Podcasts about pc12

Napsali jsme o skryté službě Ulož.to Send. Pár minut poté přestala fungovat. Je kolem toho hromada otazníků. V Las Vegas se konal jeden z posledních velkých veletrhů CES. Věnujeme se mu v Podcastu, Kuba proto jen okomentuje, že je Las Vegas ošklivé, okradli ho tam v hotelu a spíše než CES zajímá návštěvníky obrovská koule Sphere.Řeč bude také o umělé inteligenci, mirkoplastech a nanoplastech v balené vodě a také o snaze vyrobit jeden standard na rychlý a jednoduchý přenos souborů z Androidu na počítač – Quick Share. Google to tentokrát zkouší se Samsungem, který je hlavní výrobce telefonů.01:08 – Open data počasí02:52 – Mikroplasty jsou všude05:59 – Novinky v ChatGPT08:27 – Sdílení z mobilu na PC12:56 – Ulož.to Send25:39 – Trendy na CESu

Link to bioRxiv paper: http://biorxiv.org/cgi/content/short/2023.04.18.537291v1?rss=1 Authors: Sahu, B. S., Mukherjee, M., Mukherjee, C., Ghosh, V., Jain, A., sadhukhan, s., Dagar, s. Abstract: Dense core vesicles (DCVs) and synaptic vesicles (SVs) are specialised secretory vesicles (SSVs) in neurons/neuroendocrine cells harbouring cargo whose abnormal release is associated with pathophysiology. Endoplasmic Reticulum (ER) stress and inter-organellar communication are also associated with disease biology. In pursuit of investigating the cell physiological consequences arising from the crosstalk of a stressed ER and DCVs, ER stress was modelled in PC12 neuroendocrine cells using Thapsigargin (Tg). DCV exocytosis was severely compromised in ER-stressed PC12 cells, reversed by Docosahexaenoic acid (DHA). Experiments with Tunicamycin(Tm), an independent ER stressor, yielded similar results. Concurrently, ER stress caused impaired DCV exocytosis also in INS-1 cells. Molecular analysis revealed blunted SNAP25 expression, potentially attributed to augmented levels of ATF4 (a well-known CREB inhibitor) and its transcriptional regulator CREB (also known to regulate key granulogenic players Chromogranin A, Secretogranin II). Our studies revealed severe defects in DCV exocytosis in ER-stressed cells for the first time, mediated by reduced levels of key 'exocytotic' and 'granulogenic' switches regulated via the CREB/ATF4/eIF2 axis. Copy rights belong to original authors. Visit the link for more info Podcast created by Paper Player, LLC

The boys are back in the studio for a bit of a shorter episode than normal with Tyler and Ryan on the road. This week we're talking Lou's new golf obsession, Tyler's observations from the NBA All Star Game in Utah, Ryan's thoughts on the XFL becoming the NFL's minor leagues, Lou got a big PC card in the mail, the current state of the hobby / are cards getting hot again and the excitement surrounding the release of 2023 Topps Series 1.Tune in for all of that PLUS another great over/under segment and an announcement about an upcoming live Card Talk celebration!Time Stamps: :38 - What's on your mind? (Lou)4:00 - What's on your mind? (Tyler)8:33 - What's on your mind? (Ryan)11:21 - Lou adds to his PC12:18 - NBA All Star Game card recap13:29 - Ryan's market observations14:48 - 2023 Topps Series 1 vibes17:33 - Over/Under35:24 - Final thoughts & a big announcement!ALSO!!! Don't forget to submit your questions for next week's full Q&A episode!See Privacy Policy at https://art19.com/privacy and California Privacy Notice at https://art19.com/privacy#do-not-sell-my-info.

TIME STAMPS0:00 Intro Musc0:18 Intro1:30 The PopC Facebook page has been deleted!3:43 How have we been?8:24 What have we been playing?8:33 Max has been enjoying PlayStation on the PC12:04 Hogarts Legacy impressions (code provided by PlAION ANZ)35:26 And now to discuss the controversy…53:29 Inform The Players (This week in PlayStation news)53:40 The Live for PlayStation […]

Cedric "Cedi" Gitchenko ist eine fliegende Legende. Er fliegt zahlreiche Flugzeuge und ist auch Testpilot. Er lebt seine Leidenschaft immer noch wie am ersten Tag aus, sei es mit Kunden im Gulfstream 650 (oder Global 7500, oder PC12 etc) oder beim PPL-Check. Er ist auch Mitgründer der Firma Boutique Aviation mit mehr als 10 Flugzeugen.

Check out the best Protein Bars of all time https://1stphorm.com/products/vegan-power-pro-bar/?a_aid=JSiems Avination, welcome back to episode 247 of the Pilot to Pilot podcast! Todays episode is with Jace Brennan, a 19 year old PC12 pilot. This is a re release and Jace has since moved to Hawaii where he is still flying a PC12!

Invité du Podcast de l'Aviation cette semaine, Cédric Lescop, président de Jetfly.Jetfly est une société dont le modèle économique repose sur la propriété partagée davions daffaires et qui propose une flotte homogène composée de Pilatus, PC-12 et PC-24. Cédric Lescop explique que ces appareils peuvent atterrir sur tous les terrains et sont parmi les plus efficaces en termes de consommation.A ce sujet, il estime que l'aviation se défend mal contre les attaques qui lui sont portées au niveau environnemental. En revanche, il ne croit pas que les carburants durables d'aviation, non disponibles, et encore moins l'hydrogène puissent être des solutions à court terme pour décarboner le secteur et il se concentre sur l'optimisation des opérations et la compensation.Concernant la crise, il constate lui aussi que l'aviation d'affaires s'en est mieux sortie que l'aviation commerciale et qu'elle a déjà retrouvé son niveau de 2019.

