Podcasts about pamps

Today, Dr. Margaret McFall-Ngai of the California Institute of Technology, joins the #QualityQuorum to discuss her philosophy of the future of biology.  Dr. McFall-Ngai has been described as “…a recognized thought leader regarding the cornerstone role microbiology plays in the life sciences.”  I could not agree more. Host: Mark O. Martin Guest: Margaret McFall-Ngai Subscribe: Apple Podcasts, Spotify Become a patron of Matters Microbial! Links for this episode  For beginning #Micronauts, I suggest this overview of the amazing roles microbes play in the lives of animals and plants: “We are living in a bacterial world, and it's impacting us more than previously thought“. Here is the more advanced article coauthored by Dr.McFall-Ngai on this topic:  “Animals in a Bacterial World.”  A must read. A video on this topic by Dr. McFall-Ngai. Another video on this topic by Dr. McFall-Ngai. A wonderful review of the squid-Vibrio symbiosis (relevant to discussion) from the laboratory of Dr. Karen Visick. The TCT story:  how the same signal creates a light organ in squid and disease in humans. PAMPs versus MAMPs as a paradigm shift. A wonderful profile on Dr. McFall-Ngai An essay about career building by Dr. McFall-Ngai. Margaret McFall-Ngai's faculty website. The McFall-Ngai laboratory website. Intro music is by Reber Clark Send your questions and comments to mattersmicrobial@gmail.com

Jillian and Pamps are joined by Ru Pauls Drag Race all star, Detox Icunt. Detox has had it with slow walkers that just can't keep up with her gorgeous long legs in full stride. The three talk about drag queens taking over the suburbs, Gypsy Rose Blanchard's husband and how Detox was the queenpin of her cell when she was in prison. Pumps admits she lies about being pregnant (even at her advanced age) just to get better parking spots and Jennifer has had it with the fact that men can skirt the system with their beloved Viagra (and their cheesy commercials) while women are having to fight for their rights at the Supreme Court. Come see I've Had It live on the Hot Sh*t Tour! More info & tickets available at https://linktr.ee/ivehaditpodcast and subscribe to I've Had It wherever you get your podcasts. Thank you to our sponsors: Vegamour: For a limited time get twenty percent off your first subscription order by going to vegamour.com/HADIT and use code HADIT at check out. Shopify: Sign up for a $1 per month trial period at Shopify.com/hadit now to grow your business – no matter what stage you're in. SKIMS: SKIMS Bras are now available at SKIMS.com Plus, get free shipping on orders over seventy five dollars! If you haven't yet, be sure to let them know we sent you! After you place your order select "podcast" in the survey and select our show in the dropdown menu that follows. Subscribe to our Patreon: https://www.patreon.com/IveHadItPodcast Follow Us: I've Had It Podcast: @Ivehaditpodcast Jennifer Welch: @mizzwelch Angie "Pumps" Sullivan: @pumpspumpspumps Special Guest: Detox @TheOnlyDetox

The immune system works tirelessly to maintain health and well-being. This formidable defense mechanism is comprised of a vast array of cells, tissues, proteins, and molecules that work together to identify and neutralize foreign invaders. The inflammation that ensues can be recognized by redness, pain, swelling, and heat. In order for this process to take place, the body needs to be able to identify what is ‘friend or foe'. But how? There are certain molecular patterns that our immune system recognizes as part of this surveillance and recognition. Some patterns are associated with pathogens - “pathogen-associated molecular patterns” (PAMPs). Others are associated with damaged cells (DAMPs) or lifestyle choices (LAMPs).   In today's episode, we do a quick refresher on adaptive and innate immunity. We also discuss some signaling molecules that trigger the inflammatory cascade – PAMPs, DAMPs, and LAMPs. Today on The Lab Report: 3:40 Break it down – innate vs. adaptive immunity 6:40 Pathogen-associated molecular patterns (PAMPs), Pattern Recognition Receptors (PRRs) 10:25 Sterile inflammation: damage-associated molecular patterns (DAMPs) 14:30 Lifestyle-associated molecular patterns (LAMPs) & chronic inflammation 18:25 Question of the Day How do we turn off the LAMPs? Additional Resources: Genova Connect   *PROMO CODE TheLabReport20 for 20% off* Subscribe, Rate, & Review The Lab Report Thanks for tuning in to this week's episode of The Lab Report, presented by Genova Diagnostics, with your hosts Michael Chapman and Patti Devers. If you enjoyed this episode, please hit the subscribe button and give us a rating or leave a review. Don't forget to visit our website, like us on Facebook, follow us on Twitter, Instagram, and LinkedIn. Email Patti and Michael with your most interesting and pressing questions on functional medicine: podcast@gdx.net. And, be sure to share your favorite Lab Report episodes with your friends and colleagues on social media to help others learn more about Genova and all things related to functional medicine and specialty lab testing. To find a qualified healthcare provider to connect you with Genova testing, or to access select products directly yourself, visit Genova Connect. Disclaimer: The content and information shared in The Lab Report is for educational purposes only and should not be taken as medical advice. The views and opinions expressed in The Lab Report represent the opinions and views of Michael Chapman and Patti Devers and their guests.  See omnystudio.com/listener for privacy information.

What a week in golf, there is so much to discuss, so settle into your walk, drive, or chair and get ready for a big episode of Talk Birdie To Me.We start with Min Woo Lee's big win in Macau at the weekend, what a superstar he is becoming. And we discover that Mark Allen has turned into an 'FWG'....what is that? We explain on the podcast, it's not something to be proud of!Great win from Rod Pampling - a golfer with 'effortless power' according to Mark, and Nick reckons he is one of the good guys, even though Pamps beat him in a matchplay tournament, not that Nick is still bitter about it. Nick and Mark discuss Ben Eccles great win in the WA PGA Championship, an excellent win, particularly after a slow start. And Nick names one of the big earners in golf this week....and it's not a player!We discuss the decision not to award world rankings to LIV players, Mark believes that the PGA/LIV talks are not going well, although the CEO of the PGA Gavin Kirkman has told him that they are proceeding well, so Mark will take him at his word on that. We discuss some of the challenges with giving points to LIV players, and explore a couple of ways it could work, and what they'd like to see happen.This week saw Ben An suspended for doping. Nick talks about when he was drug tested on tour, how often and how it happens. He describes it as quite a humiliating process.This week Lexi Thompson played the PGA tournament in Vegas, and shot even. Nick and Mark discuss how she played, and Nick was fascinated by what she carried in her bag.Nick had a fitting at PING this week, and Mark wants to know all about it. Nick explains what he's arranged and how the fitting went.Lots of feedback this week, a great question about how US social golfers are ‘liberal' with the handicap system, and we find out who the nicest bloke on tour is.Nick takes us around the world with the PING global results. Some ripping performances from there Aussies over the weekend; and Nick then talks about playing in Madrid, and his favourite bar there called El Toro.The Top 5 this week was suggested by David Craig from Brisbane, and is the Top 5 shots that pros would like a redo on, Nick has compiled his list, thanks for all your suggestions. One of the Top 5 is from Scott Hoch in the US Masters, and Mark tells a very funny story about having dinner with Scott in New Zealand with their waiter doing something very awkward.And a great masterclass this week from Mark Allen on not moving your head when putting, but moving your eyes instead.Talk Birdie To Me, new episodes weekly wherever you get your podcasts. Subscribe and you'll never miss an episode! If you've got a friend who is into golf, we'd love you to share the podcast with them, and if you can rate and review us wherever you get your podcasts that would be great.Follow us on Facebook, Instagram, Twitter and TikTok, or send a voicemail to us here. Hosted on Acast. See acast.com/privacy for more information.

