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Love Lamp ist ein Comedy-Podcast von zwei Kölnern, Sebi und Wookie, die jede Woche über verschiedene Themen reden, die sie interessieren, amüsieren oder aufregen. Die Themen sind sehr vielfältig und reichen von aktuellen Ereignissen, Filmen, Serien, Musik, Sport, Reisen, Sex, Drogen, Politik, Kultur, Gesellschaft, Geschichte, Religion, Wissenschaft, Technik, Kunst, Literatur, Gaming, YouTube, Podcasts, Persönlichkeiten, Träumen, Ängsten, Hoffnungen, Wünschen, Plänen, Erinnerungen, Erlebnissen, Geschichten, Witzen, Rätseln, Experimenten, Produkten, Trends, Tipps, Tricks, Hacks, Challenges, Pranks, Memes, Fails, Bloopers, Outtakes, Reviews, Rankings, Umfragen, Quizzes, Wetten, Spielen, Gewinnspielen, Verlosungen, Aktionen, Events, Projekten, Kooperationen, Partnerschaften, Freundschaften, Beziehungen, Familien, Haustieren, Hobbys, Leidenschaften, Talenten, Fähigkeiten, Stärken, Schwächen, Vorlieben, Abneigungen, Meinungen, Ansichten, Einstellungen, Werten, Überzeugungen, Idealen, Zielen, Visionen, Missionen, Motivationen, Inspirationen, Innovationen, Kreationen, Fantasien, Illusionen, Halluzinationen, Paradoxien, Anomalien, Rätsel, Mysterien, Geheimnisse, Verschwörungen, Legenden, Mythen, Märchen, Sagen, Fabeln, Geschichten, Lügen, Wahrheiten, Fakten, Daten, Zahlen, Statistiken, Analysen, Interpretationen, Bewertungen, Kritiken, Kommentaren, Feedbacks, Lob, Kritik, Beschwerden, Anregungen, Fragen, Antworten, Lösungen, Problemen, Herausforderungen, Chancen, Risiken, Möglichkeiten, Optionen, Alternativen, Entscheidungen, Konsequenzen, Erfolgen, Misserfolgen, Glück, Pech, Zufall, Schicksal, Karma, Glaube, Hoffnung, Liebe, Hass, Freude, Trauer, Angst, Mut, Wut, Ekel, Überraschung, Verwunderung, Bewunderung, Respekt, Achtung, Ehre, Stolz, Scham, Schuld, Reue, Vergebung, Dankbarkeit, Genugtuung, Zufriedenheit, Glückseligkeit, Ekstase, Euphorie, Nostalgie, Melancholie, Depression, Aggression, Frustration, Stress, Burnout, Langeweile, Müdigkeit, Schlaf, Traum, Albtraum, Erwachen, Erkenntnis, Erleuchtung, Transformation, Evolution, Revolution, Rebellion, Anarchie, Chaos, Ordnung, Struktur, System, Regel, Gesetz, Norm, Wert, Prinzip, Ethik, Moral, Philosophie, Psychologie, Soziologie, Anthropologie, Biologie, Chemie, Physik, Mathematik, Logik, Rhetorik, Grammatik, Sprache, Kommunikation, Information, Wissen, Bildung, Kultur, Kunst, Literatur, Musik, Film, Theater, Tanz, Malerei, Fotografie, Skulptur, Architektur, Design, Mode, Schönheit, Gesundheit, Fitness, Wellness, Ernährung, Medizin, Pharmazie, Drogen, Alkohol, Tabak, Kaffee, Tee, Schokolade, Zucker, Salz, Fett, Eiweiß, Kohlenhydrate, Vitamine, Mineralien, Spurenelemente, Hormone, Enzyme, Antioxidantien, Probiotika, Präbiotika, Synbiotika, Bakterien, Viren, Pilze, Parasiten, Zellen, Gene, DNA, RNA, Proteine, Aminosäuren, Peptide, Polypeptide, Enzyme, Hormone, Neurotransmitter, Rezeptoren, Synapsen, Neuronen, Nerven, Gehirn, Rückenmark, Zentralnervensystem, Peripheres Nervensystem, Vegetatives Nervensystem, Sympathikus, Parasympathikus, Endokrines System, Hormondrüsen, Schilddrüse, Nebenschilddrüse, Thymus, Hypophyse, Zirbeldrüse, Nebenniere, Bauchspeicheldrüse, Eierstöcke, Hoden, Herz, Kreislauf, Blut, Lymphsystem, Immunsystem, Abwehr, Entzündung, Infektion, Allergie, Autoimmunerkrankung, Krebs, Tumor, Metastase, Apoptose, Nekrose, Zytokine, Interferone, Interleukine, Histamin, Serotonin, Dopamin, Noradrenalin, Adrenalin, Cortisol, Melatonin, Östrogen, Progesteron, Testosteron, Insulin, Glukagon, Somatotropin, Thyroxin, Trijodthyronin, Kalzitonin, Parathormon, Leptin, Ghrelin, Oxytocin, Vasopressin, Endorphine, Enkephaline, Dynorphine, Opioide, Cannabinoide, Nikotin, Koffein, Theobromin, Theophyllin, Alkaloide, Phenole, Flavonoide, Terpene, Steroide, Lipide, Fette, Öle, Wachse, Phospholipide, Glykolipide, Sphingolipide, Cholesterin, Kohlenhydrate, Zucker, Stärke, Glykogen, Cellulose, Saccharose, Glukose, Fruktose, Galaktose, Ribose, Desoxyribose, Laktose, Maltose, Polysaccharide.

References Cell. 2020 Jul 23; 182(2):404–416.e14. Nature Communications 2020. volume 11,3146 Curr Genet. 2021 Feb; 67(1):19–26 --- Send in a voice message: https://podcasters.spotify.com/pod/show/dr-daniel-j-guerra/message Support this podcast: https://podcasters.spotify.com/pod/show/dr-daniel-j-guerra/support

Reference Dr Guerra's lecture notes --- Send in a voice message: https://podcasters.spotify.com/pod/show/dr-daniel-j-guerra/message Support this podcast: https://podcasters.spotify.com/pod/show/dr-daniel-j-guerra/support