Mit einer Luftoffensive versucht die afghanische Armee im Sommer 2021 die Machtübernahme der Taliban doch noch zu verhindern. Die Streitkräfte fliegen in der Endphase des Afghanistan-Kriegs Angriff um Angriff. Nicht nur Talibankämpfer kommen dabei ums Leben, die Bomben töten auch Hunderte von Zivilpersonen. Bei den Luftattacken sind auch Pilatus-Flugzeuge aus der Schweiz im Einsatz. Welche Rolle spielen diese PC-12? Und wie kommt es, dass sie in einem Krieg im Einsatz sind? Der afghanischen Armee stehen im Kampf gegen die Taliban 18 Pilatus-Flugzeuge des Typs PC-12 zur Verfügung. Obwohl aus der Schweiz laut Gesetz keine militärischen Güter in Kriegsgebiete exportiert werden dürfen, fliegen die Pilatus-Flieger bei Attacken mit. Die USA haben die PC-12 den Afghanen 2015 und 2016 überlassen. Die Pilatus-Flugzeuge sind mit Kameras und modernen Überwachungsgeräten versehen und begleiten Luftangriffe der afghanischen Kampfbomber. Die Kameras in den PC-12 filmen die Attacken, und mit den technischen Geräten kann die Besatzung Ziele auskundschaften. PC-12 sind auch dabei, als bei einem Luftangriff vom 15. Juli 2021 nicht nur Taliban, sondern auch unbeteiligte Zivilpersonen sterben. Das lässt sich mit einer akribischen Auswertung von offenen Datenquellen nachweisen, und Augenzeugen bestätigen es. Fatal sei die mangelnde Ausbildung der afghanischen Operatoren, die in den PC-12 dabei sind, sagt der amerikanische Experte Marc Garlasco, der früher für das US-Verteidigungsministerium tätig war. Statt mit dem Auskundschaften der Ziele zivile Opfer zu verhindern, ist laut Garlasco das Gegenteil passiert. Hotspot ist ein Podcast von SRF Inhalt: Ben Heubl und Andreas Schmid (SRF Investigativ) in Zusammenarbeit mit Lighthouse Reports Produktion und Story: Eliane Leiser Archivmaterial: SRF R+A Technik: Mirjam Emmenegger

Magnoliaceae evolved flowers before the emergence of bees, how did this shape its pollination strategies? What makes the Magnolia flower perfect? In which growing conditions does Magnolia officinalis flourish? What compound gives M.officinalis essential oil its distinctive aroma? This episode is a brief introduction to a fascinating, ancient, historically significant tree. Intro tune: Botanical Berceuse by Carmen Porter (https://carmenporter.com) References: Magnolia Pollination: https://extension.illinois.edu/blogs/garden-scoop/2018-05-05-magnolia-pollination (https://extension.illinois.edu/blogs/garden-scoop/2018-05-05-magnolia-pollination) New Insights into Breeding and Propagating Magnolias. Thomas Ranney and Dominic Gillooly, Department of Horticultural Science, Mountain Horticultural Crops Research and Extension Center, North Carolina State University: https://mcilab.ces.ncsu.edu/wp-content/uploads/2017/07/ranney-and-gillooly-2014.pdf?fwd=no (https://mcilab.ces.ncsu.edu/wp-content/uploads/2017/07/ranney-and-gillooly-2014.pdf?fwd=no) Phenolic Constituents from the Stem Bark of Magnolia officinalis: https://pubs.acs.org/doi/abs/10.1021/np800494e (https://pubs.acs.org/doi/abs/10.1021/np800494e) Magnolol: A multifunctional compound isolated from the Chinese medicinal plant Magnolia officinalis, European Journal of Integrative Medicine: https://doi.org/10.1016/j.eujim.2011.09.002 (https://doi.org/10.1016/j.eujim.2011.09.002) https://www.sciencedirect.com/science/article/pii/S1876382011001405 (https://www.sciencedirect.com/science/article/pii/S1876382011001405) Neuroprotective effect of honokiol and magnolol, compounds from Magnolia officinalis, on beta-amyloid-induced toxicity in PC12 cells: https://doi.org/10.1002/ptr.3178 (https://doi.org/10.1002/ptr.3178)

Let's talk about the new book to series story and all of my thoughts on this new take of the characters, ships, and plot.

#022 - C'est avec plaisir que nous accueillons aujourd'hui Mathilde ! Pilote sur PC12 à travers l'Europe, son parcours dédié à l'aéronautique depuis son plus jeune âge est riche d'enseignements. Tu la connais peut-être déjà via sa page Instagram @pilot.mathilde où elle partage ses expériences et ses conseils ! Véritable chibane en herbe, Mathilde ne manque aucune occasion de vivre et partager sa passion au maximum. Du Tour Aérien des Jeunes Pilotes à son investissement dans l'entretien d'avions de collection, elle transmet cet amour de l'aéro avec énergie et motivation… et cet épisode ne fait pas exception… Bons vols !Notre site internet : https://www.xpchibane.com/Pour nous soutenir :Si tu apprécies ce podcast, la meilleure façon de nous soutenir est de nous mettre un gentil commentaire et 5 étoiles sur Apple Podcasts. Tu peux également parler de L'Expérience Chibane autour de toi à ta famille, tes amis et sur les réseaux sociaux !Au delà de ça, si tu souhaites nous apporter un soutien financier pour nous aider à aller à la recherche d'invités toujours plus passionnants, deux options s'offrent à toi : Patreon - Tu peux t'abonner mensuellement sur Patreon pour nous soutenir et obtenir des contreparties, comme des goodies ! https://www.patreon.com/xpchibane PayPal - Tu peux également nous faire un don ponctuel sur PayPal en cliquant sur le lien, si tu souhaites nous aider à poursuivre cette aventure palpitante !Nos réseaux sociaux :Instagram : https://www.instagram.com/xpchibane/Facebook : https://www.facebook.com/xpchibaneTwitter : https://twitter.com/xpchibane/LinkedIn : https://www.linkedin.com/company/xpchibane/L'équipe de production :Animateur : Sébastien SellePost-production : Victor Guichaoua et Paul Le Roux De BretagneIllustrations : Ilia Gerber, Léa Renault et Marek MadlRéseaux Sociaux : Maxence Le FlahecBande sonore : Thibaut MaurinInformations la protection de la vie privée : https://www.buzzsprout.com/privacySupport the show (https://www.patreon.com/xpchibane)

