Podcasts about G0

Listen to the latest oncology-focused research published in this week's issue of Oncotarget, Volume 12, Issue 12. The full issue of Oncotarget: https://www.oncotarget.com/archive/v12/i12/ COVER PAPER: “Frame-shift mediated reduction of gain-of-function p53 R273H and deletion of the R273H C-terminus in breast cancer cells result in replication-stress sensitivity” https://doi.org/10.18632/oncotarget.27975 NEWS: “LY6 gene family presents a novel class of potential biomarkers associated with overall survival outcome of pancreatic ductal adenocarcinoma” https://doi.org/10.18632/oncotarget.27968 (PDF Download) EDITORIAL: “Neoantigen evolution in head and neck cancer progression: Where do we go from here?” https://doi.org/10.18632/oncotarget.27942 (PDF Download) EDITORIAL: “Unexpected zinc dependency of ferroptosis – what is in a name?” https://doi.org/10.18632/oncotarget.27951 (PDF Download) RESEARCH PAPER: “Role for Fgr and Numb in retinoic acid-induced differentiation and G0 arrest of non-APL AML cells” https://doi.org/10.18632/oncotarget.27969 RESEARCH PAPER: “Dynamic cellular biomechanics in responses to chemotherapeutic drug in hypoxia probed by atomic force spectroscopy” https://doi.org/10.18632/oncotarget.27974 RESEARCH PAPER: “Genomic clustering analysis identifies molecular subtypes of thymic epithelial tumors independent of World Health Organization histologic type” https://doi.org/10.18632/oncotarget.27978 RESEARCH PAPER: “Deep learning with deep convolutional neural network using FDG-PET/CT for malignant pleural mesothelioma diagnosis” https://doi.org/10.18632/oncotarget.27979 Keywords - mutant p53, DNA replication, head and neck cancer, biomarker, ferroptosis, retinoic acid(RA), hypoxia, thymic epithelial tumor, mesothelioma, AML, breast cancer, pancreatic cancer, cancer, science, research, oncology About Oncotarget Oncotarget is a bi-weekly, peer-reviewed, open access biomedical journal covering research on all aspects of oncology. To learn more about Oncotarget, please visit https://www.oncotarget.com/ or connect with: SoundCloud - https://soundcloud.com/oncotarget Facebook - https://www.facebook.com/Oncotarget/ Twitter - https://twitter.com/oncotarget LinkedIn - https://www.linkedin.com/company/oncotarget Instagram - https://www.instagram.com/oncotargetjrnl/ YouTube - https://www.youtube.com/oncotargetyoutube Pinterest - https://www.pinterest.com/oncotarget/ Reddit - https://www.reddit.com/user/Oncotarget Oncotarget is published by Impact Journals, LLC. Please visit https://www.impactjournals.com/ or connect with @ImpactJrnls Media Contact MEDIA@IMPACTJOURNALS.COM 18009220957