In this episode of The Hormone Prescription Podcast, Karen Martel, a Certified Hormone Specialist & Transformational Nutrition Coach, joins us to provide insights on how to unlock the mysteries of female fat loss. In a candid conversation, Karen shares her journey of struggling with health issues and how she overcame these challenges to become a leading expert in women's hormone health and weight management.     Throughout the episode, Karen provides invaluable information on methods to balance and optimize hormones in peri and post menopause, and breakthrough weight loss resistance. Her practical tips and advice make a considerable impact on women's lives struggling with similar issues. Karen's passion for helping women improve their hormone health stems from her own experiences, and her approach to women's health is bold and practical. She is the host of the top 100 rated women's health podcast, "The Other Side of Weight Loss", where she shares her deep knowledge and expertise on female fat loss and hormone imbalance.    Episode Highlights: - Unlocking female fat loss secrets - Understanding hormonal challenges in peri and post menopause - Overcoming weight loss resistance - Karen's personal journey and experiences - Bold and practical approaches to women's hormonal health    Don't forget to subscribe to our podcast for more expert insights and breakthrough approaches in women's hormonal health!   Speaker 1 (00:00): What if you could break through weight loss resistance with a novel bioidentical hormone, would you try it? Stay tuned to find out if I would. Speaker 2 (00:10): So the big question is, how do women over 40 like us keep weight off, have great energy, balance our hormones and our moods, feel sexy and confident, and master midlife? If you're like most of us, you are not getting the answers you need and remain confused and pretty hopeless to ever feel like yourself Again. As an ob gyn, I had to discover for myself the truth about what creates a rock solid metabolism, lasting weight loss, and supercharged energy after 40, in order to lose a hundred pounds and fix my fatigue. Now I'm on a mission. This podcast is designed to share the natural tools you need for impactful results and to give you clarity on the answers to your midlife metabolism challenges. Join me for tangible, natural strategies to crush the hormone imbalances you are facing and help you get unstuck from the sidelines of life. My name is Dr. Kyrin Dunston. Welcome to the Hormone Prescription Podcast. Speaker 1 (01:03): Hi everybody. Welcome back to another episode of the Hormone Prescription with Dr. Kyrin. Thank you so much for joining me today as we're going to dive into a novel tool that's a bioidentical hormone that could help you lose up to 5% of your body weight in fat. Wanna know about it? Sure you do. Right? We always wanna look for all the natural options that could help us safely and effectively be more healthy. Right? If that includes losing weight, lose weight, regain energy, right? All the things you still gotta do all the things, but maybe this is something that can give you a little boost. We're gonna dive into that in this episode. And our guest today, Karen Martel, is gonna tell us about it and you're gonna see if I'm gonna try it or not and would you try it or not. So we're gonna have some information on that. Speaker 1 (01:51): But we're also gonna take a little tour off the main road that we usually talk about when we talk about breaking through weight loss resistance. And that has to do with the intangibles related to losing weight. The intangibles related to going from perimenopause to menopause, all the intangibles related to every symptom or disease that you might have. What are those? Well, the thoughts, feelings, and beliefs that are driving or supporting that energetic disruption. That is the symptom that you have, whether it's excess weight or you've got diabetes and your pancreas isn't functioning well, or you name the disease or symptom. There's some underlying spiritual, if you will, or energetic, cause we're gonna dive into that too. As my GE guest today, who is a weight loss resistance specialist has a lot of experience with that, as do I on my journey. It really is something that you will come to face yourself when you're healing from disease, if you're doing it right, and if you want the optimum outcomes, it really is key. Speaker 1 (03:01): So I'll tell you a little bit about Karen, then we'll get started. She's a certified hormone specialist and transformational nutrition coach and women's weight loss expert. She's got a top 100 rated women's health podcast. It's called The Other Side of Weight Loss, where she helps women to unlock the mysteries of female fat loss and hormone imbalance. She struggled with her own health issues, but she was determined to bring her knowledge to others with a bold new approach to women's hormone health and weight management. Her passion lies in helping women balance and optimize their hormones in per and post-menopause and breakthrough weight loss resistance. Yes. Please help me welcome Karen Martel to the hormone prescription. Speaker 3 (03:44): Thank you very much for having me, Dr. Kyrin. I'm very excited to be here. Speaker 1 (03:49): Yes, we talk about weight loss resistance. It's such a problem for women once we hit 40 and midlife and we age, and you really have some unique perspectives on some tools that are novel that can assist us with this weight loss resistance, and also how to address something that most people aren't aware of as an issue for them around weight loss at midlife. So maybe actually you can start by sharing a little bit about your journey and how you zeroed in on hormones and weight loss resistance and all things that we, women over 40 are concerned with . Speaker 3 (04:26): Yes, . Well, mine actually started, of course, had to start early for me in my thirties after the birth of my first child. I, you know, lost the pregnancy weight, but about a year and a half into it, I just, very suddenly, within a few months time, I rapidly started to put on weight and I had not changed anything. And I was experiencing not only weight gain, but I was having severe insomnia, really bad PMs, anxiety, depression, gut problems. And I did what every woman is told to do. I started to exercise like a mad woman . I started pretty much starving myself, trying every diet under the sun, calorie counting. I had always been a really healthy eater. So it wasn't like, it wasn't because I had changed my diet or was eating McDonald's every day and I was working out probably harder than I'd ever worked out in my life. Speaker 3 (05:25): And I only continued to gain weight. And I went to the doctor, I was put on an antidepressant, I was given a sleeping pill and kind of told to, you know, go on my way. And nobody at that time would've told me I was 33. Nobody would've said, Hey, maybe this is your hormones. And after a lot of struggling and a lot of different diets and exercise programs, I decided that I had to take matters into my own hands. And I started digging into research and decided, I think this has something to do with my hormones. And so I ended up going to a naturopath and having my hormones tested through saliva testing at the time. And it came out that I was, you know, super low on progesterone, high estrogen, and I had very low cortisol and D A T A, which I'm sure your listeners are familiar with, those two hormones and those are your stress hormones. Speaker 3 (06:24): And there I was just doing everything wrong. According to that hormonal profile, I was doing high amounts of cardio. I'm starving myself. I later found out that I also was hypothyroid and had no idea, which also those two things would've perpetuated that. So I had to go, okay, I have to make a serious shift because I didn't even think I was a stressed out person. So I started, I quit the, the hardcore CrossFitting and starving myself. And I actually started doing yoga all the time. I started asking for help. And the biggest piece of that puzzle was I started to really look at the spiritual side of what was happening. And I realized that there was a lot of things that I had not dealt with from my past. I had always had really bad self body image. I had an eating disorder as a teenager, I had sexual abuse. Speaker 3 (07:24): There was a lot of things that Karen had not dealt with. And so I went on a bit of a journey and I will tell you, it was not quick. There was no quick fixing this. And it took probably a good year of like really working on these things, slowing down, like looking at the spiritual aspect of things, dealing with some of the past stuff that the weight finally started to slowly come off. Mm-Hmm. . And I came out of that experience just going, there's gotta be more women like me. There's gotta be more women that are doing everything right that are not, you know, closet eating or eating McDonald's mm-hmm. and all of these things that, that they are eating, right. That they're exercising and they're not able to lose the weight. And so it kind of, it threw me into this world of nutrition. Speaker 3 (08:13): And then at the age of 42, I started to go into menopause and overnight Wow. Yeah. Overnight I had my thyroid crash on me and I started missing my periods. I started having hot flashes. I was, I gained two months and was like, what the heck? Because I really didn't think, I said at that point in time, I had been, you know, 10 years nutritionist, weight loss coach, like doing everything right. I don't drink, I don't, you know, dealing with the, the mental stuff, sleeping well. Like I, I really had all the boxes checked and then behold it happens again. And so that sent me on a mission to find out, okay, what's happening with my, my hormones? Why am I going into menopause so early? I was able to reverse it and I, at 47 today, I still have my period and I'm doing well. But it was a big eye-opener. Both times that when that happened to me, and I just, I know now work with so many women that very similar things have happened to them too, where they're, they are really healthy and they're trying to do things right, but they can't get that weight off. So Speaker 1 (09:22): Yeah, that period is that fifth vital sign. And when it starts going wonka donk, that is your sign emergency. Something is wrong. Just like if your pulse is sky high or your respiratory rate or you have a really high fever when your period, it really should be like clockwork. Your body has a timing mechanism where your period should come every, you know, 28 days or whatever is normal for you. And when it doesn't and it stops or it's late or it's irregular, that really should be your sign to look for the cause, not to get on a synthetic hormone. Just shut up your female system. Right? Everybody's heard me say that don't get on the birth control pill Speaker 3 (10:08): Or Speaker 1 (10:09): You know, the vaginal ring or devil shots like was done to me for decades. That's not the answer ladies. It's really not the answer. You gotta find the cause. So yeah, you hit that wall wow. At 42 and that, you know, bring drive home. Another very important point I wanna point out is that a lot of times if you go to a corporate or mainstream doctor with that scenario, they're just gonna say, oh yep, you're going into menopause. Oh well it's just biology. There's nothing we can do. And they're not going to investigate it. But you know, what you just heard Karen say was that she found the underlying cause she remediated it and then she had got her periods back and it wasn't menopause. So you don't wanna go into hormonal poverty any sooner than you have to, cuz then you have to face the consequences of them. Speaker 1 (11:01): So let's dive into, you mentioned some of the spiritual things. Let's dive into that cuz that's something we haven't talked about on the podcast that I think is super important. And then after that we'll dive into this novel tool that really is very hot and late breaking tool that you can use to help your weight loss resistance and lots of other problems in health that a lot of people don't know about. So we'll dive into that next, but what were some of your understandings that you learned through that period that you just described about the spiritual nature of this change you were going through? Speaker 3 (11:37): Yeah, I mean for my, myself personally, and I think for so many women that body self-image was a huge piece that I had to really look at. And I had self-abuse myself for a very long time. I was an alcoholic, I was a drug addict. Like I said, I had been raped, I had very bad eating disorder. I was bulimic for many years and I hadn't looked at any of that. I just literally had numbed myself and just pushed it away and thought, well I know that that's not good and that I probably am a little bit messed up from it, but I recognize that that wasn't a good thing. So I must be fine , I don't need to go back and really pick it apart. But I did and I had to. And it took a long time. I mean, like I said, it wasn't easy. Speaker 3 (12:24): I had to, I was in therapy for seven years and it was, it wasn't traditional therapy. I did a lot of what connected with me. I had started with traditional therapy and then I moved into spiritual therapy and I started seeing an energy worker. That was life-changing for me. And that's what really connected with me. And I really highly recommend that women find somebody that's gonna connect with them. Because I think a lot of the time women don't look at these problems because they don't, they immediately think if, if I'm going to go down that road, I gotta go see a therapist and I don't wanna do that. I'm not gonna go talk to somebody about my problems doesn't work, what are like, they have some issues around that. And it doesn't have to be that. It could be something completely different. E every single one of us is different. Speaker 3 (13:12): I've had, I've seen people that have gotten so much help just from body work, like going or doing yoga. Like there's so many things that you can do to connect with self. Mm-Hmm. , it doesn't have to look like that. And for some people that does work. So I think it's important to look at all of these things in, if you are continually running into the same problems over and over again, that's your sign. That's your sign that you are not looking at something that you need to look at. So if you are continually looking to the next best diet, constantly failing at that diet, constantly gaining the weight back, or maybe it's something completely different where it's just like you have this repetitive pattern of somebody wronging you constantly or getting yourself into the wrong relationships with the wrong partner. These are your big warning signs that hey, you're not listening. There's something here that you're not learning from, that you're not looking at that you have to deal with. And then, and if you don't, it's gonna keep happening over and over again and likely worse. Speaker 1 (14:22): Yes. And you know, I identify with a lot of the things that you shared. I'm wondering for everyone if you could talk about what you mean by spiritual. Because some people hear that and they're like, oh, that's not religion. I'm religious, I'm ex religion, I'm not gonna hear this, I can't hear this because spirituality is anti-religion. So maybe you could address that. So what do you mean by spiritual aspects? What does that mean? Speaker 3 (14:50): It can mean religion. I think that spirituality, I think religion is under that umbrella. So what it's whatever feeds your spirit. So if that is religion, no matter what religion it is mm-hmm. , that could be maybe what you need to connect more with. If that's what connects with you. If that's God, if it's Buddha, whatever it is, then maybe that's the road that you need to go down. Maybe you have to go talk to your priest or go talk to go get into a Bible study group wherever it might be. But if that's what is your thing, then that's great. Spirituality for me is personally my soul. What purpose am I here on earth for? I think we're all here to love and to learn. And I think that if we don't learn these things, that we're gonna have to come back in our next life and learn them again or try to learn them again. And that's my belief. I believe that there is a God, I think there was a Jesus, I think there was a Buddha. So I have this kind of like mix of all the religions put into one. But spirituality is more than what what you see here. It's that intuitiveness, it's what's going on inside. It's your heart, it's love, it's, and it can be your religion as well. So that's personally the way I see it. I don't know about you Karen, when, how do you see spirit ? Speaker 1 (16:06): Oh yeah. I mean to me it's the part of us that enlivens the human form that we're in, that we get to use while we're in this, on this earth plane. And it represents all of who we've ever been and ever will be and what we're meant to express in this lifetime. What we're meant to experience, the people we're meant to come in contact with. And to me, when I talk about the spirituality of perimenopause or menopause or the spirituality of any disease, I think there is a spiritual lesson in, in every symptom and disorder that we can experience. I'm talking about looking for the deeper meaning that isn't obvious, right? If you, when I weighed 243 pounds and I looked in the mirror, well you could, I could see I was morbidly obese. Right? But what's the meaning? What's the meaning behind that? More than, oh, I had hormonal imbalances is I had toxicities, I had gut dysfunction, I had all these physical causes contributing to it. Speaker 1 (17:08): I had energetic causes, right? Thoughts, feelings, beliefs, all garnered from past experiences that I had had that led me to certain belief systems. So it's interesting with obesity, they found in some of the studies, I think his name is Filetti, who did some of the research on this. And he just happened to notice that 80% of the women in, in the study he was doing on obesity had been sexually abused. And that's ultimately what I think led to the ACE trials, the adverse childhood experienced trials. Cuz they were starting to see this link between adverse childhood experiences, emotional psychosocial experiences, and physical health problems. And then when he, he noticed this 80% incidence of childhood sexual abuse among these women in this study that were obese. So to me it's really looking at the deeper meaning. And so how did that show up for you? Mm-Hmm. in terms of what you were going through right Here are your 42 mm-hmm. , your body's getting ready to go into menopause, which is on average nine years early, your thyroid shut down. And so what did that look like for you? Mm-Hmm. Speaker 3 (18:19): The thyroids energetically signifies your voice chakra. So we have these energy centers in our body, and this isn't woowoo, this is science. We have energy centers throughout our body. And the voice chakra sits right where your thyroid is. And I think that this is, I personally think this is why so many more women have it than men have thyroid problems, is because we don't tend to speak our mind as much as men do. We tend to stuff things down. We're not, we're taught that, you know, maybe not to voice our emotions so much. And I, looking back at my history and all the times I didn't say anything when I wanted to say something, when I wanted to stop that person from abusing me, when I had so much self-hatred for myself, I did not think that I was capable of being loved even. And that I think was really tied up in my thyroid. Speaker 3 (19:16): And I don't think it was something that I dealt with over those first few years of therapy. I don't think I, cause the thyroid thing didn't come in until later. And then I was like, oh, I'm not surprised that I have a thyroid problem. And I didn't address it. And so I really think it was my body going, n you have to pay attention to this now. This is, you have to start speaking your mind and you have to start loving yourself. And it sounds really like, oh, so cliche, oh, love yourself. But looking back at my history, I never did. I never loved myself. I had always hated my body. I always critiqued it. I didn't trust myself. I didn't, there was so many things that needed to be unpacked. And coming into my forties, going into menopause and gaining that much weight, especially as a weight loss coach, that was super hard on my ego. Speaker 3 (20:09): And I knew that no matter what I did, this was interesting. No matter what I did, I would not have lost the weight then I knew it. So every time I would like go, oh, you know, the, the old patterns would start to come up and I'd say, oh, I, I better start, you know, cutting down on the calories and we better start working out harder. And then I'd be like, Nope, stop. Don't do that. Because then that's like going back into that old pattern of dieting and thinking that that was the answer. Cause for me it's, it wasn't, and I knew that, I knew that there was something emotional, like you said, like there's always something emotional to every medical problem, every chronic ailment that you've got. I really truly believe that there is going to be an emotional component to it. Whether it's the full root cause, whether it's just a third of it, who knows. But there's always something emotional or something that, that your body is trying to get you to look at in your life. Speaker 1 (21:07): I totally agree with that 100%. And some of the things that you said, I wanna, let's go back and talk about this. So, you know, you very are very open and very freely shared some of your struggles earlier. And then you said, I hated myself. And the average per woman, if you ask her, do you hate yourself? She's gonna say no. And if you ask her, do you love yourself? She's gonna say yes. So I'm thinking we probably, because that probably would've been me at one point also. And then there was a point where I realized, no, I actually treat myself as if I hate myself, . And so what does that mean? What does, how would a woman identify, oh my gosh, I behave as if I hate myself. And the whole cliche, like you said, of self-love, we tell, we hear that so much, but really what does that mean on a day-to-day, day basis? Maybe you even take some instances, well, oh, here's something that I dealt with and here's how I treated myself with hatred around that, and here's how I now treat myself with love. So maybe we could have a little richer, more in depth discussion on that. Speaker 3 (22:11): Mm-Hmm. , I think people, women can recognize it because you have to tune into how are you speaking to yourself all day long. Right? Pay attention, like most of it is subconscious, but pay attention to, and I, and we're gonna get into this, is, I just recently lost a bunch of weight and it's so interesting to me to, to realize that I still have these like nagly little self-talk bull crap inside of me. . It's like, you know, I just go lose a bunch of weight, even though I was, you know, I, I was looking good and, but I lost weight. And then I caught myself looking in the mirror, not even realizing going, I wish I, I could lose a little bit more weight right here on my, the back of my arm. And I was like, oh, Karen, how dare you say that you just lost 10 pounds off your body and you, how dare I? Speaker 3 (23:06): And I was so ashamed and I felt so guilt. I was like, what, what? How can I even think that? And so I had to look at it and be like, where's that coming from? And I'm like, it's coming from the fact that I've done this for the last 30 years of my life. I've looked in the mirror and I have cut myself down. I, it's al it's never like, Hey, I'm looking good. Look at how great my stomach looks. I'm looking so hot. Like, it's usually like, we don't do that. We right. We pick up the one or like many things, whatever it is. But you'll, you'll nitpick. You'll be like, what's this freckle doing here on my face? , you're just, you'll find anything. And sometimes women, it's a lot, it's not one thing. It's their whole body that they look at and they hate. Speaker 3 (23:54): They're going, what? Look at my stomach, look at my legs, look at my cellulite, look at my hair, look at my wrinkles, look at my chest, look at my breasts. Like it's never ending. And so ask yourself like, how, how is your day spent talking to yourself? Are you constantly worried about your weight? Are you constantly worried about the food that's going into your mouth? Are you constantly going, Ooh, I'm gonna start this tomorrow. Ooh, I'm gonna eat better today. I'm gonna eat better tomorrow. I'm gonna start the diet next week. Because I think majority of women do this, wouldn't you say? Speaker 1 (24:28): Yes. And you know, I love the litmus test for that because it's such a habitual way that most of us think when we look in the mirror, we're not looking for what we love. We're looking for the things we don't like that we need to fix so we can be perfect. Would you say that to a friend? Would you say your friend says, oh Karen, I lost 10 pounds. Look I lost 10 pounds. And they proudly come before you and you go, you know what? Yeah. But you've got fat on the back of that arm and you need to lose that. What would your friend say to you? She would not be your friend anymore. Right. That's how you know you're hating on yourself. Speaker 3 (25:06): Yeah, exactly. And, and hate's a strong word, but yeah. Speaker 1 (25:10): , Speaker 3 (25:10): You know, like how much are you loving on yourself? Maybe ask that. How much are you actually just going like, you are rocking it. You look so fantastic. I had a friend last year who was desperately trying to lose weight. She was eating, trying to eat right. She was exercising. And then I didn't talk to her for like a month or two. And when I saw her, she had lost weight. And so I'm thinking like, oh, all my tips are working for her. Right? And she said, you know what Karen? I wasn't losing weight no matter what I was doing. And she said, so I, I stopped thinking about it. Instead I would get myself naked and I would stand in front of my mirror and I would tell myself how great I looked. And she said, I did that consistently . And she said, and the weight started coming off. And I was like, wow, . Speaker 1 (25:58): It's so true. One other thing I noticed, I just wanna share cuz I know some people listening are gonna be like, oh my gosh, I do that too is so I had been traveling and not been able to keep to my eating and exercise schedule and I gained a few pounds and then I came to a city here in Argentina where I could kind of get settled and get to a good gym and eat like I like to eat. And so I just started doing, doing the things that I do for my health. And then I started seeing that weight come off, come off. And I was very pleased. But instead of just continuing to love myself and give my body what it needed, I turned it into a project and I'm like, oh my gosh, I lost five pounds. I bet I could lose another eight pounds this month if I up my gym. Speaker 1 (26:44): And I cut my intermittent fasting to only eating four hours a day. And before I knew it, I had made myself in my progress into nothing. And I had made myself into a project and objectified myself instead of just loving myself and giving my body what it needs. Because I know she'll go to what the weight she likes. And I know there's some people listening going like, oh my gosh, I do that too. So you're not alone. But then what is the loving thing to let go of that I don't stop, I'm not, I don't usually weigh myself. Why did I even get on this scale? Right? It's not loving. So now I don't, not Speaker 3 (27:21): Not on Speaker 1 (27:21): The scale, right? I'm still in in Buenos Aries and I just go do the exercise and give my body the food it needs. That's loving. So how does a yeah. Self love look. How would you explain that? What kind of actions would it be? Speaker 3 (27:37): I think both with my, both times there when I was in my thirties and when I was in my forties, when I remember getting to a point even in my late thirties where I realized that I was still trying to lose weight even though I was at 128 pounds, that was my weight set point. It would never move from that. It would be like I would try something extreme, I'd lose some weight and it would just go right