CAPITOLI:00:00 Benvenuti su Kristal News!00:24 State of Play: a mio parere disastroso.09:43 Nuova etichetta di Sony per la distribuzione PC12:47 "News a Ruota"17:00 ISCRIVETEVI al canale e seguitemi sui social e sul Podcast!Le #KristalNews sono anche in PODCAST!https://www.spreaker.com/show/kristalnews-il-podcastDisponibile anche su SPOTIFY, APPLE PODCAST, ecc eccPLAYLIST Kristal Newshttps://www.youtube.com/playlist?list=PLiAZAlZgYAdpnFFrYRcfnqVI6WCld_E5gSECONDO CANALE:https://www.youtube.com/channel/UCO13aJlGKgOVYq7-yMe_lPASEGUIMI IN LIVE SU TWITCH:https://www.twitch.tv/kristalcrossgamingSOCIAL & COMMUNITY:FACEBOOK: https://www.facebook.com/kristalcrossgaming/INSTAGRAM: https://www.instagram.com/kristalcrossgaming/CANALE TELEGRAM: https://t.me/kristalcrossgamingGRUPPO TELEGRAM: https://t.me/joinchat/APAtYQ5DPmVA3AHTl8mBcAPLAYSTATION: Cerca Kristal Cross Gaming Community!DISCORD: https://discord.gg/yhxtdSZmYPMIXER: https://mixer.com/KristalCrossGamingTUMBLR: https://kristalcrossgaming.tumblr.com/VUOI GIOCHI SUPERSCONTATI? CLICCA QUI!https://www.instant-gaming.com/igr/kristalcrossgaming/ https://www.cdkeys.com/?mw_aref=kristalcrossgaminghttps://www.epicgames.com/store/it/ (TAG: KRISTAL-CROSS)LINK AFFILIATO AMAZON:https://amzn.to/3tXIq3dSPONSOR - ABBONAMENTO AL CANALE:https://www.youtube.com/channel/UC-isEtNtS2_30c5ANuEEyKQ/joinDONAZIONI:https://paypal.me/kristalcrossVUOI AIUTARMI ANCORA DI PIU? SUPPORTAMI SU PATREON!https://www.patreon.com/kristalcrossgamingVUOI SPEDIRMI QUALCOSA? MANDALO A QUESTO INDIRIZZO!Flow Rock Live SRLS (per Kristal)Via Fioravanti, 3 San Benedetto del Tronto, AP 63074"LA CRICCA DEL SOTTOSUOLO": Link Utili!DISCORD: https://discord.gg/BD7vqkTELEGRAM: https://t.me/LaCriccadelSottosuoloFACEBOOK: https://www.facebook.com/LaCriccaDelSottosuolo/----------------------------------------VUOI SENTIRE I MIEI LAVORI? IL SITO DEL MIO STUDIOhttp://www.kristalcrossmusicproduction.com/page0/page0.htmlLE MIE BAND:THE LOTUS: https://www.youtube.com/user/TheLotusChannelLOST RESONANCE FOUND: https://www.youtube.com/user/lostresonancefoundAttenzione, quasi tutte le musiche che sentite nei video (quando presenti) sono o fatte da me, o dalle band di cui faccio parte, o delle band registrate e/o mixate da me nel mio studio.Uso Final Cut Pro X Per editare i video e Logic Pro X per l'editing audio.Grazie per la visita da Kristal Cross!

Olá, seja muito bem-vindo ao StandardsCast EP #XX Pilatus. Nesse episódio conversamos com Fábio Stersi (chefe de equipamento do Pilatus) sobre as curiosidades da operação do Pilatus (PC12) e sua importância para a Azul. Em caso de dúvidas, críticas ou sugestões, envie um e-mail para standardscast@voeazul.com.br. Este Podcast foi produzido pela Diretoria de Operações da Azul Linhas Aéreas. Em caso de divergência entre qualquer assunto técnico abordado e os documentos oficiais, os documentos prevalecerão. Todos os direitos reservados.

Today is just a short discussion about my two best weeks in 30 years. Yes 30 years.  And it's about friendships.  When I was a Captain in the fire service and I could sense that somebody really had a like boyhood dream of going on a fire truck, I'd take him as many times as I could.  I had somebody do that for me. So help you fellow human.

Welcome back to the Pilot to Pilot podcast episode 141 featuring, Jace Brennan. To give you some perspective on how far ahead Jace is in his career. When I was 19 I didn’t even know I wanted to be a pilot and didn’t end up flying a PC12 until I was 25. Jace, has been a go getter and has made all the right decisions ever since he decided to go all in on aviation. If you are young and want to be a pilot, this is the episode for you.

Todd & Mac chat about Mac's experience with a pc12.

Episode 141 Show Notes   Topic of the show: This week we will get the details of a middle of the night incident in TRIAD airspace and discuss the troubleshooting procedures of a solo Pilatus pilot.  We also discuss the differences throughout the NAS regarding the issuance of VFR codes at Class D airports.   Timely Feedback: PATRON Mike Kilo sent in fantastic audio feedback about Class D VFR flight following codes. PATRON Bravo Sierra discusses Class D codes under a busy Bravo on the East Coast. PATRON Juliet Sierra discusses Class D codes under a busy Bravo on the West Coast. PATRON Delta Charlie discusses his experience with VFR codes at his busy Class D airport.   Feedback PATRON Charlie Hotel asks about the pilot death penalty and offers insights into the best trainers. PATRON Charlie Hotel loves the P3 Orion! PATRON Charlie Mike sent in a picture of the APG crew showing off their OB Swag! PATRON Romeo Whiskey Kilo wants to know what we mean when we say “splat T”. Have a great week and thanks for listening!  Visit our website at OpposingBases.com You can support our show using Patreon or visiting our support page on the website.  Keep the feedback coming, it drives the show! Don’t be shy, use the “Send Audio to AG and RH” button on the website and record an audio message. Or you can send us comments or questions to feedback@opposingbases.com. Find us on twitter @opposing_bases.  Music by audionautix.com.  Third party audio provided by liveatc.net.  Friends of the show and maker of bags to protect your ATC headset from dust and germs: ATCSaks.com. Legal Notice  The hosts of Opposing Bases Air Traffic Talk podcast are speaking on behalf of Opposing Bases, LLC.  Opposing Bases, LLC does not represent the Federal Aviation Administration, Department of Transportation, or the National Air Traffic Controllers Association.  All opinions expressed in the show are for entertainment purposes only.  There is no nexus between Opposing Bases, LLC and the FAA or NATCA.  All episodes are the property of Opposing Bases, LLC and shall not be recorded or transcribed without express written consent.  For official guidance on laws and regulations, refer to your local Flight Standards District Office or Certified Flight Instructor.  Opposing Bases, LLC offers this podcast to promote aviation safety and enhance the knowledge of its listeners but makes no guarantees to listeners regarding accuracy or legal applications.

Link to bioRxiv paper: http://biorxiv.org/cgi/content/short/2020.08.20.258863v1?rss=1 Authors: Barak-Broner, N., Singer-Lahat, D., Chikvashvili, D., Lotan, I. Abstract: The polybasic juxtamembrane region (5RK) of the plasma membrane neuronal SNARE, syntaxin1A (Syx), was shown by us to act as a fusion clamp in PC12 cells, making release dependent on stimulation by Ca2+. By using a Syx-based FRET probe, we demonstrated that 5RK is absolutely required for a depolarization-induced Ca+2-dependent, close-to-open transition (CDO) of Syx that involves the vesicular SNARE synaptobrevin2 and occurs concomitantly with Ca2+-triggered release. Here, we investigated the mechanism underlying the 5RK requirement, and identified phosphorylation of Syx at Ser-14 (S14) by protein kinase CK2 as a crucial molecular determinant. Following biochemical verification that both endogenous Syx and CSYS are constitutively S14 phosphorylated in PC12 cells, dynamic FRET analysis of phospho-null and phospho-mimetic mutants of CSYS and the use of a CK2 inhibitor revealed that it is the S14 phosphorylation that confers the 5RK requirement. Concomitant amperometric analysis of catecholamine release revealed that the phospho-null mutants do not support release, spontaneous and evoked. Collectively, these results identify a functionally important CK2 phosphorylation site in Syx that is required for 5RK-regulation of CDO and for concomitant Ca2+-triggered release. Copy rights belong to original authors. Visit the link for more info