染色体分配時におけるKi-67の機能性に関する論文を紹介しました。Show notes 周期ゼミ Periodical cicadas … 全米における周期ゼミの分布について 論文の縦読み … 同じオーサーの論文を時間軸で読む、論文の縦読みについてはresearchat.fmのep57にて説明しました。 Cuylen-Haering et al., Nature (2020) … “Chromosome clustering by Ki-67 excludes cytoplasm during nuclear assembly” Gerlich et al. Cell (2003) … “Global Chromosome Positions Are Transmitted through Mitosis in Mammalian Cells” Cuylen et al., Nature (2016) … “Ki-67 acts as a biological surfactant to disperse mitotic chromosomes” Samwer et al., Cell (2017) … “DNA Cross-Bridging Shapes a Single Nucleus from a Set of Mitotic Chromosomes” Daniel Gerlich 植物の細胞で染色体が分配される様子 … 1956に撮影されたAfrican Blood Lilyの細胞における染色体分配と細胞分裂の様子。植物には中心体が存在しないので少し動物細胞のものとは趣が異なる。64年前に撮影された動画だが未だ色褪せず様々な洞察を与えてくれる。無染色・ライブセルで行われた驚異的なイメージング。 上記動画に関するツイート … 海外勢もかなりコメントをくれているのがわかる。 体細胞分裂 … prophase->prometaphase->metaphase->anaphase->telophase->cytokinesis。英語だとmitosis DNA複製 遺伝情報 減数分裂 … 英語だとmeiosis HeLa細胞 染色体 核(細胞核) ミトコンドリア … パラサイト・イヴでご存知の方も多いだろう。 細胞小器官 核小体 … 仁とも呼ばれていた。 リボソーム リボソームRNA 倍数性 姉妹染色体/姉妹染色分体 コヒーシン … 姉妹染色体を接着させるために重要なタンパク質 Locke. JCS (1990) … “Is there somatic inheritance of intracellular patterns?” transvection mitophagy ヒストン ヌクレオソーム クロマチン … ヒストン、ヌクレオソーム、クロマチン、染色体についてはどこかでじっくり話します。 CFP, YFP … 蛍光タンパク質 細胞周期 … G1->S->G2->M. G1->S->G2が間期、Mが分裂期 FRAP(Fluorescence recovery after photobleaching) FISH Kimura and Cook. J Cell Biol (2001) … “Kinetics of core histones in living human cells: little exchange of H3 and H4 and some rapid exchange of H2B” 染色体テリトリー Cremer and Cremer. Cold Spring Harb Perspect Biol. (2010) … “Chromosome Territories”: ドンピシャタイトルのThomas CremerによるChromosome territoryのレビュー。 Carl Rabl Theodor Boveri Sutton-Boveriの染色体説 キネトコア セントロメア スピンドル … 紡錘体 微小管 … microtuble RNAi ki-67 G0期 核膜 核膜孔 open mitosis/closed mitosis Nuclear envelope break down … 核膜の崩壊 ノコダゾール コルヒチン FCS 両親媒性 Crasta et al., Nature (2012) … “DNA breaks and chromosome pulverization from errors in mitosis” micro nucleiに関する最近の代表的な研究 スプライシング コンデンシン Delarue et al. Cell (2018) … “mTORC1 Controls Phase Separation and the Biophysical Properties of the Cytoplasm by Tuning Crowding”: GEMs(genetically encoded multimeric nanoparticles)を開発した論文。これは激アツである！ Akita et al., JCS (2007) … “The crystal structure of a virus-like particle from the hyperthermophilic archaeon Pyrococcus furiosus provides insight into the evolution of viruses”: Delarue et al. (2018)で使われたencapsulinに関する論文。これも激アツである！ 養王田先生の科研費報告書 … “Encapsulinとは単一のシェルタンパク質で構成される内部が空洞のナノ構造体であり、一部の微生物細胞内に形成されている。” 核移行シグナル Görisch et al., JCS (2005) … “Histone acetylation increases chromatin accessibility”: クロマチンへのaccessibilityをイメージングで示そうとした論文 Dextran Flavopiridol Trichostatin A (TSA) Angelika Amon Breakthrough Prize in Life Sciences 染色体の異数性 … aneuploidy 出芽酵母 Ünal et al., Science (2011) … “Gametogenesis Eliminates Age-Induced Cellular Damage and Resets Life Span in Yeast”: 老化した出芽酵母は減数分裂に進みにくいことを示した定量解析。Amon labからの研究。 King et al., eLife (2019) … “Meiotic cellular rejuvenation is coupled to nuclear remodeling in budding yeast”: 上記のFirst authorであるÜnal labから出た論文。年老いた出芽酵母が減数分裂に進んだ場合の”Nuclear rejuvenation”(核若返り)に関するイメージング研究 エピジェネティクス(epigenetics) Conrad Hal Waddington … epigeneticsの提唱者。 Editorial notes 素人にもロジックが追いやすかった気がする (soh) 綿々と続くドイツの細胞生物学/染色体学研究の系譜はまたどこかで話せたらと思います。(tadasu) figureがモンスターボールみたいですね (coela)