JP Errico today with us, who is the co founder of a company that makes devices, specifically for vagal neuromodulation, and is an expert in this area and in many affiliated areas.LINK:JP Errico - LinkedinHealth Upgrade PodcastTruvaga - WebsiteTIMESTAMPS:00:30 - JP Errico01:41 - Deep Brain Stimulation02:58 - Vagus Nerve Stimulator03:37 - Innate Immune System05:43 - Macrophages09:16 - Pathogen Associate Molecular Patterns21:52 - 10th Cranial Nerve26:03 - Nucleus Tractus Solitarius26:37 - Nucleus Bacillus of Maynard's31:30 - TruVaga33:48 - Gammacore46:02 - Schizophrenia----------#JPEricco #neuromodulation #vagusnerve #immunesystem #PAMPs #centralnervoussystem #truvaga.com #stress #schizophrenia #inflammation #robertlufkinmd #drlufkin #robertlufkin #stephensideroff #drsideroff #stephensideroffphd #healthlongevitysecrets #pathlongevity*** CHECK OUT ROB AND STEVE'S MASTERCLASSES ***Rob's MasterclassesSteve's Masterclasses*** CONNECT WITH ROBERT LUFKIN MD ON SOCIAL MEDIA ***Web: https://robertlufkinmd.com/Twitter: https://twitter.com/robertlufkinmdYoutube: https://www.youtube.com/RobertLufkinMDInstagram: https://www.instagram.com/robertlufkinmd/LinkedIn: https://www.linkedin.com/in/robertlufkinmd/*** CONNECT WITH STEPHEN SIDEROFF PHD ON SOCIAL MEDIA ***Facebook: https://www.facebook.com/stephen.sideroffTwitter: https://twitter.com/DrSideroffInstagram: https://www.instagram.com/drstephensideroff/LinkedIn: https://www.linkedin.com/in/drstephensideroff/*** SPONSORSHIPS & BRANDS ***We do work with sponsors and brands. If you are interested in working with us for your health industry product or service, please contact us at:https://pathlongevity.com/contact-us/ NOTE: This is not intended to be a substitute for professional medical advice, diagnosis, or treatment. Always seek the advice of your physician or other qualified health provider with any questions you may have. Never disregard professional medical advice or delay in seeking it because of something you have seen here. Robert Lufkin MD may at any time and at its sole discretion change or replace the information available on this channel. To the extent permitted by mandatory law, Robert Lufkin MD shall not be liable for any direct, incidental, consequential, indirect or punitive damages arising out of access to or use of any content available on this channel, including viruses, regardless of the accuracy or completeness of any such content.

PAMPs and DAMPs, complement proteins and MAC, macrophages, Natural Killers and Interferon, MHC molecules...come get acquainted with the players in your innate immune system. For more information, visit http://www.VyvyaneLohMd.com

Dr. Anthony Cannella, Infectious Diseases Clinician and Associate Professor with the Division of Infectious Diseases at the Morsani College of Medicine, presents a primer on basic immunology for clinicians. Dr. Cannella begins by drawing a distinction between innate and adaptive immunity. He then discusses the complement cascade, antimicrobial peptides, PAMPS, and DAMPS. Next, Dr. Cannella reviews individual components of the immune system, including neutrophils, eosinophils, macrophages, dendritic cells, and natural killer cells. Lastly, the remaining components of the immune system are covered, including B cells, immunoglobulins, and T cells and their function.

This week Newsiebaby sat down with Blazar In Pamps to talk about his iconic video on putting on a diaper perfectly. We then talk about how there are different ways to enjoy a soggy pamp.

脳の中の炎症は「DJ-1」が関係していることが発覚。しかし、それをどうやって発見したのか？というお話を語っていただきました。 ゲスト 東京大学大学院 博士課程 中村 幸太郎さん (https://twitter.com/Koutarou_nkmr) 「脳梗塞に苦しまない世界をつくりたい/サイエンスコミュニケーションを通じて社会と科学の架け橋になりたい」とクラウドファンディング挑戦中 https://academist-cf.com/fanclubs/196?lang=ja 研究ピッチコンテスト GENSEKI運営中 https://www.notion.so/GENSEKI-46ac77cff54a4197a53fb348a776fd4a ▶DAMPs：damage-associated molecular patterns 壊死した細胞などから細胞外へ漏れ出ることで炎症を惹起する分子。 ▶PAMPS：pathogen-associated molecular pattern 病原体がもつ特有の構成成分、分子。 ▶尿酸結晶：血中の尿酸値が高い状態が続くと溶け切れなくなった尿酸が結晶となり、関節などに蓄積することで炎症が起こる。 ▶ATP：アデノシン三リン酸、生体内ではリン酸1分子、または、リン酸2分子が離れたり結合したりする事で、エネルギーの放出・貯蔵を行う。 ▶DJ-1：酸化ストレスのセンサーとして機能し、ニューロンを保護するタンパク質。PARK 7(パーキンソン病タンパク質7)とも呼ばれている。 ▶マクロファージ：白血球の一種で、体内に侵入した細菌などの異物を捕食し、ホルモン用のタンパク質であるサイトカインを出して他の免疫細胞を活性化する。 ▶樹状細胞：血液中の白血球の中の免疫細胞の一部で、血液によって運ばれ体の中のあらゆる場所に分布している免疫細胞。異物を発見すると、それを自分の中に取りこみ特徴を覚える。その後樹状細胞はリンパ節まで移動し自分の覚えた異物の特徴をリンパ球に教え、リンパ球にその異物を攻撃するように指示を出す。 ▶脳の城壁＝血液脳関門 (BBB, Blood-Brain Barrier) 血液と脳（中枢神経系）の組織液との間の物質交換を制限する機構。 ▶ミクログリア：中枢の免疫担当細胞として知られる細胞。細長い突起を動かし、シナプスや軸索等に接触させて機能を監視・調節している。 ▶先発部隊→自然免疫：外部から侵入した異物 「非自己」と、本来自身のもっているもの「自己」を区別し、「非自己」を速やかに処理する仕組み。主に好中球やマクロファージ、樹状細胞といった食細胞が担当。 ▶後発部隊→獲得免疫：感染した病原体を特異的に見分け、それを記憶することで同じ病原体に出会った時に効果的に病原体を排除できる仕組み。主にT細胞（細胞障害性T細胞、ヘルパーT細胞など）やB細胞といったリンパ球が担当。 ▶貪食（どんしょく）：菌やその他の固形物を取り込む作用。食作用とも呼ばれる。 ▶ポリクローナル抗体： 一つの抗原に対して様々な親和性を持つ異なる抗体の混合物。 ▶モノクローナル抗体：ポリクローナル抗体に対し、1種類のコピー(クローン)からなる抗体。 質問、感想、ゲスト出演の問い合わせなどは以下Twitter・お便りフォームまでお願いします。 Twitter　＃サイエンマニア https://twitter.com/REN_SciEnTALK 研究者レンのもう１つの番組　SciEnTALK/サイエントークはこちら→ https://lit.link/scientalk おたよりフォーム https://forms.gle/H4vg7MuN77VqbHV79 1シーズン1人のゲスト形式で、各エピソードは一区切りのチャプターとしてご利用いただけます。 流し聞きする場合は、エピソードを古い順に並べ替えていただくとシームレスに聞くことができます。 BGM Future Sky/ SAKURA BEATZ.JP Somehow/Khaim https://www.khaimmusic.com/