Link to bioRxiv paper: http://biorxiv.org/cgi/content/short/2020.07.31.229153v1?rss=1 Authors: Calo, L., Hidari, E., Wegrzynowicz, M., Dalley, J., Schneider, B., Anichtchik, O., Carlson, E., Klenerman, D., Spillantini, M. G. Abstract: synuclein aggregation at the synapse is an early event in Parkinson disease and is associated with impaired striatal synaptic function and dopaminergic neuronal death. The cysteine string protein (CSP) and synuclein have partially overlapping roles in maintaining synaptic function and mutations in each cause neurodegenerative diseases. CSP is a member of the DNAJ/HSP40 family of co-chaperones and like synuclein, chaperones the SNARE complex assembly and neurotransmitter release. synuclein can rescue neurodegeneration in CSP KO mice. However, whether synuclein aggregation alters CSP expression and function is unknown. Here we show that synuclein aggregation at the synapse induces a decrease in synaptic CSP and a reduction in the complexes that CSP forms with HSC70 and STGa. We further show that viral delivery of CSP rescues in vitro the impaired vesicle recycling in PC12 cells with synuclein aggregates and in vivo reduces synaptic synuclein aggregates restoring normal dopamine release in 1-120hsyn mice. These novel findings reveal a mechanism by which synuclein aggregation alters CSP at the synapse, and show that CSP rescues synuclein aggregation-related phenotype in 1-120hsyn mice similar to the effect of synuclein in CSP KO mice. These results implicate CSP as a potential therapeutic target for the treatment of early-stage Parkinson disease. Copy rights belong to original authors. Visit the link for more info

A Friendly discussion...as the nation attempts to continue opening, hot spots pop up throughout the country. Is there an end in sight?

pc12 True Intimacy by Douglas Dantzler

How do we eliminate flammable fuel vapor in transport aircraft fuel tanks? Equipment like the Parker fuel inerting system - OBIGGS - and Air Separation Modules fill the ullage space with inert nitrogen gas. The Pilatus PC12 was a game-changer. Is the PC24 business jet headed for the same level of success?Cirrus Aircraft introduced the airframe parachute system on their SR-20. With the new Vision Jet, Cirrus and Garmin take safety to the next level with the Safe Return System. Check out the Safe Return System in operation via YouTube - https://youtu.be/JvzPpOknjME Learn more about Weather Guard StrikeTape segmented lightning diverter strips. Follow the show on YouTube, Twitter, Linkedin and visit us on the web. Have a question we can answer on the show? Email us! 6:20 Parker Aerospace's inerting systems 17:57 The Pilatus PC12 and PC24 23:58 Lightning and turboprop engines 25:49 Cirrus parachute and Safe Return system 33:13 Garmin makes awesome products Transcript: Struck EP6 Dan: Hello, welcome back. This is the Struck podcast, episode six. I'm your cohost Dan Blewett, and here on the Struck podcast we talk about everything, lightning protection, aviation and aerospace tech. And I am here joined remotely by lightning protection expert, Allen Hall. Allen, how are you? Allen:  Great. Dan, how are you doing? Dan: Doing well, so. A lot of new things going on this week. I got a new mic on the way, so ordered some new parts for my podcasting set up here. I got this huge, like dr octopus, um, like at home set up, which is pretty intense. Um, what's going on with you up there? Allen:  Uh, well, we've been through a couple of snowstorms over the last week. Uh, everybody is still quarantined for the most part. They canceled school for the rest of the year. Uh, how's everyone feel about that? Not happy, not happy at all. Uh, all the, the kids are just missing being with the kids and the teachers are trying to work, uh, classes through zoom and, or other video formats, and it's just not the same. And so the kids. Are really struggling with the lack of sort of one-on-one or a groups with the teacher interactions. I'm just trying to get homework done. The other thing that he announced, it's, it's all pass fail. So the kids don't have grades like they would normally, and my daughter's a junior so. This is sort of like the year you need grades to start applying to schools and no one showed her and the SATs have been shoved off and it's just, everything's in chaos right now. Um, so that's, that's, uh, an unusual twist cause I didn't think the sat would get shoved off like that, but it, it did, it got pushed off until then, until the fall.  And then the other thing that seems like it's happening is a lot of the colleges and universities. Or not worrying about sat scores. So they're just saying, apply with what you have, which is going to be, it's going to be an interesting curve that's just going to be on, uh, for the next couple of years to see how that all plays out. We're not going to know, obviously for several years to see what happens there, but, um, kids not going to class all the time and, and just know a couple of teachers in different parts of the country. And. They're having trouble with the kids showing up on the, on the video, uh, video links, like someone that just don't show up and what are you going to do? There's really, you can't force them to show up. Right. So if they're not going to show up, they're not going to show up. Yeah. It's unfortunate because the kids are really missing out on a good opportunity to learn. You know, if you're sitting at home in bounds, we'll be learning something. Yeah, we'll see. Well, they're probably Googling things, if that's any consolation. Tick tock, tick tock you tubing a, yeah, yeah, yeah. Or learning how to, I mean, there's so much weird stuff on Instagram, like interesting stuff. I watch. Little clips of manufacturing and woodworking and li...

She's flying Js. high performance turbo props and if you cant find her sitting solo in the left seat of a PC12, she's probably out jeepin!© 2019 All Rights Reserved

Avination, welcome back to the Pilot to Pilot podcast episode 58 featuring Reed Chuda. Reed is a currently building his time as an FO on the mighty PC12 aka baelatus. Some of the things we talk about on today's episode are. His Grandpa flew bombers in the Army. Reed’s brothers is a fighter pilot in the Navy. How aviation skipped a generation in their family. Reed talks about his support system while training. How the University of North Dakota flying camp helped Reed realize this is what he actually wanted to do. What a flying summer camp is actually like. How a flying summer camp can help you get ahead in your flying career. Does Reed recommend future pilots going to summer camps?

