Podcasts about S100

Spring has sprung, at least it felt like it today with the beautiful weather. Breaking Away Matt joins us to talk about the upcoming Misfits Rally April 3-6. https://breakingawayadventures.com/misfits-rally-vol-3. Jim ventured out on some dirt roads on the back side of Big Sur, and shares his ride report. And of course riding on dirt roads can leave your bike dirty, so we are joined by Steve Gay from S100, who makes cleaning products engineered for motorcycles. He shares the history of S100, starting with Doctor Wack, who created the formula to clean his kids dirtbikes. He also talks about how the formula is designed to not be harmful to alloys, rubber, plastic, paint and all the other surfaces typically found on motorcycles. If you'd like to try it yourself, enter our contest to win an S100 cleaning kit. Send us a pic of your dirty bike to Motorcyclesandmisfits@gmail.com. Next up, we talk about the latest Common Tread video, which deserves and award of some kind. And lastly, Neil talks about a dark subject, and then we finish with listener emails. With Liza, Miss Emma, Neil, Scottie, Jim and Bagel. https://www.s100.com/ www.leodescapes.com/ breakingawayadventures.com/misfits-rally-vol-3 motorcyclesandmisfits.com/shop Join our Discord at discord.gg/hpRZcucHCT www.motorcyclesandmisfits.com motorcyclesandmisfits@gmail.com www.patreon.com/motorcyclesandmisfits www.zazzle.com/store/recyclegarage www.youtube.com/channel/UC3wKZSP0J9FBGB79169ciew

The sounds of the S100 part 2.  Björn and Andreas keep exploring the sounds of the S100. Björn explains the effect loop and they play through both the lead and normal Chanel's of the amp. This episode is divided into  three segments: 1. Using the loop with reverb and the amps echo. 2. Normal chanel sounds and different echo settings. 3. Using the amp as a pedal platform and stacking drive pedals for different gain textures

Nobles out of capacity for S100 , Douglas Prom tram plan, our Ørsted offshore wind farm, Allinson on Assisted Dying Bill & wildlife bounty of cash and cartridges. It's Mannin Line with Andy Wint #iom #manninline #manxradio

The Spanish TEMSA Metallurgical Group is one of the world's leading manufacturers of special tools for cold forming. The company also uses cylindrical grinding machines from STUDER in its production facility in Barcelona. Alfonso Vivar walks through a large hall that radiates a sense of order, despite its multitude of machinery, workstations with monitors, and focused employees. Everything has its place, every work step is carefully planned, and at its heart is a yellow robot arm working away methodically in an 8,000sq/m plant to the west of Barcelona, Spain. Production Manager Alfonso Vivar knows every corner of the plant. His shirt bears the bright red logo of his employer, TEMSA Metallurgical Group, a leading global specialist in the production of high-precision tools for cold forming and an expert in powder metallurgy, sintering and fine cutting. With cold forming, metal below the recrystallisation temperature is forced into a specific shape using high compressive and tensile forces. Compared to metal-cutting operations, this allows for shorter processing times per workpiece, thereby reducing costs in series production. Cold forming also allows for high strength, complex geometries, and excellent surface properties of components for high-tech industries such as aerospace and automotive.  “Our team can manufacture special tools in a tolerance range of just a micron,” explains Vivar, proudly. TEMSA's site in Barcelona has around one hundred employees, all working to manufacture tools to exacting customer requirements. “Our expertise also helps us to accomplish short lead times. But that's only because we use the very best machinery.”  Confidence in the Technology “We acquired a new STUDER S100 earlier this year, together with a favoritCNC.” Vivar points to the CNC universal internal cylindrical grinding machine, painted white with blue accents. This colour combination is typical of STUDER's cylindrical grinding machines and is a familiar sight at the TEMSA plant. The company operates several STUDER machines for grinding tasks, including the S131, a new generation CNC universal machine for internal cylindrical grinding. “We have confidence in the technology, and value the positive relationship we have with the manufacturer,” explains Vivar of the decision to invest. As an example, the S100 is a great all-rounder offering maximum precision thanks to its numerous options for internal, face, and external grinding. The machine facilitates the production of a wide range of workpieces up to 550mm in length. The favoritCNC is a CNC universal cylindrical grinding machine for individual and batch production of medium-sized workpieces with a length of up to 680mm. Both machines have an exceptional price-performance ratio, while at the same time offering premium technology, such as the machine bed of solid mineral cast Granitan® and optimal hardware-software interplay for ease of operation. The S131 for internal cylindrical grinding with a patented StuderGuide® guide system, turret wheel head with up to four grinding spindles, and an additional C-axis is ideal for high-precision manufacture of flanged parts and smaller workpieces in a wide range of applications. In addition to the quality level of the machinery, Vivar cites another important reason for opting for STUDER: “The customer service is outstanding, and we have several contact persons who can speak Spanish,” he explains. This helps to ensure easy and direct communication. He also values what STUDER has to offer in terms of preventive maintenance. This includes regular and routine inspections of the machinery by STUDER's technical personnel, designed to minimise the risk of production failures and improve their operational durability. Riccardo Delai, Sales Manager for Latin Europe at STUDER visits the plant in Barcelona at regular intervals and is often on the phone with TEMSA employees. “Personal contact is extremely important as it helps me to know what is working well and what needs attention. We are delighted with the international success of TEMSA as a fully Spanish company, and it demonstrates what can be achieved with our machinery,” emphasises Delai. Investing in the Future Safeguards Success TEMSA's success story goes back more than 30 years when the fledgling company produced high-precision tools from tungsten carbide and steel. Through consistent investment, the Spanish company has quickly been able to establish an excellent reputation around the world. Today, the company is a world leader in special tools for cold-forming processes. “I am extremely proud of our team. We have many second-generation employees working with us, and at Christmas, we always have a meal together with the first generation,” explains Vivar, a TEMSA veteran of 25 years.  This appreciation for tradition is paired with foresight for the future, and TEMSA was an early adopter of automation to make production more efficient. One such example is the yellow, 360-degree operating robot arm here in the plant, which automatically sorts numerous tools and allocates them to the machine according to requirements and processing cycles. “We want to move automation up a level in the future, which is a real challenge,” explains Vivar. This is another area in which STUDER is a valuable partner, having ample experience in customised and standardised automation solutions. As an example, the S100 can be equipped with a loader interface and automatic sliding door and thus integrated into an automated production line. The S131 also has a standardised interface for a loader and peripheral equipment.

