to the native states for this kind of ribo-land case. We analyzed the experiment data and then we found that about 100 of the ATP has to be consumed for the collection of the misforded states in the native states. Okay? And, okay, as I pointed out, unlike protein chaperone, RNA chaperone actually, if you add more and more RNA chaperone because kappa equal non-zero, it actually reduced the native population into non-zero value, actually, non-reality value. So, if you add in more and more chaperone, the native yield actually decreases. That's kind of puzzling because then, why on earth you are using the RNA chaperone to fold the RNA, actually unfold the RNA and then put it into the misforded states? But, if you think about this as a kind of molecular machine, you have to think about the yield per unit time. So, if you multiply this native steady state yield by the time scale of this entire process, actually, both processes increase as a function of chaperone concentration. Okay? So, molecular chaperone, basically maximizes the native yield on a biological time scale by driving the substrate out of equilibrium. So, that's the mechanism, okay? So, all this, even though I just multiply this lambda from some number we obtained from the analysis, this lambda can be actually obtained by, I mean, analytically from the three states of the magnetic model. So, you just multiply this lambda and then this entire expression for the small phi and the large phi. And, I mean, as you see, if you increase the chaperone action, lambda PSS increase whatever kappa is. I mean, it always increased this yield per unit time. To summarize, so, I talked about the logarithm, it's folding on the logarithm landscape. In that kind of logarithm landscape, protein or RNA has to fold via kinetic partitioning mechanism and then chaperone basically helps the folding of protein or RNA by iterative annealing mechanism, which is essentially non-equilibrium. And then iterative annealing mechanism, the previous iterative annealing mechanism has to be generalized by considering the unfolding of a native RNA, native states. And so the final conclusion is that the chaperone, which expands a lot of the ATP, drives the population of protein or RNA out of the equilibrium and making the native population more accessible on a biological time scale. Okay, so I think time is, I think I have a little more slide, but the time is up. So, this many of the work was done this Union's home. He was actually, in my room now, he's working, doing some experiments in Harvard. And these are the collaborator and then these are the paper. I mean, just I, some of the contents of the paper and then put it into this slide. Okay, thank you very much. The degradation of proteins. Yes. Should also play into this, right? Because there's a lot of ubiquitinization that... Right. So that they're also going down the ubiquitinase, the, you know, the degradation funnel at the same time. Yeah, yeah, yeah. So I haven't actually, but the duration rate is actually compared to other rates. It's very small. Maybe this, maybe the duration rate is comparable to this transition, but as I described this rate, directed, direct the transition from native to Missfulton state or vice versa is a kind of a, not, it's a kind of a negligible in my story and the degradation is also very small. So, but in case we can write down all the effects. So I haven't actually explained the situation in vivo, but you can actually write down all the equation and then get this kind of expression. So which take into account to the effect of degradation. Yeah. But the story just remained the same. Of all message, I convey it remained the same. Yeah. Professor Hyun, thanks for great talk. So can you tell what would be another benefit having a non-equilibrium system compared to the, the steady state? The left side example was sort of fully steady state but right side example, RNA, chaperon, what is non-equilibrium dynamics? What is the biological? Everything is non-equilibrium here. Yeah, yeah. Here is pseudo, what are the benefit of having this flux in this, yeah, yeah. In direct sense. So benefit, maybe I can, I can talk about the benefit in this way. So what happens is that even though the yield of the relative state is smaller, what, if you have a larger flux, then even though that population is smaller, you can make that accessible to the cellular condition. That's very important. So even if your, let's say your yield is like 100%, but if this yield is, yield getting to 100%, take like one hour, then what's the benefit? Right? If this process is very, very slow process, then the biological system will be dead after 10 minutes. So you have to make the things very accessible to the system even though yield is smaller. That's very important, I think. That's the role of Chaperone, I would say. That's kind of not well appreciated by the experimental community, at least the biochemical community. This point is not appreciated very well, right. So when you are considering the ATP consumption, 100 ATP, I think, then you are also considering the amount of ADP that is produced due to the ATP hydrolysis. Right, right, right. Means ATP versus ATP to ADP ratio is fixed in your system. Right, it's there, yeah. So the thing is, this is kind of a local situation, right? The concentration of ATP and ADP is maintained to be one million more than 70 micromole in the cellular condition. This is taken care by other machineries, like ATP, F0, ATPs, all this kind of stuff. They actually take care of the homeostasis of the cellular environment. So concentration is kind of fixed. There are some, you can consider there are chemical baths. External chemical baths is actually working on this system so that you don't need to worry about the production of ADP, but eating with degraded. Or I mean, putting it back to the ATP by the ATP synthase. Yeah, so entire concentration remain constant. Okay, thank you. Other questions? Or comments? I think there can be a multiple state of misfolded. So my question is that, is it okay to consider all that multiple misfolded states into the one state? Say that again. I cannot hear, can you? Can you say without mask? I think there can be a multiple states. Much what? Multiple misfolded states. Multiple misfolded states, yeah, yeah, yeah, sure, sure. There can be several misfolded. I see. So you explained it as one state. There are multiple misfolded states, but I, okay, so you are referring to this state, right? This is like a many state, but I just said that this is ensemble of M state, which is somehow separated by the time scale from the native states. You can actually have a transition between misfolded states. Maybe I don't know, but I can consider this as ensemble of the misfolded states, which is separated far away from the native state by the large barrier. So this is the situation. Thank you. I have another question. It seems our cells make a lot of mistakes and errors, but Penny, for you, you show us on some theoretical viewpoint. How can you confirm that this calculation is correct in real cells? This calculation is correct for the, I would say for molecular shaperone, but not all the resetting process. I mean error correction process, but general underlying idea is that baseline is that in order to make some error correction, you have to consume ATP, and error correction means you are resetting things into the, and then give another chance for this system to evolve again, right? So this is the idea of resetting. It sounds very pleasurable. All right, all right, but it's not purely theoretical. I never, I didn't show any experimental data, but there are a lot of data which is supporting this, and this whole, like let's say, graph I showed you, I mean, this graph is the data point I didn't actually specify, but these are the, these numbers are all from the experimental feeding. And then after feeding that, after determining many, many parameters, I actually draw this line, actually. So it's not just a thought experiment. I see. I was confused because you showed a clean theoretical. Yeah, yeah, yeah, there are many, I mean, noisy data points, but I just removed everything. Thank you. Another question? Okay, if not, let's send to Professor Heng. Thank you.


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