What is going on Avination, welcome to episode #52 featuring Stan Kuliavas from Levaero Aviation. In this episode, Stan and I talk about what it is like to go to Embry Riddle without wanting to become a pilot, how he had zero interest in aviation even though his dad was involved in aviation and much more. How Stan took his first intro flight. How Stand is thankful his dad didn’t push aviation on him. Why you have to love the process of becoming an airline pilot. How your first few hundred hours really mold you into the pilot you will be later in your career. We talk about what General aviation in Canada is like. How Stan learned quite quickly that he didn’t want to be an Air Canada pilot. How the “aviation bug” can be different for everyone. Stan figured out that he wanted to fly when he wanted to, not when he had too. Why Stan chose to go to Embry Riddle. You can go to Riddle and not want to be a pilot. How Ohio State went from wanting to get rid of their Aviation Program to wanting to keep it and build on it. Did you know Embry Riddle had athletics? How Stan had the bright idea to move back to Canada from Florida. We talk about what it is like to work in Sales for a fractional company. How it is important to always try to learn new things. How you are either getting better or getting worse  We talk about when the podcast started getting some traction. Why it is important to put yourself out there and be vulnerable. Stan talks about ocean crossing with a PC12. Thank you for listening to today's episode. Make sure to check out our sponsor Cadence Aviation they make premium headsets at a great price. Avination, I hope you enjoyed this episode if you did. Leave us a review on all social media, iTunes and check out our Patreon page for early access and Pilot to Pilot swag.

What?  We’ve all been told for years, “winners never quit and quitters never win” Learn to ignore that adage and start quitting to win!

There's a science behind everything. Whether it's health based, thought based, or even the science behind a belief, we now have the resources to find out why these things occur, and how they can be reprogrammed. An example of this is the clump of cells that make up a belief. There's a literal neurological response that occurs leading to us producing this belief any way we can, hence our constant run ins with repetitive situations. This is why we recreate our circumstances based off that singular belief of "I'm not good enough" or "this person will leave me for someone else". These beliefs are our truths, and we go to the depths of actually manifesting situations to prove us right. This episode is about reprogramming the subconscious neurological patterns that occur and why it's important that we do so. Something as simple as a new experience creates a new neurological pathway in the way. Then new science showing mindfulness activities improve grey matter in areas of the brain that host our decision making, focus, emotional regulation and so many other mechanics, shows us how we can alter the brain's literal way of working. Everything learnt through this episode in the brain region are from the workings of brain researchers Dr. Daniel Amen and John Assaraf. --- Send in a voice message: https://anchor.fm/pocketcoach/message

80 Fast Track to an Airline Pilot Career, Piper M-series, Pilatus PC-12 + GA News Your Cirrus Specialist. Call me if you're thinking of buying a new Cirrus SR20 or SR22. Call 1-650-967-2500 for Cirrus purchase and training assistance. Send us an email - http://www.sjflight.com/Forms/inquiry.htm If you have a question you'd like answered on the show, let listeners hear you ask the question, by recording your listener question using your phone. COPA Las Vegas Pilot Interviews80 Max interviewed pilots while attending the Cirrus Owner Pilots Association (COPA) annual meeting in Las Vegas. One future airline pilot who is on track to fly 1500 hours in one year, and she talks about how she’s accomplishing that. Max also interviews pilots about using flight simulators and Angle of Attack indicators, and two dealers about the Piper M350, M500, and M600, and the Pilatus PC-12. If you love the show and want more, visit my Patreon page to see fun videos, breaking news, and other posts  in the Blog section. And if you decide to make a small donation each month,  you can get some goodies! Check out our recommended ADS-B receivers, and order one for yourself. Yes, we'll make a couple of dollars if you do.  Check out our recommended Aviation Headsets, and order one for yourself! Get the Free Aviation News Talk app for iOS or Android. Mentioned in the ShowAOPA Region Fly-In Gulf Shores, AL FAASTeam Facebook Group Turning Fledglings into Fliers - Three Challenges for Flight Instructors News Stories Operation Airdrop sends planes with supplies after Hurricane Michael Chinese Investor Rescues Eclipse, to Buy Company NetJets deal with Cessna could be worth almost $10 billion Sheriff: Helicopter pilot stopped in car with 2 kilos of cocaine Sydney seaplane crash pilot may have been knocked out Pressurized cabin factor in pilot fatality during opening of G150 door MagniX tests new electric motor on Cessna Ironbird Scalewings reveals secret hybrid with multiple redundancy Airlines impose cannabis ban on pilots and cabin crew Max Trescott is a participant in the Amazon Services LLC Associates Program, an affiliate advertising program designed to provide a means for sites to earn advertising fees by advertising and linking to amazon.com.

Bienvenue dans le septième épisode de ce podcast !Aujourd'hui nous allons nous intéresser à un sujet orienté vers la formation de pilote professionnel. La première étape d'une telle formation après l'obtention de la licence de pilote privé (PPL) est le passage de l'ATPL théorique. Il s'agit d'un examen théorique qui couvre un large panel de sujets théoriques allant de la réglementation aérienne aux systèmes techniques d'un avion en passant par les techniques de navigation.En introduction, je présente notre invité de la semaine, Alexandre. Alexandre est un élève en cours de formation de pilote professionnelle qui vient d'obtenir récemment son ATPL théorique.ActualitésCette semaine, nous parlons de la certification du nouvel avion de Pilatus, le PC-24. Le PC-24 est une grande nouveauté pour Pilatus car il s'agit de leur premier avion à turbo-réaction. Ils ont essayé de reproduire le succès qu'a été le PC12 mais en version équipés de turbo-réacteurs.Cette approche est un pari audacieux afin de percer dans le marché très compétitif des avions d'affaire d'entrée de gamme. De nombreux programmes d'avions destinés à ce marché ont échoué ces dix dernières années. La proposition de Pilatus est néanmoins radicalement différente grace à une polyvalence exceptionnelle et devrait donc réussir à se démarquer de ses compétiteurs. Pour plus d'informations, vous pouvez consulter la fiche technique sur le site de Pilatus et la brochure du PC-24.Photo du PC-24La seconde actualité de la semaine est une analyse d'un incident impliquant une perte de contrôle d'un Airbus A321 Lufthansa à cause d'un problème de commandes de vol. Cet incident n'est pas récent car il date de 2014 mais sa résolution n'est que très récente.Lors de la montée au départ de Bilbao, l'avion a rencontré un givrage des sondes d'angle d'attaque ce qui a causé une activation erronée des protections induisant une descente incontrôlée de l'avion. Suite à une discussion avec la maintenance, ils ont réussi à reprendre le contrôle de l'avion. La logique de fonctionnement du fly-by-wire de la famille Airbus A320 a été remise en cause.Suite à cet incident, Airbus a publié un avis urgent afin de rendre obligatoire la modification de la documentation pour y insérer une nouvelle procédure palliative. Vous trouverez la procédure complète dans le lien ci-dessous.Document Air France détaillant la procédure AirbusVous trouverez plus d'informations sur cet incident sur Aviation Herald.Rubrique thématiqueCette semaine, nous nous intéressons à l'ATPL théorique en discutant avec Alexandre. Alexandre est un élève pilote professionnel en cours de formation qui vient juste d'obtenir son ATPL théorique.Nous discuterons de son parcours jusque là avant de s'intéresser aux raisons qui l'ont amené à se lancer dans une telle aventure. Nous passerons en revue le contenu de l'ATPL théorique sans oublier d'évoquer les choix importants tels que le choix de l'école et de la langue. Nous parlerons également des techniques de révision et des difficultés de cet examen.Pour conclure, nous proposerons quelques conseils pour les personnes souhaitant se lancer dans un ATPL théorique.Ma pile de livres de l'ATPL théoriqueConclusionAinsi se conclut donc le septième épisode. J’espère qu’il vous a plu et je vous invite à vous abonner sur votre application de podcast favoris. Egalement, n’hésitez pas à laisser un avis 5 étoiles sur iTunes ce qui permettra à d’autres personnes de découvrir ce podcast.Si vous avez des questions, des remarques ou des suggestions, n’hésitez pas à utiliser le formulaire de contact.Si vous voulez recevoir des notifications lors de la sortie des nouveaux épisodes, vous pouvez vous inscrire à la newsletter dans la barre latérale droite de notre site. Vous pouvez également nous suivre sur Twitter sur @ParlonsAviation et sur Facebook.CréditsPhoto PC-24 : PilatusCe podcast est proposé sous licence Creative Commons BY-NC-ND 3.0.