Like the ‘Sound of Music' that has won its place in movie folklore, the 2024 ‘Sound of Studer' themed event that took place at the company's Steffisburg headquarters has won its place in the hearts and minds of the 65 journalists from over 20 countries that attended. The annual Fritz Studer AG event is always packed with the latest innovations in grinding, an insight into the company's ongoing evolution and an overview of the global trends in the manufacturing industry - and the February event once again lived up to its billing. By Rhys Williams Located in the Swiss Alps, in a region more stunning than the backdrop to the legendary movie, the ‘Music Motion Meeting 2024' kicked off with presentations from the management team and was followed by technical presentations on the latest innovations and an insight into what we can expect to see as we move through 2024.  Studer CEO Jens Bleher kicked off proceedings with a presentation that discussed the company's performance over the last 12 months, the market turbulence, new updates to the CORE interface, product enhancements and the pride in the success the Swiss manufacturer continues to enjoy with its success at the World Skills event.  “Despite the challenging investment environment, we again increased our sales, especially in key markets like the USA and China. Asia was the largest single region followed by Central Europe and North America and consistent investments in product development and site infrastructure have paid off. We have turned our announcements into reality and improved our market position to strengthen ourselves for the future in the long term,” the Studer CEO announced.  Studer once again won market share in many regions around the world, and it hit a new sales record in its Customer Care segment. Bleher took it as a positive sign for the 2024 fiscal year that the order situation developed exceptionally well towards the end of 2023. Whilst confident that order intake will continue in a positive direction, the CEO was particularly bullish about the expected performance in the second half of 2024 – a period when the manufacturer of cylindrical grinding machines will present new technologies.  Although the development of incoming orders was weaker in individual markets, such as Germany, China and some Asian countries, other parts of the world saw positive results. At the 2023 event, Turkey and several Eastern European countries were the surprise performers. At this year's event, Sandro Bottazzo, CSO at Studer said: “We achieved good results in many countries and even posted a new record order intake in some.” Some of the standout performers were announced as Spain, Austria and Holland, a welcome boost with the headwinds of global uncertainty and inflation faced by some of the manufacturing powerhouse nations.  Looking at the industry by customer segments, a point that should be marked as a good barometer of worldwide manufacturing performance for MTDCNC readers was that the aerospace industry once again generated the largest increase in new orders. This industry has now overtaken the declining automotive industry for Studer. The ‘tool' manufacturing segment remained below expectations with the largest individual segment for the second consecutive year being ‘precision engineering'. “I am very pleased that our grinding machines are so highly trusted by small and medium-sized enterprises, and that this segment remains strategically important for us,” explained Bottazzo. The ‘mechanical engineering' and ‘mould & die' segments also maintained their critically important positions. The increasing demand for precision cylindrical grinding machines in the semiconductor industry has been marked as very pleasing for Studer with Bottazzo confirming robust semiconductor growth in the Americas and Asia. The performance of specific precision cylindrical grinding machines and maintaining a broad portfolio remained of particular importance in 2023. Incoming orders for CNC universal cylindrical grinding machines were reported as very solid. The machine with the highest sales volume was the S33, followed by the favoritCNC, the S31, the S41 and the favorit. “When it comes to internal cylindrical grinding machines, it was the third-best year for incoming orders in the company's history for the S131,” said Bottazzo. Orders for the new S100 internal cylindrical grinding machine were also very pleasing and exceeded targets. Whilst 2023 sales volumes didn't exceed the 2022 numbers that were reported as the company's third-best year on record, the order value was almost on par. This was due to higher-value turnkey solutions and the Customer Care packages reaching record numbers. Alluding to the Customer Care division, Bottazzo added: “Here we were able to set a sales record for the second consecutive year. This applies to all business areas from maintenance and service to spare parts and overhauls. Particularly pleasing was the development in Eastern Europe, where Studer now performs service with its skilled personnel in the Czech Republic, Slovakia, Poland, and Hungary.” This expansion and the incredible growth have lit the way for the company to continue investing in the Customer Care division.  In 2023, Studer participated in more than 30 trade shows. At EMO 2023 the company presented three automation solutions for the first time. The arrival of the new insertLoad loader as well as the roboLoad and uniLoad was a paradigm shift for the company and these were prominent during the press tour of the facility.  Throughout 2023, the focus remained on the development and advancement of grinding technologies. The S36 external cylindrical grinding machine introduced in May 2022 has proven particularly suitable for components in the e-mobility sector since its arrival. Alluding to this, CTO Daniel Huber added: “Now, our customers can use all spindle variants and automation solutions without restrictions on the S36. Thanks to the new, powerful grinding spindle with 25kW, it is possible to use particularly wide grinding wheels up to 160mm.” The S36 fills a gap between the existing S11 and S22 machines and it offers greater spindle and automation variants. This includes integration with the easyLoad and uniLoad systems that permit customers to automate production on the S36. Additional options include the ability to add high-speed grinding with CBN or diamond grinding wheels and the Smartjet system. Huber also mentioned the success of the new S100 internal cylindrical grinding machine that was introduced in October 2022. Since this introduction, new spindle options have already been added such as the powerful Ø58mm dressing spindle with more additions planned for further models in the future. As the company moves through 2024, sales are expected to remain strong with a projected upturn in the second half of the year. This will be set in motion by major exhibition appearances and the arrival of new models for the respective events. The new arrival will be a successor to the popular favoritCNC that will reveal new features. Some features in the pipeline include an angle display of the wheel head, the popular QuickSet setup function and a conventional mode as a manual grinding cycle that permits manual grinding without any programming. A particular focus for development in 2023 was given to the company's C.O.R.E. hardware and software architecture from the UNITED GRINDING GROUP. “The C.O.R.E. touch operating panel already allows an unprecedented level of intuitive use. Soon there will be a customisable interface where operators can define and save their workspace,” explained Huber. Data collection and analysis, as well as new sensors and instruments, were also a focus. Looking to the future of the business, the CTO added: “The issue of sustainability continues to be important. The SmartJet® cooling system developed by Studer already sets new industry standards with its ability to reduce the need for coolant and energy in the grinding process. Demand for high-performance semiconductors is also rising due to e-mobility and photovoltaics. Studer's S41 CNC universal cylindrical grinding machine with in-situ X-ray measuring head now sets the standard in wafer manufacturing. It is extremely successful in the market. In the long term, only the machine manufacturers who offer intelligent and efficient machines incorporating the latest technology can be successful,” summarised the CTO as he reflected on the extensive development activities over the past 12 months. Obviously, the success of a business is not purely based upon its product lines, but also its very foundations. To reflect upon the internal business activities, COO Stephan Stoll provided an overview, saying: “The production mix of the machines manufactured has shifted to more complex systems through 2023. With active procurement management and the normalisation of global supply chains, orders were completed on time.” Stoll positively assessed the implementation of the joint production strategy within the UNITED GRINDING Group, which provided Studer with significantly improved utilisation. Major operational projects included investments in automated test stands and manufacturing tools. As one of the very few grinding manufacturers producing high-quality spindles, the expertise for these strategically important machine components has been expanded. Comprehensive structural and logistical measures were further implemented in Steffisburg. This has been bolstered in the internal grinding competence centre in Biel with plans for further growth. Referring to the additional growth, the COO said: “After the considerable investments of the past years, the redesign of our logistics processes and warehouse infrastructure is now imminent. The centrepiece of this project is a central logistics hub with a directly connected container warehouse. The higher efficiency of the fully automated warehouse system will benefit machine production.”  The press was shown a roadmap of the planned changes to the manufacturing facility and how it will streamline production, service and logistics. In conclusion, Jens Bleher emphasised the great importance of well-trained employees. He was very pleased with the recent successes at the prestigious professional championships, SwissSkills. Last year, Studer apprentices Luis Salzmann (1st place, design engineer EFZ) and Noah Rossel (2nd place, automation engineer EFZ) won the gold and silver medals with their outstanding performances. “STUDER is represented for the third time in a row at the WorldSkills showcase. We are very proud of this. It confirms our extensive commitment to vocational training,” concludes CEO Bleher. This commitment to education was further strengthened with an award presentation at the press event for its research prize, the ‘Fritz Studer Award'.  Applicants from several European countries submitted their work and it was Dr. Emil Sauter who won the award and the prize of CHF 10,000. The Fritz Studer Award is aimed at graduates from European universities and technical colleges. “The objectives of the research prize are to boost innovation in the machine tool industry with feasible solutions whilst also promoting young technical and scientific talents,” says Dr Frank Fiebelkorn, Head of Research and Technology at Studer.  Numerous theses and dissertations were submitted to Studer and evaluated by a panel of experts consisting of Prof. Konrad Wegener, Inspire Institute for Machine Tools and Manufacturing at ETH Zurich, Dr Hans-Werner Hoffmeister, former head of the Manufacturing Technology department at the Institute for Machine Tools and Manufacturing Technology of the TU Braunschweig, and Dr Frank Fiebelkorn, Head of Research and Technology at Fritz Studer AG. Criteria for evaluating the works included the feasibility of the findings in the mechanical engineering industry, the degree of innovation and quality of the research idea, scientific content, form, and the accuracy of statements, and the results and findings.  The Fritz Studer Award 2023 was presented to Dr. Emil Sauter from the Institute for Manufacturing Technology and Machine Tools at ETH Zurich. He impressed the entire jury with his topic. ‘Detection and avoidance of thermal damage for high-performance metal grinding processes using hybrid machine learning models.' His work deals with the development of an innovative condition monitoring system for external cylindrical grinding with metal-bonded CBN tools, which recognises thermal damage in situ and predicts the remaining useful life of grinding tools with relative accuracy. Process parameters such as structure-borne noise, spindle current and force characteristics can be identified. With time-frequency transformations, the research identified features of a process to detect different stages of thermal damage. The remaining tool life can also be estimated. In general, this work with its many industry-oriented practical tests also shows that machine learning methods can lead to higher productivity and improved component quality. Whilst MTD magazine extends its congratulations to Dr Emil Sauter, we also applaud Studer for creating a platform for the engineers of tomorrow.

Install Nexus Wars for Free ✅ : https://nwen.onelink.me/gb8n/Blitz! and get over S100 dollars worth of prizes by using code: NWHeroPack! Come join me on this adventure! Support Inside Games on Patreon: https://www.patreon.com/insidegamesYT Hosted by: Lawrence: http://twitch.tv/sirlarr | Bruce: http://twitch.tv/brucegreene Edited by: ShooklynTV: https://twitter.com/ShooklynTV Written by: Lawrence Sonntag & Brian Gaar: https://www.twitch.tv/briangaar Sources -- [Metacritic] Marvel's Spider-Man 2 - https://www.metacritic.com/game/marvels-spider-man-2/critic-reviews/ [OpenCritic] Marvel's Spider-Man 2 - https://opencritic.com/game/15052/marvels-spider-man-2 [GamesRadar] Marvel's Spider-Man 2 review: "Quite simply the best superhero game yet" - https://www.gamesradar.com/marvels-spider-man-2-review/ [IGN] Marvel's Spider-Man 2 Review - https://www.ign.com/articles/marvels-spider-man-2-review [Destructoid] Review: Marvel's Spider-Man 2 - https://www.destructoid.com/reviews/review-marvels-spider-man-2/ [Eurogamer] Marvel's Spider-Man 2 review - cluttered but no less lovable action - https://www.eurogamer.net/marvels-spider-man-review-1 [Kotaku] Spider-Man 2 Fast Travel Is So Quick Players Think It's A Trick - https://kotaku.com/spider-man-2-fast-travel-ps5-instant-1850938656 [ResetERA] Marvel's Spider-Man 2 – Review Thread - https://www.resetera.com/threads/marvels-spider-man-2-%E2%80%93-review-thread.775337/page-22#post-113553404 [The Washington Post] ‘Marvel's Spider-Man 2' offers a familiar story and pristine gameplay - https://www.washingtonpost.com/entertainment/video-games/2023/10/16/marvels-spider-man-2-ps5-review/ [Kotaku] Spider-Man 2: The Kotaku Review - https://kotaku.com/marvel-spider-man-2-review-ps5-venom-miles-morales-1850929271 [PlayStation Blog] New look for PS5 console this holiday season - https://blog.playstation.com/2023/10/10/new-look-for-ps5-console-this-holiday-season/ [IGN] PS5 Slim U.S. Release Date and Spider-Man 2 Bundle Leaked Online - https://www.ign.com/articles/ps5-slim-us-release-date-and-spider-man-2-bundle-leaked-online [Dealabs] PS5 Slim : Le tout premier pack s'apprête à sortir, on vous révèle le jeu qui l'accompagne ! - https://www.dealabs.com/magazine/ps5-slim-le-tout-premier-pack-sapprete-a-sortir-on-vous-revele-le-jeu-qui-laccompagne-31953 Music — Switch It Up - Silent Partner https://youtu.be/r_HRbXhOir8 Get Back - Silent Partner https://youtu.be/iQYmgOrPEvs Kula - Topher Mohr and Alex Elena https://youtu.be/0bywp0qTVNo Funk Down - MK2 https://youtu.be/SPN_Ssgqlzc