back to 1 28. And I just kept doing this for years. And I was constantly like, I wanna get down, I wanna get down. I shouldn't be this weight. I should be 1 25, I should be one 20, whatever it might be. And then I remember this one day, it just dawned on me, I'm like, this is where my body wants to be . It's not a super skinny little person. Speaker 3 (28:20): And I never will be. I never have been, I never will be. So quit trying to get down to this unrealistic weight. And I'm like, what, Karen, are you really willing to do what it would take to be 120 pounds? Like I would have to starve myself. I would have to, I I, even if I did, I still don't think I would've gotten down to that. And I let it go cuz I was like, I can sit here and spend the rest of my life trying to get down to this fricking number on the scale because I feel like that's how I should look. Or I can accept where I'm at right now and work on just accepting that. And so I did and I did great. And for many years I was, I was very pleased with where I was at till I hit my forties and I, and I gained the 15 pounds and then I immediately started to hate on myself. Speaker 3 (29:11): But I did the same thing where I, I knew if no matter what I was gonna do at that time, I wouldn't have been able to lose the weight. And so I worked on loving myself and I worked on sharing that message. And I think that that really helped because menopausal women, perimenopausal women and menopausal women, I really believe that it's very normal for us to soften a little bit. It just is men do. So why can't women? And it's not realistic to think we're gonna look like we did when we were 20, when we're 50. It's just not, and it's okay to soften a little bit and we need to start accepting that and having that be the normal where we can look at women that are in this midlife go and go, they look beautiful because I do, I look at women my age and I think they're beautiful when they're healthy. Speaker 3 (30:10): And so I just kept telling myself that is, I'm gonna be healthy. I'm gonna still eat right, I'm gonna still exercise, I'm gonna still do yoga and I'm gonna be healthy. And I had super clear skin and I, and I looked good. And yes, I was more overweight than I wanted to be. And I knew that that would come off eventually. And it did take a couple of years and it was slowly started to come off. And I still was though accepting. Cause I still wasn't down to 1 28. I still accepted that I needed to, that it was okay for me to soften a little bit mm-hmm. because I was getting older and that's okay. I wasn't overweight. Mm-Hmm. . I, but I had softened. Speaker 1 (30:52): I mean, I think if you do want to be the same as at 60 as you were at 20, you can do it. It's just, do you want to do it? I will say, I wanna ask you this cuz I, I don't know how old you are now. I'm in my fifties 47. Speaker 1 (31:07): Okay. So my generation, I guess that's your generation. I don't know if we're, I'm a baby boomer. I think we had some pretty unrealistic expectations of what weight we were supposed to be at any age. Yeah. Literally we had, you know, those little Barbie dolls where you could wrap your little pinky finger around their waist. And for my whole life, I thought because my thighs touched and I didn't have a gap thigh gap, there was something wrong with me. But now the, there were earlier, more recent generations, they really accept themselves. And I love some of these kind of TV and internet stars where they're big and beautiful and bold and they love themselves and nobody judges them and they're just accepted for who they are and their beauty. And I, you know, I don't know how you overcome the conditioning we went through, but any thoughts on that you'd like to share? Speaker 3 (31:56): Yeah, you could look like you did when you were 20, but what is it gonna take to get there? Like, how much suffering are you going to to have to go through and, and, and it's just not realistic. And I always tell women like whatever their numbers that they're trying to get down to, I asked them, when was the last time you were at that number and how long were you there for? Because usually it's like, oh, well I did this really extreme diet when I was 25 and I got down to 115 pounds, so I think that's what I'd like to be at because I love the way I looked. I'm like, yeah, I bet you did. But is that realistic? No, that's not your set point. So I just think be realistic about it. Be okay with softening a little bit. I'm not saying you have to accept being, you know, 300 pounds or two. Speaker 3 (32:40): Like I'm not saying that. I'm just saying that there is a place where you have to begin to accept where you're at because it likely will not change. Like you can only get down to that set point. Maybe that a healthy set point that is still maybe in your eyes overweight, even though it's only like five to 10 pounds or 15 pounds overweight maybe for you. But could you just work on accepting that and loving yourself for that? And just focusing on being healthy and feeling really good. And that's, I think, a really hard thing to ask women really, really hard. Speaker 1 (33:18): And what about the cultural norms that we grew up with, with very skinny barbies and a very skinny twiggy kinda expectation of what the definition of beauty was and what we should look like? Yeah. Speaker 3 (33:30): Well I, I ask you then, all of you, how many women do you know that look like that? Especially in their forties, even in their thirties, how many women did you know that were stick thin and had a perfect model figure? I can count like two friends out of all my friends that looked like that. Most didn't. So it's more normal to have curves than not to have curves. And we just see it in the magazines as these, this 1% of society of women that ha that were literally born genetically superior and that have those shredded bo like, they just come out looking fantastic and never have to worry about their weight. So it's more normal to be curvaceous and we have to tell ourselves this, that, that, that is what's normal. It's not normal. And, and look at men, geez, the discre, like between men and women. Speaker 3 (34:24): I just, it drives me nuts that men, all of my guy friends right now, they're in their forties and fifties. They've got the bellies. They're not getting Botox, they're not doing plastic surgery, they're not concerned . Like, they're just riding along, you know, and we look at them and oh, they're handsome with their gray hair. But yeah, women, we start to age and we're looked at like, Ooh, no, no, you better get your Botox. You better. Not that I'm saying anything wrong with Botox, I do get myself, but you know, like the pressure on us is far more right. And you have to remind yourself, like, I remind myself all the time, like, my husband's not doing all of these things. So take it easy on yourself. Don't feel like you have to too. Mm-Hmm. , this is the time in our life, like talking about spirituality. Speaker 3 (35:11): This is a time in our life where we, it's all about us finally. You know, like we in our twenties and thirties, it's all about our career, having children, getting married, and a lot of the time, right? We, we've, at this point, we've figured out if we're keeping the man or we're getting rid of 'em or, or, or your wife, whatever it is. And as we go into these years, our hormones start to shift. And I really think that this is is a calling to start paying attention to yourself, to start paying attention to your health mm-hmm. and let it be about you. You know, your kids are growing up, they're getting older. If you have children, you know, your ki your ki you figured out your job at this point. So let these years be about you because these can be the best years of your life. You know, we talk a lot about the bad, about the weight gain, about the hot flashes, the, the low sex drop, all the things that come with losing our hormones. But there's so much good that can come out of these years. These can be the best years of your life if you can start to take care of yourself and treat yourself with love. Like, it, it is, it's the time to do this and to work on these things. I really, really believe that. Speaker 1 (36:21): So one of the things that you shared with me before we started, I said, oh, we have to talk about that, that you can do to love yourself and really help yourself be your best as you age is peptide therapy, particularly if you're experiencing weight loss resistance. And I thought that is something that we should cover and talk about in this episode. So tell me how you became introduced with peptide therapy to help weight loss resistance. Yes. Speaker 3 (36:49): So we have actually a, a peptide weight loss program now because it started last year when I had a client who had Hashimotos. She was one of those women that was doing everything right, eating right, exercising, could not lose the weight no matter what she did. She had optimized her hormone, she optimized the thyroid, could not get the weight off. And we have, we, I hadn't seen her for a couple months, we have this session together and she just lost 17 pounds, . I said, how the heck did you do that? She said, Ozempic. And I'm like, what's Ozempic ? I hadn't even heard of it. So she tells me what it is and it's a weight loss peptide. And so then I went on the search of what the heck is this? And I started googling it and finding out for myself and thought, wow, this is amazing. Speaker 3 (37:38): I'm gonna give this a go. So I try it for myself and it was a complete disaster, to be honest, . I actually ended up getting severe migraines from it. I felt super sick, didn't lose any weight the first couple weeks. So I was like, well of course this isn't gonna work for Karen. Lo lo and behold, like, not shocking at all. Right? So I then started to recommend it to other client, other perimenopausal, menopausal women that I knew that had tried everything. You know, like they had done all the work and every single one of them were got results. And it was, it's, it was like, wow, this is amazing. So then fast forward to now, we've created our own, we like weight loss peptide program because it, I, it really truly just blows my mind how well these peptides work. So I have now gone to the other one, which is known as Manjaro. Speaker 3 (38:34): So I do it just the the peptide. I don't do the name brand Manjaro. So that's called Tze Peptide. So there's Semaglutide and there's Tpat. Tpat is the newest Semaglutide, which is also known as ozempic has been around for 10 years. So we've got a lot of research now on that originally developed for type two diabetics and they realized that, oh my goodness, people are losing a ton of weight, but they're on average in a year's time, or 72 weeks I think it was, they were losing 15 to 20% of their weight. Which just so everybody knows bef prior to this drug, if a drug could help a person lose 5% of their weight, that was considered amazing. Cuz that's all we had seen. That was how good it got. Mm-Hmm. , these were 15 to 20%. And it wasn't just that people were reversing all of these metabolic diseases. Speaker 3 (39:29): You know, cholesterol was getting better, their fatty liver was disappearing. It was helping there, there's research now showing that it helps to prevent Alzheimer's disease, plaque formation on the brain. All of these little metabolic markers were getting substantially better. People were getting off blood pressure mega medications and so many things were getting better mm-hmm. and plus they were effortlessly losing weight. These, these medications not only help to suppress appetite, but they work on your brain and they help it so that you don't crave sugary foods anymore. You just kind of just disappears . So you don't care to you, you're not struggling that whole time. Right? The whole, any other diet, you're struggling. We all know that. You're, you're like, I can't, I shouldn't eat that, but I really want to, you know, that's like why most diets fail is because our willpower runs out and we end up eating the highly palatable foods that are all around us. Speaker 3 (40:31): Well, this drug eliminates that need. So weight loss becomes easier than it's ever been before in your entire life. It's nuts how it works. So I went on Tze peptide, I was at 138 pounds and no side effects. In six weeks I lost 10 pounds and it was mm-hmm. . And it's been easy, fantastic. Like, I have no problem with it. This is though something that you have to typically stay on. So that's the downfall is you come off of it and a some people will, will keep the weight off, but because most people will blow past their set point, then you will likely, if you come off of it, you may go back to that normal set point, right? So for a menopausal woman, if you've gained, let's say you were always 130 pounds, but in menopause you've gained 15 pounds and you cannot get it off. Speaker 3 (41:30): Well, you could go on this drug and it's gonna gonna get you down to that 130 pounds. But if you wish that you could go actually down even farther. So if you came off of it, you would likely, as long as you were eating right and still exercising, you could go back. You may gain back, let's say right back up to maybe the hundred and 30, because that's your normal set point when most women I find have to stay on a maintenance dose. So they have to have seen everything from once every couple weeks to every six weeks. They have to do an injection, their once a week injection. So they, you start to spread them apart and then you can maintain that weight loss as long as you continue doing a shot every four to six weeks. Sometimes did everybody's individual. Mm-Hmm. . Yeah. Speaker 1 (42:17): So can you talk a little bit about how semaglutide works? So everybody knows like, is this an appetite suppressant? Is this uppers? What is it? Speaker 3 (42:27): No, definitely not. Uppers. So both semaglutide and tze peptide are hormones actually. And they're gut hormones. We produce them in our gut and then they tell your brain when you're full. So when you're eating these, because we're increasing, so they're 90, you can think of them as, I think of 'em as like a bioidentical hormone. They're 94% bioidentical to something you already make in your body. So it's gonna create more of this hormone that's gonna be telling you that you're full a heck of a lot faster. So there's no stimulation to it. It slows gastric emptying as well. It does affect the dopamine centers of the brain and the pleasure centers. So you don't get the same payoff from food, drugs, alcohol. Mm-Hmm. . So anything that is addictive to you that you are getting a dopamine hit from, it will bring that down. So there were mm-hmm. Speaker 3 (43:29): , there's actually some PubMed research showing like how it helps with co for cocaine addicts. So, interesting enough, I was just looking at that last night. So it works on different, a couple different mechanisms. There's two different, so Semaglutide is a glucagon-like peptide is what it's called. And then the tze peptide is a dual agonist, not that anybody needs to know this, it's a G I P. So glucose dependent, insulinotropic probably not saying that, right? Polypeptide. So these are peptides that we've already got in our body that are mimicking these peptides when we're taking them. So mm-hmm. , it's the best way to explain it. . Speaker 1 (44:11): And then everybody's gonna wanna know, oh my gosh, I want this, what does it cost? ? Yeah, that's the next question. Speaker 3 (44:18): Unfortunately, if you're not diabetic, you cannot get it covered through, if you've got insurance, you will not get it covered if you're not diabetic. Ozempic has now been approved for weight loss for obesity, but you have to be, I think it is over 30% or 35% bmi. So if you were to just go and get yourself a prescription, manjaro is gonna cost you about $1,500 a month. Semaglutide is about 1200 a month, but there's now compounding pharmacies that are making it. So compounding pharmacies can make semaglutide. And you're looking at about 200, 2 50 a month for that through a compounding pharmacy. Mm-Hmm. . And then with Tpat, it's a little bit of more gray area that they're not allowing compounding pharmacies to do it. There's still some that are, and I think eventually we'll start to see it more mm-hmm. . But right now, the company that made it is coming down on compounding pharmacies if they're trying to compound it. Speaker 3 (45:23): So then you can get it through peptide sites. So I, I don't know how much you've talked about peptides on here, but you know, there's growth hormone peptides, b BPC 1 57, all of these very familiar peptides that people are using now, and they're ordering them online from reputable peptide sites. Mm-Hmm. . And there you can get, we've got Tze peptide that starts at 170 a month, and the semaglutide is like 75 a month to start mm-hmm. . And then you increase your dose as you go along. So then it starts to, then the price goes up the farther along you go. So that's not too bad considering what, what us women will do to lose weight in the money that we spend on it. , Speaker 1 (46:05): I mean, I'm still a proponent of addressing the underlying root cause, but if something is natural and can be an aid to assist you, hey, why not? In fact, as you're talking about it, I've never, I always have to try everything first. You know, that's how I developed same everything that I I do with women is what I learned from losing a hundred pounds and getting off all prescription medications and healing from all the chronic medical conditions I had. I said I was gonna see if it worked, and then if it worked, I was gonna spend the rest of my life teaching other women about it. So that's what I do. And when I hear about something that could help us, that isn't gonna harm us, and you know, with it being a peptide, that's just a protein and it is fairly bioidentical. I'll tell everyone right now, I'm gonna get it. I'm gonna try it and I'll give you my feedback. But it sounds like you've had a, some good results with this. Do you wanna share any patient stories that come to mind? Oh, Speaker 3 (47:01): I've got so many patient stories. It's, it's so gratifying. It's just, we've been, we've been brain, we've been, not brain, we've been, it has been very, very hard. Our whole lives to lose weight. Like weight loss is really hard. And we all know this. And I have spent my life helping women to lose weight. Like, it's something I'm very passionate about. And I feel the same way as you do Dr. Kiran, that you have to look at all of these other factors. You know, it's, you gotta look at your hormones for all of you perimenopausal, menopausal women. You've gotta address the hormone loss, right? Well, you're gonna be, if you go take these peptides, you have to building, you're building a house on sand. That's what I'm saying. It's like you're building a house on sand, you gotta have a foundation. And so this is why we've created a program to go with it, because you could go out, buy these peptides on your own, go for it. Speaker 3 (47:52): But you know, if you're not, if you're still not eating right and you're not exercising, you're not replacing those hormones and working on your spiritual self like we've talked about here today. Then I just feel like, why, why? Like, why don't do that to your health? Don't use it as a, well, I'm gonna go eat my donut and I'm not gonna lose any weight. That's not good for your health then. Mm-Hmm. . So I really think you need to prioritize and you have to have those foundational pieces be part of it and take the time because this, this drug works so well take the time to change your eating habits, then if it's always been a struggle for you to eat, well then use this as a tool to work while to take, while you work on your eating habits, because it does make it so much easier. Speaker 3 (48:41): So work on the eating habits, work on the time that you eat, work on how much you eat because it's very hard to overeat on this drug. Use this as a positive thing. And it is very exciting because you just, the weight just falls off and it's like, oh my gosh. Like I felt, I, I've gone through this like battle within myself of guilt and shame and like, oh, it's been nuts. I've just been like, oh my. And like, and I've had to turn and look at like, some of these things that have arisen because I feel like if it's not hard, then I have no right to be thin. Speaker 1 (49:16): Wow. That's a belief system. Do you like believing that Speaker 3 (49:21): ? No. So I've been like, really try, I've been speaking about it on my own podcast and sharing my journey because these are things that may come up for other people where you do start to feel guilty or when people ask you like, oh, you've lost a lot, a lot of weight very quickly they think there's something wrong with you. Or they're like, oh, what she's doing a peptide or, you know, and I've just been like, no, let me educate you on this. Like this is actually the most astounding weight loss show we've ever seen to date. And if it can be easy, why not? I say, let's, let's embrace it. . Speaker 1 (49:59): Yes. Anne, I just wanna reiterate, you gotta address the underlying causes. Otherwise you are building a house on sand. If you don't fix your hormones and detoxify and get rid of the inflammation and fix your gut and do all the things, you still gotta do that. Well feel Speaker 3 (50:14): Good. You may be thin, but you're gonna feel like garbage. Speaker 1 (50:17): Right? And I, I will reiterate, I know we have a difference of opinion. It was pretty effortless for me to lose the hundred pounds when I address the underlying causes. And that's my experience with the women I work with. So I just wanna incorporate both of our experiences for everyone listening. It can be effortless, sometimes it's harder, sometimes it's easier. But why not take advantage of a tool that could help you? So I'm for sure gonna try it. I'll share my experience. If anybody listening decides to try it, maybe you'll decide to do Karen's program. She's got amazing programs, which she's gonna share some in, in information on right now. But you share these amazing quotes with me, Karen, before we start started, and we haven't offered any. So I will, before we wrap up, I just wanna offer two of my favorites that you shared. So one is one of the most courageous things you can do is identify yourself. Know who you are, what you believe in, and where you want to go. Sheila Marie Bethel, I don't know who she is. Can you talk a little bit about this quote and what it means to you? Speaker 3 (51:20): What it means to me is, is that's been my life mission as really being who I truly am. And that's all the work that I've done has led me to that. And it's, I I, I want that for everybody. I do. Speaker 1 (51:34): Amen. Yes. It be courageous. Identify yourself, know who you are, what you believe in and where you wanna go. And then Tony Robbins quote, the path to success is to take massive determined action. Action. You know, sometimes we women, we can just get stuck in our heads believing not true things. And you really deserve to have amazing health. You deserve to have a great sex life and great sexuality. You deserve to have great hair. Yes, you deserve to feel amazing in your skin. Look amazing in your skin if that matters to you. And do all the things that you want in life, right? Those dreams that you have we're given to you to fulfill. They're not some pipe dreams. So any last words you wanna share with everybody? Speaker 3 (52:22): Yeah, just off of, right off of what you just said there, like, don't settle for subpar. This is, like I said, this is the time in your life that could be the best time of your life. So go after it. Be courageous. Take massive action, but do what you, whatever it is that you have to do to be where you wanna be and look how you wanna look and that's fantastic. And embrace it and just, just don't be afraid and don't think that you have to suffer. I think poor, perimenopausal, menopausal when we have it in our head that there's gonna be some suffering or a lot of suffering. And we take this on like a badge of honor and it's like, no, we don't have to suffer and we can look fantastic and we can be fantastic and we can thrive. We don't have to suffer. Speaker 1 (53:11): Yes. So get about that business and you know, I think that's really part of loving yourself and maybe this is where we'll end. Mm-Hmm. self-love is honoring your dreams. That your dreams are meant to be fulfilled and taking massive action to make them come true. So where can everybody find you online? You have a free gift, a hormone quiz you can tell everyone about. We'll have all her links in the show notes. So if you're driving, please don't try to write these down, but share with everyone about the hormone quiz and all the places to find you. Speaker 3 (53:44): So karen martel.com. You can take my hormone quiz to find out, you know, which hormones could be stopping you from losing weight. It's just a quiz. So, but it's a great place to start and you'll get a little free meal plan from it. And besides that, you can find me on my podcast, the other side of weight loss, or we talk a ton about hormones and weight loss for women. We just hit 1 million downloads, so that's very exciting. So we've got 260 episodes on there. And then on social media, you can find me at Karen Martel hormones, Speaker 1 (54:14): Karen Martel hormones, and, and your podcast. Absolutely, definitely check it out. We'll have links to all of that in the show notes. Thank you so much for your journey, your wisdom and your passion for helping women. Thank you for being here, Karen. Speaker 3 (54:29): Thanks for having me. Speaker 1 (54:31): And thank you for joining me for another episode of The Hormone Prescription with Dr. Kirin. So glad that you joined us today for this insightful discussion with maybe some things you hadn't thought about when it comes to your health, whether you have a weight problem or not. Looking at the deeper message of the malady or illness that you're experiencing can really help give you some insights that you can work towards healing and may just be the root cause that's keeping you stuck and you learned about a novel tool that you might want to consider using to help you. If you are struggling with weight loss, I'm gonna try it too and we'll share notes and see how it works, and I look forward to seeing you next week on another episode of The Hormone Prescription. Until then, peace, love, and Hormones. Y'all talk to you soon. Speaker 2 (55:20): Thank you so much for listening. I know that incredible vitality occurs for women over 40 when we learn to speak hormone and balance these vital regulators to create the health and the life that we deserve. If you're enjoying this podcast, I'd love it if you'd give me a review and subscribe. It really does help this podcast out so much. You can visit the hormone prescription.com where we have some free gifts for you, and you can sign up to have a hormone evaluation with me on the podcast to gain clarity into your personal situation. Until next time, remember, take small steps each day to balance your hormones and watch the wonderful changes in your health that begin to unfold for you. Talk to you soon.   ► Take Karen Martel's Hormone Quiz and find out which hormones could be stopping you from losing weight - CLICK HERE.   ► Feeling tired? Can't seem to lose weight, no matter how hard you try? It might be time to check your hormones. Most people don't even know that their hormones could be the culprit behind their problems. But at Her Hormone Club, we specialize in hormone testing and treatment. We can help you figure out what's going on with your hormones and get you back on track. We offer advanced hormone testing and treatment from Board Certified Practitioners, so you can feel confident that you're getting the best possible care. Plus, our convenient online consultation process makes it easy to get started. Try Her Hormone Club for 30 days and see how it can help you feel better than before. CLICK HERE.   If you enjoyed this episode, don't forget to subscribe, rate, and review The Hormone Prescription Podcast on your preferred podcast platform. Your support helps us reach even more listeners, allowing them to benefit from our expert advice and knowledge. ✨  