W tym odcinku cyklu Maciej Bennewicz opowiada, czym jest kognitywistyka i jakie są wzajemne relacje między tą szybko rozwijającą się nauką a coachingiem i dlaczego dzięki niej coaching może być jeszcze bardziej skuteczny. Zapraszam!

W tym odcinku cyklu Maciej Bennewicz opowiada, czym jest kognitywistyka i jakie są wzajemne relacje między tą szybko rozwijającą się nauką a coachingiem i dlaczego dzięki niej coaching może być jeszcze bardziej skuteczny. Zapraszam!

Bienvenue dans le cinquième épisode de ce podcast !Aujourd’hui nous allons discuter d’un sujet un peu plus technique qu’à l’habitude. Nous allons nous intéresser aux commandes de vol et plus particulièrement aux commandes fly-by-wire. C’est un sujet plus orienté technique et aviation commerciale que d’habitude mais je le trouve particulièrement intéressant.En introduction de cet épisode, je présente le sujet de cette semaine.ActualitésCette semaine, nous parlons d’un nouveau programme de chez Cessna, le Denali. Le Denali est un nouvel avion avec un unique turbopropulseur. Cette motorisation est un nouveau développement de chez General Electrics. La ressemblance avec le PC12 est frappante et cet avion en sera, sans aucun doute, un concurrent direct.Photo de comparaison Denali vs PC12Illustration de comparaison turbopropulseur vs turboréacteurVous trouverez plus d’informations sur le site de Cessna au sujet de ce nouvel avion.Nous parlerons également de l’introduction par easyJet de vols par correspondance sous le produit « Worldwide by easyJet ». Cette nouvelle offre n’est pas novatrice de manière générale car les compagnies traditionnelles le proposent depuis toujours avec leurs alliances telles que Sky Team ou Star Alliance. La nouveauté est que ces vols en correspondance s’effectuent entre des low-costs ne proposant traditionnellement que du point-à-point. Nous détaillerons l’implémentation de ces nouveaux services et les différences avec les compagnies classiques.Rubrique thématiqueQuelles sont les commandes et les gouvernes de vol d’un avion ? Quels sont les différents systèmes permettant de relier les commandes et les gouvernes ? Comment fonctionne le système fly-by-wire d’un avion comme l’A320 ?Nous nous intéresserons au commandes de vol et aux différents systèmes historiques avant d’étudier le système fly-by-wire de l’Airbus A320. Nous détaillerons les différentes lois de commandes de vol ainsi que les protections associées.Les deux schémas ci-dessous récapitulent la première partie de cette rubrique thématique. Le premier présente les axes de rotation d’un avion alors que le second présente les différentes gouvernes.Schémas récapitulatifs commandes et gouvernes de volDocument Airbus détaillant les commandes de vol de l’A320 Le document détaillant les statistiques d’accident par type d’avion est disponible sur le site d’Airbus. Le graphique ci-dessous détaille les taux d’accidents par génération d’avions.Les avions de troisième génération sont les avions à commandes hydrauliques classiques disposant de système d’autopilote dits « intégrés ». Cela inclue par exemple le Boeing 737, 747, 757 et 767 ainsi que les Bombardier CRJ et Embraer ERJ. Les avions de quatrième génération sont les avions à commande fly-by-wire. Cela inclue tous les Airbus à l’exception des A300 & A310 ainsi que les Boeing 777 et 787 et les Bombardier C-Series.Graphique comparatifs statistiques d'accidents AirbusOn voit bien une différence non-négligeable entre ces deux générations d’avion.Vidéo de la semaineLa vidéo de la semaine est une vidéo de la chaine « High Pressure Aviation Films » tenue par un copilote de chez Air France prénommé Hugo Possamaï. On y voit une approche de nuit en Boeing 777 au dessus de Los Angeles.Cette vidéo ainsi que les autres vidéos proposées par Hugo sont vraiment exceptionnelle. La clarté et la précision de l’image sont impressionnantes. Je ne peux que vous regarder d’activer la 4K afin d’en prendre plein les yeux.Lien vers la vidéoConclusionMerci d’avoir écouté le cinquième épisode de Parlons Aviation. J’espère que cet épisode vous a plu. Si vous souhaitez nous envoyer vos questions, remarques ou suggestions, n’hésitez pas à utiliser le formulaire de contact.N’hésitez pas à laisser un avis sur iTunes afin d’aider d’autres personnes à trouver ce podcast. Si vous pensez que ça mérite 5 étoiles, alors c’est encore mieux !CréditsSchémas d’explication des commandes de vol et motorisations : L’avionnaire. Images reproduites avec l’autorisation de l’auteur.Diagramme des statistiques d’accident : AirbusDocument commandes de vol A320 : smartcockpit.comCe podcast est proposé sous licence Creative Commons BY-NC-ND 3.0.