Link to bioRxiv paper: http://biorxiv.org/cgi/content/short/2022.12.05.518134v1?rss=1 Authors: Sullivan, M. A., Lane, S. D., Ball, S. R., Sunde, M., Neely, G. G., Moreno, C., Werry, E. L., Kassiou, M. Abstract: Background: Widescale evidence points to the involvement of glia and immune pathways in the progression of Alzheimers disease (AD). AD-associated iPSC-derived glial cells show a diverse range of AD-related phenotypic states encompassing cytokine/chemokine release, phagocytosis and morphological profiles, but to date studies are limited to cells derived from PSEN1, APOE and APP mutations or sporadic patients. The aim of the current study was to successfully differentiate iPSC-derived microglia and astrocytes from patients harbouring an AD-causative PSEN2 (N141I) mutation and characterise the inflammatory and morphological profile of these cells. Methods: iPSCs from three healthy control individuals and three familial AD patients harbouring a heterozygous PSEN2 (N141I) mutation were used to derive astrocytes and microglia-like cells and cell identity and morphology were characterised through immunofluorescent microscopy. Cellular characterisation involved the stimulation of these cells by LPS and A{beta}42 and analysis of cytokine/chemokine release was conducted through ELISAs and multi-cytokine arrays. The phagocytic capacity of these cells was then indexed by the uptake of fluorescently labelled fibrillar A{beta}42. Results: AD-derived astrocytes and microglia-like cells exhibited an atrophied and less complex morphological appearance than healthy controls. AD-derived astrocytes showed increased basal expression of GFAP, S100{beta} ; and increased secretion and phagocytosis of A{beta}42 while AD-derived microglia-like cells showed decreased IL-8 secretion compared to healthy controls. Upon immunological challenge AD-derived astrocytes and microglia-like cells show exaggerated secretion of the pro-inflammatory IL-6, CXCL1, ICAM-1 and IL-8 from astrocytes and IL-18 and MIF from microglia. Conclusion: Our study showed, for the first time, the differentiation and characterisation of iPSC-derived astrocytes and microglia-like cells harbouring a PSEN2 (N141I) mutation. PSEN2 (N141I)-mutant astrocytes and microglia-like cells presented with a primed phenotype characterised by reduced morphological complexity, exaggerated pro-inflammatory cytokine secretion and altered A{beta}42 production and phagocytosis. Copy rights belong to original authors. Visit the link for more info Podcast created by Paper Player, LLC

Navaは、Jamesが行けなかった銚子の滝に再びナーバーダビットソン（YAMAHA BW'S100）で様子見に。駐車場から銚子の滝まで徒歩20分程度の山道散策コースを確認しましたよ。台風14号、15号が過ぎた後だったので、川の水量も滝の水量はかなり増えていました。滝からの帰り道、ガマガエルに遭遇。踏みそうになりました。その後、ナーバーダビットソンでの帰り道、道のど真ん中で居眠りする？（車と接触して脳震盪で倒れていた？）うりぼうと遭遇。そんな話をしています。そして、初のレギュ..

Davey Todd and his performance coach, Sam Yassin return to the show to chat about the season so far and just how demanding the schedule has been for both of them from the NW200 to the TT to National Superstock to the S100, Armoy and a look ahead to the return of Olivers Mount. Look out for a cameo from Junior Superstock rider, Jack Roach, too! Hosted on Acast. See acast.com/privacy for more information.

In the last half of the 70s there was one gold standard in home computing: S100. This was a standardized bus that was the heart of many computers. It allowed for the interchange of parts from different manufacturers. Best of all, the S100 bus was simple. This made for a wonderful platform for hobbyists, and helped jump start the home computer revolution. And then... it disappeared. Where did the S100 bus go, and would we have been better off if it stuck around? This episode we tackle these questions and more.   Selected Sources:   https://archive.org/details/IoNewsVolume1Number1/page/n5/mode/2up?view=theater - The Cromemco Story   https://www.digibarn.com/stories/MITS/mholley-images/Ed_Roberts_Oct_1984_ME.pdf - An interview with Ed Roberts   https://mirrors.apple2.org.za/Apple%20II%20Documentation%20Project/Books/W.%20Gayler%20-%20The%20Apple%20II%20Circuit%20Description.pdf - Circuit Description of the Apple II

晚报 | 茅台冰淇淋与顺丰同城、顺丰冷运达成合作；钉钉发布S100战略生态伙伴计划

Chris is restoring this vintage IMSAI computer with a Z80 CPU and memory cards connected to the S100 bus, and some things are starting to work! Here, Chris uses the front panel to start a ROM program that can read hex code from a paper tape. The loaded code then prints "HELLO WORLD", as one must do. Visit the Adafruit shop online - http://www.adafruit.com ----------------------------------------- LIVE CHAT IS HERE! http://adafru.it/discord Adafruit on Instagram: https://www.instagram.com/adafruit Subscribe to Adafruit on YouTube: http://adafru.it/subscribe New tutorials on the Adafruit Learning System: http://learn.adafruit.com/ -----------------------------------------

All of us have become more reliant on technology and software to solve problems and improve our lives.  However, many of us understand less and less about the programming that goes into these solutions.  Further, we forget or ignore the human element of this problem solving.   In this episode, I speak with ANDY HUNT who has devoted his life to these issues. Andy Hunt is a programmer turned consultant, author and publisher. He's authored a dozen books including the best-selling “THE PRAGMATIC PROGRAMMER,” was one of the 17 authors of the AGILE MANIFESTO and founders of the Agile Alliance, and co-founded the PRAGMATIC BOOKSHELF, publishing award-winning and critically acclaimed books for software developers. He's currently writing science fiction (see conglommora.com) and experimenting with The GROWS Method®. We talk about his early days of programming and the power of community and process in the world of software design as we go into his experience with broader consulting at the enterprise level. Further on, Andy dives into what he considers to be important in the future of programming and advice for young programmers. Finally, as a consummate Renaissance Man, Andy discusses how his hobbies in writing science fiction, music production and woodworking excite his brain and inform his problem solving ability. Andy started in the do-it-yourself days of CP/M and the S100 bus, of Heathkits and Radio Electronics. Andy wrote his first real program, a combination text editor and database manager, for an Ohio Scientific Challenger 4P. It was a great era for tinkering. Andy started hacking in 6502 assembler, modifying operating systems, and wrote his first commercial program (a Manufacturing Resources Planning system) in 1981. He taught himself Unix and C, and began to design and architect larger, more connected systems. Working at large companies, Andy kept an ear on Usenet, and started his early email habit via a direct bang-path to ihnp4. Next he settled into electronic pre-press and computer graphics, and worked on that wondrous eye-candy that was Silicon Graphics machines. By now a firm command of several flavors of Unix, from BSD to System V, led Andy to try consulting in the early 1990's. His knack for stirring things up really began to come in handy, and it soon became obvious that many of his clients each suffered similar problems—problems that Andy had already seen and fixed before. Andy joined up with Dave Thomas and they wrote the seminal software development book, The Pragmatic Programmer, followed a year later by the original Programming Ruby: The Pragmatic Programmer's Guide, which introduced the Western world to this new language from Japan. Together they founded The Pragmatic Programmers and are well known as founders of the agile movement and authors of the Agile Manifesto, as well as proponents of Ruby and more flexible programming paradigms. They founded the Pragmatic Bookshelf publishing business in 2003, helping keep developers at the top of their game. Andy is a founder of the Pragmatic Programmers, founder of the Agile Alliance and one of the 17 authors of the Agile Manifesto, and author of a dozen or so books on programming, agile methods and learning, as well as science fiction and adventure. He is an active musician and woodworker, and continues looking for new areas where he can stir things up. Comments from Andy on his early career- -Where did your initial interest in programming develop? (The Do-It-Yourself days) -What were the types of problems that attracted your attention? -What languages did you gravitate toward?  Working for larger, more complex situations -What were the big lessons you pick up in those work environments? -Programming considered by laymen as a solitary pursuit- was there an adjustment in delegating work or collobroating?  When does management of strategy get in the way of execution?