Link to bioRxiv paper: http://biorxiv.org/cgi/content/short/2023.03.15.532834v1?rss=1 Authors: Howell, J. A., Gaouette, N., Lopez, M., Burke, S. P., Perkins, E., Bidwell, G. L. Abstract: Inflammatory processes are activated following ischemic strokes and lead to increased tissue damage for weeks following the ischemic insult, but there are no approved therapies that target this inflammation-induced secondary injury. Here, we report that SynB1-ELP-p50i, a novel protein inhibitor of the nuclear factor kappa B (NF-{kappa}B) inflammatory cascade bound to drug carrier elastin-like polypeptide (ELP), is able to enter both neurons and microglia, cross the blood-brain barrier, localize exclusively in the ischemic core and penumbra in Wistar-Kyoto and spontaneously hypertensive rats (SHRs), and reduce infarct volume in male SHRs. Additionally, in male SHRs, SynB1-ELP-p50i treatment improves survival for 14 days following stroke with no effects of toxicity or peripheral organ dysfunction. These results show high potential for ELP-delivered biologics for therapy of ischemic stroke and other central nervous system disorders and further support targeting inflammation in ischemic stroke. Copy rights belong to original authors. Visit the link for more info Podcast created by Paper Player, LLC

Listen to a press release about a new research paper published by Oncotarget, entitled, “Targeted elastin-like polypeptide fusion protein for near-infrared imaging of human and canine urothelial carcinoma.” _______________________________ A new research paper was published in Oncotarget on September 6, 2022, entitled, “Targeted elastin-like polypeptide fusion protein for near-infrared imaging of human and canine urothelial carcinoma.” Bladder cancer (BC) is the 10th most common malignancy, affecting more than half a million people worldwide each year, and accounts for 4.6% of the total new cancer cases in the United States. With urothelial carcinoma (UC), the most common form of BC, the 5-year BC recurrence rate is nearly 78%, necessitating life-long surveillance, making it one of the costliest cancers to treat and manage. Cystoscopic visualization of bladder cancer is an essential method for initial bladder cancer detection and diagnosis, transurethral resection, and monitoring for recurrence. Researchers Aayush Aayush, Saloni Darji, Deepika Dhawan, Alexander Enstrom, Meaghan M. Broman, Muhammad T. Idrees, Hristos Kaimakliotis, Timothy Ratliff, Deborah Knapp, and David Thompson from Purdue University and Indiana University sought to develop a new intravesical imaging agent that is more specific and sensitive using a polypeptide based NIR (near-infrared) probe designed to detect cells bearing epidermal growth factor receptors (EGFR) that are overexpressed in 80% of urothelial carcinoma (UC) cases. The NIR imaging agent consisted of an elastin like polypeptide (ELP) fused with epidermal growth factor (EGF) and conjugated to Cy5.5 to give Cy5.5-N24-EGF as a NIR contrast agent. In addition to evaluation in human cells and tissues, the agent was tested in canine cell lines and tissue samples with naturally occurring invasive UC. “Dogs with naturally-occuring UC are an emerging option for a suitable large animal model of BC, where the cancer displays similar microscopic anatomy, histological appearance, biological behavior, heterogeneity, and molecular subtypes and markers to human invasive BC.” Full press release - https://www.oncotarget.com/news/pr/oncotarget-targeted-elastin-like-polypeptide-fusion-protein-for-near-infrared-imaging-of-human-and-canine-urothelial-carcinoma/ DOI: https://doi.org/10.18632/oncotarget.28271 Correspondence to: David Thompson – davethom@purdue.edu Keywords: bladder cancer, elastin-like polypeptide, NIR imaging, epidermal growth factor receptor (EGFR), translational studies About Oncotarget: Oncotarget is a primarily oncology-focused, peer-reviewed, open access journal. Papers are published continuously within yearly volumes in their final and complete form, and then quickly released to Pubmed. To learn more about Oncotarget, visit Oncotarget.com and connect with us: Twitter – https://twitter.com/Oncotarget Facebook – https://www.facebook.com/Oncotarget YouTube – www.youtube.com/c/OncotargetYouTube Instagram – https://www.instagram.com/oncotargetjrnl/ LinkedIn – https://www.linkedin.com/company/oncotarget/ Pinterest – https://www.pinterest.com/oncotarget/ LabTube – https://www.labtube.tv/channel/MTY5OA SoundCloud – https://soundcloud.com/oncotarget For media inquiries, please contact: media@impactjournals.com

Voyage dans le temps et l'espace mondial pour la séquence d'orthopédie musicale. Après ce dimanche pluvieux, nous sommes fiers que notre opération Sieste Géante n'ait pas trop abimé votre brushing de télé achat et que vous ayez recouvré une motricité décente. Vous voilà donc prêt à accueillir la meilleure nouvelle depuis la découverte de Vénus […] L'article Crossover – Cosinus et Polypeptide est apparu en premier sur Radio Campus Tours - 99.5 FM.