The Practical Catholic welcomes Daniel Conigliaro, an old friend from UF who is now a Postulant with the Oblates of the Blessed Virgin Mary. Daniel has moved in with the guys for the summer, so for his first episode as a guest star we interviewed Daniel about how he discerned religious life, what that means, and also got into some practical tips to help you discover what calling God is asking of you. Tune in for lots of jokes, lots of making fun of each other, and welcome the newest member of our team (AKA Summer Intern)

Hosts: Tom Hering & Mike Mathews Today's Stories: We’re lucky if we get to be old, physician and professor believes - Dr. Bill Thomas, 5 Timeless Ways To Turn Your Life Around at Any Age, Portland Auto Show, Super Bowl 50, and more…. For additional show notes, visit BoomerMale.com. Download or subscribe to this show at BoomerMale.com, the podcast for men 50+

Hosts: Tom Hering & Mike Mathews Today's Stories: We’re lucky if we get to be old, physician and professor believes - Dr. Bill Thomas, 5 Timeless Ways To Turn Your Life Around at Any Age, Portland Auto Show, Super Bowl 50, and more…. For additional show notes, visit BoomerMale.com. Download or subscribe to this show at BoomerMale.com, the podcast for men 50+

Background: Epidemiological studies have suggested that long-term use of nonsteroidal anti-inflammatory drugs (NSAIDs) is associated with a reduced incidence of Alzheimer's disease (AD). Several mechanisms have been proposed to explain these findings including increased shedding of the soluble ectodomain of the amyloid precursor protein (sAPP), which functions as a neurotrophic and neuroprotective factor in vitro and in vivo. Objective: To clarify whether NSAIDs consistently stimulate sAPP secretion. Methods: 293-EBNA cells with stable overexpression of an APP-alkaline phosphatase fusion protein (APP-AP), SH-SY5Y and PC12 cells or primary telencephalic chicken neurons were treated with ibuprofen or indomethacin. APP shedding was then determined by measuring AP activity in conditioned media, Western blot analysis with antibodies against total sAPP or specific for sAPP-alpha, or in a pulse-chase paradigm. Results: AP activity in conditioned media was not increased after NSAID treatment of 293-EBNA cells whereas it was elevated by phorbol ester. Surprisingly, ibuprofen or indomethacin treatment of SH-SY5Y and PC12 cells expressing endogenous APP did not cause changes in sAPP or sAPP-alpha secretion or downregulation of cellular APP. These findings were further corroborated in primary chicken neuronal cultures. Conclusions: Using various experimental settings, we were unable to confirm sAPP or sAPP-alpha stimulation with the NSAIDs ibuprofen and indomethacin in transfected and nontransfected cells of neuronal and nonneuronal origin. Importantly, these findings seem to rule out chronic sAPP stimulation as an alternative mechanism of NSAID action in AD. Copyright (C) 2008 S. Karger AG, Basel

In this podcast I deliver the long promised review of FSX. It is a good game to be sure, however, I gave it a ratting of 7.5/10 due to its lack of play progression and poor engine performance. In the process of reviewing FSX I also talk a bit about its third party dependent content system and the way games of its type have been so successful from a business stand point. I also talk about the events of Fancon 2007. Below you will find the text of the report I made for Sky Blue Radio. Thanks guys for helping me attend such a great event! Pics are on the way got to get them off my camera phone. -Nicolas Friday November 9th, 2007 I landed in SEA for Fancon @ 4:46 PST yesterday on rwy 16l. The route used was SILOW1.SILOW FLG GCN BERYL ELY REO IMB.CHINS5 with a take off from rwy 7l @ PHX. Overall it was a good flight. The show starts bright and early on Saturday so I'll be ingesting some of Seattle's best coffee. If there is any info you folks want me to retrieve please let me know. I'll be posting updates on this thread as often as I can. If anyone happens to be going to the show look for a long haired leaping gnome. I'd love to say hi:P That's all for now. Update #1 The show is now underway, and what better way to start than free food and alcohol! I got to talk shop with the AVSIM staff writer, as well as one of Boeings flight simulation developers not to mention a certain international third party dev team working on a very extensive A320 series sim product. Will report more tomorrow. For now nicnacjak is at the gate and shutting down. Update #2 Saturday November 10th, 2007 The main day of Fancon has now come to a close. But, oh what a day it was! I spent most of my day at the conference sessions learning as much as I could from the shows presenters. P.I. Engineering was the first up with recommendations on how to make ground based scenery look more realistic. Although most of their work is more applicable to train sim, the FS modeler can also gain a lot from their insight. Next up was Raynald Bedard of Embry-Riddle University. His talk was based around the FSX mission system and the way that he and a team of his students have been able to add an advanced scoring system to the missions. Although the original intent of this system was to assist in training students as the university, the technology developed for this is being brought into the flight simulation community. During the show visitors were able to play missions that qualified them to be part of the FanCon Fly-Off. The winner of the event received a $25,000 scholarship to Embry-Riddle. The next talk saw PMDG premiering an early version of their MD11 aircraft. It goes without saying that this was a very exciting presentation. The crowds went wild and the complexity of the simulation was plainly evident. After the premier, the software was demoed at PMDGs booth. Although I didn't get a chance to play with it, it's on the top of my to do list for day 3! Following these talks I went to spend sometime on the show floor. The impending release of the PC12 for FSX courtesy of Flight1 has got me excited. There is also AirSimmer, who is currently working a very realistic model of the A320. Although getting the fly-by-wire- system right is clearly a big hurdle, the demo I saw this weekend was very promising. I also stopped to have a chat with Captain Mike Ray, one of two former 747-400 pilots that I talked to today! Captain Ray has a new manual out covering the A320, a handy asset to have aboard. Next it was back to the talks to hear from Aces themselves. It would appear as if they are putting a very heavy emphasize on feeding the community of payware add on developers. Their eventual goal is to make both flight and --- Send in a voice message: https://anchor.fm/nicnacjak/message

The adhesive properties of neural cell adhesion molecules (NCAMs) can be modified by alternative splicing of the primary transcript or posttranslational modifications. In the present study, we describe distinct forms of alternative splicing and posttranslational modification of the extracellular domain of NCAM of various endocrine tissues and derived tumor cells of the rat. Using an antiserum detecting the immunoglobulin-like domains of NCAM as well as a monoclonal antibody recognizing the NCAM-specific polysialic acid (PSA), we observed a similar staining pattern in adrenals, pituitary, and neoplastic endocrine cells. In endocrine tumor cells [pheochromocytoma (PC12), insulinoma (RINA2), and pituitary tumor cells (GH3)], NCAM immunoreactivity was most intense at contact sites between the cells. The immunocytochemical data were substantiated by results of in situ hybridization histochemistry. Specifically, higher levels of NCAM mRNA were detected in the adrenal cortex than in the medulla. In the pituitary, NCAM mRNA was more abundant in the anterior and intermediate lobes than in the neural lobe. The sequence of NCAM mRNAs in endocrine cells was analyzed by polymerase chain reaction and S1 nuclease protection assays. We found that major exons 4-13 of the NCAM mRNA in endocrine tissues and related tumor cell lines were homologous to those in the brain. However, PC12, RINA2, and GH3 tumor cells; normal rat pituitaries; and adrenals contained different amounts of NCAM mRNA with an alternative extra exon, termed VASE (also called pi in mouse) between constitutive exons 7 and 8. In addition, in pituitaries, we detected an alternative exon in splice site a between the constitutive exons 12 and 13, termed a15, with or without an AAG triplett. These sites are thought to be important for the adhesive properties of NCAM. Therefore, these results suggest that modifications of NCAM may be important for adhesive interactions in normal and neoplastic endocrine cells.

