Podcasts about flk

durée : 01:04:25 - First Mike Radio Show - Juicy P est de retour en avril 2025 avec un nouveau projet « de Player à Player ». Avant sa sortie, il était de passage dans le First Mike Radio Show avec Dinax, TDC, Killer, S2, Myssa, FLK, Jody, DMC, Taro OG, Mo Thug, Chakib, Trafiq, S-Pion, Le Foulala et Pash Corleone.

On this episode #22 Professional trainer and ex FLK staff member Eric Teson of Show-Me Retrievers joins the show to talk about this background of training dogs and his time spent with Chris Jobman

Flüssigkunststoff (FLK) für An- und Abschlüsse an Fenstern, Türen und Brüstungen hält jahrzehntelang dicht – wenn er korrekt verarbeitet wird! Leider kommt es hier aber immer wieder zu unnötigen Bauschäden: Es geschehen Fehler in der Planung, in der Ausführung oder bereits bei der Materialwahl. Unsere neueste Podcast-Folge will dem entgegenwirken. Experte Rico Wolf, Produktmanager Flüssigkunststoffe bei der Soprema AG, erklärt unter anderem… …worauf du bei An- und Abschlüssen mit FLK achten musst, ...was die SIA-Normen dazu sagen,  ...welche Materialien dir zur Auswahl stehen und ...was deren Vor- und Nachteile sind.   Mehr Infos und Kontakt Soprema AG: https://www.soprema.ch/de/liquids    Weitere Podcastfolgen und Blogartikel findest du unter: https://marcofehr.ch/   

On this episode Chris and Elliott talk about recent events at FLK and Elliott's journey to title Georgie. Then Chris goes into describing his method for obedience training and introducing dogs to the E-Collar. Learn more about your ad choices. Visit megaphone.fm/adchoices

Llega a quien llamamos la enciclopedia del rave en la isla, el gran FLK con un sinnúmero de flyers e info de eventos/raves que sucedieron hace décadas. Como co-host, nos acompaña BZ, la figura detrás de PR_Undrgrnd. Comparamos varias décadas de música Electrónica, incluyendo la actual, mencionando detalles pal' de interesantes.

Welcome to episode two of a miniseries focusing on the zooarchaeology of various world regions. This episode is centred around African zooarchaeology, focusing on the natural history and anatomy of the most prominent wild and domesticated species. Find out more about African Giant Rats, how loud Guinea fowls can be and what a zebrinny is. Interested in learning about how to use X-Rays and similar technology in archaeology? Check out the linked PaleoImaging course from James Elliot! Connect with James on Twitter: @paleoimaging Interested in sponsoring this show or podcast ads for your business? Zencastr makes it really easy! Click this message for more info. Start your own podcast with Zencastr and get 30% off your first three months with code ANIMALS. Click this message for more information. For rough transcripts of this episode go to www.archpodnet.com/animals/46 Links Beja-Pereira, A., et al. (2004). African origins of the domestic donkey. Science, 304, 1781. Boeyens, J. C., & Van der Ryst, M. M. (2014). The cultural and symbolic significance of the African rhinoceros: a review of the traditional beliefs, perceptions and practices of agropastoralist societies in southern Africa. Southern African Humanities, 26(1), 21-55. Marshall, F. (1989). Rethinking the role of Bos indicus in sub-Saharan Africa. Current Anthropology, 30(2), 235-240. Parkinson, J. A. (2018). Revisiting the hunting-versus-scavenging debate at FLK Zinj: a GIS spatial analysis of bone surface modifications produced by hominins and carnivores in the FLK 22 assemblage, Olduvai Gorge, Tanzania. Palaeogeography, Palaeoclimatology, Palaeoecology, 511, 29-51. Pikirayi, I. (2018). The demise of Great Zimbabwe, AD 1420–1550: an environmental re-appraisal. In A Green and R Leech (eds) Cities in the World, 1500-2000.Routledge, 31-47.. Potts, R. (1984). Home Bases and Early Hominids: Reevaluation of the fossil record at Olduvai Gorge suggests that the concentrations of bones and stone tools do not represent fully formed campsites but an antecedent to them. American Scientist, 72(4), 338-347. Rossel, S. et al. (2008). Domestication of the donkey: Timing, processes, and indicators. Proceedings of the National Academy of Sciences, 105(10), 3715-3720. Shen, Q. et al. (2021). Genomic analyses unveil helmeted guinea fowl (Numida meleagris) domestication in West Africa. Genome biology and evolution, 13(6). Stiner, M. C. (2004). Comparative ecology and taphonomy of spotted hyenas, humans, and wolves in Pleistocene Italy. Revue de Paléobiologie, 23(2), 771-785. Wylie, D. (2009). Elephant. Reaktion Books Contact Alex FitzpatrickTwitter: @archaeologyfitz Simona FalangaTwitter: @CrazyBoneLady Alex's Blog: Animal Archaeology Music "Coconut - (dyalla remix)" https://www.youtube.com/watch?v=_2UiKoouqaY Affiliates Wildnote TeePublic Timeular Motion