This month on Episode 16 of the Discover CircRes podcast, host Cindy St. Hilaire highlights four featured articles from the August 28 and September 11 issues of Circulation Research. This episode features an in-depth conversation with Drs Andrew Murphy and Michelle Flynn from The Baker Heart and Diabetes Institute at Monash University in Melbourne, Australia regarding their study Transient Intermittent Hyperglycemia Accelerates Atherosclerosis By Promoting Myelopoiesis.   Article highlights:   Fish, et al. KRAS Mutations Cause Arteriovenous Malformations   Ehling, et al. B55a in Vascular Biology   Barrett, et al.  Platelet Activity and Vascular Health in COVID-19   Furmanik, et al. Nox5 in VSMC Phenotype and Calcification     Cindy St. Hilaire:  Hi. Welcome to Discover CircRes, the podcast to the American Heart Association's journal, Circulation Research. I'm your host, Dr Cindy St. Hilaire, from the Vascular Medicine Institute at the University of Pittsburgh. And today I'm going to share with you four articles selected from our late August and early September issues of Circulation Research. I'm also going to speak with Drs Andrew Murphy and Michelle Flynn from The Baker Heart and Diabetes Institute at Monash University in Melbourne, Australia regarding their study Transient Intermittent Hyperglycemia Accelerates Atherosclerosis By Promoting Myelopoiesis. So first, the highlights. The first article I'm sharing with you is titled Somatic Gain of KRAS Function in the Endothelium is Sufficient to Cause Vascular Malformations that Require MEK but not PI 3-Kinase Signaling. First authors are Jason Fish, Carlos Perfecto Flores-Suarez, and Emily Boudreau. And the corresponding authors are Jason Fish and Joshua Wythe, and they're from University of Toronto and Baylor College of Medicine. Arterial venous malformations, or AVMs, are tangles of blood vessels in which the arteries are directly connected to the veins without going through the capillary bed. These are thought to be present from birth and when they occur in the brain, they can cause an array of symptoms such as headaches or seizures, but they are also the leading cause of hemorrhagic stroke in children and young adults. This is because the venous system is not muscularized to respond to the pressure forces that are exerted on arteries. These pressure forces cause distension and eventual leakage at the site of AVMs. Vessel tissue recovered from patients undergoing AVM repair has been shown to contain sematic gain of function mutations in the protein RAS GTPase, which is encoded by the gene, KRAS. However, whether these gain of function mutations directly cause AVMs has not been established. This study now shows that endothelial cells with constitutive expression of gain of function KRAS mutants in mice and zebra fish causes vascular malformations and cranial hemorrhages. Inhibiting a MEK kinase, which is a downstream mediator of RAS signaling, prevented hemorrhages in the mutant KRAS carrying fish. In vitro studies also showed that overactive RAS GTPase protein caused excessive angiogenic behavior of endothelial cells. Together, this work confirms the link between gain of function KRAS mutations and brain AVMs, and suggests that MEK inhibition could be a potential strategy for nonsurgical treatment. The second article I want to share with you is titled B55a/PP2A Limits Endothelial Cell Apoptosis During Vascular Remodeling: A Complimentary Approach To Kill Pathological Vessels. The first author is Manuel Ehling and the corresponding author is Massimiliano Mazzone. And the work was completed at Leuven Center for Cancer Biology in Belgium. Building a mammalian vascular system is a dynamic process that is dependent on both growth of new vessels, as well as the pruning of unwanted ones. But while much is known about molecular mechanisms underlying angiogenesis, comparatively little is understood about the mechanisms regulating vascular pruning. This study discovered that suppression of the protein phosphatase 2 subunit, B55A, is a key protein regulating the pruning process. They found that in mouse vascular development, B55a is widely expressed. However, in adult mice expression is restricted only to sites of active angiogenesis. Deletion of B55a in mice caused death in mid to late stages of embryogenesis as a result of vascular problems that appeared to be due to excessive vessel pruning. Switching off B55a in adult mice when the vascular development is for the most part complete did not cause any apparent problems. They did find though, that inhibition of B55a significantly delayed growth of tumors that form from the injection of cancerous cells. Inhibition of B55a produced tumors with less dense vasculature and reduced metastatic potential. Thus, the author suggests that ramping up blood vessel pruning, be it inhibition of B55a, could be a novel strategy for limiting tumor growth. The next article I want to share is titled Platelet and Vascular Biomarkers Associated With Thrombosis and Death in COVID-19. The first author is Tessa Barrett and the corresponding author is Jeffrey Berger, and they're from New York University. Our knowledge of the complications of COVID-19 is evolving every day. Laboratory testing done to date suggests that approximately 30% of hospitalized COVID-19 patients go on to develop thrombotic events. Platelets are central characters in both arterial and venous thrombosis, and it is known that virus platelet interactions can stimulate a pro-coagulant and inflammatory state during a viral infection. Further, recent studies have reported COVID-19 patients have hyperactive platelets and autopsies of COVID-19 patients exhibit micro and macro thrombi across vascular beds, even in patients without clinical thrombosis. This group then hypothesized that biomarkers of platelet activation are associated with incident thrombosis or death in COVID-19 patients. To test this, they randomly selected 100 COVID-19 positive patients and analyzed banked samples collected on the day of the COVID-19 diagnosis to investigate in vivo platelet activity, as well as vascular health biomarkers. They show for the first time that biomarkers of platelet activation at the time of diagnosis are associated with thrombosis or death in patients hospitalized with COVID-19. Their findings suggest platelet activation mechanisms may contribute to adverse events and highlight the potential role of antiplatelet therapy in this disease. The last article I want to share with you before we switch to our interview is titled Reactive Oxygen-Forming Nox5 Links Vascular Smooth Muscle Cell Phenotypic Switching and Extracellular Vesicle-Mediated Vascular Calcification. The first authors are Malgorzata Furmanik and Martijn Chatrou. And the corresponding author is Leon Schurgers from Maastricht University in The Netherlands. Vascular calcification is an active process regulated by several mechanisms, including vascular smooth muscle cell apoptosis, osteochondral genic transdifferentiation, extracellular vesicle release, and cellular senescence. In healthy adult arteries, smooth muscle cells maintain a contractile phenotype. However, various insults such as oxidative or mechanical stress, can induce smooth muscle cells to lose their contractility and this process of de-differentiation is termed phenotypic switching. And phenotypic switching is thought to precede the development of vascular disease. Patients with conditions such as chronic kidney disease have mineral imbalances in their circulation and also exhibit higher levels of vascular calcification. However, the mechanisms behind these observations are not well defined. This group found that extracellular calcium can enter the smooth muscle cells via extracellular vesicles and this increased cytosolic calcium concentration. Increased calcium induces expression and activity of Nox5 in NADPH oxidase. Activation of Nox5 increased production of reactive oxygen species, which in turn decreased contractile marker expression, and also promoted calcification in vitro. Intracellular calcium signaling also further enhanced extracellular vesicle secretion, and decreased extracellular vesicle uptake. This then promoted the accumulation of extracellular vesicles in the extracellular matrix, which is a mechanism that promotes calcification. Together, these data suggest that mineral imbalances, such as those seen in chronic kidney disease patients, contribute to loss of smooth muscle cell contractility, which promotes osteochondral genic transdifferentiation. For the interview portion today, I have with me Drs Andrew Murphy and Michelle Flynn from the Baker Heart and Diabetes Institute and Monash University in Melbourne Australia. And we're going to be discussing their manuscript titled Transient Intermittent Hyperglycemia Accelerates Atherosclerosis by Promoting Myelopoiesis. But really I like the running title, which is Hyperglycemic Spikes Accelerate Atherosclerosis. So thank you both very much for joining me today. Michelle Flynn: Thanks for having us. Cindy St. Hilaire: So before we start to ‘stalk a bit about what the details of this study is, could you maybe give us a little primer on what you've done that led up to this study? Andrew Murphy: Yeah, so this really was a continuation of a study that began actually when I was in my postdoc in Allan Tall lab and working with Ira Goldberg’s lab with the postdoc  Prabhakara R Nagareddy there. We've shown along with Ed Fisher’s group at NYU, that mice that had established atherosclerotic lesions that were then made diabetic, failed to have lesion regression compared to those that were non-diabetic with normalized plasma cholesterol levels. We showed that if we gave an SGLT-2 inhibitor to normalize glucose that regression then started to occur. And then we found that this was primarily driven by myelopoiesis, suddenly increased production of monocytes, which through that entered the plaque. And so from that, that was in the hyperglycemic model which is sort of a very rare patient group these days, because most people are on well-controlled glucosteroid drugs. And really the SGLT-2 inhibitors have been a game changer in that scenario. And what we were trying to do with this study was bring it into a more clinically relevant setting that might show the potential importance of glucose on a much larger population. Cindy St. Hilaire: Excellent. Maybe you could give us an introduction to the link between what's known about diabetes and cardiovascular disease and the interplay? Michelle Flynn: So diabetic and pre-diabetic patients actually account for 65% of all cardiovascular deaths, which really indicates that diabetes itself plays a major factor alongside other things like obesity and hypercholesterolemia. And so we've previously shown that hyperglycemia was actually driving atherosclerosis in a chronic hyperglycemic setting. So given that kind of vascular disease actually affects both diabetic and pre-diabetic patients, we suspected that it may not just be chronic hyperglycemia or really intense hyperglycemia that could be driving this issue. And so what we were actually looking at in this paper was how more transient levels of hyperglycemia, which actually occur quite often in both diabetic patients and pre-diabetic patients, how much this can contribute to cardiovascular disease. Andrew Murphy: I guess this link between poor glucose control and cardiovascular disease is obviously very well established. The interesting part is that HbA1c only predicts part of the risk. If you look at fasting blood glucose, again, that's only partially responsible, but if you look at postprandial or two hour glucose loads, you'll see that that is more predictive of cardiovascular events than the other two measures. And it seems to be a continuum. So even if you are a healthy or non-diabetic individual, you obviously still have those postprandial events and depending how high that goes up is thought to be a predictive of future cardiovascular outcomes. And so obviously that's worse than people with pre-diabetes and then again worse with people that have actual, full blown diabetes. Cindy St. Hilaire: And what is a transient hyperglycemic event? What would do that in maybe you and me who don't have diabetes versus someone who has diabetes or is pre-diabetic? Michelle Flynn: So essentially what we're modeling with this transient hyperglycemia is that postprandial increase in glucose after you have a meal, which in people who have impaired glucose tolerance is going to be more pronounced than in someone who has a normal glucose tolerance. Cindy St. Hilaire: Got it. And so how did you test this in the mice? Michelle Flynn: We did this by developing a novel model of transient hyperglycemia. So we used ordinary wild type mice that weren't diabetic, and we injected them with glucose intraperitoneally, which then increased blood glucose levels in the plasma after about 15 minutes up to about 15 to 20 millimolar. And then after about two hours, this decreased back down to baseline levels. So this was very similar to what you actually see in a postprandial event. And by doing this four times throughout the day, we were able to mimic what you might see in a patient who has had several meals across the day who has impaired glucose tolerance. Andrew Murphy: One other advantage with the model that we used was that we were trying to really isolate the effects of glucose. And so by injecting glucose intraperitoneally in otherwise healthy mice, it bypasses the incretin response, which we know loses efficacy, I guess, in people that are diabetic. And so we were just really mimicking acute glucose rises that would occur after a meal. And then obviously in this wild type mouse the insulin response would then kick in to clear the glucose so it really tests that glucose hypothesis. Cindy St. Hilaire: So it's really digging in deeply on the actual sugar component, not just eating in general or other aspects. So in some of your experiments, or I guess in actually most of them, you show that the injection of glucose, it increased the plaque size in these mice, but it didn't alter the cholesterol levels. So can you explain a bit what's going on there? A little bit about the mechanism you discovered and kind of specifically introducing RAGE and the S100A8 and A9 axis? Michelle Flynn: Yeah, so what we showed was that regardless of cholesterol levels, we were seeing an increase in clot plaque size, and this was actually driven by the monocytes and neutrophils which were increased in the circulation of these mice. And then these are able to infiltrate into the plaque where they promote plaque progression. And what we found was that the increase in monocytes and neutrophils was due to an increase in their production within the bone marrow. And this was in turn due to the signaling by a protein heterodimer of S100A8 and A9, which signals via the receptor RAGE in the bone marrow on the progenitors of these cells, which induces their proliferation and differentiation. And then that produces an increase in the production of those immune cells, which promote plaque progression. Cindy St. Hilaire: Interesting. So it's really independent of kind of the basic thing that everyone thinks about, or I guess as non-scientists think about, is cholesterol. The public really focus on cholesterol, but what your study's showing is there's this whole other glucose mediated immune arm to it. What else does this S100A8-A9 regulate? Andrew Murphy: So S100A8 and A9 has some intracellular roles, which may direct the development of the model itself, but really a lot of its extracellular roles and so on is promoting sterile inflammation, chemotaxis, so activation of local immune cells. And in the context of diabetes and obesity, many of other diseases, it can signal via RAGE, as Michelle said, but it can also signal by TLR4. And so it seems as though in those diseases driven mainly by glucose, such as the modeling of postprandial hyperglycemia or all kinase in general, it will signal via RAGE, but we've also shown in the setting of obesity that it will signal via TLR4 to stimulate things like interleukin 1 beta. We've also had a paper just recently in Circulation  with Prabhakara Nagareddy’s group where we've shown post myocardial infarction that prime neutrophils in the heart to eventually release IL1-beta and cause myelopoiesis in that way. Cindy St. Hilaire: Wow, so this is really kind of an early activator of a much bigger immune response, whether it's in atherosclerosis or MI or probably, I don't know, a handful other things, I guess, right? Andrew Murphy: It seems to be really important when neutrophils are involved. So in a setting of an MI, we know that they come into the heart very early and become activated and it really makes them about 40% of the cytosol proteins of the cells. So when it degranulates or lyses, they are kind of neutral, at least in the predominant protein. Cindy St. Hilaire: Okay. So this is released in NETs in NETosis then? Andrew Murphy: That's what we're sort of discovering so far. So I guess all I can say is, stay tuned, this is a story for another day. Cindy St. Hilaire: Okay. That's really interesting though. Andrew Murphy: We haven’t looked in gglucose driven events yet. Cindy St. Hilaire: Yeah and actually one of the interesting things I've learned from your study, I had known about GLUT1 and that GLUT1 was I guess the constituently active of the glucose transporters, but I didn't realize it was so high on neutrophils and that neutrophils were so dependent metabolically on glucose. Can you maybe tell a little bit more about that story? Michelle Flynn: Yes. So the neutrophil itself is actually very highly dependent on glycolysis because it doesn't actually have many mitochondria. So compared to most cells, they have very few mitochondria so they can't really rely upon the oxidative phosphorylation for their general metabolism. And so they predominantly rely on glucose coming into the cell and then being shuttled through glycolysis to generate their energy. And yeah this does seem to be predominantly due to uptake of glucose through GLUT1. Cindy St. Hilaire: And then that excess glucose, the byproduct, is reactive oxygen species and upregulation and this cascade of- Michelle Flynn: Yes, yes that's correct. Cindy St. Hilaire: Okay, great. So currently we use HbA1c as a biomarker for overall kind of glucose regulation in diabetic patients. And based on your studies and perhaps the studies of others, would neutrophil numbers or even S100A8 or A9 be a better metric to figure out where a pre-diabetic or even a healthy patient is in terms of their glucose tolerability? Michelle Flynn: Yeah. That could actually be an interesting marker to look at. Given that neutrophils and S100 are also associated with obesity and diabetes in general, and as well as the risk for cardiovascular disease. So with the progression of diabetes, you could expect that the levels of these would increase as well. Andrew Murphy: We've shown previously when we first discovered that the S100 was important in diabetes, that in the Pittsburgh study with Trevor Orchard's group, he had followed people with type one diabetes for 20 years, that those that did develop a cardiovascular event had a higher S100A8 and A9 levels and that correlated with neutrophils. And so it certainly seems to be a marker of predictive outcomes. And so those that do have poorer glycemic control will have higher neutrophils. That's well known. And so perhaps you're right that probably in combination with HbA1c or things like two hour post glucose challenges, S100A8 and A9 and perhaps neutrophil counts would also be a nice predictive measure of potential cardiovascular outcomes of that person. Cindy St. Hilaire: Wow. That'd be really great because you could then maybe kind of more fine tune and predict which patients might be more or less susceptible to cardiovascular events. Andrew Murphy: That's right. Yeah. I think one other important aspect would be if HbA1c is deemed to be relatively well under control, yet you still have a high level of S100A8 and A9, that perhaps those transient spikes are contributing. You're not picking that up in the HbA1c, which looks like the average over approximately a month. And so that could be a nice way to add value onto that score. Cindy St. Hilaire: Interesting. I didn't realize it was that stable about over a month. All right. So I'm relatively healthy. I'm not pre-diabetic, but if I eat a whole bunch of cake or a whole bunch of ice cream or drink a lot of beer, does that create un me a transient hyperglycemic event that is of the same range we're talking about and what do your findings suggest for people who are relatively healthy and things we should be aware about regarding eating habits and things like that? Andrew Murphy: Yeah. I think it's a really good question. And it's sort of hard to give you an exact answer to that right now. We need to look at that in people, model these sort of same spikes in people, but what we I guess don't know yet, even in the preclinical models is how high and how long does that glucose have to be? And I think that's one of the most important questions first. So is there a danger zone where these neutrophils start be the innate senses of hyperglycemia that start to then release S100A8 and A9 to cause these downstream events? But what our data does show is that if you're doing this, having a binge night or a binge day once a week for your life, then that's probably not going to be a great thing. Cindy St. Hilaire: Yeah. All right. So you need to figure out is one scoop of ice cream okay, but two not so great. Andrew Murphy: Maybe if it's two different flavors it'll be okay. Cindy St. Hilaire: Maybe, right? That's great. So, I mean, is there a way we could potentially therapeutically target this signaling axis or is it too ubiquitous in terms of what it regulates? Is there a way to harness what you've found potentially in the clinic? Michelle Flynn: Yeah, so there's an inhibitor of S100A8 and A9 that prevents its binding to RAGE. It's currently approved as an Orphan Drug for systemic sclerosis in both the US and the UK. And that drug itself, we tested in our preclinical mouse model, and we found that it was in fact able to prevent their production of these immune cells, as well as prevent the accelerated atherosclerosis in response to these transient hypoglycemic spikes. Andrew Murphy: So another sort of line of thinking that we're exploring is that we could actually target neutrophil metabolism itself. And so now we're sort of understanding, are there certain proteins that are more abundantly expressed in neutrophils and not other cells in the body that would regulate glycolysis? I know that might sound a bit of a pie in the sky sort of idea, because glycolytic pathway's quite regulated, but there we have found some proteins that are rich in neutrophils and not other cells that may be responsible for the early steps of glycolysis. And so whether that can be harnessed or not, we'll have to see in the future, but it might be a way of more directly targeting neutrophils rather than approaching that's important in sterile inflammation. Cindy St. Hilaire: That makes sense. That is such a cool idea and this is really such a beautiful story. It's one of those papers that you just read it and it's just such a logical progression, but it's also really interesting and I really appreciated all those bone marrow transplants. I did those in grad school, so well done. It's a beautiful story. And then I'm just really happy that you published it with us. So thank you so much for joining me today. Andrew Murphy: Yeah thanks for having us. Michelle Flynn: Thank you. Cindy St. Hilaire: That's it for highlights from the late August and early September issues of Circulation Research. Thank you so much for listening. Please check out the Circulation Research Facebook page and follow us on Twitter and Instagram with the handle @CircRes and #DiscoverCircRes. Thank you to our guests, Drs Andrew Murphy and Michelle Flynn. This podcast is produced by Rebecca McTavish and Ashara Ratnayaka, edited by Melissa Stoner, and supported by the Editorial Team of Circulation Research. Some of the copy text for the highlighted articles is provided by Ruth Williams. I'm your host, Dr Cindy St. Hilaire, and this is Discover CircRes, your on-the-go source for the most exciting discoveries in basic cardiovascular research.  