EP 140 with Rossco, Rocket & Magic - Herbies gets tour card heads straight to Georgia. - Tony breaks the curse - Minjee & Steph close as Anna gets it done - Crans sur Sierre. Can Magic win some More Min Woo $$ or will Wade Win - Pamps wins - BMW Championship - Gabrielle Macdonald wins on LET ACCESS Thank you so much for the continued support. Hit rocket up for his Rabbit Hole info ROCKETS INSTA Hit Magic Mike up at MAGIC MIKE INSTA Hit Rossco at ROSSCO INSTA   Don't forget to jump into the other podcasts in the family. Improve your GOLF Mental Game INCITEGOLF Academy - Build an Unbreakable Mental Game ( Free 1st Program) https://incitegolfacademy.teachable.com/   The My Love of Golf Podcast Insta: Instagram Facebook: Facebook Twitter: Twitter Rocket: Rocket Insta The Mental Mastery Golf Podcast Want to improve your Mental Game? Then check out The Mental Mastery Golf Podcast hosted by Rossco and Jamie from Dare2Dream www.dare2dream.com.au Mental Mastery Golf Podcast Mental Mastery Facebook Group   The GolfRules Questions Podcast Facebook: GolfRules Questions FB Insta: Insta YouTube: GRQ Youtube PODCAST: The GolfRules Questions Podcast        

Dr. Cannella, Assistant Professor at the University of South Florida Morsani College of Medicine, presents an introductory lecture on immune system function. He begins by discussing the first line body defenses such as the skin, mucous membranes, and lymphatic system. He then differentiates the innate versus the adaptive immune system. He then discusses other components of the immune system, including the complement pathway, antimicrobial peptides, PAMPS, DAMPS, phagocytosis, neutrophils, macrophages, dendritic cells, and NK (Natural Killer) cells. A second lecture is planned for later in the fall of 2021.

In this episode, Eugenio, Natalie, and Jatin sit down with Dr. Claudia Jakubzick from Dartmouth University. They discus Claudia's 2018 publication that elucidated the role of natural IgM in the recognition of neoantigens in absence of PAMPs. Check out our memes on Facebook (@antibuddies), Twitter (@antibuddiesP), and Instagram (@AntibuddiesPodcast). Join us on our monthly journal club at our YouTube channel: https://www.youtube.com/channel/UCxyrHotyyY3sSwcp1zigeCw Send us your queries/questions/suggestions at antibuddies1@gmail.com. Article of discussion: Immune Surveillance by Natural IgM Is Required for Early Neoantigen Recognition and Initiation of Adaptive Immunity (nih.gov)

293: On this episode of CCNT, birds playing basketball, trucker Flippy, #JewishSpaceLasers goes viral, #GameStop story, Yellen conflicts abound, Elon PAMPS #Bitcoin and Dogecoin, Davos capitalism, Biden EO is American Davos (Biden EO history and debunk attempt), Biden family issues, HHS busted (Newsom busted too), funny Brian Williams news blunder, 33 sports, 33 deaths, 666 Cuba, 11666 India, waccine death rising, China USA waccine New World Order, WHO says no vax for pregnant women, BMI nihilism, TALEN beats CRISPR, and a Nephilim Update!    SHOW NOTES: https://bit.ly/3p77uBU  COMMUNITY: https://bit.ly/2I9nTVC   Paypal: https://bit.ly/2JKbBE0  Patreon 1: https://bit.ly/3k5P9ll  Patreon 2: https://bit.ly/3k4uxcY  Crypto:

Programa transmitido todos los lunes a las 10 pm (Centro de México), a través de mixlr.com/vallis-mortem/ Conducido por: El Papalou y Dani V. Black 1. Austero - Ego 2. Mountain Creed - Credo de la Montaña 3. Valley of the Sun - Old Gods 4. Seum - Saliva Bath (Fatima Cover) 5. Fatima - Raining Brick (Seum Cover) 6. Snow - Escape From Brasil 7. Merlock - Prolapse 8. Loot the Body - Expedition to the Barrier Picks 9. Gas Giant - electric Maze 10. PAMPS! Les Pamplemousses Ethyliques - Caracas

Here is our episode with OG Pamps and the rest of his artists.Support the show (https://www.paypal.com/cgi-bin/webscr?cmd=_s-xclick&hosted_button_id=JALRZC89YNEEG&source=url)

Hoje é a Mari Pamps que chega nesse podcast, calçando Havaianas e um vestido meio destruido depois do festerê de formatura dela. E temos um Lado B, vem nessa! Pra quem quiser mandar algum feedback, falar qualquer p... q seja, só vem. Instagram: @alexandrejapa_mwa Twitter: @alexandre_ojapa Contato: K7@meuwalkmanamarelo.com    

La pancreatitis, tanto aguda como crónica, es una de las entidades más interesantes desde el punto de vista de su fisiopatología. Discutiremos brevemente su etiología, poniendo mayor énfasis en los cuadros obstructivos y relacionados con el consumo de alcohol, pero mencionando otros factores que pueden actuar como desencadenantes. Nos enfocaremos luego en el proceso fisiopatológico propiamente dicho, abordando los cambios agudos tempranos que ocurren a nivel del acino pancreático, la activación inapropiada de la tripsina, el origen de la lesión microcirculatoria, entre otros. También entraremos en lo que realmente justifica el daño sistémico y local severo, a saber, la activación del sistema inmune y la traslocación bacteriana intestinal. Haciendo énfasis en el rol de los DAMPs y los PAMPs y la respuesta inflamatoria en general. ¿Quieres enterarte semana a semana de las nuevas publicaciones, y acceder a contenido exclusivo? Únete a la lista de correo de Leucocitos isotópicos. Para suscribirte al Podcast de Medicina, estas son las opciones más recomendadas: Apple Podcasts Google Podcasts Spotify Si prefieres explorar más alternativas, haz clic aquí. ¿Te gustó el episodio? Seguro disfrutarás este también: Inflamación I (039) Además, puedes acceder a la lista curada y actualizada de los episodios con mayor aceptación. Este show es para ti. Puedes apoyarlo entrando a Apple Podcasts y dejando allí una calificación positiva. Encuentra las notas de este episodio y dirige a tus amigos a isotopicos.com/062 El objetivo de Leucocitos isotópicos es entretenerte mientras complementas lo que recibes en tu Escuela o Facultad de Medicina. Soy Médico Internista, y comprendo lo demandante que puede ser nuestra Carrera. Por eso decidí crear el Podcast como un curso de Medicina ameno y sin una estructura rígida, que despierte tu interés y curiosidad por esta maravillosa Ciencia. Nunca reemplazará a la Universidad, ni a los libros, pero cumplirá con la misión que lo fundamenta: Ser el lugar donde descansamos de leer, sin dejar de aprender. No olvides que la mejor manera de ayudar a que el proyecto crezca, es contarle a todos de él. ¡Gracias por compartir este episodio con alguien!

