Podcasts about y6

La Chine enregistre une hausse des voyages interrégionaux et du débit de marchandises en octobre ;Plus de 90 universitaires chinois et français ont échangé sur la modernisation;Une version multilingue du théâtre d'ombres "Bussière au mont Xishan" présentée à Beijing ;La Chine vise un taux de pénétration de la 5G supérieur à 85% d'ici 2027 ;Huawei lance des téléphones portables utilisant son propre système d'exploitation;Les autorités chinoises s'attaquent à l'exploitation algorithmique des livreurs;La fusée porteuse chinoise Lijian-1 Y6 achève le test d'assemblage final;La Chine prévoit de récupérer des échantillons de Mars vers 2031 ;Le box-office chinois atteint 40 milliards de yuans en 2024;La population mondiale de pandas géants évaluée au nombre de 757

Uit een nieuw rapport blijkt dat de wetenschappelijke geletterdheid van studenten in Australië in bijna twintig jaar nauwelijks is verbeterd. Uit gegevens van de Australian Curriculum Assessment and Reporting Authority blijkt dat in 2023 slechts 57% van de Y6 leerlingen de wetenschappelijke vaardigheidsnormen haalden. Bijna twintig jaar geleden, in 2006, lag dat percentage op 54%.

Year 6-7 Transition. Two experienced SENCOs - Katie Hall (secondary) and Emily Smith (primary) share their thoughts on what works well in two very different schools. Hear from Emily about primary schools inviting secondary SENCOs in to meet students in Y5 as well as Y6 and Katie explains how she has set up a bespoke provision to support Y7s with EHCPs.  From both, some great tips and advice around the power of relationships and  collaboration plus my usual wellbeing word for the weekend.

Welcome to Episode Four of Y6 of The Academy of Esports podcast! I'm your host, James O'Hagan. Thank you for joining me as I welcome a giant of the medical device field who is now using decades of experience— and his own personal journey— to build bold, collaborative mental health roadmaps within responsible, therapeutic video game environments in his new venture with Mike Wilson. It's an honor to speak with Ryan Douglas, co-founder & co-chairman of DeepWellDTx. Ryan takes us on a journey that goes far beyond the “one and done” attitude of many developers. His is a manifesto rooted in science, fed by sincerity and best practices, and allowed an ever-evolving timeline that strives not to be the first across a short-term finish line but rather the best at serving the long-term individual needs of human beings of all ages. It's quite a ride. Sit back, relax, and enjoy a conversation that will likely redefine the way you look at video games and mental health. Connect with Ryan Douglas: LinkedIn Connect with DeepWellDTx: LinkedIn Facebook Instagram YouTube Twitter Connect with James O'Hagan, Founder & Host of The Academy of Esports: LinkedIn Twitter Contact The Academy of Esports LinkedIn Twitter Facebook Episode References: Groundbreaking new study says time spent playing video games can be good for your well being— University of Oxford Gaming | Federal Trade Commission The Academy of Esports podcast is powered by LeagueSpot. Subscribe to Watch⁠ ⁠Subscribe to Listen⁠ --- Send in a voice message: https://podcasters.spotify.com/pod/show/taoesports/message Support this podcast: https://podcasters.spotify.com/pod/show/taoesports/support

048 - This is the Berkhamsted Spotlight, the podcast showcasing a remarkable community. And today we're speaking to some of the cast of the Prep Musical, Beauty and the Beast.Now, there was a cast of more than 100 pupils in this, so speaking to them all was never going to be an option, but we do have with us seven of the cast including Chip, Madame de la Grande Bouche, Belle, Cogsworth, Lumiere, Maurice and Gaston.We're going to hear about the rehearsals, the stressful parts of the preparation, what it feels like for our pupils to be on stage and how they cope when things don't quite go as planned.So let's dive into this episode with our Y6 pupils and also with the school's very own, Mrs Kate Turnley.Production Photoshttps://www.berkhamsted.com/prep-musical-beauty-and-the-beast/Berkhamsted onlineWebsite: www.berkhamsted.comFacebook: BerkhamstedSchoolTwitter: @berkhamstedschInstagram: berkhamstedschools 