Link to bioRxiv paper: http://biorxiv.org/cgi/content/short/2020.07.09.190777v1?rss=1 Authors: Dimitri Ryczko, Maroua Hanini-Daoud, Steven Condamine, Benjamin J. B. Bréant, Maxime Fougère, Roberto Araya, Arlette Kolta Abstract: AbstractThe most complex cerebral functions are performed by the cortex which most important output is carried out by its layer 5 pyramidal neurons. Their firing reflects integration of sensory and contextual information that they receive. There is evidence that astrocytes influence cortical neurons firing through the release of gliotransmitters such as ATP, glutamate or GABA. These effects were described at the network and at the synaptic levels, but it is still unclear how astrocytes influence neurons input-output transfer function at the cellular level. Here, we used optogenetic tools coupled with electrophysiological, imaging and anatomical approaches to test whether and how astrocytic activation affected processing and integration of distal inputs to layer 5 pyramidal neurons (L5PN). We show that optogenetic activation of astrocytes near L5PN cell body prolonged firing induced by distal inputs to L5PN and potentiated their ability to trigger spikes. The observed astrocytic effects on L5PN firing involved glutamatergic transmission to some extent but relied on release of S100β, an astrocytic Ca2+-binding protein that decreases extracellular Ca2+ once released. This astrocyte-evoked decrease of extracellular Ca2+ elicited firing mediated by activation of Nav1.6 channels. Our findings suggest that astrocytes contribute to the cortical fundamental computational operations by controlling the extracellular ionic environment.Key Points SummaryIntegration of inputs along the dendritic tree of layer 5 pyramidal neurons is an essential operation as these cells represent the most important output carrier of the cerebral cortex. However, the contribution of astrocytes, a type of glial cell to these operations is poorly documented.Here we found that optogenetic activation of astrocytes in the vicinity of layer 5 in the mouse primary visual cortex induce spiking in local pyramidal neurons through Nav1.6 ion channels and prolongs the responses elicited in these neurons by stimulation of their distal inputs in cortical layer 1.This effect partially involved glutamatergic signalling but relied mostly on the astrocytic calcium-binding protein S100β, which regulates the concentration of calcium in the extracellular space around neurons.These findings show that astrocytes contribute to the fundamental computational operations of the cortex by acting on the ionic environment of neurons.Competing Interest StatementThe authors have declared no competing interest.View Full Text Copy rights belong to original authors. Visit the link for more info

We are blessed with the pleasure to speak with Courtney great friend of the show, ties deeply into our S100 roots. She talks about her company “The Urban & The Mystic “ her one women show “No I won’t take you to the airport “ and all the fun stuff between. We also have Cliff from our CC collective to talk current music and hang out for a lil.