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  Episode 2 features Dr. Adam Matson, attending neonatologist at Connecticut Children’s Medical Center-Newborn Intensive Care Unit (Hartford, CT) and Assistant Professor of Pediatrics and Immunology at the University of Connecticut School of Medicine (Farmington, CT). During this episode, Dr. Matson provides a comprehensive overview of NEC as it relates primarily to very low birth weight babies, those weighing less than 1500 grams (3 pounds 4.91 ounces) and who have the greatest risk for developing the disease. He discusses: * The early warning signs of NEC, what steps are taken when NEC is suspected, and how X-rays are used to diagnose NEC * How a premature baby’s immune response to the microbiome (bacterial communities) of the intestine appears to play a role in the development of NEC * Known risk factors of NEC, and how they may affect the intestinal microbiome * His current research focused on innate immune signaling in the developing intestine as it pertains to the development of NEC * Current prevention strategies for NEC * Additional research trends in NEC, and the importance of efforts to prevent prematurity Copyright © 2015 The Morgan Leary Vaughan Fund, Inc. This episode was produced in part by the TeacherCast Educational Broadcasting Network. [powerpress] STEPHANIE VAUGHAN, HOST: Welcome to Episode 2 of Speaking of NEC—a free, audio podcast series about Necrotizing Enterocolitis. Produced by The Morgan Leary Vaughan Fund, and funded by The Petit Family Foundation, Speaking of NEC is a series of one-on-one conversations with relevant NEC experts—neonatologists, clinicians and researchers—that highlights current prevention, diagnosis, and treatment strategies for NEC, and the search for a cure. For more information about this podcast series or The Morgan Leary Vaughan Fund, visit our website at morgansfund.org. Hello, my name is Stephanie Vaughan. Welcome to the show. I’m the Co-founder and President of The Morgan Leary Vaughan Fund. Today, my guest will be Dr. Adam Matson, attending neonatologist at Connecticut Children’s Medical Center-Newborn Intensive Care Unit in Hartford, CT, and the Assistant Professor of Pediatrics and Immunology at the University of Connecticut School of Medicine in Farmington, CT. Dr. Matson will share with me today a comprehensive overview of NEC as it relates primarily to very low birth weight babies, those weighing less than 1500 grams or 3 pounds 4.91 ounces, who have the greatest risk for developing the disease. During our conversation, he will discuss in varying degrees: Early warning signs, Steps that are taken when NEC is suspected, Diagnosis, Risk factors, Prevention, Current areas of research, and The importance of efforts to prevent prematurity He will also discuss how a premature baby’s immune response to the microbiome or bacterial communities of the intestine appears to play a role in the development of NEC, and his current research focused on innate immune signaling in the developing intestine as it pertains to the development of NEC. With that in mind, let me introduce my guest today. Welcome, Dr. Matson, thank you for joining us today. I’m very excited to talk to you. DR. ADAM MATSON, GUEST: Thanks for having me here. STEPHANIE: As you know, we’re talking about Necrotizing Enterocolitis, but I’d love for you to tell me about your experience in the NICU and then in the NICU in relation to your experience with NEC. DR. MATSON: Okay, well, I am an attending neonatologist at Connecticut Children’s Medical Center, which is located in Hartford, Connecticut, and there I’m involved with taking care of premature babies and infants with other types of medical problems. And unfortunately, Necrotizing Enterocolitis is one of the disease processes that does affect premature babies in our unit as like many other NICUs around the world. In our NICU, we average probably about 14 cases of Necrotizing Enterocolitis, or I’ll refer to it as NEC, per year, so it’s a major medical problem for these infants. As I mentioned before, it’s unfortunate that I do have experience in managing these infants. STEPHANIE: So what can you tell me as a parent about I guess signs and symptoms and what you guys as the doctors and clinicians and nurses are looking for that’s, I guess raises a red flag for you that this baby might have NEC? DR. MATSON: Sure, so NEC is most common in the very small premature babies, particularly those that are with birth weights less than 1500 grams (3 pounds 4.91 ounces). So these are infants that are typically being fed by a feeding tube that’s introduced into the nose and goes down to the stomach, or into the mouth and goes down to the stomach. Usually these babies are too small or weak to eat on their own. And it’s a gradual process. We start with small volumes of feeds and increase them gradually. And the types of symptoms that babies can start to develop when this process begins can sometimes be nonspecific. They can have decreased activity, they may have increased apnea spells (moments when the baby stops breathing) is something that we’ll see. Their abdomens can become more distended. One of the things that we’ll frequently check for are something that is referred to as aspirates. This is when a nurse is going to give a feed with feeds being given every three hours. They will check the stomach to see how much of the prior feed has actually gone out of the stomach and into the intestines. So often times if the intestine is starting to not feel too happy, that feed can sort of back up and that’s called an aspirate. If the volume becomes excessive, one of the measurements that we’ll use in our unit is more than 50% of the prior feed, that’s a red flag for us. STEPHANIE: Okay, actually that was the first symptom that Morgan had was his aspirate they said was tinge green which was an immediate red flag and x-rays were taken bedside and that’s—rapidly they discovered that he had NEC and that’s when he had his surgery. So that was definitely a red flag with him. DR. MATSON: Sure, those signs occur particularly when the aspirate turns green, as you had mentioned for your son that indicates that bile that’s being emptied into the intestine is not emptying down into the more distal portions of the intestines. So for his bile to start backing up, that’s absolutely a warning sign. STEPHANIE: Okay, thank you. So is there anything else that would be a good warning for parents or questions that they should ask if something’s maybe not looking right? DR. MATSON: Well, as I had mentioned, many of the signs can be nonspecific and they can actually often occur very fast as well. You know, we do monitor as I had mentioned for those things, bloody stools as well. And if those sort of warning signs come up, typically we’ll end up holding some feeds for a while to not overwhelm the stomach or the intestine with additional food, and as you had mentioned, we’ll end up doing x-rays and that’s the primary way that Necrotizing Enterocolitis is diagnosed. Really what we’re looking for with those x-rays is a finding referred to as pneumatosis intestinalis. And what that is is part of the pathophysiology of NEC is as bacteria are starting to invade through the intestinal wall, they can start to produce gas and make gas bubbles, and when we do x-rays looking for NEC, if we visualize those gas bubbles in the walls of the intestine, that’s diagnostic that the process is indeed happening. STEPHANIE: Okay, so can you tell me a little bit on the flip side of your experience with NEC on the research side? DR. MATSON: Sure, you know, perhaps I should talk a little bit about in that regard on what we think actually causes NEC. And I think that the answer to that right now is that we don’t know exactly. But it appears to be a rather complex interaction between bacteria that are inside the intestine, and exaggerated or overactive immune response that’s happening inside the intestine. The whole hypoxia or decrease in oxygen within the intestine also probably plays a role in some cases. But studies have indicated at least in many cases of NEC it’s not—it doesn’t appear to be attributable to a single bacterial species like E. coli or Salmonella. But it appears to be more related to bacterial communities or what we would say is the microbiome of the intestine which can be influenced by certain things that we know to be risk factors for Necrotizing Enterocolitis as well such as formula feeding, where breast milk—human milk is protective, excessive use of antibiotics, antacids, those sorts of things are thought to disrupt the microbiome and result in overgrowth of different species, particularly gram negative bacteria. And when there’s an overgrowth of those types of bacteria in the intestine, those appear to activate certain receptors that are inside the intestine— this is getting into a little bit of the research that I’m involved with, because these receptors primarily in premature infancy appear to be very sensitive to a large number of these gram negative bacteria, and as they start to become activated, they start to break down the intestinal epithelial lining and this results in trans-location of bacteria through the intestinal mucosa—the protective barrier, and then activation of immune cells in the deeper layers. Another feature of the premature infant is that they’re really not able to control that immune response in their intestine very well, so they end up with a very profound inflammatory response in their intestine. That’s really what Necrotizing Enterocolitis is. It’s the most common gastrointestinal emergency in premature babies. It occurs primarily in premature infants. It’s characterized by diffuse inflammation and necrosis, or tissue death inside the intestine. And it’s also associated with very significant morbidity and mortality. About 15 to 30 percent of infants who develop NEC may ultimately die. So it’s a major problem for this population. STEPHANIE: And can you tell me, I guess a little bit more about what the hospital’s doing in their research? And more specifically, what other areas you’re researching? DR. MATSON: Sure, so our hospital, we have a number of different projects that we’re involved in. We have a very active lactation program where we’re looking at different aspects of human milk. I had mentioned before that one of the main risk factors for Necrotizing Enterocolitis is diet and formula feeding, and we do know that providing human milk reduces the risk of NEC by about 50 to 90 percent providing a diet of exclusive human milk. So we are currently looking at factors inside of breast milk, macronutrients and how they affect the bacterial populations inside of the intestine and how that may ultimately contribute to infants developing this process. More specifically in terms of laboratory work, we’re now working with some collaborators at UConn Storrs as well and we’re doing a preemie poop project where we’re collecting a lot of fecal samples from babies inside our NICU. And we’re doing a real detailed analysis, molecular analysis where we sequence out basically all the different microbial species or bacterial species inside the intestine. And one of our hopes with this study is that we’re able to identify how diet and exposure to medications affect the bacterial populations inside the intestine, which we know has a very strong role in Necrotizing Enterocolitis. I also have a laboratory at UConn Health Center in the department of pediatrics and we’re looking a little bit deeper at some of the receptors inside the intestine. There’s a group of receptors that I refer to as toll-like receptors, and these recognize molecules that we refer to as pathogen associated molecular patterns or PAMPs. So these are the receptors that are on the surface layer of the cells that line the intestine and respond to these different bacteria. And I think this is the type of research that tying in aspects of clinical care with breast milk to knowing what’s actually growing inside the intestines in terms of bacterial populations, and then looking at more detailed molecular aspects of immune signaling inside the intestine and what’s ultimately controlling the inflammatory process. STEPHANIE: That’s very interesting. Is there anything else that you would like to add about research specifically? I know one of our major goals is to help the doctors and researchers advance research through funding. So can you talk to me a little bit about funding for research within the NEC community? DR. MATSON: Sure, well I think that one of the areas that would likely help the most is more funding to look at causes of premature birth. This continues to be a major problem in the United States and elsewhere. Up to ten to eleven percent of infants are born premature. And a significant number of those babies are the very premature infants that are at the highest risk for developing NEC. So I think that I need to mention that as really one of the primary areas because there’s a lot of different challenges that these babies face, and the more that we can prevent preterm birth, I think that would be advantageous for them. The other aspect I think would be important to look at is in terms of diagnosis or earlier diagnosis. Being able to identify which babies are starting to develop some changes in their intestine earlier. I have a colleague that I work with who often says that it’s when we’re diagnosing by x-ray, it’s almost like arriving at the crime scene after the crime has already been committed. STEPHANIE: Mm-hmm. DR. MATSON: The care that we implement at that stage is really is very supportive in terms of holding feeds, antibiotics, bringing the suction tube into the stomach, getting frequent x-rays, getting the surgeons involved to help follow the infants, and in many ways, the time that we’re diagnosing these infants at this point is the process is already much too far ahead. STEPHANIE: It’s definitely a complex disease, and I know that with Morgan, I think within a span of five hours or so he was diagnosed and in and out of surgery and in recovery, so I know that it’s a rapid time frame. But I appreciate all of the information that you shared with us today—I think you’ve given a really good perspective on causes and signs and symptoms, and if there is anything else that you’d like to add in any area for parents that might be listening to this from your perspective as a doctor talking to parents, please feel free. DR. MATSON: Sure, so I could mention just a little bit more about prevention of Necrotizing Enterocolitis. In some diseases, an ounce of prevention’s worth a pound of cure. When we’re looking at certain populations in the NICU, we often classify premature infants according to their weight. Those at highest risk of developing Necrotizing Enterocolitis are what we would refer to as very low birth weight infants, and those are less than 1500 grams at birth. STEPHANIE: And that’s about three pounds? DR. MATSON: Yes, pretty close to that. And I had mentioned efforts to prevent prematurity is a major goal, also diet. The American Academy of Pediatrics came out with a statement in 2012 really encouraging the provision of human milk to all of these babies. We do know that human milk does help protect against Necrotizing Enterocolitis. And if mom’s milk is not available for these infants, many units including ours are now using pasteurized donor human milk. It’s a very safe product, and that has been shown to help as well. Other potential preventative measures is—one would be using a standardized feeding protocol. There is very good data on that. That means really sort of having a very strict protocol for each size baby and how much milk you start with with the feeds, how rapidly you advance them, and what sort of warning signs that the healthcare team should be observing for. So that has been shown to be very important. Limited use of antibiotics appears to be very important. It’s a difficult task for us while we’re inside the Newborn Intensive Care Unit because these babies are at such high risk for infection. But one of the things that data has shown is that the more antibiotics, the more unnecessary antibiotics, that these babies receive increases their chances of getting Necrotizing Enterocolitis, so that probably relates to overgrowth of gram negative and other bacteria inside the intestine that activate the inflammatory cascade. There’s a few interesting other preventative measures that are topics of conversation within our field and one is using probiotics. There is good data out of other countries. So, I should say that probiotics are live bacteria. They’ve been using older children and adults for some time for various reasons. Bifidobacterium and Lactobacillus are the most common probiotics. Those are bacteria that are typically found in the stool of breastfed infants. And many units outside of the United States are now giving these probiotics, which they’re giving them to extremely premature infants in an effort to prevent NEC from happening. And the thought is that these help to prevent some of the pathogenic bacteria from growing, they also help to mature the intestinal barrier inside the intestine. At this point in time in the United States, however, there has not been a—at least to my knowledge—there has not been a properly randomized, controlled trial to study these here. And also another major issue using probiotics in the United States is how are they regulated by the FDA as they’re considered a food. So really you can go to GNC or CVS to buy probiotics over the counter. So with that type of designation by the FDA, they don’t have the same oversight as a drug would, and one of the concerns with many of the NICUs in using a product like that is it doesn’t have the same consistent quality oversight, meaning that we don’t know how pure it is or how consistent the actual dose would be that we’re giving to premature infants, so hopefully some research down the line will help answer those questions. STEPHANIE: Well, I think you’ve given us a lot of information, a lot of really good information I think, and a lot of really relevant information for parents that will be listening. So I really appreciate you sharing your time with us, and joining us today. And so with that, I will let you go. And… DR. MATSON: Okay, well thank you very much. STEPHANIE: we will talk again. DR. MATSON: Sounds great. STEPHANIE: Thank you. DR. MATSON: Okay, take care Steph. STEPHANIE: Thank you. STEPHANIE: For more information about Dr. Matson and his research in NEC, visit: connecticutchildrens.org. A direct link can also be found in this episode’s show notes: http://www.connecticutchildrensfoundation.org/document.doc?id=402 In closing, I’d like to share a few thoughts about today’s conversation with Dr. Matson. One of Morgan’s former doctors described NEC to me as “an inflammatory response gone haywire.” That simple, but vividly descriptive, phrase gave me pretty quick understanding of the disease that nearly took my son’s life. The inability of a very premature baby to regulate their immune response, and in turn their inflammatory response, appears to be a crucial factor in the development of NEC. And as Dr. Matson mentioned, understanding not only how diet and exposure to medications affect the bacterial populations inside the intestine, but also understanding the immune signaling inside the intestine and what’s ultimately controlling the inflammatory process are critical to fully understanding, and preventing, NEC. Show your support for our smallest and most fragile babies, those who have the greatest risk for developing NEC. Show your support for continued research in NEC. And join our effort to raise awareness about, and funds for research in NEC by making a donation to Morgan’s Fund at morgansfund.org/donate. If you’ve had a personal experience with NEC and would like to share your story, or have a question or topic that you’d like to hear addressed on our show, e-mail us at feedback@morgansfund.org. We’d love to hear from you! Additional Information You can make a donation directly to Dr. Matson’s research in NEC at Connectiut Children’s Medical Center by visiting https://www.connecticutchildrensfoundation.org/giving/nec Copyright © 2015 The Morgan Leary Vaughan Fund, Inc. The opinions expressed in Speaking of NEC: Necrotizing Enterocolitis (the Podcast series) and by The Morgan Leary Vaughan Fund are published for educational and informational purposes only, and are not intended as a diagnosis, treatment or as a substitute for professional medical advice, diagnosis and treatment. Please consult a local physician or other health care professional for your specific health care and/or medical needs or concerns. The Podcast series does not endorse or recommend any commercial products, medical treatments, pharmaceuticals, brand names, processes, or services, or the use of any trade, firm, or corporation name is for the information and education of the viewing public, and the mention of any of the above on the Site does not constitute an endorsement, recommendation, or favoring by The Morgan Leary Vaughan Fund.