Welcome to Episode Three of Y6 of The Academy of Esports podcast! I'm your host, James O'Hagan. Thank you for joining me as I welcome a friend, a mentor, and a global legend in esports and education— Mark “Garvey” Candella, former founder & director of Twitch Student and current Chief Business & Strategy Officer of AcadArena. Garvey joined me from his home base in the Philippines to share a journey that has taken him from a challenging start on the streets of Brooklyn to becoming one of the most well-known and respected figures in esports education and community-building.  Just one example of a chapter along the way: his work ethic as a young UPS driver led him to a spot at power lunches over the famous red sauce of Forlinis in Manhattan– delivering financial prowess through osmosis as a side order– and established him as a founding innovator of day trading.  Without question, this is and will remain one of my favorite podcast conversations of all-time. No summary could do justice to the wisdom shared, how much I learned about someone I thought I already knew, and how grateful I am to Garvey for his openness.  Sit back, relax, and listen to a story that will captivate you, educate you, and inspire you.  Connect with Garvey: LinkedIn: http://linkedin.com/in/mark-garvey-candella-a9946bb3 Twitter: https://twitter.com/garveynyc Connect with AcadArena: https://linktr.ee/acadarena CONQuest Festival: https://www.conquestph.com/ Connect with James O'Hagan, Founder & Host of The Academy of Esports: LinkedIn: https://www.linkedin.com/in/jamesohagan  Twitter: https://www.twitter.com/jimohagan  Contact: https://hihello.me/p/b1241eeb-bf95-476b-a258-6d80ef2a79fe  The Academy of Esports Site: https://taoesports.com LinkedIn: https://www.linkedin.com/company/taoesports Twitter: https://twitter.com/taoesports Facebook: https://www.facebook.com/profile.php?id=100063689103336 Subscribe to Watch: https://www.youtube.com/@TheAcademyofEsports Subscribe to Listen: https://podcasters.spotify.com/pod/show/taoesports Episode References: Welcome Back Kotter Opening: https://youtu.be/Mmm3KTa601s D&D Paladin: https://www.dndbeyond.com/classes/paladin 2018 TAO Esports Interview: https://youtu.be/1HFdlJM7-Z4 Keeping Up With the Dow Jones; From Suites to Streets, All Eyes Are on Stock Market (NYT Aug 1998):  https://www.nytimes.com/1998/08/23/nyregion/keeping-up-with-dow-jones-suites-streets-all-eyes-are-stock-market.html A Pocket Monitor for Stock Quotes (NYT Apr 1984): https://www.nytimes.com/1984/04/28/business/a-pocket-monitor-for-stock-quotes.html Investors Business Daily: https://get.investors.com/ibd/ibd-digital/ The Academy of Esports podcast is powered by LeagueSpot. https://leaguespot.gg/ --- Send in a voice message: https://podcasters.spotify.com/pod/show/taoesports/message Support this podcast: https://podcasters.spotify.com/pod/show/taoesports/support

Photometric Properties of Jupiter Trojans detected by the Dark Energy Survey by DES Collobration et al. on Tuesday 22 November The Jupiter Trojans are a large group of asteroids that are co-orbiting with Jupiter near its L4 and L5 Lagrange points. The study of Jupiter Trojans is crucial for testing different models of planet formation that are directly related to our understanding of solar system evolution. In this work, we select known Jupiter Trojans listed by the Minor Planet Center (MPC) from the full six years dataset (Y6) of the Dark Energy Survey (DES) to analyze their photometric properties. The DES data allow us to study Jupiter Trojans with a fainter magnitude limit than previous studies in a homogeneous survey with $griz$ band measurements. We extract a final catalog of 573 unique Jupiter Trojans. Our sample include 547 asteroids belonging to L5. This is one of the largest analyzed samples for this group. By comparing with the data reported by other surveys we found that the color distribution of L5 Trojans is similar to that of L4 Trojans. We find that L5 Trojans' $g - i$ and $g - r$ colors become less red with fainter absolute magnitudes, a trend also seen in L4 Trojans. Both the L4 and L5 clouds consistently show such a color-size correlation over an absolute magnitude range $11 < H < 18$. We also use DES colors to perform taxonomic classifications. C and P-type asteroids outnumber D-type asteroids in the L5 Trojans DES sample, which have diameters in the 5 - 20 km range. This is consistent with the color-size correlation. arXiv: http://arxiv.org/abs/http://arxiv.org/abs/2211.10719v1