Peptide sind nichts anderes als kleine Eiweiße (Proteine). Es gibt wieder Unterteilungen wie z.B. Oligopeptide, bis hin zu Polypeptide. Allen gemeinsam ist aber , dass sie aus Aminosäuren bestehen. Wie werden Peptide in der Kosmetik gewonnen oder hergestellt? Peptide werden oft synthetisch hergestellt. Aber es gibt auch Natürliche (wie Thymus- oder Colostrum-Peptide) oder naturidentische Peptide. Die technologische Verarbeitung in Form von einem Trägersystem ist nämlich besonders wichtig. Peptide können wesentlich besser in die spezifischen Hautschichten ein penetrieren und Ihre Wirkung entfalten, wenn die hauteigenen Abwehrmechanismen erst gar nicht aktiv werden müssen. Hier eignet sich z.B wenn ein Pflegeprodukt Liposomal verkapselt ist. Die Freisetzung ist sehr gleichmäßig, ein Grund dafür, dass in Liposomen verkapselte Wirkstoffe für die Haut auch sehr verträglich sind. In der Idealkonstellation werden Peptide noch mit weiteren wertvollen Inhaltsstoffen (wie Vitamin A, C, E, Pro-Vitamin B5, Hyaluronsäure u.ä.) kombiniert.

My AP Biology Thoughts  Episode #41Welcome to My AP Biology Thoughts podcast, my name is Arthur and I am your host for episode 41 called Unit 1: Monomers and Polymers of the Macromolecules. Today we will be discussing dehydration and hydrolysis reactions in regards to macromolecules. Segment 1: Introduction to Monomers and Polymers of the MacromoleculesThe formation and breaking of macromolecules are essential for complex life to function. We will be discussing the chemical mechanisms by which macromolecules both form and break down.   Monomers: The unit components of the larger macromolecules. Polymers: What is formed when the monomers bond together, which are known as macromolecules. Dehydration synthesis: A water molecule being ejected in order to allow for a monomer to covalently bond to another monomer or polymer. Hydrolysis: A polymer splitting apart after reacting with a water molecule.   Activation energy: An energy threshold, which must be met in order for a reaction to proceed.   Segment 2: Example of Monomers and Polymers of the MacromoleculesMonosaccharides are the monomers of carbohydrates. In the presence of the necessary enzyme, a hydroxyl group is ripped off one of the monosaccharides and a hydrogen off the hydroxyl group of the other. This results in the two monosaccharides bonding together via an ether (glycosidic) bond as well as the formation of a water molecule. Conversely, this reaction can happen in reverse via hydrolysis.   The formation of triglycerides involves dehydration synthesis. An ester linkage is created between a fatty acid and a glycerol and a water molecule is released. A hydrogen is ripped of the carboxylic acid from the fatty acid and the hydroxyl is ripped off the glycerol. Polypeptide chains form via dehydration synthesis between two amino acids. An OH is ripped off the carboxylic acid and a hydrogen is ripped off the amine group, allowing the amino acids to covalently bond as well as allowing the formation of a water molecule. This occurs many times allowing for long polypeptide chains to form, which eventually leads to proteins.   Segment 3: Digging Deeper Monomers and Polymers of the MacromoleculesThe creation of macromolecules from monomers gives complex life its structure (ex. Cellulose). Additionally, the breaking down of macromolecules is essential for metabolism and the operations of cells (ex. Breaking down polysaccharides down to glucose for cellular respiration).   Thank you for listening to this episode of My AP Biology Thoughts. For more student-ran podcasts and digital content, make sure that you visit http://www.hvspn.com/ (www.hvspn.com). See you next time! Music Credits:“Ice Flow” Kevin MacLeod (incompetech.com) Licensed under Creative Commons: By Attribution 4.0 License http://creativecommons.org/licenses/by/4.0/ Subscribe to our Podcasthttps://podcasts.apple.com/us/podcast/my-ap-biology-thoughts/id1549942575 (Apple Podcasts) https://open.spotify.com/show/1nH8Ft9c9f6dmo75V9imCk?si=IvI4iQV-SSaFb0ZmvTabxg (Spotify) https://podcasts.google.com/feed/aHR0cHM6Ly9mZWVkcy5jYXB0aXZhdGUuZm0vbXlhcGJpb2xvZ3l0aG91Z2h0cw (Google Podcasts  ) https://www.youtube.com/channel/UC07e_nBHLyc_nyvjF6z-DVg (YouTube)   Connect with us on Social MediaTwitterhttps://twitter.com/thehvspn ( )https://twitter.com/thehvspn (@thehvspn)

This episode covers vasoactive intestinal polypeptide!

Link to bioRxiv paper: http://biorxiv.org/cgi/content/short/2020.11.11.378026v1?rss=1 Authors: Suvlu, D., Thirumalai, D., Rasaiah, J. C. Abstract: Water-mediated interactions (WMIs) play diverse roles in molecular biology. They are particularly relevant in geometrically confined spaces such as the interior of the chaperonin, at the interface between ligands and their binding partners, and in the ribosome tunnel. Inspired in part by the geometry of the ribosome tunnel, we consider confinement effects on the stability of peptides. We describe results from replica exchange molecular dynamics simulations of a system containing a 23-alanine or 23-serine polypeptide confined to non-polar and polar nanotubes in the gas phase and when open to a water reservoir. We quantify the effect of water in determining the preferred conformational states of these polypeptides by calculating the difference in the solvation free energy for the helix and coil states in the open nanotube in the two phases. Our simulations reveal several possibilities. We find that nanoscopic confinement preferentially stabilizes the helical state of polypeptides with hydrophobic side chains, which is explained by the entropic stabilization mechanism proposed on the basis of polymer physics. Polypeptide chains with hydrophilic side chains can adopt helical structures within nanotubes, but helix formation is sensitive to the nature of the nanotube due to WMIs. We elaborate on the potential implications of our findings to the stability of peptides in the ribosome tunnel. Copy rights belong to original authors. Visit the link for more info

Transcription and Translation Importance of mRNA and tRNA Function/importance of polypeptide synthesis  Genes/environment affect phenotypic expression  Structure/function of proteins Thanks to STEM Reactor for sponsoring this podcast. They provide everything you need to do biotechnology at school, check them out at www.stemreactor.com.au

Serie EVOLUTION - WISSENSCHAFT? mit Dr. med. univ. Klaus Gstirner Ist die Evolutionstheorie wissenschaftlich begründbar? Was sind die Aussagen der modernen Wissenschaft des 21. Jahrhunderts zur Evolutionslehre? 5.Die Entstehung von Eiweißen Für die Entstehung von Leben, benötigt es Eiweiße (Polypeptide). Diese entstehen in der Stufe zwei der Entstehung des Menschen. Aber wie entstand der Herzkreislauf oder die Atmung wirklich? Kann es eine langsame Entwicklung von einfachen zu komplexen Strukturen geben? Im kommenden Video finden sie Antworten. Für Videoaufnahme: https://vimeo.com/396303035

Juggling a relationship and a STEM degree in college can be a daunting task. If you succeed, then come to the hard decisions of managing duel careers. Join me as Miguel and Natalie Sanchez share their insights on how they managed to get through school and careers, at Chevron and PolyPeptide group respectively, in the mayhem of southern California.  Miguel is a Chemical Engineer and Natalie an Environmental Scientist by degree. They live in southern California where they also work. They attribute MAES for having a huge impact on their success as both students and now professionals  Miguel Sanchez:https://www.linkedin.com/in/miguel-sanchez-60b2b426/ Chevron:https://www.linkedin.com/in/natalie-ambriz-sanchez-ab3a71167/ Natalie Sanchez: https://www.linkedin.com/in/natalie-ambriz-sanchez-ab3a71167/ PolyPeptide Group:https://www.polypeptide.com/ MAES:http://mymaes.org/ MUSIC Song: (Royalty Free Music) Utopia - Ikson Music promoted by Tyler Montgomery. Video Link: https://youtu.be/GXcUofHwOkc  --- Send in a voice message: https://anchor.fm/manny-de-la-cruz3/message

The cover for issue 2 of Oncotarget features Figure 5, "MASPIN can prevent the formation of UPA - UPA-receptor complex by a single step, and thus decrease the possibility of the abnormal degradation of the ECM, the development metastasis and angiogenesis," by Fodor, et al. In the present study, the research team investigated AEZS-108 induced cytotoxicity and the altered mRNA expression profile of regulatory factors related to angiogenesis and metastasis in LHRH receptor-positive OCM3 cells. Their results show that AEZS-108 upregulates the expression of MASPIN/SERPINB5 tumor suppressor gene, which is downregulated in the normal uvea and UM specimens independently from the LHRH receptor-ligand interaction. In order to investigate the mechanism of the induction of MASPIN by AEZS-108, OCM3 cells were treated with free DOX, D-Lys6 LHRH analog, or AEZS-108. Dr. Gabor Halmos from the University of Debrecen, Department of Biopharmacy, in Debrecen, Hungary as well as the Veterans Affairs Medical Center, Endocrine, Polypeptide and Cancer Institute, in Miami, FL, USA said, "Although uveal melanoma (UM) is a rare disease, it is the most prevalent lethal ophthalmological tumor." The discovery of specific receptors for peptide hormones on cancer cells has led to the development of cytotoxic or radiolabeled hormone analogs that are appropriate for tumor localization and targeted therapy. AEZS-108 guides the chemotherapeutic agent specifically to those tumors that express LHRH-receptors, which could result in targeted cytotoxicity and less damage to healthy tissues. The authors reported that previously OCM3 UM cell line expresses the receptor of LHRH localized on the cell membrane and in the cytoplasm, rendering them susceptible to AEZS-108 uptake and the detection of the LHRH receptor in OCM3 cells has led to the use of AEZS-108 for targeted therapy of the tumor. Moreover, that the OCM3 UM cell line expresses the LHRH receptor and LHRH rendering them susceptible to AEZS-108 uptake. The Halmos Research Team concluded, "In summary, our data confirmed previous results showing LHRH receptor expression in OCM3 cells, a UM in vitro model. Furthermore, we report for the first time that AEZS-108 causes changes in the expression of genes that are involved in angiogenesis and ECM degradation and which might inhibit cell proliferation and induce apoptosis in OCM3 cells. These findings suggest that AEZS-108 plays a pivotal role in the regulation of angiogenesis and tumor suppression. Taken together, targeted cytotoxic LHRH analogs, such as AEZS-108, might serve as an effective treatment for patients with LHRH receptor-positive uveal melanoma.” Keywords - uveal melanoma; luteinizing hormone-releasing hormone (LHRH) receptor; angiogenesis; MASPIN/SERPINB5; AEZS-108 (AN-152/Zoptarelin Doxorubicin Acetate DOI - https://doi.org/10.18632/oncotarget.27431 Full text - http://www.oncotarget.com/index.php?journal=oncotarget&page=article&op=view&path%5B%5D=27431&path%5B%5D=89358 Correspondence to - Gabor Halmos, email: halmos.gabor@pharm.unideb.hu About Oncotarget Oncotarget is a weekly, peer-reviewed, open-access biomedical journal covering research on all aspects of oncology. Oncotarget is published by Impact Journals, LLC. To learn more please visit http://www.oncotarget.com or connect with: Facebook - https://www.facebook.com/Oncotarget/ Twitter - https://twitter.com/oncotarget LinkedIn - https://www.linkedin.com/company/oncotarget Pinterest - https://www.pinterest.com/oncotarget/ Reddit - https://www.reddit.com/user/Oncotarget/

Sat, 20 Jul 2013 12:00:00 +0100 https://edoc.ub.uni-muenchen.de/16855/ https://edoc.ub.uni-muenchen.de/16855/1/Streckel_Elisabeth_Therese.pdf Streckel, Elisabeth Therese

Hosts: Vincent Racaniello, Rich Condit, and Kathy Spindler Vincent, Rich and Kathy discuss mechanisms of protein synthesis and regulation in virus-infected cells. Links for this episode: Slides for this episode (pdf) T7 ribosomal binding sites (CSH Symp Quant Biol) Tinkering with translation (CSH Persp) Translation at nexus of infection and immunity (Cell Host Microbe) Viral subversion of protein synthesis machinery (Nat Rev Micr) Weekly Science Picks Kathy - Vaccine-preventable outbreaksRich -  To Catch a Virus by John Booss and Marilyn J. AugustVincent - This is Water by David Foster Wallace Listener Pick of the Week Jon - Confessions of a converted lecturer by Eric MazurPeter - FameLab Send your virology questions and comments (email or mp3 file) to twiv@twiv.tv

MDS presents the latest research and findings from the field of Movement Disorders. Abstracts of articles from the Society Journal, Movement Disorders, are taken from the April 2013 (Vol. 28, Issue 4) issue.