Welcome to episode two of a miniseries focusing on the zooarchaeology of various world regions. This episode is centred around African zooarchaeology, focusing on the natural history and anatomy of the most prominent wild and domesticated species. Find out more about African Giant Rats, how loud Guinea fowls can be and what a zebrinny is. Interested in learning about how to use X-Rays and similar technology in archaeology? Check out the linked PaleoImaging course from James Elliot! Connect with James on Twitter: @paleoimaging Interested in sponsoring this show or podcast ads for your business? Zencastr makes it really easy! Click this message for more info. Start your own podcast with Zencastr and get 30% off your first three months with code ANIMALS. Click this message for more information. For rough transcripts of this episode go to www.archpodnet.com/animals/46 Links Beja-Pereira, A., et al. (2004). African origins of the domestic donkey. Science, 304, 1781. Boeyens, J. C., & Van der Ryst, M. M. (2014). The cultural and symbolic significance of the African rhinoceros: a review of the traditional beliefs, perceptions and practices of agropastoralist societies in southern Africa. Southern African Humanities, 26(1), 21-55. Marshall, F. (1989). Rethinking the role of Bos indicus in sub-Saharan Africa. Current Anthropology, 30(2), 235-240. Parkinson, J. A. (2018). Revisiting the hunting-versus-scavenging debate at FLK Zinj: a GIS spatial analysis of bone surface modifications produced by hominins and carnivores in the FLK 22 assemblage, Olduvai Gorge, Tanzania. Palaeogeography, Palaeoclimatology, Palaeoecology, 511, 29-51. Pikirayi, I. (2018). The demise of Great Zimbabwe, AD 1420–1550: an environmental re-appraisal. In A Green and R Leech (eds) Cities in the World, 1500-2000.Routledge, 31-47.. Potts, R. (1984). Home Bases and Early Hominids: Reevaluation of the fossil record at Olduvai Gorge suggests that the concentrations of bones and stone tools do not represent fully formed campsites but an antecedent to them. American Scientist, 72(4), 338-347. Rossel, S. et al. (2008). Domestication of the donkey: Timing, processes, and indicators. Proceedings of the National Academy of Sciences, 105(10), 3715-3720. Shen, Q. et al. (2021). Genomic analyses unveil helmeted guinea fowl (Numida meleagris) domestication in West Africa. Genome biology and evolution, 13(6). Stiner, M. C. (2004). Comparative ecology and taphonomy of spotted hyenas, humans, and wolves in Pleistocene Italy. Revue de Paléobiologie, 23(2), 771-785. Wylie, D. (2009). Elephant. Reaktion Books Contact Alex FitzpatrickTwitter: @archaeologyfitz Simona FalangaTwitter: @CrazyBoneLady Alex's Blog: Animal Archaeology Music "Coconut - (dyalla remix)" https://www.youtube.com/watch?v=_2UiKoouqaY Affiliates Wildnote TeePublic Timeular Motion