Drogo presents Gyromancer 001. Techno all the way, featuring: Tin Man, Edit Select, Advanced Human, Mathew Jonson, UZB, Exercise One, Eomac, Planetary Assault Systems, S100, HD Substance, Marcel Dettman, Luke Slater, Trevino, Exium, Reeko, Markus Suckut and MTD

In my thesis 25 feline ovaries (Felis catus) were studied using histological, glycohistochemical, immunhistochemical and ultrastructural methods. Additionally to the different follicle stages, the ovarian stroma and the thecal glands were also evaluated. For the glycohistochemical investigations, an appropriate panel of lectins was used, including Concanavalin Agglutinin (Con A), Wheat germ Agglutinin (WGA), Wheat germ Agglutinin succinylated (WGAs), Sambucus nigra Agglutinin (SNA) , Pisum sativum Agglutinin (PSA), Ricinus communis Agglutinin (RCA), Viscum album Agglutinin (VAA), Phaseolus vulgatis Erythroagglutinin (PHA E), Maackia amurensis Agglutinin I (MAA I), Phaseolus vulgaris Leukoagglutinin (PHA L), Sophora japonica Agglutinin (SJA) and Griffonia simplicifolia Agglutinin I (GSA I). The most interesting glycohistochemical staining was the strong reaction of WGA in the zona pellucida and the surrounding corona radiata cells. The staining with WGA-FITC demonstrates N-acetylglucosamine and sialic acids in the zona pellucida and the surrounding corona radiata. The immunhistochemical examination using antibodies against cytokeratins, vimentin, laminin, desmin, synemin, tubulin, SMA, S100, connexin 43, ERα and progesterone receptors showed the localization of cytoskeletal components within the different compartments of the feline ovary and the distribution of steroid hormone receptors. The ovarian surface epithelium contains not only cytokeratins but also synemin and nuclear estrogen receptors. The interstitial gland cells show a strong immunohistochemical staining with antibodies against vimentin, S100 and connexin 43. A similar immunohistochemical staining pattern was also observed in cells of the theca interna of tertiary follicles. The immunohistochemical staining pattern differs between the luteal cells derived from granulosa cells (granulosa lutein cells) or from thecal cells (theca lutein cells). The small theca cells only showed a distinct reaction with tubulin and S100 antibodies, in contrast to the large luteal cells which reacted much more strongly. In the rete ovarii the expression of cytokeratins, tubulin, progesterone receptors and ERα could be immunohistochemically demonstrated. Oocytes of the follicles of different developmental stages only showed a positive reaction with the synemin antibody. At all developmental stages, the follicle cells showed a strong immunohistochemical staining with the tubulin antibody. The intensity of connexin immunostaining increased during follicular development within the follicular epithelium from primordial to tertiary follicles. This proves the increase in the number of gap junctions in the follicular epithelium during follicle growth. In contrast the staining intensity with the vimentin antibody decreased in granulosa cells from primordial to tertiary follicles. This may indicate the remodelling of the cytoskeleton of the granulosa cells in growing follicles to obtain steroidogenic potential. The formation of nuclear ERα and progesterone receptors varies under the influence of different hormones, depending on the ovarian cycle. The rete ovarii is the only structure in which both receptor types are expressed. ERα can be detected in granulosa cells of some primary follicles, as well as in the ovarian surface epithelium. Progesterone receptors are localized in the theca interna of tertiary follicles and in small luteal cells and fibrocytes of the corpus luteum. In conclusion the feline ovary shows, in comparison to other mammals, an early differentiation of cytoskeletal and glykan containing elements. The zona pellucida is already formed in primary follicles and the granulosa cells contain vimentin filaments and gap junctions, which can already be observed in the stage of primordial follicle. The distribution of the S100 protein in the ovary of the cat differs fundamentally from all other species investigated so far. S100 was predominantly found in steroidogenic cells and may indicate its involvement in steroidogenesis, in conjunction with the expression of vimentin and connexin.

The pathognesis of psoriasis still remains not fully elucidated. Recent advances favor the idea that interactions between innate and adaptive immune response drive inflammatory process in this disease. Innate antimicrobial peptides and proteins (AMPs) are diverse group of small molecules that provide the first line of defense against invading pathogens. In recent years, the novel functions ofAMPs have been identified. There are three subclasses among AMPs that have gained the special interest as a potentially important player in the pathogenesis of psoriasis: cathelicidin, S100 proteins, and defensins. These AMPs have been shown to modulate and trigger host immune response in psoriasis acting as interplayer between innate and adaptive immune mechanisms. Overexpressed in psoriatic lesions, they prime immune cells for enhanced production of proinflammatory mediators and act as chemoattractant for leukocytes. Therefore, the novel term describing AMPs alarmins has been suggested. As multifunctional player in pathogenesis of psoriasis, AMPs may constitute potential target for therapeutic interventions. However, further investigations are required to establish the methods of downregulation of the aberrant proinflammatory functions of AMPs without increasing the risk of infections.

Background: The usefulness of S100 as a prognostic marker and aid in follow-up care in patients with malignant melanoma as well as in individuals with various neurological pathologies is well known. The aim of this study was to investigate its release and clinical relevance in benign and malignant disorders beyond these indications to elucidate tumor and organ specificity of S100. Methods: S100 levels were studied in serum samples of 1856 untreated patients, among them 59 healthy individuals, 358 patients with benign disorders, and 1439 patients with malignant tumors. Results: Healthy individuals had low S100 levels reaching a median of 0.041 ng/mL and 95th and 100th percentiles of 0.096 ng/mL and 0.144 ng/mL, respectively. The medians of patient groups with benign diseases ranged from 0.030 to 0.057 ng/mL, patients with malignant diseases from 0.020 to 0.059 ng/mL, and thus were comparable to healthy individuals. Only 2% of patients with benign diseases, mainly suffering from infectious, autoimmune, or benign gastrointestinal diseases and 1% of patients with malignant diseases showed slightly higher values than healthy individuals, in most cases up to 0.5 ng/mL. Conclusions: In contrast to many other oncological biomarkers, S100 is only rarely released in elevated levels from most benign and malignant diseases apart from malignant melanoma and neurological diseases, resulting in superior organ and tumor specificity. As potentially influencing factors, severe infectious diseases have to be considered.

Although many aspects of ovarian differentiation have been established,comparatively little is known about prenatal follicle formation anddifferentiation of bovine ovaries. The objective of this investigationwas to study the role of the surface epithelium during the developmentof germ cell nests, germ cell cords and follicle formation in the fetalbovine ovary. Associated important proliferation and apoptotic featureswere further investigated. Additionally, the expression pattern of theS100 protein was detected. A strong increase of mitotic figures wasdetected in the surface epithelium, germ cell nests and germ cell cordsof ovaries with a crown-rump length (CRL) of 13.0-58.0 cm. Oocytes werepositively stained with S100 in bovine ovaries from fetuses with a CRLof 21.0 cm. The staining intensity enhanced parallel to increasingoocyte and follicle sizes during the ovary development. In later stages,a strong staining for S100 was observed in healthy oocytes incontradistinction to atretic oocytes where no expression of the S100protein could be found. In conclusion, increasing mitosis index ofsurface epithelium cells, as well as oogonia directly beneath thesurface epithelium, in combination with open surface connection duringstages from a CRL of 11.0-94.0 cm of bovine fetal ovaries could play animportant role in the period of time of ongoing folliculogenesis andderivation of granulosa cells. Additionally, S100-positive oocytes inprimordial and later follicle stages joined by a high rate ofKi67-positive index in surrounding granulosa cells indicate that in theoocytes the S100 protein can perhaps be a useful marker for intactoocytes in bovine ovaries.