Hosts: Vincent Racaniello, Rich Condit, Dickson Despommier, Alan Dove, and Kathy Spindler The complete TWiV team reviews identification of the cell receptor for hepatitis B and D viruses, and the cell enzyme that cleaves the genome-linked protein from picornaviral RNA. Links for this episode: HBV and HDV cell receptor (eLIFE) Getting to grips with hepatitis (eLIFE) Sodium taurocholate Poliovirus hijacks DNA repair enzyme (PNAS) Letters read on TWiV 210 Weekly Science Picks Rich - Vendée GlobeDickson - Earth at nightAlan - Trophée Jules VerneKathy - Science cookiesVincent - Trilobite Glassworks Listener Pick of the Week Ken - Spanish flu pandemic model (teacher's guide)Jim - Santa Cruz Science Communication Program (also see SHERP) Send your virology questions and comments (email or mp3 file) to twiv@twiv.tv

This Unit studies 'proteins'. Starting with a simple analysis of the molecular make up, the Unit moves on to look at the importance of protein and how they are digested and absorbed. This study unit is just one of many that can be found on LearningSpace, part of OpenLearn, a collection of open educational resources from The Open University. Published in ePub 2.0.1 format, some feature such as audio, video and linked PDF are not supported by all ePub readers.

The incretin hormones GIP (glucose-dependent insulinotropic polypeptide) and glucagon-like peptide-1 (GLP-1) mediate the so-called incretin effect, which describes the phenomenon that glucose given orally causes a higher insulin response compared to an isoglycemic intravenous glucose load. The insulinotropic action of GIP is reduced to almost absent in type 2 diabetes patients, while the action of GLP-1 is vastly preserved. GIPRdn transgenic pigs were generated by lentiviral genetransfer to establish a large animal model to investigate the effects of an impaired insulinotropic action of GIP on glucose homeostasis. At the age of 5 months GIPRdn transgenic pigs showed a disturbed oral glucose tolerance going along with reduced insulin secretion. Eleven-month-old GIPRdn transgenic pigs exhibited an impaired intravenous glucose tolerance and reduced insulin secretion as well as a significantly reduced total β-cell volume compared to controls. In this work different age classes (11 weeks, 5 months, 1-1.4 years) of GIPRdn transgenic were investigated to obtain detailed data about physiological and morphological characteristics. To proof specifity of the GIPRdn GIP/Exendin 4 stimulation tests were carried out in 11 week-old GIPRdn transgenic pigs. The insulinotropic action of intravenously injected porcine GIP was impaired, while this of Exendin-4, a GLP-1 mimetic, was enhanced in GIPRdn transgenic pigs compared to controls. Marked alterations in the expression profile of the GIPR and the GLP-1R were excluded as no apparent differences of immunohistochemically stained pancreas sections for GIPR and GLP-1R were detectable between GIPRdn transgenic pigs and controls at any age group. The effects of the impaired insulinotropic action of GIP on glucose metabolism were investigated by oral and intravenous glucose tolerance tests. Eleven-week-old GIPRdn transgenic pigs exhibited significantly reduced oral glucose tolerance with a delay in insulin secretion compared to controls. The area under the insulin curve (AUC insulin) during the first 45 minutes following glucose load was 31% smaller in transgenic pigs compared to controls. The total amount of insulin secretion was not different between the two groups indicating that GIPRdn expression initially only interferes with the incretin effect. This was supported by the fact that intravenous glucose tolerance and insulin secretion in transgenic pigs were not different from controls. Five-month-old GIPRdn transgenic pigs exhibited a tendency towards reduced intravenous glucose tolerance and reduced insulin secretion in response to an intravenous glucose challenge. To determine the reason for the alterations in glucose metabolism quantitative stereological analyses of the pancreas were performed. In 11-week-old pigs, transgenic and control groups showed similar β-cell mass. However, pancreatic -cell mass was reduced by almost 40% in 5-month-old and by 60% in adult (1 1.4 years) GIPRdn transgenic pigs compared to controls. Furthermore, the cellular composition of the islets was analyzed by quantitative stereological investigations. The relative volumes of α- and -cells in the islets were increased in 1-1.4-year-old GIPRdn transgenic pigs but the absolute volumes of these non-β-cell populations were not different from those of age-matched controls. To investigate the reason for the reduced pancreatic β-cell mass in GIPRdn transgenic pigs, β-cell proliferation and apoptosis rate was determined performing a double-immunohistochemistry for insulin and the proliferation marker Ki67 and cleaved caspase-3, respectively. Eleven-week-old GIPRdn transgenic pigs showed significantly less Ki67 positive cell nuclei compared to controls, whereas proliferation rates in 5 month-old and 1-1.4-year-old GIPRdn transgenic pigs reached no statistical significance. No differences were shown in the apoptosis rates of GIPRdn transgenic pigs compared to controls at any age, although a trend of higher numbers of cleaved caspase-3 positive β-cells was visible in 1-1.4-year-old GIPRdn transgenic pigs. In conclusion, GIPRdn transgenic pigs exhibit a comparable situation as in type 2 diabetes mellitus patients like impaired insulinotropic action of GIP, disturbed glucose tolerance and reduced β-cell mass. Moreover, the results of this work demonstrate an essential role of GIP for the physiological expansion of β-cell mass. In this context GIPRdn transgenic pigs represent a valuable model for further investigations on type 2 diabetes mellitus including diet studies and therapeutic trials.

Wed, 21 Jul 2010 12:00:00 +0100 https://edoc.ub.uni-muenchen.de/11936/ https://edoc.ub.uni-muenchen.de/11936/1/Denschlag_Robert.pdf Denschlag, Robert ddc:530, ddc:500, Fakultät für Physik

Transcript -- A more detailed look at transcription, showing how messenger RNA is formed.

Transcript -- A more detailed look at transcription, showing how messenger RNA is formed.

A more detailed look at transcription, showing how messenger RNA is formed.

A more detailed look at transcription, showing how messenger RNA is formed.

Vorangegangene Studien zeigten, dass das Prion-Protein (PrP) an der ER-Membran in verschiedenen topologischen Isoformen synthetisiert wird und teilweise sogar im Zytosol vorliegen kann. Sowohl die ER-Signalsequenz als auch die hydrophobe Domäne von PrP wurden dabei als Domänen identifiziert, die eine Rolle in der Translokation spielen. Die hier durchgeführte Analyse des ER-Imports von PrP und verschiedenen chimären Proteinen hat nun erstmals gezeigt, dass auch der Faltungszustand von Polypeptiden Einfluss auf deren Translokation ins ER-Lumen haben kann. Die vorliegende Studie ergab, dass - unabhängig von der Primärstruktur - ein gewisses Maß an α-helikalen Bereichen notwendig für einen produktiven ER-Import ist. Sowohl die Länge der Polypeptide als auch posttranslationelle Modifikationen wie die GPI-Verankerung, die N-Glykosylierung oder die Ausbildung einer Disulfidbrücke beeinflussen die Translokation nicht. Darüber hinaus deuten die Ergebnisse der vorliegenden Arbeit darauf hin, dass Proteine mit ausgedehnten unstrukturierten Domänen am N-Terminus einer kotranslokationalen Qualitätskontrolle unterliegen und noch vor der Translokation ins ER-Lumen einer proteasomalen Degradierung im Zytosol zugeführt werden. Die in dieser Doktorarbeit dargestellten Ergebnisse legen daher die Vermutung nahe, dass die Ausbildung von Sekundärstrukturen vor oder während der Translokation die weitere Biogenese des naszierenden Polypeptids beeinflusst. Die gewonnenen Erkenntnisse können dazu beitragen die physiologischen aber auch die möglichen pathophysiologischen Konsequenzen der Regulation der Translokation besser zu verstehen. Der zweite Teil der Arbeit erbrachte erstmals experimentelle Evidenzen, dass trotz sehr geringer Sequenzhomologie zwischen den PrP-Homologen im Zebrabärbling (Danio rerio) und Säugetier-PrP die charakteristischen posttranslationalen Modifikationen, wie beispielsweise die komplexe Glykosylierung und der C-terminale GPI-Anker, konserviert sind. Die neu etablierten Zellkulturmodelle zur Analyse von PrP-homologen Proteinen deuten auf eine evolutionär konservierte Funktion von PrP hin und könnten dazu beitragen, neue Einsichten in die physiologische Aktivität von PrP zu gewinnen.

Background: Gastric inhibitory polypeptide ( GIP) is postulated to be involved in type 2 diabetes mellitus and obesity. It exerts its function through its receptor, GIPR. We genotyped three GIPR SNPs (rs8111428, rs2302382 and rs1800437) in German families with at least one obese index patient, two case-control studies and two cross-sectional population-based studies. Methods: Genotyping was performed by MALDI-TOF, ARMS-PCR and RFLP. The family-study: 761 German families with at least one extremely obese child or adolescent (n = 1,041) and both parents ( n = 1,522). Case-control study: ( a) German obese children ( n = 333) and (b) obese adults ( n = 987) in comparison to 588 adult lean controls. The two cross-sectional population-based studies: KORA ( n = 8,269) and SHIP ( n = 4,310). Results: We detected over-transmission of the A-allele of rs2302382 in the German families (pTDT-Test = 0.0089). In the combined case-control sample, we estimated an odd ratio of 1.54 (95% CI 1.09; 2.19, pCA-Test = 0.014) for homozygotes of the rs2302382 A-allele compared to individuals with no A-allele. A similar trend was found in KORA where the rs2302382 A-allele led to an increase of 0.12 BMI units (p = 0.136). In SHIP, however, the A-allele of rs2302382 was estimated to contribute an average decrease of 0.27 BMI units (p-value = 0.031). Conclusion: Our data suggest a potential relevance of GIPR variants for obesity. However, additional studies are warranted in light of the conflicting results obtained in one of the two population-based studies.

Das kleine Hitzeschockprotein αB-Crystallin ist in vielen verschiedenen Gewebetypen u.a. in der Niere nachweisbar. Es wird sowohl unter physiologischen als auch unter pathophysiologischen Bedingungen exprimiert. Viele verschiedene Stressfaktoren induzieren in den betroffenen Zellen die Expression von αB-Crystallin, das – wie viele andere Hsps - als molekulares Chaperone die Zellen vor sonst letalen Umgebungseinflüssen schützt. Dies wird bewerkstelligt, indem es partiell oder komplett entfaltete, fehlgefaltete oder aggregierte Polypeptide bindet und deren native Konformation wiederherstellt, oder bei irreversibler Schädigung dem lysosomalen Abbau zuführt. Im Nierenmark stellen hohe extrazelluläre Konzentrationen an NaCl und Harnstoff, die im Rahmen der Harnkonzentrierung das interstitielle Milieu prägen, die größte Bedrohung für die Zellen der medullären Nephronanteile dar. Wie vorliegende Untersuchungen zeigen, exprimieren die Zellen der medullären Sammelrohre und der dünnen Schenkel der Henle-Schleife αB-Crystallin konstitutiv sehr stark und es macht im Innenmark etwa 2% des gesamten Zellproteins aus. Im Nierenkortex ist αB-Crystallin hingegen kaum nachweisbar. Da die αB-Crystallin Expression parallel zum osmotischen Gradienten vom Nierenkortex zum Innenmark ansteigt und in Diurese im Nierenmark signifikant abfällt, kann man davon ausgehen, dass die αB-Crystallin Expression wesentlich durch die interstitielle Osmolalität reguliert wird. Diese Annahme wird durch die vorliegenden in-vitro-Versuche mit MDCK-Zellen unterstützt, die zeigen, dass αB-Crystallin durch hypertonen Stress induzierbar ist. Interessanterweise führt auch eine erhöhte extrazelluläre Harnstoffkonzentration zur αB-Crystallin Induktion. αB-Crystallin wird daher vermutlich über andere Regulationsmechanismen induziert als beispielsweise Hsp72, ein gut untersuchtes Hsp in der Niere und TonEBP-Zielgen, welches durch erhöhte Harnstoffkonzentrationen nicht induzierbar ist. Dafür spricht auch, dass bei der Induktion von αB-Crystallin die JNK MAP-Kinase eine Rolle spielt und nicht die p38 MAP-Kinase, die an der Expression vieler anderer durch Hypertonizität induzierbarer Gene, u.a. Hsp72, beteiligt ist. Interaktionen zwischen αB-Crystallin und Stukturen des Zytoskeletts konnten im Rahmen dieser Arbeit nicht nachgewiesen werden. Die Ergebnisse lassen vermuten, dass αB-Crystallin im Rahmen der Harnkonzentrierung eine bedeutende Funktion für die Zellen im Innenmark der Niere zukommt und dass dessen Expression nicht nur durch hypertonen Stress, sondern auch durch hohe Harnstoffkonzentrationen und vermutlich mehrere andere Stressfaktoren im Nierenmark reguliert wird.