Málokdy se poštěstí, aby manželka děkana byla sama děkankou. Lukáš si popovídal s doc. Ing. Zuzanou Tučkovou, Ph.D., děkankou Fakulty logistiky a krizového řízení UTB, která sídlí v Uherském Hradišti. Jak hodnotí své působení na fakultě za poslední rok v jejím čele? Jakou další funkci si s sebou neodmyslitelně nese post děkana FLKŘ? Jak se žije rodině děkanů? Jaké otázky spojené se změnou klimatu jí nedávají smysl? A co si myslí o podnikatelských dovednostech našich studentů?

EPISODE 56 Introductions Karla - K-Rae on Ravelry and Karlacrafts on Instagram, Twitter and Pinterest Emily -  EmGemKnits, EmilyTheTechEditor - Instagram, and  Emily Tech Edits Ravelry.   Shout outs Kimberly sent us a sweet email telling us how much she loves the podcast! She has been loving the things she ordered from Sarah from RIV Creative and suggested we check out Sweet Skein O Mine in NS. We did, and the yarn looks AMAZING! Definitely worth a trip to the ole etsy store! Also Hello to Ravelers Steph, Allison and Carla!   Introduce yourself in the Ravelry group or leave us a review on itunes or google play music or send us an email with feedback at mail@relentlessknitting.com We’d also love to hear from you in our “Meet a Knitter” thread.   What's knitting now Emily -  Smock Madness by elisabeth white in West Coast Colour sock yarn in an orange. Playdate Cardigan by Tin Can Knits in Flamingo Lounge colour way by TINK Friendship Shawl by Silivia McFadden out of Berocco Vintage chunky. A joyful thread by silvia mcfadden in sweet fiber merino lace rose gold   Karla - Fidra - Gudrun Johnston Copy.Cat C.C beanie by Emily Ingrid Mini Cooper by Ginny Sturdy for my Dad Soooo many Swatches!   Hibernating Woolly Lamb, Aeon Vest by Hillary Smith-Calis, Tulip bag, Fougasse by Bristol Ivy - 2 sleeves & almost at raglan decreases. My cup of tea socks - Robyn Lynn for my Mom Fintry - Knox Mountain Knit Co Harlow by Andrea Mowry Graphic Socks by Melissa Morgan Oakes   FO’s - Emily -Red Heart it’s a wrap knit for Edie by Isabell Kramer Vanilla socks in Sweet Georgia with a FLK heel Karla -  Casa Loma Vest as a shop sample for Kelowna Yarn, by Casa Loma Jacket by Sachiko Burgin, West Yorkshire Spinners Re:treat Chunky Roving Design features Emily-  Gauge smage! Playdate cardigan needs to restart. Forgot a few smocks in Smock madness. A joyful thread - Frog pond Karla - moss stitch   Knit-lit Audiobook - Sally Goldenbaum   Talky Talk Emily - 5 winners for CommitToKnit so everyone will get something, they’ll just need to be patient… :D Karla - Thinking a lot about my knitting to do list : write a gauge blog post to share with the guild, lesson plan for the sorrento retreat, tech editor course, yarn okanagan planning, cool arts collaborations.   Taught a coworker to knit!   KAL! Thinking about 2019... #committoknit2018 - I’ll close the thread on my birthday, jan 23. I’m done! I took an extra 3 days #revelinknit2018/9 - keep up with your selfcare knitting!   Signed up for the Wet Coast Wools year long bingo.   Spinning Emily -. Ordered 3D bobbins for my sweater spin. Karla  -Finished the Rambouillet spin from knitmas 2019. Another spin for a shawl set ongoing.   Best laid plans Emily - SOCK MADNESS, Canal Rings, Exploration Station. Karla - get back to the hibernation list, last things to organize for YO   Crafternoon/Sew-what Emily - When warping goes wrong. Karla - 4/6 project bags finished and a sock snake   Stash Enhancement Emily- Vegas Yarn from Hubs. Karla -   Adventures in Mommyhood Emily- Sleeping better. All the sicknesses. Karla - bunkbed   Inquiring minds Tell us about you! Join us on Ravelry or our Facebook page