Toll-like receptors (TLRs) are in the front-line during the initiation of an innate immune response against invading pathogens. TLRs are type I transmembrane proteins that are expressed on the surface of immune system cells. They are evolutionarily conserved between insects and vertebrates. To date, 13 groups of mammalian TLRs have been identified, ten in humans and 13 in mice. They share a modular structure that consists of a leucine-rich repeat (LRR) ectodomain, a single transmembrane helix and a cytoplasmic Toll/interleukin-1 receptor (TIR) domain. Most TLRs have been shown to recognize pathogen-associated molecular patterns (PAMPs) from a wide range of invading agents and initiate intracellular signal transduction pathways to trigger expression of genes, the products of which can control innate immune responses. The TLR signaling pathways, however, must be under tight negative regulation to maintain immune balance because over-activation of immune responses in the body can cause autoimmune diseases. The TLR ectodomains are highly variable and are directly involved in ligand recognition. So far, crystal structures are missing for most TLR ectodomains because structure determination by X-ray diffraction or nuclear magnetic resonance (NMR) spectroscopy experiments remains time-consuming, and sometimes the crystallization of a protein can be very difficult. Computational modeling enables initial predictions of three-dimensional structures for the investigation of receptor-ligand interaction mechanisms. Computational methods are also helpful to develop new TLR agonists and antagonists that have therapeutic significance for diseases. In this dissertation, an LRR template assembly approach for homology modeling of TLR ligand-binding domains is discussed. To facilitate the modeling work, two databases, TollML and LRRML, have been established. With this LRR template assembly approach, the ligand-binding domains of human TLR5-10 and mouse TLR11-13 were modeled. Based on the models of human TLR7, 8 and 9, we predicted potential ligand-binding residues and possible configurations of the receptor-ligand complex using a combined procedure. In addition, we modeled the cytoplasmic TIR domains of TLR4 and 7, the TLR adaptor protein MyD88 (myeloid differentiation primary response protein 88) and the TLR inhibitor SIGIRR (Single immunoglobulin interleukin-1 receptor-related molecule) to investigate the structural mechanism of TLR negative regulation.