Die allogene Knochenmarks- bzw. Stammzelltransplantation wird seit den späten 70er Jahren als kurativer Behandlungsansatz bei myeloproliferativen Syndromen wie Leukämien und Lymphomen etabliert. Eine schwere und häufige (25-45%) Komplikation dieser Transplantation ist die Wirt-gegen-Spender-Erkrankung bzw. Graft-versus-Host Disease (GvHD). Die Erkrankung ist mit einer hohen Letalität von etwa 30% unter moderner Therapie verbunden und manifestiert sich häufig zunächst an der Haut. Eine zuverlässige und rasche Diagnosesicherung ist für die Früherkennung und adäquate Therapie der GvHD entscheidend. Leider ist die akute, das heißt binnen 100 Tagen nach Transplantation auftretende GvHD (aGvHD) von akuten Arzneimittelreaktionen (AR) klinisch und histologisch schwer zu unterscheiden. Etablierte Kriterien für diese Differentialdiagnostik existieren nicht. Die Feststellung des histologischen Schweregrads der aGvHD ist bislang eher untersucherabhängig, die des klinischen Schweregrads ist dermatologisch sehr grob und zur Verlaufskontrolle eher ungeeignet. Diese Punkte zu optimieren und einen Beitrag zur Aufklärung der Immunpathologie der aGvHD zu leisten waren die Hauptziele der vorliegenden Dissertation. Zwanzig Patienten mit klinisch gesicherter aGvHD nach allogener Knochenmarks- oder Blutstammzelltransplantation und dreizehn Patienten mit klinisch verifizierter AR wurden in die Studie aufgenommen. Die klinischen Befunde wurden nach dem etablierten Glucksberg-Score sowie dem neu entwickelten klinischen GvHD-Schweregrad-Score (GvHSco) klassifiziert. Zusätzlich wurden Hautproben entnommen und histopathologisch sowie immunhistochemisch (Expression von CD1a, CD2, CD11c, CD20, CD25, CD34, CD68, CD197, CD206, CD207, CD 208, CD209, CD303 und S100) analysiert. Klinische und histologische Ergebnisse wurden einzeln analysiert und miteinander korreliert. Zur besseren Beschreibung des klinischen Schweregrades der kutanen GvHD wurde der klinische GvHSco a priori entwickelt. Er bietet durch die Standardisierung und die hundertteilige Skala im Vergleich zum Glucksberg Score Vorteile bezüglich der individuellen Verlaufskontrolle. Als histologische Schweregradkriterien korrelierten epidermotrope lymphozytäre Infiltration und Kontinuitätsverluste der Basalmembran (Epidermolyse) am deutlichsten mit dem klinischen Schweregrad. Aufgrund dieser Ergebnisse wurde auf der Basis des histologischen Scores nach Lerner durch Ergänzung des Kriteriums Epidermolyse und durch besondere Gewichtung des Kriteriums Lymphozyteninfiltration der Modifizierte Histologische Score zur Abschätzung des Schweregrads akuter GvHD (GvHiScore) entwickelt. Die Vorteile dieser modifizierten Klassifikation sind die genaue, Untersucher-unabhängige Definition und die feinere Stratifizierung der Schweregrade. So wird eine bessere inter- und intraindividuelle Differenzierbarkeit erreicht. Als differentialdiagnostische Parameter sprachen hohe Zahlen reifer T-Zellen (CD2+, CD45RA+) und Makrophagen (CD68+), Epidermolyse, Basalzellballonierung, junktionales lymphozytäres Infiltrat differentialdiagnostisch für aGvHD, eosinophiles Infiltrat jedoch gegen eine aGvHD. Basierend auf diesen neuen Erkenntnissen wurde der differentialdiagnostische Test DSHIG („Differentialdiagnostischer Score mittels Histopathologie und Immunhistochemie für akute Graft versus Host Disease“) entwickelt. Der Test errechnet sich aus der Addition sieben dichotomer Kriterien. Die retrospektive Analyse des DSHIG ergibt eine Testspezifität und -sensitivität von 95% für die Differentialdiagnose „Akute GvHD“ versus „Akutes Arzneiexanthem“. Der differentialdiagnostisch vielversprechende DSHIG sollte prospektiv validiert werden. Bei der Lupusband-positiven akuten GvHD zeigte sich ein histologisch besonders schweres Bild mit ausgeprägter Epidermolyse. Ein Einfluss quoad vitam oder auf den klinischen Schweregrad ließ sich nicht zeigen. Die Lupusband-positiven Fälle traten bevorzugt in der späteren Phase von aGvHD auf. Für den klinischen Schweregrad und das Ein-Jahres-Überleben bei aGvHD günstig waren hohe Zellzahlen von IDEC (CD206+/CD11c+), plasmazytoiden Dendritischen Zellen (BDCA-2+) und Mastzellen. Diese Zusammenhänge wurden bislang nicht an Hautbiopsien gezeigt und könnten klinisch bedeutsam sein. Die in dieser Arbeit an Hand einer kleineren Fallzahl retrospektiv erstellten Scores sollten in zukünftigen Untersuchungen mit höherer Patientenzahl unabhängig prospektiv validiert werden. Die Dynamik der kutanen GvHD könnte darüber hinaus mit weitern Methoden wie durchflußzytometrischer Analyse und Gewinnung von sequentiellen Hautproben im zeitlichen Verlauf analysiert werden.

Unter den Prognosefaktoren beim kolorektalen Karzinom hat das Tumorstaging den größten Stellenwert und dient der weiteren Therapieplanung. In den letzten Jahren wurden zunehmend auch Tumormarker hinsichtlich der prognostischen Aussagekraft untersucht, es existieren aber kaum Studien, die an großer Fallzahl mehrere Tumormarker im Vergleich zu den etablierten Prognosefaktoren multivariat evaluiert haben. In der vorliegenden Arbeit wurden die Tumormarker CEA, CA 19-9, CA 242, CA 72-4, CYFRA 21-1, hCGß, S100 und HGF hinsichtlich ihrer prognostischen Wertigkeit beim kolorektalen Karzinom analysiert. Die präoperativen Tumormarkerwerte von CEA und CA 19-9 lagen bei 1089 Patienten (Kollektiv I) vor. Bei einem Teil dieser Patienten (n=450) waren Restproben vorhanden, so dass retrospektiv zusätzlich die Tumormarker CA 242, CA 72-4, CYFRA 21-1, hCGß, S100 und HGF analysiert werden konnten (Kollektiv II). Es zeigte sich bei allen Tumormarkern eine höhere Freisetzung mit zunehmender Tumorinfiltrationstiefe. Bei CEA und CA 19-9 wurde außerdem ein signifikanter Zusammenhang zwischen der Serumkonzentration und dem Lymphknotenstatus festgestellt. Bezüglich der Korrelation der Tumormarker untereinander wurde der höchste Koeffizient für CA 19-9 und CA 242 errechnet (rs=0.79). Basierend auf den aktuellen Empfehlungen für adjuvante Therapien beim kolorektalen Karzinom wurde eine Unterteilung in Prognosegruppen vorgenommen: In der günstigen Prognosegruppe (GPG) befanden sich Patienten mit einem Kolonkarzinom Stadium I-II oder Rektumkarzinom Stadium I, in der ungünstigen Prognosegruppe (UPG) befanden sich Patienten mit einem Kolonkarzinom Stadium III oder einem Rektumkarzinom Stadium II oder III. In Anlehnung an eine Studie von Harrison et al wurden zusätzlich alle Patienten mit einem Kolonkarzinom ohne Lymphknotenbefall gesondert betrachtet. Ziel unserer Studie war es zu untersuchen, ob durch Berücksichtigung von Tumormarkern innerhalb der Prognosegruppen eine detailliertere prognostische Einschätzung mit entsprechender therapeutischer Konsequenz erfolgen kann. Dabei war unser primäres Zielkriterium das rezidivfreie Intervall, zusätzlich untersuchten wir das tumorbedingte Gesamtüberleben. 161 In der statistischen Auswertung wurde zunächst überprüft, ob eine lineare Abhängigkeit zwischen der Serumkonzentration der Tumormarker und dem Rezidivrisiko besteht. Eine solche Linearität konnte nur für CEA gezeigt werden, es ging daher in logarithmierter Form in die multivariate Analyse ein. Bei den übrigen Tumormarkern musste ein Cut-Off gesucht werden, wobei in der vorliegenden Arbeit weder die herstellerüblichen Cut-Off-Werte verwendet wurden, noch nach dem für das Kollektiv optimalen Cut-Off gesucht wurde. Vielmehr wurde versucht, mittels Bootstrapverfahren reproduzierbare Cut-Offs zu ermitteln. Nach unseren Ergebnissen ist CEA zu Recht als Kategorie I Prognosemarker klassifiziert worden. Dies konnte an großer Fallzahl und unter Berücksichtigung verschiedener anderer Tumormarker bestätigt werden. Auch in der Gruppe der Patienten mit einer günstigen Prognose sowie der N0-Kolontumoren erwies sich CEA als unabhängiger Prognosefaktor. Die Patienten dieser Gruppen erhalten laut dem aktuellen Konsensus keine adjuvante Therapie. Basierend auf den Ergebnissen dieser Arbeit sollte CEA jedoch in den Leitlinien der American Society of Clinical Oncology als prognostisch relevanter Parameter berücksichtigt werden und auch in die Therapieplanung mit eingehen. In der multivariaten Analyse hat sich darüber hinaus beim Kollektiv I gezeigt, dass neben CEA auch CA 19-9 ein stadienunabhängiger Prognosefaktor ist, jedoch nur für die ungünstige Prognosegruppe. Innerhalb des Kollektivs II erreichten die Tumormarker CEA, CA 19-9, CA 242, CA 72-4, S100 und HGF Signifikanz. Die beiden neuen und wenig erforschten Tumormarker S100 und HGF waren starke unabhängige Prädiktoren in der günstigen Prognosegruppe dieser Auswertung, sogar stärker als CEA. Dieses Ergebnis muss durch prospektive große Studien bestätigt werden. Im Gegensatz zu S100 erreichte HGF auch in der ungünstigen Prognosegruppe Signifikanz, war aber bezüglich der prognostischen Aussagekraft dem CA 242 bzw. CA 19-9 unterlegen. Gerade im Rahmen kommender Therapiestudien, bei denen der Einfluss neuer Medikamente auf die Angiogenese untersucht wird, könnte HGF von grossem Interesse sein. Der Tumormarker CA 242 kann als äquivalenter Marker zum CA 19-9 gesehen werden, so dass die parallele Bestimmung beider Tumormarker primär nicht sinnvoll ist. CA 72-4 erreichte in der uni- und multivariaten Analyse nur für das Überleben innerhalb der ungünstigen Prognosegruppe Signifikanz. 162 Die beiden Tumormarker hCGß und CYFRA 21-1 zeigten in dieser Arbeit keinerlei prognostische Bedeutung. Um unsere Ergebnisse für die klinische Anwendung zusammenzufassen, wurde für alle prognostisch relevanten Faktoren ein Prognoseindex entwickelt und zwar zunächst anhand des großen Kollektivs I. In der günstigen Prognosegruppe beinhaltet dieser Score derzeit lediglich T und logCEA, in der ungünstigen Prognosegruppe T, N, Lokalisation des Primärtumors, logCEA und CA 19-9 (Cut-off 55 U/ml). Eine Einteilung der Patienten bezüglich dieser Prognose-Scores zeigt eine deutliche Überlappung der ursprünglichen Prognosegruppen im Bezug auf das rezidivfreie Intervall. Daraus geht eindeutig hervor, dass die Bestimmung und Bewertung beider Tumormarker, also von CEA und von CA 19-9, sinnvoll ist und eine sehr viel genauere Prognoseabschätzung ermöglicht.