In Episode 42, Greg and Pam talk about life, the universe, and everything. Sponsors We are a Knitcrate Ambassador! Are you the kind of knitter who would enjoy receiving a surprise package of yarn in the mail every month? Then you need to sign up for a Knitcrate subscription. Every month, you will receive a selection of curated yarn, a pattern specifically paired to use with the yarn, and a treat. Use code KD20at checkout for 20% off your first package when you sign up for a subscription. Notes Thank you to Tiffany for meeting with Pam and giving her a notions pouch. Visit her etsy shop: TheProjectBag. Sara Bauer for sending Greg a copy of the Humblebeesock pattern.  Projects Greg is working on Lana Grossa Socks (making up basic pattern), yarn is Lana Grossa Meilenweit Ashbury. Greg snagged his Blue Box Exploding Socks. Pam is working on Stormy Sky Shawl by lifeiscozy. Yarn is Indigo Dragonfly Chameleon Sock. (Project page) Pam is working on Mini Bubblesby Kieran Foley. Yarn is Wolle's Yarn Creations Color Changing Cotton. (Project page) Pam is working FLK socks. Yarn is Lang Yarns Jawoll Cotton Stretch Color. (Project page) Pam is looking for an asymmetrical triangular shawl. Here is the bundle of ideas she started. She also mentioned the Vamping Shawl. If you have any suggestions, leave them in the episode thread on Ravelry for her. Greg and Pam reviewed the book Wool and Wine(affiliate link). Podcasts We Love Down Cellar Studio Yarns At Yin Hoo Actually Knitting (PAL KAL) Upcoming Events Rare Find Farms Central Carolina Yarn Crawl FiberCrafty Pam talked about the Uptown Bag from Baa Berry Farms. Debi is offering 20% off until July 31. Use code TDF2018 Pam's Youtube Channel Affiliate Link Disclosures This post contains affiliate links. That means that if you click on a link to Craftsy, Cooperative Press, Amazon, or Knitcrate and subsequently make a purchase, we'll receive a small commission from the sale. You pay the same, and the commissions will help cover our podcasting expenses. Our opinions are always our own, and we're never compensated to endorse any particular product or service. Find us all over the Internet Subscribe in iTunes: The Unraveling Podcast Podcast RSS Feed: Unraveling Podcast Twitter: @UnravelingPod Facebook: Unraveling Podcast Instagram: @UnravelingPodcast Periscope: @UnravelingPodcast Ravelry Group: Unraveling Podcast Greg is KnittingDaddy on Ravelry, @KnittingDaddy on Instagram, and also writes the KnittingDaddy blog. Pam is pammaheron Ravelry, @FiberCraftyand @pammaheron Instagram, and owner of the FiberCraftymarketplace.