Toll-like receptors (TLRs) recognize pathogen-associated molecular patterns (PAMPs) on invading organisms and are the first line of defense in innate immunity. To date, much has been learned about TLRs and their roles in autoimmune diseases are being unraveled. The autoimmune disease systemic lupus erythematosus (SLE) progresses as a consequence of the inappropriate recognition of self nucleic acids by TLRs. For the development of therapeutic approaches of SLE it is necessary to understand possible negative regulation mechanisms of TLR. Single immunoglobulin interleukin-1 receptor-related molecule (SIGIRR) is the best characterized TLR signaling inhibitor. It can interfere with the receptor complexes and attenuate the recruitment of downstream adaptors to the receptors. So far, the mechanisms of structural interactions between SIGIRR, TLRs and adaptor molecules are unknown. To develop a working hypothesis for these interactions, we constructed three- dimensional models for these single molecules based on computational predictions. Then, models of essential complexes involved in the TLR signaling and the SIGIRR inhibiting processes were yielded through protein-protein docking analysis. With the high-throughput genome sequencing projects, a central repository for the growing amount of TLR sequence information has been created. However, subsequent annotations for these TLR sequences are incomplete. For example, the indicated numbers and positions of leucine-rich repeat (LRR) motifs contained in individual TLR ectodomains are greatly distinct or missing in established databases. In this vein, we have developed a database of TLR structural motifs called TollML (http://tollml.lrz.de). It integrates all TLR protein sequences that have been identified to date. These sequences were semi-automatically partitioned into three levels of structural motif categories. The manual motif identification procedure provided TollML with the most complete and accurate database of LRR motifs compared with other databases that contain TLR data. LRR motifs are present not only in TLRs, but also in many other proteins. To date, more than 6,000 LRR protein sequences and more than 130 crystal structures of them have been determined. This knowledge has increased our ability to use individual LRR structures extracted from the crystal structures as building blocks to model LRR proteins with unknown structures. Because the individual LRR structures are not directly available from any protein structure database, we have developed a conformational LRR database called LRRML (http://lrrml.lrz.de). It collects three- dimensional LRR structures manually identified from all determined crystal structures of LRR-containing proteins and thus provides a source for the structural modeling and analysis of LRR proteins. With the help of TollML and LRRML, we constructed models of the human/mouse TLR5-13 ectodomains and suggested some potential receptor-ligand interaction residues based on these models.