Date: May 31, 2020Series: Joy No Matter, Part 10Scripture: Phil 2:19-30Title: “The Example"Today's Bottom line:Follow the Example, Be the Example as You Worship the Example, no matter what.IntroYour Trial Will Become Your Triumph by Nicky Gumbel ‘Houston, we've had a problem,' were the words of Jim Lovell on the evening of 13 April 1970. Nearly fifty-six hours into the mission to the moon, an explosion aboard the spacecraft plunged the crew into a fight for their survival. Within less than a minute there was a cascade of systems failures throughout the spacecraft. ‘It was all at one time – a monstrous failure,' said NASA's flight controller.The spacecraft looped around the moon, using its gravity to return to earth. Millions of people followed the drama on television. Eventually, the capsule splashed down in the Pacific Ocean near Tonga.In an article headed ‘Apollo 13: From Disaster to Triumph' the BBC science reporter wrote, ‘Although the mission was not a success from a conventional perspective, it was a triumph of ingenuity and determination.'Jim Lovell said it showed the people of the world that even if there was a great catastrophe, it could be turned into a success.The supreme example of triumph coming out of apparent catastrophe is the cross. What seemed to the world to be the ultimate defeat was in fact the ultimate triumph.As we transition towards what's next, knowing that God can leverage evil for good, why would we care about anyone else? Shouldn't we just go into survival mode and get things back in order at all costs and forget everyone else?Paul shows us in this letter that that's not the path Jesus would take. He takes "the road less traveled” - the way of “others first.”The OfficeKurt Cooper was a co-worker at my first job. He'd been out of school and in the workplace about 4 years and was doing a great job. He was a great example to me because he showed me how to live out my faith in the workplace. I didn't have many examples. And I wasn't looking for any either.He invited me to lunch that first week.Later he would invite us to church where we eventually would serve as volunteer youth leaders.He and his wife would model for us our first small group experiences (besides Sunday school) and well as parenting.They would invite us to a life-changing retreat weekend called the Emmaus Walk.All of these hugely shaped my faith-walk in my early years of marriage. I'd never had someone pour themselves into me like that. And I really didn't even realize it was happening until I looked back later.Today I'm going to call you to follow The Example, be an example as you worship The Example as Lord and Savior. Paul says it this way, “Follow me as I follow Christ.”Follow the Example Paul describes.Be the Example Jesus called you to be.(As you) Worship the Example who empowers us to follow and be the example.Series bottom line:God pours out his grace and peace in and through us as we show and tell the good news no matter what.One way we do this seen in the examples of Timothy and Epaphroditus in Phil 2:19-30.The Example leads to ExamplesTruth 1: Jesus will show and tell his example to the world through his examples by their example.Truth 2: Jesus' true followers, by definition, follow his words, ways and works. To not follow Jesus' example is to not be a follower of Jesus.OutlineI. Follow the Example Paul has described already. (2:5-8) REVIEWII. Be the example Jesus called you to be. (2:19-30) 2 Examples:A. Timothy - An example of 2:19-24Lives sent (19, 23) or on mission 24/7 - Jesus sends us out tooNo one else like him; Paul sees him as special family like a faithful son in the Lord (20) - Jesus sees us as family; brothers and sistersGenuinely cares about others; compassionate and merciful (20) - Jesus calls us to “love our neighbor as ourself.”“Others first” (21) mentality c.f. 2:3 and Jesus' example”Proved himself” faithful son in the work “as a son with his father” (22) “served with me in the work of the gospel” - Jesus calls us to this great work 24/7 too. “Seek first his kingdom and his righteousness and all these things will be added to you” (Matt 6:33)B. Epaphroditus - Another example of 2:25-30Also lives sent (25, 28)Also lives as family (“brother”) (25)Paul calls him a “co-worker” or partner c.f. (1:5)- Jesus calls us to work from rest (semi-circle; abiding; fruitfulness)Paul calls him a “fellow soldier” (25) because he went to battle with Paul- Jesus calls us to go to war (sheep among wolves) fighting on our knees; prayer + armor of God (Eph 6:10-18)Cares for his flock back home even as he cares for Paul on their behalf;“Love is showing compassion for other people's reality.” - Neal McGlohon. In other words, love does for others in need. Think Good Samaritan parable.III. (As you) Worship the Example with your lips and your lives. (2:9-11; Heb 13:15-16)A. Jesus is exalted as worthy of our allegiance and worship. (2:9-11)B. We worship Jesus with (Heb 13:15-16)1. Our lips2. Our livesConclusionThe OfficeKurt Cooper was a co-worker at my first job. He'd been out of school and in the workplace about 4 years and was doing a great job. He was a great example to me because he showed me how to live out my faith in the workplace. I didn't have many examples. And I wasn't looking for any either.He invited me to lunch that first week.Later he would invite us to church where we eventually would serve as volunteer youth leaders.He and his wife would model for us our first small group experiences (besides Sunday school) and well as parenting.They would invite us to a life-changing retreat weekend called the Emmaus Walk.All of these hugely shaped my faith-walk in my early years of marriage. I'd never had someone pour themselves into me like that. And I really didn't even realize it was happening until I looked back later.The best example I've had has been my wife. She's changed my life. Since high school I've known that I wanted live life like her.She's been an example to me, our four girls and so many others. She has done that by following the Example, being an example as she worships the Example, no matter what.What about you?Notes{Keep in mind to share the application or calling for the day throughout the message. Share it in the intro, explain it in the message body, and call for it in the conclusion. Really, call for it throughout the whole message.}IllustrationsChris K - That we all learn something from the changes we've recently experienced; it makes him a little sad that maybe nothing has changed as some churches prepare for the “big show”; like coming home from camp on fire and then nothing changes.“We are the sum of our actions, therefore habits make a difference.” - Aristotle$3 worth of gospel (DA Carson's book)To estimate how easily a virus spreads, scientists calculate its "basic reproduction number," or R0 (pronounced R-nought). R0 predicts the number of people who can catch a given bug from a single infected person, Live Science previously reported. Currently, the R0 for SARS-CoV-2, the virus that causes the disease COVID-19, is estimated at about 2.2, meaning a single infected person will infect about 2.2 others, on average. By comparison, the flu has an R0 of 1.3.Not R0 but G0!What if we calculated our personal, gospel R0. We could call it our G0. (G nought) Will we “infect” at least 1 person in our life? 2?Why don't we? For the same reasons we don't minimize spreading the virus we have now:We don't want to inconvenience ourselves (appetites, comfort idol)We don't love people enough to sacrifice for them.We are afraid. (See Phil 1:12-14 for enCOURAGEment to live and share the gospel boldly)Living like you “have it"Good news right now might be that there is a cure and vaccine for COVID-19! And that we could begin the end of the lockdown very soon.Isn't that what we sometimes do when we withhold the good news the Bible gives us?Paul presents Christ to believers as: (by HL Willmington, Outline Bible)Our life-purpose.Our life-pattern.Our life-prize.Our life-power.References/Bibliography:“The Bible Knowledge Commentary” by Walvoord, Zuck (BKC)“the Bible Exposition Commentary” by Warren Wiersbe (BEC)“Exalting Jesus in The Book of Acts” by Tony Merida (CCE)“Exalting Jesus in The Book of Philippians” by Tony Merida & Chan (CCE)Let's Study Philippians by Sinclair FergusonOutline Bible, D WillmingtonDA Carson Basics for Believers commentaryFF Bruce NIBC Commentary on PhilippiansTo Live is Christ To Die is Gain by Matt ChandlerNIV Study Bible (NIVSB)ESV Study BibleESV Gospel Transformation Bible (GTB)J Vernon McGee Commentary