  Thank you for tuning in to Episode 122 of the Down Cellar Studio Podcast. This week's segments included:   Off the Needles On the Needles From the Armchair Crafty Adventures KAL News Events Contest, News & Notes Life in Focus On a Happy Note Quote of the Week   Thank you to this episode's sponsors: Imagined Landscapes and Fibernymph Dyeworks   Off the Needles New Crescent Shape Shawl Needles: US 7 (4.5 mm) Yarn: Main Color: Baah Ja Lolla, Gradient set from Woolbearers, Border in Periwinkle Sheep Pattern: a new design I have out to testers now. I hope to release it in October. Dragon Belly Shawl Pattern: Dragon Belly by Johanna Lindahl Hook: J-6.0 mm Yarn: Jill Draper Makes Stuff- Hudson (worsted) in the Raspberry Beret colorway (2 skeins- 480 yards) My 1st crochet shawl I need to wash an block it and will do a citric acid soak. Details on how to use Citric Acid to set your yarn on the Knitmore Girls website. On The Needles Scrappy Halloweeny Socks Yarn: Leftovers of black, white, gray, orange, yellow, tan Needles: US 1.5 (2.5 mm) Pattern: TBD   Spring Leaf Stripes Socks Yarn: Paton's Kroy Spring Leaf Stripes Needles: US 1.5 (2.5 mm) Pattern: OMG Heel by Megan Williams   Christmas Ribbon Socks Yarn: Sunsoaked Yarns Sock 80/20 Superwash Merino/Nylon in the Christmas Ribbon Colorway Needles: US 1.5 (2.5 mm) Pattern: Fish Lips Kiss Heel by Sox Therapist Holly Jolly Christmas Socks Yarn: Dancing Dog Dyeworks Twist Sock in the Holly Jolly colorway Needles: US 1.5 (2.5 mm) Pattern: Fish Lips Kiss Heel by Sox Therapist   Poppy Troll Hat x2 Carina's Hat + Ella's Hat Pattern: Poppy Troll Hat by  Tawnya Myers (free crochet pattern) Hook: H Yarn: Red Heart Super Saver in 2 colors of pink: Carina: 5-10 year old child zie Ella:1-4 year old child size. Both hats done. Earflaps on. Ears made. Need to sew on. Wash them. then will add eyes and hair From the Armchair I talked about this article: What The Health Review Bottom line: the What the Health Documentary cites sources that aren't reliable research studies and is, if nothing else, at least cause for a second look. Check out this great new audio podcast: (mis)adventures in knitting- Audio Podcast hosted by Ashley + Tara, based out of Baltimore, MD In the next episode, I am going to talk about the Four Tendencies Book by Gretchen Rubin and nd knowing our own tendency can help us accomplish our crafty goals... Invite you to take the quiz on Gretchen Rubin's website before the next episode so you know your tendency. Crafty Adventures I bought a dress and petticoat from Goodwill for $20 to make Emelyn a Marie Antoinette halloween costume for her last year of Trick or Treating.   KAL News   12 Months to Christmas KAL Click here to find about more about this KAL. Check out: Chat Thread & FO Thread  The 12 Months to Christmas KAL is a craft-a-long that helps you prepare for the busy winter holiday season. Complete gift projects each month (or just in the months that you choose) so you don’t have to scramble in December. Find some cool new gift ideas, cheer each other on, and we’ll provide some motivation in the form of the potential to win prizes!     WIP’ing 2017 in the Butt Declare a WIP that you started June 30th or earlier but haven't yet finished within the first 5 days of the month, posting a photo of it in the thread. Finish the project by the end of the month, update your post with your FO photo and you'll be eligible to win a Ravelry downloadable pattern of your choosing up to $8.   