The objective of this work was to systematically review and discuss recent studies and articles dealing with the subject of the immunology of female genital tract mucosal tissue. The emphasis hereby lies on the evaluation of studies concerning the basics of female reproductive immunology, research on immunology of the most important genital infections and vaccination strategies, immunologic principles at the fetomaternal interface during normal pregnancy and its complications as well as on immunologic data on infertility and immunocontraception. It is now established that the mucosal immune system is a distinct and separate component of the host`s immune apparatus and differs from the lymphoid tissues in peripheral sites. Furthermore, despite some common features, the female genital tract mucosal system displays some distinct characteristics which outlines its special role. Analysis of the female genital tract indicates that the key cells of the innate and adaptive immune systems are present and functionally responsive to antigens; however, there is a certain degree of compartmentalization within the tract. The identification of TLRs in the fallopian tubes, uterus, cervix, and vagina and the presence of ECs, macrophages, DCs, NK cells, and neutrophils throughout the reproductive tract along with their responsiveness to selected PAMPs indicate that the female reproductive tract has evolved to meet the challenges of STDs, while at the same time supporting an immunologically distinct fetal placental unit. To meet these diverse challenges, innate and adaptive immune system in the female genital tract are precisely regulated not only by a network of cytokines and chemokines, but also by the sex hormones estrogen and progesterone. Understanding the specialty of the genital tract immune system is of critical importance, because STDs are and will be a major worldwide health problem. Despite extensive efforts, only limited success has been achieved in dealing with a growing list of STDs. The role of immune factors in the control of genital viral and bacterial infections appears complex and needs further studying, also with respect to creating vaccines. Despite the recognition that innate immunity as the first line of defense and adaptive immunity, especially Th1 immune responses, play a critical role in preventing infection and in limiting viral replication, factors such as antimicrobials and TLRs that contribute to the mucosal response in the female genital tract have only recently begun to receive attention. Further studies are also needed to elucidate the relationship between mucosal immunity, the hormonal environment, and response to pathogen challenge. More data must be collected on the mechanisms of immune evasion by several pathogens such as HSV, N. gonorrheae or Chlamydia. While considerable information can be obtained from animal experiments, important differences in the physiology of reproduction and the immune sytem result in the need for studies in humans. Further knowledge on female tract immunology will also impact on immunological approaches to contraception, immunological infertility and the immunological aspects of pregnancy. This does not only involve new options for diagnostics but also for treatment of pregnancy complications such as preeclampsia, preterm birth and early pregnancy loss as well as for infertility. Pregnancy involves maternal tolerance of the semiallogenic histoincompatible fetus and is characterized by the enhancement of the innate immune system and suppression of the adaptive immune response, probably with progesterone as the important regulator. In opposite to normal pregnancy, improper immune responses and an unbalanced cytokine network may characterize implantation failures, pregnancy loss and obstetric complications. These are the presence of elevated Th1/Th2 cell ratios, high concentrations of Th1 cytokines, elevated NK cell cytotoxicity and levels, and emergence of various autoantibodies. These immunological approaches needs to be investigated and evaluated further with respect to widening of treatment options by modification of immune responses.

Die beiden Pathogen-assoziierten molekularen Muster, CpG-Oligonukleotide, als Imitate bakterieller DNA, und LPS, sind in der Lage, das menschliche Immunsystem zu stimulieren. Vor Entdeckung der Toll-like Rezeptoren konnte nicht zwischen direkten und indirekten Effekten von CpG-Oligonukleotiden und LPS auf Zellen des menschlichen Immunsystems unterschieden werden. Durch die Entdeckung der Toll-like Rezeptoren und vor allem die Charakterisierung von TLR9 als Rezeptor für CpG und TLR4 als Rezeptor für LPS entstand die Möglichkeit, die Zielzellen von PAMPs an Hand der Expression der dazugehörigen Rezeptoren zu definieren. Dendritische Zellen sind im Immunsystem des Menschen essenziell für die Auslösung einer Immunantwort. Sie sind in der Lage, eindringende Pathogene an Hand von PAMPs schnell und sicher zu erkennen, und daraufhin eine passende Immunantwort zu initiieren. Zwei Subpopulationen von dendritischen Zellen konnten kürzlich im peripheren Blut identifiziert werden: Plasmazytoide dendritische Zellen (PDC) und myeloide dendritische Zellen (MDC). In der vorliegenden Arbeit wurden die Unterschiede zwischen PDC und MDC in ihren Reaktionen auf CpG-Oligonukleotide, LPS und CD40 Ligand charakterisiert. Funktionelle Untersuchungen zeigten, dass nur PDC und nicht MDC direkte Zielzellen von CpG-ODN im humanen Immunsystem darstellen, während LPS MDC aktiviert, jedoch nicht PDC. Damit konsistent konnte auf molekularbiologischer Ebene nachgewiesen werden, dass PDC TLR9 exprimieren, jedoch nicht TLR4, während MDC den für die Erkennung von LPS notwendigen Rezeptor TLR4 besitzen, aber TLR9 nicht exprimieren. In gemischten Populationen reagierten auch myeloide dendritische Zellen auf CpG-Oligodesoxynukleotide, was auf eine indirekte Aktivierung durch plasmazytoide dendritische Zellen hinweist. PDC reagierten nach Stimulation mit ODN 2006 mit einer Hochregulation von Reifemarkern und kostimulatorischer Moleküle, der Expression von Chemokinrezeptoren, der Produktion proinflammatorischer Chemokine und einer verminderten Apoptoserate. Nach Stimulation mit ODN 2216 sezernierten sie große Mengen IFN-α, während ODN 2006 für die Induktion von IFN-α eine Kostimulation mit CD40 Ligand benötigte. Weder ODN 2006, ODN 2216 oder CD40 Ligand alleine waren in der Lage, IL-12 in PDC zu induzieren, die synergistische Stimulation von PDC mit CpG-ODN und CD40 Ligand führen jedoch zur Produktion großer Mengen an IL-12. Unter optimaler Stimulation mit ODN 2006 und CD40 Ligand können PDC damit gleichzeitig IL-12 und IFN-α produzieren. Das Verhältnis der produzierten Zytokine ist dabei abhängig vom Differenzierungsgrad der PDC. Durch zunehmende Ausreifung der PDC mit IL-3 verschiebt sich nach der Stimulation das Produktionsverhältnis an sezernierten Zytokinen zugunsten von IL-12. Ausreifung mit ODN 2006 ermöglicht PDC, zudem naive allogene CD4 Zellen zu aktivieren, und induziert in Kokulturen IL-12-abhängig ein Th1-gerichtetes Zytokinprofil in den CD4 T-Zellen. IFN-α schien dabei eine geringe Rolle zu spielen. Durch die Charakterisierung der PDC als TLR9-tragende Zielzelle für CpG-DNA, trägt diese Arbeit entscheidend dazu bei, die PDC als Schlüsselzelle für die physiologischen Wirkungen von TLR9-Liganden zu identifizieren und zu verstehen. Dies ist von hoher Relevanz für die Entwicklung therapeutischer Anwendungen von CpG-ODN in der Tumortherapie, Asthmabehandlung, Infektionsprophylaxe und als Adjuvans bei Impfungen.