#방금뉴스 이 시각 가장 핫!하고 중요한 뉴스 - 박찬형 / KBS 보도본부 기자 #각설하G0! ① 180석 ‘슈퍼여당’ 탄생, 의미는? ② 민주 호남, 통합 TK ‘싹쓸이’…왜? - 현근택 / 더불어민주당 선대위 대변인 - 조대원 / 미래통합당 전 고양시 당협위원장 #4.15 총선 당선인 연결 21대 국회 입성…당선 소감과 각오 - 김 웅 / 미래통합당 서울 송파갑 당선인 - 강선우 / 더불어민주당 서울 강서갑 당선인 - 정진석 / 미래통합당 공주·부여·청양 당선인 - 이용호 / 무소속 전북 남원·임실·순창 당선인

1957 - Eerste werkdag. Maarten krijgt opdracht een commentaar bij de kaart van de kabouters te maken. Als hij thuiskomt zit zijn schoonmoeder in de kamer. Tweede werkdag. Beerta vertrekt naar een vergadering in Arnhem. Telefoongesprek met 't Mannetje over een vergadering van de Boerenwagenclub. Juffrouw Haan valt uit omdat Maarten de telefoon heeft aangenomen. Nijhuis geeft Maarten fiches en een kaartenbak. Van Ieperen probeert gemene zaak met Maarten te maken. Moedeloos. Maarten vraagt Beerta wat zijn positie is. Beerta over juffrouw Haan. Met: Yvonne van den Hurk (Nicolien), Krijn ter Braak (Maarten Koning), Joop Keesmaat (Beerta), Hanneke Riemer (Dé Haan), Arthur Boni (Teun Nijhuis), Jack Vecht (Geert Meierink), Gijs de Lange (Koert Wiegel), Titus Muizelaar (Veerman), Huub van der Lubbe (Cor de Bruin), Kitty Courbois (moeder van Nicolien), en Adriaan Olree ('t Mannetje).