Pigskin Party KAL Wondering what this crazy KAL is? Check out this page which gives you the best overview with all necessary links. There's also a helpful "Start Here" Thread in the Ravelry Group Check out the Rules but if you still have questions, come over to the Questions thread and ask. We’re happy to help. Put your Name on the Roster then claim your spot in the End Zone Dance Thread where you'll track your points. For every 100 points, you'll earn an entry into the drawing for the Grand Prizes Continue to update this with all of the points you earn during the KAL Check out our amazing Sponsors; if you use their products, you'll earn more points per FO Some of our Pro Shop Level Sponsors will be creating exclusive items for Pigskin Party participants. They may come out at different times and will likely be limited in quantities, so keep your eyes on this Exclusive Items thread in the group for all the latest news Scope out the prizes! Some will be given out during the KAL; some will be part of the Grand Prizes. Don’t forget to use #DCSPigskinParty17 on Instagram so we check out what you’re doing and you can be eligible for participation prizes. Thank you to our Pro Shop Sponsors! Colorful Eclectic Daisy Girl and Company Fancy Image Yarn Fibernymph Dye Works Inner Yarn Zen Joy in Rugging Knit Style Yarns Knitted Wit Knitters Brewing Company Knitty Kitty Bags Mekamika Nekozuki Knits Plum Deluxe Tea Prairie Bag Works Queen City Yarn Sunsoaked Yarns Woodsy and Wild Yarn Love   Key Details Mentioned in this Episode: Monday Night Scavenger Hunt Reminder about September Interception sponsored by Yarn Love Shop Announced the October Interception sponsored by Knitters Brewing Company. Details can be found here. Life in Focus Happiness Project: September--- Learn new things I learned a lot from making my first crochet shawl. Trying the FLK heel was a great learning moment for me. I want to try another heel in the near future. What do you recommend? Vanilla is the New Black? October: Time & Energy Goal: Look at ways I'm wasting time and energy & reset. Think hard about what I do. Ask "why" more often. When I have an automatic thought- don't just act on instinct. See what really makes sense. Get off auto pilot. On a Happy Note Took a Friday off to go Outlet shopping with my best friend, Kris! I brought my niece Emelyn to Stitch House during the Greater Boston Yarn Crawl. She knit on her cowl for a little bit before we headed north to go apple picking. Met Kris, Matt, Aila and Will for an unseasonably warm day of Apple picking at Applecrest in Hampton Falls NH. Every year they add something- this year it was a sunflower maze. So beautiful. Only regret was not taking photos earlier in the day. They started turning away from the sun as the day went on. Congrats to Mom and Dad on their 40th anniversary! Finished my first session of Hiit Classes at the gym near work. It was brutal but I definitely feel stronger. I was able to sign up for the 3 out of the next 8 I'm able to go to. Ordered 12x12 lay flat photo book Picaboo and love it! My grandmother is on Facebook and sent me this cute message the other day! Made my day! I got a haircut. Loving this shorter look. My niece Aila is going to be performing in the Boston Ballet's The Nutcracker this winter as a lamb. Quote of the Week “Children think not of what is past, nor what is to come, but enjoy the present time, which few of us do.” -- Jean de La Bruyère   ________________ Thank you for tuning in!   Contact Information: Ravelry: BostonJen & Down Cellar Studio Podcast Ravelry Group Twitter: Instagram & Periscope: BostonJen1 Facebook: https://www.facebook.com/downcellarstudio Sign up for my email newsletter to get the latest on everything happening in the Down Cellar Studio!    