Photometric Properties of Jupiter Trojans detected by the Dark Energy Survey by DES Collobration et al. on Tuesday 22 November The Jupiter Trojans are a large group of asteroids that are co-orbiting with Jupiter near its L4 and L5 Lagrange points. The study of Jupiter Trojans is crucial for testing different models of planet formation that are directly related to our understanding of solar system evolution. In this work, we select known Jupiter Trojans listed by the Minor Planet Center (MPC) from the full six years dataset (Y6) of the Dark Energy Survey (DES) to analyze their photometric properties. The DES data allow us to study Jupiter Trojans with a fainter magnitude limit than previous studies in a homogeneous survey with $griz$ band measurements. We extract a final catalog of 573 unique Jupiter Trojans. Our sample include 547 asteroids belonging to L5. This is one of the largest analyzed samples for this group. By comparing with the data reported by other surveys we found that the color distribution of L5 Trojans is similar to that of L4 Trojans. We find that L5 Trojans' $g - i$ and $g - r$ colors become less red with fainter absolute magnitudes, a trend also seen in L4 Trojans. Both the L4 and L5 clouds consistently show such a color-size correlation over an absolute magnitude range $11 < H < 18$. We also use DES colors to perform taxonomic classifications. C and P-type asteroids outnumber D-type asteroids in the L5 Trojans DES sample, which have diameters in the 5 - 20 km range. This is consistent with the color-size correlation. arXiv: http://arxiv.org/abs/http://arxiv.org/abs/2211.10719v1

Photometric Properties of Jupiter Trojans detected by the Dark Energy Survey by DES Collobration et al. on Monday 21 November The Jupiter Trojans are a large group of asteroids that are co-orbiting with Jupiter near its L4 and L5 Lagrange points. The study of Jupiter Trojans is crucial for testing different models of planet formation that are directly related to our understanding of solar system evolution. In this work, we select known Jupiter Trojans listed by the Minor Planet Center (MPC) from the full six years dataset (Y6) of the Dark Energy Survey (DES) to analyze their photometric properties. The DES data allow us to study Jupiter Trojans with a fainter magnitude limit than previous studies in a homogeneous survey with $griz$ band measurements. We extract a final catalog of 573 unique Jupiter Trojans. Our sample include 547 asteroids belonging to L5. This is one of the largest analyzed samples for this group. By comparing with the data reported by other surveys we found that the color distribution of L5 Trojans is similar to that of L4 Trojans. We find that L5 Trojans' $g - i$ and $g - r$ colors become less red with fainter absolute magnitudes, a trend also seen in L4 Trojans. Both the L4 and L5 clouds consistently show such a color-size correlation over an absolute magnitude range $11 < H < 18$. We also use DES colors to perform taxonomic classifications. C and P-type asteroids outnumber D-type asteroids in the L5 Trojans DES sample, which have diameters in the 5 - 20 km range. This is consistent with the color-size correlation. arXiv: http://arxiv.org/abs/http://arxiv.org/abs/2211.10719v1

Photometric Properties of Jupiter Trojans detected by the Dark Energy Survey by DES Collobration et al. on Monday 21 November The Jupiter Trojans are a large group of asteroids that are co-orbiting with Jupiter near its L4 and L5 Lagrange points. The study of Jupiter Trojans is crucial for testing different models of planet formation that are directly related to our understanding of solar system evolution. In this work, we select known Jupiter Trojans listed by the Minor Planet Center (MPC) from the full six years dataset (Y6) of the Dark Energy Survey (DES) to analyze their photometric properties. The DES data allow us to study Jupiter Trojans with a fainter magnitude limit than previous studies in a homogeneous survey with $griz$ band measurements. We extract a final catalog of 573 unique Jupiter Trojans. Our sample include 547 asteroids belonging to L5. This is one of the largest analyzed samples for this group. By comparing with the data reported by other surveys we found that the color distribution of L5 Trojans is similar to that of L4 Trojans. We find that L5 Trojans' $g - i$ and $g - r$ colors become less red with fainter absolute magnitudes, a trend also seen in L4 Trojans. Both the L4 and L5 clouds consistently show such a color-size correlation over an absolute magnitude range $11 < H < 18$. We also use DES colors to perform taxonomic classifications. C and P-type asteroids outnumber D-type asteroids in the L5 Trojans DES sample, which have diameters in the 5 - 20 km range. This is consistent with the color-size correlation. arXiv: http://arxiv.org/abs/http://arxiv.org/abs/2211.10719v1