Fri, 8 Feb 2008 12:00:00 +0100 https://edoc.ub.uni-muenchen.de/11089/ https://edoc.ub.uni-muenchen.de/11089/1/Renner_Simone.pdf Renner, Simone ddc:590, d

Die vorliegende Arbeit wurde in zwei Bereiche gegliedert. Der erste Themenbereich beschäftigte sich mit der Transformation von pflanzlichen Mitochondrien bzw. mit den Voraussetzungen, die erfüllt sein müssen, um Mitochondrien höherer Pflanzen zu transformieren. Im zweiten Themenbereich wurde die Regulation der Lysinbiosynthese erstmals mit Hilfe der Plastidentransformation modifiziert sowie ein universeller plastidärer Selektionsmarker entwickelt und etabliert. Mitochondrientransformation- In dieser Arbeit wurden zwei Strategien zur Transformation von Mitochondrien höherer Pflanzen entwickelt. Zum einen wurde ein mitochondrienspezifischer Ansatz gewählt, d.h. es wurden Selektionsmarker verwendet, die eine hemmende Wirkung auf die Atmungskette der Mitochondrien besitzen. Zum anderen wurden in Anlehnung an seit die seit 1990 erfolgreich durchführbare Plastidentransformation generelle Inhibitoren der Proteinbiosynthese verwendet, welche durch die Produkte der entsprechenden in die Mitochondrien eingebrachten Resistenzgene detoxifiziert werden sollten. Markergenstrategie- Geeignete Selektionsbedingungen zur Identifizierung von mitochondrialen Transformanten wurden bei Tabakblättern und Tabakprotoplasten mit den Antibiotika Blasticidin, Chloramphenicol und Hygromycin als Hemmstoffe ermittelt. Aus Transformationen mit den mitochondrialen Transformationskassetten pBMhph II, pBMhph III und pBMhph IV konnten über 200 Hygromycin-resistente Linien selektiert und charakterisiert werden. PCR- und Southern-Analysen lassen bei mindestens 7 Linien eine zielgerichtete mitochondriale Integration des Transgens vermuten. Möglicherweise sind aber nur wenige Kopien des Transgens in das Mitochondriengenom inseriert bzw. repliziert worden, was eine eindeutige Charakterisierung erschwert. Bei einer großen Anzahl resistenter Linien handelt es sich wahrscheinlich um funktionelle zielortfremde Integrationen der Transformationskassetten in die Plastiden-, Kern- oder Mitochondriengenome. Mitochondrienspezifischer Transformationsansatz- Die in dieser Arbeit entwickelten mitochondrienspezifischen Transformationsvektoren pUMmyx II, pUMglu II, pUMarg II und pUManti II enthalten ein Fragment des mitochondrialen cob-Gens. In dieses cob-Fragment wurden Punktmutationen eingeführt, die zu einer Veränderung der Sekundär- bzw. Tertiärstruktur des Apocytochroms b führen. Bei einer korrekten homologen Integration dieses modifizierten cob-Fragmentes in das Mitochondriengenom sind die Inhibitoren Antimycin A, Myxothiazol und Moa-Stilben nicht mehr in der Lage, an den Komplex III der mitochondrialen Atmungskette zu binden. Zur Bestimmung optimaler Selektionsbedingungen von mitochondrialen Transformanten mit den Hemmstoffen Antimycin A, Myxothiazol und Moa-Stilben wurden Testreihen mit Tabakblättern, Tabakprotoplasten sowie Tabak- und Arabidopsis-Suspensionskulturen durchgeführt. Bei „particle-gun“-Transformationen von Tabakblättern mit der Transformationskassette pUMarg II wurden 23 Moa-Stilben-resistente Linien selektiert. Bei 18 Linien konnte eine Integration des mutierten cob-Fragmentes eindeutig per PCR und Sequenzierung nachgewiesen werden. Ob es sich dabei um eine mitochondriale Integration handelt, konnte nicht eindeutig belegt werden. In diesem Fall wäre jedoch zu klären, auf welchem Mechanismus der offensichtliche selektive Vorteil der mit dem Transformations-vektor transformierten Pflanzen beruht, da die Integration des cob-Gen-Fragmentes nur innerhalb des mitochondrialen Zielortes funktionell sein dürfte. Plastidentransformation- Bei dem plastidären Transformationsansatz dieser Arbeit wurden zwei Ziele verfolgt. Zum einen wurde der „feedback“-Regulationsmechanismus der DHDPS innerhalb des Aspartat-Stoffwechsels durch die Integration eines insensitiven dhdps-Gens in die Plastiden von Tabak und Kartoffel beeinträchtigt. Der Gehalt der essentiellen Aminosäure Lysin wird somit gesteigert. Zum anderen wurde das insensitive dhdps-Gen als Markergen genutzt, um eine Selektion von plastidären Transformanten auf dem Lysinanalogon AEC zu ermöglichen. Erhöhung des Lysingehaltes- Es konnten 34 Spectinomycin-resistente Tabakklone selektiert werden, die mit der Transformationskassette pHoDh2b transformiert wurden. Die molekularbiologische Charakterisierung der Transformanten konnte bei 8 Linien eindeutig eine korrekte plastidäre Integration der Transgene belegen. Biochemische Analysen zeigen eine bis zu 32-fache Erhöhung an freiem Lysin im Blattgewebe von plastidären Transformanten. Es war damit erstmals mit der Plastidentransformation möglich, regulatorisch die Lysinbiosynthese zu verändern. AEC als plastidärer Selektionsmarker- AEC (S-Aminoethyl-L-Cystein) konkurriert als Lysinanalogon mit Lysin um den Einbau in Polypeptide. Wird die AEC-Konzentration im pflanzlichen Gewebe im Vergleich mit Lysin zu hoch, kommt es zum Erliegen des Stoffwechsels und damit zum Absterben der Zelle. Durch die Überproduktion von Lysin in den Transformanten verschiebt sich das Verhältnis von AEC → Lysin. Es wird somit kompetitiv mehr Lysin in die Polypeptide eingebaut. Dies lässt sich als Selektionsvorteil nutzen. Mit dem „Markergen“ dhdps-r1 und AEC als selektivem Hemmstoff wurden insgesamt 49 Tabaklinien selektiert. Bei 3 selektierten Linien konnte eindeutig die korrekte plastidäre Intergration der Transformationskassette nachgewiesen werden. Es ist damit erstmals gelungen, einen antibiotikumfreien universell einsetzbaren plastidären Selektionsmarker zu etablieren. In weiteren Transformationsexperimenten konnten mit dem gleichen „Markergen“ plastidäre Tomatentransformanten identifiziert und charakterisiert werden (Diplomarbeit L. Schaeffer). Damit ist es bereits bei 2 Spezies gelungen, AEC als selektiven Hemmstoff einzusetzen. Bei weiterer Optimierung der Selektionsbedingungen dürfte es möglich sein, auch bei weiteren Pflanzenspezies diesen universellen Selektionsmarker zu etablieren.

Im Unterschied zu den HLA-Klasse-Ia-Molekülen ist das nicht-polymorphe Klasse-Ib-Molekül HLA-G v.a. in der Plazenta exprimiert. Es wird postuliert, daß HLA-G die Immuntoleranz des mütterlichen Immunsystems gegen den „semiallogenen“ Fetus mitreguliert. In allen anderen Zellen und Geweben, in denen es gefunden wurde, ist die Menge im Vergleich zu klassischen Klasse-I-Molekülen um Größenordnungen geringer. Auch in der Genexpression unterscheidet sich HLA-G drastisch von allen übrigen Klasse-I-Molekülen. Am auffallendsten ist dabei das Auftreten verschiedener alternativer Spleißformen. Neben dem Volle-Länge-Transkript G1m gibt es eine Reihe verkürzter Isoformen: G2 (G2m, Da2-Domäne), G3 (Da2/Da3-Domäne), G4 (Da3-Domäne) sowie zwei Formen, die für lösliche Proteine kodieren, da die nicht-entfernte Intron-4-Sequenz zu einem vorzeitigen Translationsstop führt, G5 (G1s), G6 (G2s, Da2-Domäne, + In4). Die schwache Expression von HLA-G bestätigte sich auch für die im Rahmen dieser Arbeit untersuchten Haut- und Muskelbiopsien sowie Gehirnproben. Transkripte für HLA-G und die verkürzten Isoformen waren in fast allen Proben nachweisbar, wobei das Volle-Länge-Transkript die dominante Form war. Bei den nach Krankheitsgruppen eingeteilten Hautbiopsien und den Muskelbiopsien mit definierten Diagnosen konnte keine Korrelation eines bestimmten Expressionsmusters mit einer bestimmten Krankheitsgruppe bzw. Diagnose festgestellt werden. Diese Heterogenität des Expressionsmusters sowie die selektive Hochregulation der Volle-Länge-Isoform G1m auf Transkriptions- und Proteinebene in Glioblastomzellinien und Myoblasten, die nur eine äußerst schwache konstitutive Expression von HLA-G aufweisen, nach Behandlung mit IFNg deutet auf eine differentielle Regulation hin. Um die einzelnen Isoformen getrennt voneinander untersuchen zu können, wurden Transfektanten für jede Form in der Klasse-I-negativen B-Zellinie 721.221 etabliert. Nur die Volle-Länge-Isoform G1m sowie deren lösliche Variante G1s konnten auf der Zelloberfläche bzw. im Kulturüberstand nachgewiesen werden. Von den übrigen Isoformen konnten nur EndoH-sensitive Polypeptide gefunden werden, und auch Immunofluoreszenzfärbung mit einer Reihe von Klasse-I-Ak zeigte keine Zelloberflächenexpression. Es muß daraus geschlossen werden, daß die verkürzten Isoformen in der Zelle zurückgehalten werden. HLA-G kann auf zwei Wegen die Aktivität von Immuneffektorzellen regulieren: direkt über ILT2 und indirekt über HLA-E. Das MHC-Klasse-I-Molekül HLA-E wird durch Bindung eines Nonamers (P3-11) aus dem Signalpeptid verschiedener Klasse-I-Moleküle auf der Oberfläche von Zellen stabilisiert. Aus der Interaktion dieses funktionellen HLA-E/Peptid-Komplexes mit dem inhibitorischen Rezeptorkomplex CD94/NKG2A auf NK-Zellen resultiert ein Schutz dieser Zellen vor der NK-Lyse. Auch das entsprechende Peptid aus der Signalsequenz von HLA-G ist ein Ligand für HLA-E. Allerdings wurde gezeigt, daß die Stabilisierung von HLA-E auf der Zelloberfläche durch das Peptid G3﷓11 schwächer als mit anderen Klasse-I-Peptiden und auch weniger stabil ist. Effektive Inhibition der Lyse der NK-Zellinie NKL über diese Interaktion von HLA-E mit CD94/NKG2A findet man nur bei HLA-G1m-Transfektanten. Diese werden auch durch die direkte Interaktion von HLA-G1m mit einem weiteren inhibitorischen Rezeptor auf NKL - ILT2 - geschützt. In den 721.221-Transfektanten der verkürzten HLA-G-Isoformen war eine unphysiologisch hohe Konzentrationen an HLA-G-Polypetid notwendig, um die kritische Menge an HLA-E-Ligand für den Schutz dieser Zellen vor der Lyse durch NK-Zellen liefern zu können. Das war nur für eine äußerst stark exprimierende G3-Transfektante der Fall. Der Grund dafür liegt wahrscheinlich in einer ineffizienteren Prozessierung des Signalpeptids von HLA-G und einer im Vergleich mit anderen HLA-E-Liganden geringeren Bindungsaffinität des Peptids G3-11 für HLA-E. Eine Funktion der verkürzten HLA-G-Isoformen durch die direkte Interaktion mit Rezeptoren auf NK-Zellen konnte nicht nachgewiesen werden und ist wegen ihrer intrazellulären Expression auch unwahrscheinlich. Vielmehr deuten erste Daten, die zeigen, daß die Isoformen, direkt oder indirekt, mit dem TAP-Komplex assoziiert sind, auf eine mögliche Funktion im Rahmen der Antigenpräsentation hin. Worin diese besteht, müssen weiterführende Untersuchungen zeigen. Daher ist anzunehmen, daß HLA-G in vivo hauptsächlich über HLA-G-bindende KIR immunregulatorische Funktionen wahrnimmt und durch indirekte Wirkung über HLA-E-CD94/NKG2A modulierend eingreift.