Hello Fellow Travelers! In our Season 2 Finale, Dr J gathers Dr's Santhosh, Ward, Susana, and newest cohost Dr Praz to talk about our favorite medical slang. Along the way they discuss their favorite medicalese, High-Fives, status hispanicus, Aye Tach, or Ooh Brady, The O sign and the O face, spanish cpr, GOMERs, frequent fliers, Hollywood Codes, cows, anaesthesia nicknames, pneumonia smith, FLK syndrome, Adult humane society, the cabbage patch and vegetable garden, getting hammered, british and brazilian medical slang, and more! So Sit back and prepare to learn a whole new language in the season finale of TMP. We will be back in October with new shows, topics, and guests, see you then! Contact Us! Twitter: @doctorjcomedy @toshyfro @travelnmedicine Facebook: facebook.com/travelmedicinepodcast Squarespace: travelmedicinepodcast.squarespace.com Patreon: https://www.patreon.com/travelmedicinepodcast Find Us! itunes: https://itunes.apple.com/us/podcast/episodes-travel-medicine-podcast/id914407095 stitcher: http://www.stitcher.com/podcast/travel-medicine-podcast?refid=stpr

Enterobacteria have evolved several strategies to survive the acidic environment of the gastrointestinal tract. One of the acid stress resistance systems is the Cad system in Escherichia coli, which is induced by low pH and in the presence of external lysine. It consists of CadA, which catalyzes the decarboxylation of lysine to cadaverine, the lysine/cadaverine antiporter CadB and the pH sensing transcriptional regulator CadC. Moreover, the lysine permease LysP inhibits the induction of cadBA expression when lysine is absent, and the small histon-like molecule H-NS acts as repressor for both cadBA and cadC transcription. Additionally, a random mutagenesis approach revealed that a deletion in yjeK leads to highly reduced cadaverine production. YjeK acts as 2,3-lysine aminomutase (LAM) while catalyzing the isomerization of (S)-α-lysine to (R)-β-lysine. The truncated lysyl-tRNA synthetase YjeA uses (R)-β-lysine as substrate to post-translationally modify and to activate the translation elongation factor EF-P at a conserved lysine residue (K34). EF-P and its ortholog eukaryotic initiation factor 5A (eIF5A) have been investigated for more than thirty years, but their roles in translation remained enigmatic. In this work the role of active EF-P in the Cad system was investigated in more detail. Reduced cadBA expression in ΔyjeA, ΔyjeK642-1029 and Δefp mutants was linked to impaired CadC translation. As the translation of cadA and cadB was EF-P independent, a general role of EF-P in translation could be excluded. The identification of CadC as first direct target for EF-P in E. coli allowed further investigations on the role of EF-P in translation. Determining the β-galactosidase activities of CadC´-LacZ translational fusions of increasing CadC length in efp- and efp+ cells revealed that EF-P is required for translation of the sequence found between codon 108 and 158 in cadC. This region comprises a cluster of three consecutive prolines (Pro120-Pro121-Pro122). Substitution of these prolines by alanines diminished EF-P dependency. Remarkably, cells harboring the CadC-PPPIP/AAAIS variant revealed cadBA expression even under non-inducing conditions. Thus, EF-P tightly controls the CadC copy number, which is crucial for stress dependent regulation of the Cad system. In order to investigate the work mechanism of EF-P in more detail, EF-P independent CadC´-LacZ hybrids were employed to artificially introduce prolines. Three consecutive prolines were sufficient for EF-P dependency, regardless of the codon or the context. The proline-rich proteins AmiB, FlhC, Flk, NlpD, RzoR, TonB and UvrB also showed EF-P dependent expression. Thus, the recognition of three consecutive prolines by EF-P is a general mechanism and not limited to CadC. Dr. Agata Starosta of the group of Dr. Daniel Wilson (Gene Center, LMU Munich) confirmed ribosomal stalling at polyproline-stretches in samples lacking EF-P with in vitro translation assays. Finally it was investigated, if EF-P expression and modification could be stress-dependently regulated. In this work first hints are given that the efp promoter contains a repressor site, and that expression of yjeA and yjeK is dependent on the pH of the medium and the presence of the small RNA binding protein Hfq. This leads to the suggestion that small regulatory RNAs are also involved in regulation of the EF-P modification enzymes. In conclusion, the results obtained in this work reveal a new regulatory mechanism by EF-P dependent translation. 100-1000´s of polyproline rich proteins exist in bacteria, archaea and eukaryotes. Therefore, EF-P and its orthologs aIF5A and eIF5A most likely play an important role in the adjustment of copy numbers of proteins with different functions in all kingdoms of life.

AIMS: The proliferative potential of pluripotent stem cell-derived cardiomyocytes is limited, and reasonable yields for novel therapeutic options have yet to be achieved. In addition, various clinical applications will require the generation of specific cardiac cell types. Whereas early cardiovascular precursors appear to be important for novel approaches such as reseeding decellularized hearts, direct cell transplantation may require ventricular cells. Our recent work demonstrated that MesP1 represents a master regulator sufficient to induce cardiovasculogenesis in pluripotent cells. This led to our hypothesis that 'forward programming' towards specific subtypes may be feasible via overexpression of distinct early cardiovascular transcription factors. METHODS AND RESULTS: Here we demonstrate that forced expression of Nkx2.5 similar to MesP1 is sufficient to enhance cardiogenesis in murine embryonic stem cells (mES). In comparison to control transfected mES cells, a five-fold increased appearance of beating foci was observed as well as upregulated mRNA and protein expression levels. In contrast to MesP1, no increase of the endothelial lineage within the cardiovasculogenic mesoderm was observed. Likewise, Flk-1, the earliest known cardiovascular surface marker, was not induced via Nkx2.5 as opposed to MesP1. Detailed patch clamping analyses showed electrophysiological characteristics corresponding to all subtypes of cardiac ES cell differentiation in Nkx2.5 as well as MesP1 programmed embryoid bodies, but fractions of cardiomyocytes had distinct characteristics: MesP1 forced the appearance of early/intermediate type cardiomyocytes in comparison to control transfected ES cells whereas Nkx2.5 led to preferentially differentiated ventricular cells. CONCLUSION: Our findings show proof of principle for cardiovascular subtype-specific programming of pluripotent stem cells and confirm the molecular hierarchy for cardiovascular specification initiated via MesP1 with differentiation factors such as Nkx2.5 further downstream.