Iona Jackson is Head of Research at Edurio, managing a team of survey experts and data analysts through projects relating to stakeholder feedback in schools. New Edurio research reveals half of children feel stressed and a quarter feel lonely. Edurio has published their latest research examining pupil wellbeing, support systems in school and how pupils feel about school. The study drew on responses from 45,000 children of which 15,000 were from primary. Children feel progressively less well as the move through primary school – 76% in year 1 feel well but this drops by 17 percentage points in Y6 when 59% report feeling well. Children feel more stressed in Y6 (36%) than in Y1 (22%) More primary aged children feel overworked in Y2 and Y3 than at any other time during primary school. The research shows that the transition to secondary school has a negative impact on children's wellbeing and the drop is greater than at other times during school. Children's overall wellbeing drops from 59% feeling well in Y6 to 46% in Y7. More students often feel stressed – rising from 36% in Y6 to 43% in Y7 More children report not sleeping well in Y7 (30%) than in Y6 (28%)   A survey of 45,000 school-aged children conducted by Edurio reveals that less than half (47 per cent) of pupils reported that they had been feeling well in the period leading up to the survey. A similar number (46 per cent) often felt stressed. Added to this, a quarter (24 per cent) of pupils admitted feeling lonely. This is much higher than a previous study by the Office for National Statistics in 2018 which found 11 per cent of 10-15 year olds felt lonely. It suggests that the pandemic has had a significant impact on pupils. The research, which was conducted during the summer term, found that pupils in the latter stages of their schooling fared worst for overall wellness, sleep and overwork. A third (31 per cent) of pupils in their final year of GCSEs reported feeling well; four in 10 (41 per cent) slept badly; two-thirds (66 per cent) felt overworked and over half (63 per cent) of year 11 pupils felt stressed.  The research found significant differences in wellbeing between girls and boys, with girls reporting lower wellbeing scores. More than half (55 per cent) of girls feel quite or very often stressed compared to just a third (36 per cent) of boys. Almost half (48 per cent) of girls feel frequently overworked compared to just over a third (38 per cent) of boys. Furthermore, less girls (43 per cent) felt well than boys (54 per cent) and less girls (40 per cent) slept well than boys (47 per cent). Interestingly, the research points to a correlation between pupil wellbeing and a school's Ofsted rating. Pupils at schools judged ‘outstanding' have consistently higher stress levels, problems sleeping and feel overworked: Almost half of pupils (49 per cent) have felt stressed lately compared to 44 per cent at ‘good' schools and 45 per cent at ‘requires improvement (RI)' schools. Under half (41 per cent) reported having good sleep compared to 44 per cent at ‘good' schools and 45 per cent at RI schools. Just under half (46 per cent) feel overworked compared to 42 per cent at 'good' schools and 44 per cent at RI schools. Edurio found that 71 per cent of students feel safe in class. However, eight per cent reported feeling unsafe in class which means, in an average-sized class, two to three pupils feel unsafe. Furthermore, when there is an issue, less than half (41 per cent) feel they have an adult at school whom they can trust and talk to. Similarly, just over a third (37 per cent) feel that they would rarely or never have an adult at school that they can trust and talk to. So, who do pupils turn to when they feel sad or worried? Almost half (48 per cent) choose to speak to their parents, 41 per cent to their classmates and just 29 per cent choose to speak to teachers. Most concerning is that less than a quarter (15 per cent) do...

New Edurio research reveals half of children feel stressed and a quarter feel lonely. Edurio has published their latest research examining pupil wellbeing, support systems in school and how pupils feel about school. The study drew on responses from 45,000 children of which 15,000 were from primary. Children feel progressively less well as the move through primary school – 76% in year 1 feel well but this drops by 17 percentage points in Y6 when 59% report feeling well. Children feel more stressed in Y6 (36%) than in Y1 (22%) More primary aged children feel overworked in Y2 and Y3 than at any other time during primary school. The research shows that the transition to secondary school has a negative impact on children's wellbeing and the drop is greater than at other times during school. Children's overall wellbeing drops from 59% feeling well in Y6 to 46% in Y7. More students often feel stressed - rising from 36% in Y6 to 43% in Y7 More children report not sleeping well in Y7 (30%) than in Y6 (28%) Mark Taylor chats to Iona Jackson co-author of the Edurio report about her findings. Full details can be found at: https://home.edurio.com/pupil-learning-experience-and-wellbeing-report (https://home.edurio.com/pupil-learning-experience-and-wellbeing-report) To keep up to date with all the work by National Association for Primary Education please visit: https://nape.org.uk/ (https://nape.org.uk/)