Die LQYLWUR Selektion ermöglicht es aus kombinatorischen Nukleinsäurebibliotheken Oligonukleotidsequenzen zu identifizieren, die verschiedenste Zielmoleküle mit hoher Affinität und Spezifität binden können. Dadurch haben sich Aptamere zu einer potenten Alternative zu den in der Diagnose, Therapie und als Forschungsreagentien etablierten Antikörper entwickelt. Mit Hilfe der SELEX-Technologie (6ystematic (volution of /igands by (;ponential Enrichment) ist es in dieser Arbeit gelungen, 2' amino-stabilisierte RNA-Aptamere gegen das Neuropeptid Y und ein ausgewähltes, funktionell relevantes Prionproteinepitop zu generieren. Die Anreicherung funktioneller Sequenzen erfolgte durch einen affinitätschromatographischen Prozess. Zudem sollten bereits vorliegende RNA-Aptamere, die gegen das rekombinante Prionprotein in früheren Arbeiten selektiert wurden, charakterisiert werden. Das Neuropeptid Y (NPY), bestehend aus 36 Aminosäuren, gehört zur Familie der pankreatischen Polypeptide und ist bei der Steuerung einer Vielzahl physiologischer und pathophysiologischer Prozesse von Bedeutung. Es wird angenommen, daß durch selektive Bindung unterschiedlicher NPY-Konformationen an die einzelnen GProtein gekoppelten NPY-Rezeptorsubtypen (Y1, Y2, Y3, Y4, Y5 und Y6) unterschiedliche Signale vermittelt werden können. Dieses differentielle Bindungsverhalten von NPY an seine Rezeptorsubtypen ist bisher unvollständig verstanden. Die in dieser Arbeit generierten Anti-NPY-Aptamere binden ihr Zielmolekül -NPY- mit einer Affinität von 370 nM und sind durch eine hohe Spezifität innerhalb der pankreatischen Polypeptidfamilie charakterisiert. Die Bindungsregion des Aptamers an den C-Terminus des Neuropeptid Y wurde durch Kartierungs-Experimente mit NPY-Analoga LQ YLWUR bestimmt. Die NPY-Analoga stellen sowohl verschiedene Untereinheiten von NPY, als auch Modifikationen des Peptides, die zu Rezeptorsubtypspezifitäten führen, dar. Durch Punktmutationen im C-terminalem NPY-Bereich konnte u.a. gezeigt werden, daß die Aminosäure Arginin an Position 33 für die Komplexbildung von NPY und Aptamer essentiell ist. In den Bindungsstudien in Gegenwart selektiver Agonisten zeigte sich, daß die Bindungseigenschaften von NPY am Y2 Rezeptor weitgehend mit denen an das Aptamer übereinstimmen. Die Kompetition des Aptamers mit den Rezeptoren um 3H-NPY wurde an Zellen, die die Rezeptoren NPY-Y1, NPY-Y2, bzw. NPY-Y5 exprimieren, untersucht. Das Aptamer verdrängte NPY mit besonders hoher Affinität am Y2 Rezeptor im Vergleich zur Verdrängung am Y1- bzw. Y5-Rezeptor. Die Anti-NPY-Aptamere weisen ein Bindungsverhalten am NPY vergleichbar zum Y2-Rezeptor auf und stellen damit ein wertvolles Werkzeug zur selektiven Charakterisierung der Interaktion zwischen NPY und seinen Rezeptoren dar. Von entscheidender Bedeutung für die Pathogenese der übertragbaren spongiformen Enzephalopathien ist die infektiöse Form des Prionproteins (PrPSc). Es wird angenommen, daß PrPC durch einen posttranslationalen Prozeß in PrPSc konvertiert werden kann. Trotz identischer Primärstruktur unterscheiden sich die beiden Prionproteinisoformen (PrPC und PrPSc) grundlegend in ihren biochemischen und biophysikalischen Eigenschaften. Die in früheren Arbeiten selektierten Prionprotein-Aptamere sollten im Hinblick auf ihr diagnostisches Potential charakterisiert werden. Erste strukturelle Untersuchungen führten zu der Annahme, daß die RNA-Aptamere ein G-Quartett als stabilisierendes Sekundärstrukturmotiv ausbilden können. Sowohl Kartierungsstudien mit unterschiedlichen Prionproteinpetiden als auch Bindungsstudien mit N-terminal trunkiertem PrPSc zeigten, daß der N-Terminus für die Bindung der Aptamere essentiell ist. In Gelshiftexperimenten mit verschiedenen Hirnhomogenaten konnte die spezifische Bindung der Aptamere an authentisches PrP gezeigt werden. Aufgrund der fehlenden PrPSc-Isoformspezifität der untersuchten Aptamere ist eine diagnostische Anwendung kaum denkbar. Die Bindung der Aptamere in der pKsensitiven, N-terminalen Prionproteindomäne läßt eine Anwendung in Kombination mit Proteinase K-Verdau in Analogie zu den derzeit benutzten BSE-Testverfahren nicht zu. Im letzten Teil der Arbeit sollten RNA-Aptamere gegen einen für die Konversion wichtigen Bereich des Prionproteins (AS 90-129) generiert werden. Es konnte gezeigt werden, daß die in einer vorgeschalteten Prionpeptidselektion (AS 90-129) identifizierten Aptamere in der Lage sind, ihr Zielmolekül im Gesamtkontext des Prionproteins zu erkennen. In funktionellen Studien in persistent Prion-infizierten Neuroblastomzellen wurde eine statistisch signifikante und spezifische Reduktion der Akkumulation von GHQRYR synthetisiertem PrPSc zu hochmolekularen Aggregaten in Gegenwart einer ausgewählten Aptamersequenz beobachtet. Im Verlauf der Pathogenese von spongiformen Enzephalopathien korreliert die PrPSc- Aggregatbildung mit Infektiosität und Neurodegeneration. Damit bieten die selektierten Aptamere möglicherweise eine Ausgangsbasis um Therapeutika zu entwickeln, die den Verlauf der Prionerkrankungen beeinflussen.

Intraduodenal (i.d.) application of bile or Na-taurodeoxycholate (TDC) dose dependently enhances basal exocrine pancreatic secretion. The hydrokinetic effect is mediated at least in part by secretin. This study should show, whether vasoactive intestinal polypeptide (VIP), a partial agonist of secretin, may also be involved in the mediation of the hydrokinetic effect. Furthermore, plasma concentrations of somatostatin-like immunoreactivity (SLI) were measured in order to check whether this counterregulating hormone is also released by bile and TDC. Twenty investigations were carried out on 10 fasting healthy volunteers provided with a double-lumen Dreiling tube. Bile and TDC were intraduodenally applied in doses of 2-6 g and 200-600 mg, respectively, at 65-min intervals. Plasma samples were withdrawn at defined intervals for radioimmunological determination of VIP and SLI. Duodenal juice was collected in 10-min fractions and analyzed for volume, pH, bicarbonate, lipase, trypsin, and amylase. I.d. application of bile or TDC dose dependently stimulated hydrokinetic and ecbolic pancreatic secretion. Bile exerted a slightly stronger effect than TDC. Pancreatic response was simultaneously accompanied by a significant increase of plasma VIP and SLI concentrations. The effect of bile on integrated plasma VIP and SLI concentrations seems to be dose dependent; the effect of TDC on integrated SLI, too. For the increase of integrated plasma VIP concentrations after TDC no dose-response relation could be established. We conclude that VIP may be a further mediator of bile-induced volume and bicarbonate secretion. The release of plasma SLI indicates that inhibitory mechanisms concomitantly are triggered by i.d. bile and TDC, as already shown during digestion for the intestinal phase of pancreatic secretion.

Sat, 1 Jan 1994 12:00:00 +0100 http://epub.ub.uni-muenchen.de/2451/ http://epub.ub.uni-muenchen.de/2451/1/2451.pdf Gottschalk, Lothar; Lottspeich, Fritz; Scheer, Hugo Gottschalk, Lothar; Lottspeich, Fritz und Scheer, Hugo (1994): Reconstitution of an allophycocyanin trimer complex containing the C-terminal 21-23 kDa domain of the core-membrane linker polypeptide Lcm. In: Zeitschrift für Naturforschung C: Journal of Biosciences, Vol

We have purified bleomycin hydrolase from yeast (molecular mass 55 000 Da). Using protein sequence-derived degenerate oligonucleotide primers and amplification by polymerase chain reaction, the yeast gene BLH1 was isolated and characterized. The deduced amino acid sequence (483 amino acids) exhibits surprisingly high homology to vertebrate bleomycin hydrolase (43% identical residues and 22% conserved exchanges). It contains three blocks of sequences found conserved in other members of the thiol proteinase family and thought to be associated with the catalytic centre. BLH1 is non-essential under all growth conditions tested. However, in the presence of 3.5 mg bleomycin/ml medium wild-type cells have a slight growth advantage compared to blh1 mutant cells.

The core linker polypeptide Lc 8.9 was isolated from Mastigocladus laminosus and purified on a preparative scale. A method for the reconstitution of allophycocyanin (AP)—linker complexes from isolated polypeptides was developed. The complex (αAP(βAP)3 Lc 8.9 was reconstituted and compared to (αAPβAP) and (αAPβAP)3 by sucrose density gradient ultracentrifugation, absorption, fluorescence emission and circular dichroism spectroscopy. Differences in the spectra of reconstituted and of directly isolated AP complexes are discussed.

DURING photosynthetic CO2 fixation, fixed carbon is exported from the chloroplasts in the form of triose phosphate by the chloroplast phosphate translocator, which is the principal polypeptide (E29) from spinach chloroplast envelopes1. We have sequenced this nuclear-coded envelope membrane protein from both spinach and pea chloroplasts2,3. An envelope membrane protein, E30, has been identified as a possible receptor for protein import into pea chloroplasts using an anti-idiotypic antibody approach4–6; antibodies raised against purified E30 inhibited binding and import of proteins into chloroplasts7. The amino-acid sequence of E30 deduced from its complementary DNA7 turned out to be highly homologous to that of E29, assigned by us as the spinach phosphate translocator2, and was identical to the corresponding polypeptide from pea chloroplasts3. Differences in the binding properties to hydroxylapatite of £30 and the phosphate translocator suggested that E30 was not responsible for the chloroplast phosphate-transport activity but was the chloroplast import receptor7. Here we present evidence that argues against this and which identifies E30 as the chloroplast phosphate translocator.

A chloroplast outer envelope membrane protein was cloned and sequenced and from the sequence it was possible to deduce a polypeptide of 6.7 kDa. It has only one membrane-spanning region; the C terminus extends into the cytosol, whereas the N terminus is exposed to the space between the two envelope membranes. The protein was synthesized in an in vitro transcription-translation system to study its routing into isolated chloroplasts. The import studies revealed that the 6.7-kDa protein followed a different and heretofore undescribed translocation pathway in the respect that (i) it does not have a cleavable transit sequence, (ii) it does not require ATP hydrolysis for import, and (iii) protease-sensitive components that are responsible for recognition of precursor proteins destined for the inside of the chloroplasts are not involved in routing the 6.7-kDa polypeptide to the outer chloroplast envelope.

The effect of intraduodenally administered cattle bile (CB) and Na-taurodeoxycholate (TDC) on basal pancreatic secretion and plasma levels of secretin, pancreatic polypeptide (PP), and gastrin were investigated on two separate days in 10 fasting volunteers. Doses of 2-6 g CB and 20&600 mg TDC were given intraduodenally at 65-min intervals. Volume, bicarbonate, lipase, trypsin, amylase, and bilirubin were measured in 10-min fractions of duodenal juice, and GI peptides determined by radioimmunoassay. CB and TDC enhanced significantly and dose-dependently volume, bicarbonate and enzyme secretion, and plasma secretin and PP levels. In contrast, plasma gastrin showed only a marginal increase. We conclude that the hydrokinetic effect of intraduodenal CB and TDC is at least partially mediated by secretin. Gastrin could be ruled out as a mediator of the ecbolic effect, whereas other GI peptides, primarily CCK, and/or neural mechanisms must be considered possible mediators. Both pathways may also play a role in the PP release.

Reaction centers from Rhodobacter sphaeroides have been modified by treatment with sodium borohydride similar to the original procedure [Ditson et al., Biochim. Biophys. Acta 766, 623 (1984)], and investigated spectroscopically and by gel electrophoresis. (1) Low temperature (1.2 K) absorption, fluorescence, absorption- and fluorescence-detected ODMR, and microwave-induced singlet-triplet absorption difference spectra (MIA) suggest that the treatment produces a spectroscopically homogeneous preparation with one of the ‘additional’ bacteriochlorophylls being removed. The modification does not alter the zero field splitting parameters of the primary donor triplet (TP870). (2) From the circular dichroism and Raman resonance spectra in the1500–1800 cm-1 region, the removed pigment is assigned to BchlM, e.g. the "extra" Bchl on the "inactive" M-branch. (3) A strong coupling among all pigment molecules is deduced from the circular dichroism spectra, because pronounced band-shifts and/or intensity changes occur in the spectral components assigned to all pigments. This is supported by distinct differences among the MIA spectra of untreated and modified reaction centers, as well as by Raman resonance. (4) The modification is accompanied by partial proteolytic cleavage of the M-subunit. The preparation is thus spectroscopically homogeneous, but biochemically heterogenous.

The precursor of the mitochondrial inner membrane protein ADP/ATP carrier is cytoplasmically synthesized without an amino-terminal peptide extension. We constructed a truncated precursor lacking the 103 amino acids from the amino terminus (about a third of the protein). Import of the truncated precursor into mitochondria showed the import characteristics of the authentic precursor, including nucleoside triphosphate dependence, requirement for a protease-sensitive component on the mitochondrial surface, two-step specific binding to the outer membrane, and membrane potential-dependent translocation into the inner membrane. We conclude that, in contrast to all other mitochondrial precursor proteins studied so far, domains of the ADP/ATP carrier distant from the amino terminus can carry specific targeting information for transport into mitochondria.

We have constructed target cells by cotransfection of the MHC gene Ld and fragments of murine cytomegalovirus (MCMV) DNA coding for nonstructural immediate-early (IE) proteins. Transfectants were tested by using CTL clone IE1 with specificity for an IE epitope presented in association with Ld. Data show that clone IE1 recognizes a product of the ie1 transcription unit of MCMV, and that its specificity is shared by approximately 25% of polyclonal IE-specific CTL. The results provide the first definite evidence that expression of a herpes virus IE gene encoding a regulatory protein gives rise to antigen expression detectable by specific CTL

Thu, 1 Jan 1976 12:00:00 +0100 https://epub.ub.uni-muenchen.de/7306/1/7306.pdf Wachter, Elmar; Neupert, Walter; Michel, Rainer; Machleidt, W.

Wed, 1 Jan 1975 12:00:00 +0100 https://epub.ub.uni-muenchen.de/7284/1/7284.pdf Neupert, Walter; Otto, J.; Machleidt, W.; Liebl, A.; Michel, Rainer

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