Aim of the study was the description of the transcriptional profile of different vasculogenic, angiogenic and haematogenic growth factors (Flk-1, Flt-1, VEGF, c-kit, SCF, VE-Cadherin, FGF-2, Ang-1, BMP2, BMP4) in the tissues of early vessel- and blood cell formation of the chick embryo. For this purpose the quantitative real-time RT-PCR was used, with which the amount of defined transcripts in previously processed tissue samples can be determined. In the course of the first seven days (E0 – E7) of embryologic development tissue samples of the two extraembryonic haemangiopoetic organs, the yolk sac and the allantois/chorioallantois, as well as of the embryo itself were taken and the mRNA-quantity of the numbered factors determined as mentioned above. By this means a comparison of the transcriptional levels in the different samples was achieved and enabled relative quantification. So the transcriptional activity and the approximate time course of gene expression of the investigated growth factors in the first third of chick development could be determined. It was shown that the transcriptional activity of the genes of interest was generally higher in the allantois/chorioallantois compared to the yolk sac. This imbalance even grew constantly from embryonic age E4 to E7. Thus in the allantois/chorioallantois a more intense vasculogenesis/angiogenesis compared to the yolk sac was taking place and according to that the higher amount of VE-Cadherin suggested a higher fraction of endothelial cells in the tissues of the allantois/chorioallantois. The target genes Flk-1, Flt-1_cons, VEGF und VE-Cadherin showed only small fluctuations of the cellular mRNA content over the course of the observed development, i.e. they were transcribed with a quite stable intensity. For the target genes Flt-1_ms, c-kit, SCF, BMP2 und BMP4 minor increases as well as minor decreases of the cellular mRNA content were observed, which varied from one day of embryologic development to the next with a factor in the range of 0,2 to 5. Differential transcriptional intensities higher than that were only observed for FGF-2 and Ang-1. For FGF-2 in the yolk sac, respectively Ang-1 in the allantois/chorioallantois, an approximate sevenfold increase of the transcriptional level from E4 to E5 could be shown. This may underline the importance of the correlated proteins for the ontogenesis of these organs at that point of time. In the embryo from E1 to E2 an approximate 40fold increase of the cellular mRNA content of Ang-1 was observed, which was followed by an about 20fold increase up to E3. From E1 to E3 these increases summed up to an almost 1000fold increase, which was by far the strongest regulational activity observed in this study. In the yolk sac a similarly high amount of Ang-1 mRNA was detected, suggesting an effective translation, great importance of the correlating angiogenic proteins for the maturation of the primary capillary plexuses in the embryo as well as in the yolk sac after three days of extrauterine development can be postulated. The determination of the individual amplification efficiency of each reaction of the real-time RT-PCR with the help of computer-based linear regression analysis resulted for the first cycles with detectable amplification and according increase in fluorescence-intensity in a multiplication to far over the double up to the fourfold RFU-value from one cycle to the next. This contradicts the theoretical maximum of an exponential amplification to the base of two and could be the result of a quantitative imbalance in the limited matrix-molecules compared to the other, prominent at the beginning of the reaction in extense present, PCR-educts. An intense observation of the reaction kinetics of exponential phase PCR-amplification seems helpful for an even more exact nucleotide-quantification.