Louise Howlett has not been teaching long, but she is already an Assistant Headteacher in an outstanding school. What has made her so good, so fast? Probably it's her forensic attention to detail, because Louise had always wanted to be a forensic scientist before catching the teaching bug when helping an excluded boy see that school could really change his life. And that is what has motivated Louise to be the very best teacher she can be for every child. She knows if she does her job really well it changes children's lives, and, believe me, she does it really well. In this podcast we talk about ... The similarities between forensic science and teaching. How helping an excluded, vulnerable boy change his life was the moment Louise realised she must become a teacher. Why children remember the people who have made a real difference to their lives. The fact that it feels like your training hasn't prepared you for day 1 minute 1 with your first class. How a dragon fly helped to break the ice with her first class. Telling children they would be fine performing their Y6 play on a very hot day as it was going to be really fanny in the hall. How relationships must be built consciously and unconsciously through every action that you do as a teacher. Being sick in the classroom bin before her first parents evening as she was concerned she wouldn't know what to say. The value of making a folder full of the many positive comments made by children and parents over the years and looking at it after a tough day to remember all the good things that have happened. Being a perfectionist is ok but it is not attainable every moment of every day, so accept this and aim to be the best you can be. Accepting you can't know everything and know when to stop working. Becoming very skilled at prioritising the things that really make a difference to children's learning.  

#每日一經濟學人 #Linda老師商用英文班 #職場英文 #英文學習 #線上課程

This week we park the Highwind and hop aboard the Gummi Ship to take a look at the Final Fantasy representation within Kingdom Hearts. Join us as we travel from world to world discussing what is and what could have been.Trivia question of the day: Name an American sitcom from the early 2000s that makes reference to Final Fantasy. (Answer at the bottom of the episode summary). Episode SummaryIntroduction / TriviaOur week in Final FantasyFinal Fantasy in Kingdom Hearts discussionTrivia answer: Two and a Half Men Recommended ResourcesTwo and a Half Men clip: https://www.youtube.com/watch?v=Y6-d-YcYJWAFollow us on Twitter! https://twitter.com/HighwindHeraldJoin our Discord! https://discord.gg/CK8mCMJ

Dear DCIS Parents, With Christmas just around the corner, and at the end of another busy term, it seems the perfect time to reflect on what has been happening in the Primary School. We returned from our restful half term break straight into our busy UN Day preparations which saw the children busily researching the country assigned to their class, as well as making and collecting artefacts for their displays. The culmination of the week was UN Day itself with our annual parade and spectacular country displays down on the field. A big thank you to the teachers who supported to the children with their work, the parents who also helped to set up the displays and of course the Mrs Gray and Mrs Hyland who once again came up with the grand plan for the day! We have had several other celebrations this term. Our Deepavali cultural dress-up day saw 2RB and 2SL, and 3CD and 3AS present their assemblies to staff, children and parents, sharing some interesting facts and stories with us all. Thank you to the students and staff for their collaboration and hard work on this. The DCA treated us to a spooky affair for Halloween, the school looked truly terrifying thanks to the creative decorations and gruesome costumes around the place. Thank you to the DCA for frightening us even more than last year! This year, we raised awareness of Remembrance Day across the school too. You may have noticed the sea of poppies in the corridors; our first ever red and black day was a huge success and we were proud to hand over a financial contribution to the British Legion. Kindness Week, which linked with House Day, got us all thinking of ways to look after each other and was a thoroughly thought-provoking week for us all. Thank you also to Miss Sanderson and Miss Lindars for planning World Children’s Day for us. It was fantastic to see the children take a lead with their learning as they became the teachers and shared skills with their friends in other classes. Our trips continued this term with Year 1 taking part in drama workshops at Centre Stage and Year 2 visiting the National Gallery. There was great excitement as Year 5 headed off to Telunas for their annual residential, with Mr Odey and the Year 5 team. We were delighted to also send a group of children off to Ho Chi Min City for the South-East Asia NAE games led by Mr Carswell and Miss Hughes. As well as this, Mr Baines and Mr Northridge chaperoned a party of Y6 and Y7 students to the NAE STEAM festival in Bangkok. I know the children loved it – they were praised for their innovation, creativity and their impeccable behavior. Trips such as these take a great deal of preparation and planning – a special mention goes to Mr Odey for his work on the Telunas trip and to Mr Carswell for planning the NAE games and to Mr Baines for all the prep with the STEAM trip. A great number of staff have given up their time to support the children – a great big thank you to each and every one of them for their dedication to the school. We were delighted to invite our Year 6 parents into school for an Open Morning in which they found out about the transition to Secondary School. Our Year 6 children thoroughly enjoyed a day in Secondary, meeting the teachers and having a taste of secondary lessons and life as a secondary student. Christmas crept up on us and once again the DCA sprang into action with reindeer, elves and some very familiar faced gingerbread people popping up around the campus! The lower primary nativities were amazing and well-enjoyed by children, parents and some lucky grandparents too. Both nativities were absolute bonanzas and certainly got us all into the Christmas spirit! Wishing happy holidays to you all and for those who celebrate Christmas, I hope you enjoy the festivities.