The role of Mg2+ during the final steps of exocytosis was investigated using rat pheochromocytoma cells (PC12) permeabilized with bacterial pore forming toxins. Concentrations of free Mg2+ between 0.2 and 2 mM slightly lowered the basal but greatly enhanced the [3H]dopamine release elicited by 8 μM free Ca2+. Maximal effects were obtained at approximately 1 mM free Mg2+. At higher concentrations Mg2+ was less potent. Similar effects of Mg2+ were obtained in cells permeabilized either for small molecules (by α-toxin) or for large ones (by streptolysin O). It is concluded that millimolar concentrations of cytoplasmic Mg2+ play an important role in Ca2+ triggered exocytosis.

The inositol 1,4,5-trisphosphate (IP3)-sensitive Ca2+ compartment of endocrine cells was studied with alpha-toxin- and digitonin-permeabilized rat insulinoma (RINA2) and rat pheochromocytoma (PC12) cells. The Ca2+ uptake was ATP-dependent, and submicromolar concentrations of IP3 specifically released the stored Ca2+. Half-maximal Ca2+ release was observed with 0.25-0.5 mumol of IP3/l, and the amount of Ca2+ released due to IP3 could be enhanced by additional loading of the Ca2+ compartment. Consecutive additions of the same concentration of IP3 for 1-2 h always released the same amount of Ca2+ without desensitization, providing an ideal basis to further characterize the IP3-induced Ca2+ release. Here we describe for the first time a reversible inhibitory effect of decavanadate on the IP3-induced Ca2+ release. Among the vanadium species tested (decavanadate, oligovanadate and monovanadate), only decavanadate was inhibitory, with a half-maximal effect at 5 mumol/l in both cell types. The effect of decavanadate could be overcome by increasing the amount of sequestered Ca2+ or added IP3. Decavanadate did not affect the ATP-driven Ca2+ uptake but oligovanadate was inhibitory on Ca2+ uptake. p-Hydroxymercuribenzoate (pHMB) at concentrations between 10 and 30 mumol/l also inhibited the Ca2+ release due to IP3. Thiol compounds such as dithiothreitol (DTT; 1 mmol/l) added before pHMB removed all its inhibitory effect on the IP3-induced Ca2+ release, whereas the inhibition caused by decavanadate was unaffected by DTT. Thus, the decavanadate-dependent inhibition functions by a distinctly different mechanism than pHMB and could serve as a specific tool to analyse various aspects of the IP3-induced Ca2+ release within endocrine cells.

Conditions are described for controlled plasma membrane permeabilization of rat pheochromocytoma cells (PC12) and cultured bovine adrenal chromaffin cells by Streptolysin O (SLO). The transmembrane pores created by SLO invoke rapid efflux of intracellular 86Rb+ and ATP, and also permit passive diffusion of proteins, including immunoglobulins, into the cells. SLO-permeabilized PC12 cells release [3H]dopamine in response to micromolar concentrations of free Ca2+. Permeabilized adrenal chromaffin cells present a similar exocytotic response to Ca2+ in the presence of Mg2+/ ATP. Permeabilized PC12 cells accumulate antibodies against synaptophysin and calmodulin, but neither antibody reduces the Ca2+-dependent secretory response. Reduced tetanus toxin, although ineffective when applied to intact chromaffin cells, inhibits Ca2+-induced exocytosis by both types of permeabilized cells studied. Omission of dithiothreitol, toxin inactivation by boiling, or preincubation with neutralizing antibodies abolishes the inhibitory effect. The data indicate that plasma membrane permeabilization by Streptolysin O is a useful tool to probe and define cellular components that are involved in the final steps of exocytosis.

Two possible cellular pathways of catecholamines from the chromaffin vesicles of PC 12 cells to the surrounding medium are explored in this study. The direct one circumventing the cytoplasm can be activated in a-toxin-permeabilized cells with micromolar levels of free Ca2+. Catecholamine metabolites formed in the cytoplasm (i.e., 3,4-dihydroxyphenylacetic acid and 3,4-dihydroxyphenylethanol) are neither formed nor released from the cells under these conditions. However, when vesicular catecholamines were discharged into the cytoplasm by addition of the ionophore nigericin, such metabolites are formed and released into the medium independent of Ca2+. Both types of experiments provide direct evidence for the operation of Ca2+-induced exocytosis of dopamine and noradrenaline in permeabilized PC12 cells. The Ca2+ dependence of dopamine or noradrenaline release, as measured by the determination of the endogenous catecholamines using the high-performance liquid chromatography technique, exhibits two different phases. One is already activated below 1 pM free Ca2+ and plateaus at 1-5 pM free Ca2+, while a second occurs in the presence of larger amounts of free Ca2+ (10-100 pM). Ca2+-induced catecholamine release from the permeabilized cells can be modulated in different ways: It is enhanced by the phorbol ester 12-0-tetradecanoylphorbol 13-acetate and the diacylglycerol 1 -oleyl-2-acetylglycerol provided Mg*+/ATP is present, and it is inhibited by guanosine 5’-0-(3-thiotriphosphate). The latter effect is abolished by pretreatment of the cells with pertussis toxin but not by cholera toxin. Thus, it appears that Ca2+-induced exocytosis can be modulated via the protein kinase C system, as well as via GTP binding proteins.

Crude alpha-toxin was produced by Staphylococcus aureus, strain Wood 46. The amount of exotoxin was monitored during growth and all subsequent purification steps by determination of its hemolytic activity against rabbit erythrocytes. The culture supernatant was treated with ammonium sulfate (75% saturation). The resulting precipitate was dialyzed and subjected to cation-exchange chromatography. The fractions containing the hemolytic activity were further purified by gel chromatography. The final product was enriched by a factor of 8.5 compared to the crude toxin. In sodium dodecyl sulfate-polyacrylamide gel electrophoresis the purified toxin exhibited one major band. It caused the release of 86Rb+ and ATP from rat insulinoma (RIN A2) as well as pheochromocytoma cells (PC12) in culture, indicating efficient permeabilization of their plasma membranes for small molecules.