Background: The neuroprotein S100 released into the circulation has been suggested as a reliable marker for primary brain damage. However, safe identification of relevant traumatic brain injury (TBI) may possibly be hampered by S100 release from peripheral tissue. The objective of this study was to measure early S100 levels using the Elecsys((R)) S100 immunoassay for real-time assessment of severe TBI in multiple trauma. Methods: Consecutively admitted multiple trauma patients (injury severity score >= 16 points) were stratified according to the results of the initial cerebral computed tomography (CCT) examination. S100 serum levels were determined at admission and at 6, 12, 24, 48 and 72 h after trauma. Data were correlated to creatine phosphokinase (CK) and lactate dehydrogenase (LDH) serum levels. Using receiver operating characteristic (ROC) analysis, the discriminating power of S100 measurement was calculated for the detection of CCT+ findings. Results: Median S100 levels of CCT+ patients (n=9; 37 years) decreased from 3.30 mu g/L at admission to 0.41 mu g/L 72 h after trauma. They revealed no significant differences to CCT- patients (n=18; 44 years), but remained elevated compared to controls. Median CK and LDH levels correlated with the corresponding S100 levels during the first 24 h after trauma. ROC analysis displayed a maximum area under the curve of only 0.653 at 12 h after trauma. No significant difference was calculated for the differentiation between CCT+ and CCT- patients. Conclusions: Measurements of S100 serum levels using the Elecsys((R)) S100 immunoassay are not reliable for the real-time detection of severe TBI in multiple trauma patients. Due to soft tissue trauma or bone fractures, S100 is mainly released from peripheral sources such as adipocytes or skeletal muscle cells.

Ziel der vorliegenden Arbeit war es, ein chronisches Modell zur minimal-invasiven Organperfusionsmessung am Kaninchen vorzustellen. Hierzu musste als Voraussetzung für die chronischen Messungen die Implantation eines Portkathetersystems in den linken Ventrikel etabliert werden. Mit Hilfe der Portkatheter wurde der regionale Blutfluss zu verschiedenen Zeitpunkten bei gesunden Kontrolltieren und in einer Pilotstudie bei Tieren mit experimentell induzierter Peritonitis bestimmt. Die Messung der Perfusion erfolgte mit fluoreszenzmarkierten Mikrosphären (Latexkugeln mit 15 mm Durchmesser). Aus der Anzahl der im präkapillären Stromgebiet arretierten Mikrosphären kann der regionale Blutfluss in verschiedenen Organen qualitativ und, bei gleichzeitiger Gewinnung einer Referenzprobe, quantitativ in ml pro g Organgewebe pro Minute erfasst werden. Die Implantation des Portsystems wurde unter perioperativer Antibiotikaprophylaxe bei weiblichen weißen Neuseeland-Kaninchen (n = 30, 3,8 ± 0,3 kg KG) in Medetomidin/Ketamin-Anästhesie durchgeführt. Speziell entwickelte Portkatheter wurden über die Arteria carotis communis mit der Katheterspitze in den linken Ventrikel eingeführt. Perioperativ erfolgte die kontinuierliche intraarterielle Blutdruckmessung sowie eine Bestimmung der Herzfrequenz und der Sauerstoffsättigung. Prä- und postoperativ wurden Blutproben zur Bestimmung der S100-b-Serumkonzentration als Marker einer cerebralen Ischämie entnommen. Nach einem Erholungszeitraum von 2 bis 4 Wochen wurden zwei Versuchsgruppen untersucht. Zunächst wurde bei einer Versuchsgruppe (n = 16, 3,7 ± 0,4kg) zu sieben Zeitpunkten (0, 2, 24, 26, 48, 72 und 96 Stunden nach Versuchsbeginn, t1 – t7) je eine Mikrosphäreninjektion durchgeführt. Bei einer zweiten Versuchsgruppe, der Peritonitisgruppe (n = 4, 3,5 ± 0,4kg) wurde zu den gleichen Zeitpunkten unter den gleichen Narkosen bzw. Sedierungen je eine Mikrosphäreninjektion durchgeführt, darüber hinaus wurde zwischen den Zeitpunkten t1 und t2 eine „cecal ligation and puncture“ zur Auslösung einer kotigen Peritonitis mit nachfolgender septischer Allgemeinerkrankung durchgeführt, welche dann zwischen den Zeitpunkten t3 und t4 revidiert, die Bauchhöhle gespült und der Peritonitisherd saniert wurde. Die Anlage der linksintraventrikulär inserierten Portkatheter war bei 29/30 (97%) Tieren innerhalb von 71 ± 9 Minuten problemlos möglich. Weder intra- noch postoperativ kam es zu signifikanten, katheterassoziierten Rhythmusstörungen, Blutdruckabfällen (MAP präop. 73 ± 2 mmHg vs. postop. 71 ± 2) oder Hypoxieereignissen (SaO2 präop. 84 ± 2% vs. postop. 95 ± 2). Durch eine speziell modifizierte mikrochirurgische Technik war das Einbringen des Katheters im Bereich der Vorderwand der Arteria carotis communis unter Aufrechterhaltung der Durchgängigkeit des Gefäßes und somit unter Erhalt der zerebralen Perfusion möglich. So war klinisch bei keinem der Tiere eine postoperative zerebrale Ischämie nachweisbar. Die S100-b-Serumkonzentration zeigte postoperativ keinen signifikanten Anstieg (präop. 1,6 ± 0,4 ng/dl vs. postop. 1,8 ± 0,4). Das Ausgangsgewicht der Tiere wurde innerhalb weniger Tage wieder erreicht. Durch Sektion wurde die korrekte Katheterlage bei 26/29 Tieren (90%). In der Kontrollgruppe konnte gezeigt werden, dass minimal-invasive Messungen der Perfusion gut toleriert werden. Es war keine Beeinflussung des Blutflusses durch die Mikrosphäreninjektionen und die damit verbundenen notwendigen Narkosen bzw. Sedierungen zu beobachten. Die Perfusion der paarigen Organe Lunge, Gehirn und Niere war im Rechts-Links-Vergleich nicht unterschiedlich. Auch die Analyse der Werte über den gesamten Zeitraum zeigte eine gleichmäßige und nicht signifikant unterschiedliche Perfusion. So betrug die Durchblutung beispielsweise im Gehirn zum Zeitpunkt t1 rechts 1,11 ± 0,31 ml/g/min, links 1,25 ± 0,34, zum Zeitpunkt t7 rechts 0,97 ± 0,44 ml/g/min, links 1,04 ± 0,52, in der Niere bei t1 1,33 ± 0,21 ml/g/min (rechts) vs. 1,53 ± 0,23 (links), bei t7 1,11 ± 0,23 ml/g/min (rechts) vs. 1,05 ± 0,22 ml/g/min (links). Bei der Peritonitisgruppe ließ sich zunächst im Rechts-Links-Vergleich zu den einzelnen Zeitpunkten eine gute Korrelation der Perfusion nachweisen, so dass die vorliegenden Werte reliabel erschienen. In der Lunge war die Durchblutung bei t2 rechts 0,59 ± 0,19 ml/g/min, links 0,66 ± 0,20. Im Vergleich mit der Kontrollgruppe zeigte sich bei stabiler Hämodynamik ein signifikanter Abfall der Durchblutung der von dem septischen Geschehen betroffenen Organe (Niere, Leber, Magen, Lunge), welche sich zum Versuchsende nur langsam wieder erholte. Die Perfusion des Magens fiel zum Beispiel von anfänglich (t1) 0,63 ± 0,14 ml/g/min auf 0,35 ± 0,12 (t3) ab. Die Muskeldurchblutung war jedoch über den gesamten Zeitraum vergleichbar (z.B. t1 0.04 ± 0,01 ml/g/min vs. t4 0,06 ± 0,02). Die hier beschriebene Technik erlaubt somit erstmals die minimal-invasive Messung der Organperfusion beim leicht sedierten Versuchstier über mehrere Tage. Dadurch wird zum einen das bisher erforderliche erhebliche operative Trauma einer intrakardialen Injektion bzw. einer Thorakotomie vermieden und zum anderen die Notwendigkeit einer repetitiven Allgemeinanästhesie. Somit wird die Belastung für die Tiere sowie die unerwünschte Beeinflussung der Untersuchungsergebnisse durch die erwähnten Prozeduren vermindert. Die Insertion des Portkatheters unter der Aufrechterhaltung der zerebralen Perfusion trägt zur Verminderung des Risikos zerebraler Ischämien und kardiozirkulatorischer Dysregulationen bei. Die in diesem Modell notwendige Applikation von Sedativa hatte in der Kontrollgruppe per se keinen Einfluss auf die Organdurchblutung. Bei der experimentell induzierten Peritonitis fand sich eine Umverteilung der Perfusion zu Ungunsten der von der Sepsis betroffenen Organe bei stabiler Makrohämodynamik. Die repetitive Messung des regionalen Blutflusses kann in Zukunft für chronische Untersuchungen zur Perfusionsänderung, z.B. bei der Wundheilung oder in Sepsismodellen, eingesetzt werden.