Broadcasted on Friday 21 June on Vintage Radio as part of the Radio Radio morning show with Cliff Howarth. Y5 and Y6 students from Green Park School in Maghull feature on some radio jingles.

Reinvesting dividends! It sounds boring.   Just wait ‘till you see the compounding cash flow numbers on this strategy, you'll understand why it's the only reason Ryan cares about dividend paying stocks at all.   Are you looking for an investment workhorse that will build a nest egg for your future? Pick up a good habit and put 10% of your earnings into this long term strategy.   Key Takeaways [2:55] Ryan starts off by sharing his personal minimum criteria: a stock that pays a minimum of 4% and has raised its dividend every year for at least 10 years.   [5:35] As an example we'll use AT&T whose stock trades at about 40$ a share.   Let's buy a hypothetical 300$ worth at a 6% dividend a year for a measly 18$ a year.   [7:08] AT&T have raised their dividend for at least 10 years: let's posit a 10% annual raise. Y1 — 6% Y2 — 6.6% Y3 — 7.3% Y4 — 8% Y5 — 8.8% Y6 — 9.8% Over 10 years these numbers become really interesting.   [9:03] The next step is the dividend reinvestment plan: Y1 — 10 shares at 6% reinvested Y2 — 10.5 shares at 6.6% reinvested Y3 — 11 shares at 7.3% reinvested Y4 — 12 shares at 8% reinvested Y5 — You see where Ryan is going with this...   [11:35] There is a double compounding effect over time which means that at some point when you stop reinvesting, you still have cash flow, and you still own the underlying stock which means you can sell it, borrow against it, etc.   You liked this content? Comment, subscribe and share!

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Today was a very special first for Westwood Radio as we broadcast our first live outside broadcast from Wiltshire Heights Residential Care Home in Bradford on Avon. Our team of Y6 broadcasters spent the show entertaining and even interviewing the residents to find out about their work and living locally - all part of our Oral History Project. It was a wonderful afternoon and a real privilege to meet such wonderful people. Our children were so respectful and such a credit to our school.

Hello from a rather noisy back garden in the North East of England! In this episode, I get over excited about all thinks Olympic. Forgive me. Finished Objects In reverse order, I finsihed the weaving and made the cusion from the fabric my class were weaving. Rav details here. I also assembled the green woven cushion that's been woven for over two months. Rav details here. And here are front (above)n and back(below) pictures of the cushion worked on during the Y6 residential that I mentioned last episode. Rav details here. Something I really like My brand new Olympic venue Top Trumps set! Links Woolsack Music: Rondopolska by Barry Philips, from the album Tråd, available from Magnatune. We have a listeners' map. Please go on over to pop in a pin - we're covering 4 continents now but I'd still love to see where you all are! Feel free to leave a comment here or at http://www.yarnsfromtheplain.blogspot.com/, or email me at yarnsfromtheplain AT googlemail DOT com. We have a Ravelry group here, so come on over to chat. You can find me on Ravelry as talesfromtheplain and on Twitter as talesfromplain (although Tweeting can be sporadic!). TTFN, Nic

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