Der opportunistisch humanpathogene Hefepilz Candida albicans gehört bei vielen gesunden Menschen zur mikrobiellen Schleimhautflora, kann jedoch bei abwehrgeschwächten Patienten oberflächliche Infektionskrankheiten sowie lebensbedrohliche Organmykosen verursachen. Obwohl der Immunstatus des Wirtes für eine Infektion mit diesem Erreger von entscheidender Bedeutung ist, trägt auch eine Reihe von Virulenzfaktoren, insbesondere die sekretorischen Aspartatproteasen (Saps), zur Pathogenität von C. albicans bei. Die angeborene Immunität ist in der Lage, derartige Pathogene schon beim Erstkontakt zu erkennen und zu bekämpfen. Haupteffektoren dieser schnellen, angeborenen Immunantwort sind Makrophagen und neutrophile Granulozyten. Mitglieder der Toll-Proteinfamilie, sogenannte Toll-like Rezeptoren (TLRs), wurden kürzlich als Rezeptoren auf diesen Immunzellen in Säugern identifiziert. Sie erkennen unterschiedliche Erreger anhand von in der Evolution hoch konservierten Strukturen, den Pathogen-assoziierten molekularen Mustern (PAMPs), was zur Aktivierung des Transkriptionsfaktors NF-кB und zur Freisetzung von Zytokinen führt. Sowohl TLR2 als auch TLR 4 wurden kürzlich für die Erkennung von C. albicans diskutiert. Zielsetzung dieser Arbeit war es, die Interaktion von Makrophagen mit C. albicans im Hinblick auf die Aktivierung von Toll-like Rezeptoren und die nukleäre Translokation von NF-кB zu untersuchen. Neben dem lebenden C. albicans-Isolat wurden zudem drei weitere Präparationen untersucht: Mit den Antimykotika (AM) Amphotericin B, Nystatin und Itraconazol vorbehandelte Keime, durch Hitze inaktivierte Keime sowie Sap-inaktivierte Keime. Die Zellstimulationsexperimente wurden mit murinen Wildtyp-Makrophagen, TLR2- bzw. TLR4- defizienten Einzelknockoutmutanten und mit TLR2/4-Doppelknockoutmutanten durchgeführt. Die TLR-vermittelte Aktivierung von NF-кB wurde mit Gelshifts (EMSA) nachgewiesen. Mit Western Blots wurden die intrazellulären Signaltransduktionswege untersucht. Der Hitze-inaktivierte Stamm bewirkte keine Translokation von NF-кB in Wildtyp-Makrophagen. Eine Inhibition der Saps bewirkte keine Abschwächung der NF-кB Induktion, so dass im Umkehrschluss dieser bedeutende Virulenzfaktor die TLR-vermittelte NF-кB Aktivierung nicht beeinflusst. Der lebende Stamm benutzte sowohl TLR2 als auch TLR4 für die Induktion von NF-кB. Nach Vorstimulation der Makrophagen mit Interferon-γ ließ sich jedoch eine klare TLR2-Abhängigkeit – unabhängig von TLR4 – in der Aktivierung von NF-кB und in der Induktion von TNF-α zeigen. In beiden Fällen wurden die Makrophagen erst ab einer Candida-Dichte von 106 Zellen pro 100 µl PBS stimuliert. Für den AM-vorbehandelten Stamm ergab sich eine deutliche TLR2-Abhängigkeit in der Regulation von NF-кB, welche durch die Präinkubation der Makrophagen mit IFN-γ nicht beeinflusst wurde. AM-vorbehandelte Keime konnten NF-кB in den Makrophagen erst ab einer Dichte von 107 Zellen pro 100 µl PBS aktivieren. Zusammenfassend zeigen diese Ergebnisse, dass lebende und AM-vorbehandelte Keime im Gegensatz zur Hitze-inaktivierten Präparation und zu den Saps relevante PAMP-Strukturen für eine TLR-vermittelte NF-кB Hochregulation besitzen. Die Beteiligung beider Rezeptoren, TLR2 und TLR4, belegt beim lebenden Stamm das Konzept, dass immunkompetente Zellen sich mehrerer TLRs bedienen, um die Immunantwort möglichst spezifisch und fein zu regulieren. Beim AM-vorbehandelten Stamm scheint den Antimykotika Amphotericin B, Nystatin und Itraconazol eine besondere Rolle zuzukommen, da diese die Integrität der Pilzmembran stören und somit TLR2-aktivierende PAMPs aus Zellwand und/oder Zytosol freisetzen. Neben dem direkten Effekt auf die Pilzmembran kommt es somit zusätzlich zu einer indirekten, TLR2 vermittelten Stimulation der Makrophagen. Untersuchungen der Signaltransduktion (Stimulation von Wildtyp-Makrophagen mit dem AM-vorbehandelten C. albicans-Isolat) ergaben eine vorübergehende, zeitlich eng begrenzte Induktion von NF-кB, die durch den Inhibitor IкB-α reguliert wird. Gleichzeitig wurden im zeitlichen Verlauf der Stimulation auch MAP Kinasen (ERK, p38, JNK) und c-Jun, eine Subeinheit des Transkriptionsfaktors AP-1, phosphoryliert. Diese simultane Aktivierung beider Transkriptionsfaktoren weist auf eine feinregulierte Immunantwort der Makrophagen gegenüber C. albicans hin und legt zudem einen Cross-Talk zwischen NF-кB und AP-1 nahe.