#방금뉴스 이 시각 가장 핫!하고 중요한 뉴스 - 박찬형 / KBS 보도본부 기자 #각설하G0! ① 이해찬 “열린민주, 文 정부 참칭 말라” ② 공천 4곳 긴급 취소…두 번 살아난 민경욱 ③ 4.15 총선 격전지 판세 분석 - 최민희 / 전 더불어민주당 의원 - 김용남 / 전 자유한국당 의원 #시사법정 ① '신림동 CCTV 사건' 또 강간미수 무죄 ② 한화손보, 초등생에 구상금 청구 논란 ③ '민식이법' 시행…무엇이 달라지나 - 신유진 / 변호사

#각설하G0! ① 문 대통령, 오늘 첫 '비상경제회의' 주재 ② 윤호중 “성소수자 문제, 소모적 논쟁” 발언 논란 ③ 공병호 “윤주경 비례대표 3번 배치…조수진 1번 유지” - 최민희 / 전 더불어민주당 의원 - 김용남 / 전 자유한국당 의원 #시사법정 ① 법원, 바른미래당 '셀프제명' 무효 ② 수임제한 ‘1년→3년’…전관예우 근절될까 - 신유진 / 변호사

#방금뉴스 지금 이 시각 가장 핫!하고 중요한 뉴스 - 박찬형 / KBS 보도본부 기자 #각설하G0! ① 민주당 오늘 비례연합정당 참여 투표 ② 안철수, 미래한국당 통합 제의 거절 ③ 미래통합당 공천 진통…화약고 된 대구 수성갑 - 최민희 / 전 더불어민주당 의원 - 김용남 / 전 자유한국당 의원 #시사법정 ① 코로나가 앞당긴 '스마트법원' 분위기는? ② ‘사법농단’ 임종헌 재판, 9개월만에 재개 ③ 코로나19에 여행·예식 등 위약금 분쟁 급증 - 신유진 / 변호사

#방금뉴스 지금 이 시각 가장 핫!하고 중요한 뉴스 - 박찬형 / KBS 보도본부 기자 #각설하G0! ① 마스크 수급 논란 · 신천지 강제조사, 어떻게 보나? ② ‘범진보 연합정당’ 창당 논란…각 당 입장은? ③ 박근혜, 왜 갑자기 옥중편지 썼나? - 임병식 / 전 국회대변인 - 김용남 / 전 자유한국당 의원 #시사법정 ① '타다 금지법' 국회 법사위 통과 ② 대학 등록금 '부분 환불' 가능할까? ③ 서울시, 신천지 법인허가 취소 돌입 - 신유진 / 변호사

#방금뉴스 지금 이 시각 가장 핫!하고 중요한 뉴스 - 박찬형 / KBS 보도본부 기자 #각설하G0! ① ‘코로나 3법’ 통과…중국 입국금지 놓고는 설전 ② ‘비례민주당’ 창당 논란…통합당은 ‘컷오프’ - 최민희 / 전 더불어민주당 의원 - 김용남 / 전 자유한국당 의원 #시사법정 ① '징역 17년 선고' MB…6일 만에 재석방 ② 이재명, 신천지 긴급 강제 조사 문제없나? - 신유진 / 변호사

#방금뉴스 지금 이 시각 가장 핫!하고 중요한 뉴스 - 박찬형 / KBS 보도본부 기자 #각설하G0! ① 확진자 31명 추가…방역 대응 어떻게 보나? ② 김남국 결국 공천 신청…경선 전망은? ③ 이언주 전략공천 논란…김무성 반발 - 최민희 / 전 더불어민주당 의원 - 김용남 / 전 자유한국당 의원 #시사법정 ① MB, 2심서 징역 17년…구치소 재수감 ② '타다' 불법 아니다…1심 무죄 선고 - 신유진 / 변호사

#방금뉴스 지금 이 시각 가장 핫!하고 중요한 뉴스 - 박찬형 / KBS 보도본부 기자 #각설하G0! ① 남대문 간 文…2월 임시국회 17일부터 ② 선관위 “비례대표 전략공천은 불법”…각당 전략은? ③ 김두관·홍준표 ‘낙동강 대전’ 예고 - 최민희 / 전 더불어민주당 의원 - 김용남 / 전 자유한국당 의원 #시사법정 ① 최서원, '국정농단 파기환송심' 내일 선고 ② ‘아내 살해’ 치매노인에 ‘치료적 사법’ 첫 적용 - 신유진 / 변호사

#방금뉴스 지금 이 시각 가장 핫!하고 중요한 뉴스 - 박찬형 / KBS 보도본부 기자 #각설하G0! ① 법무부 공소장 비공개 결정, 어떻게 보나? ② 미래한국당 정식 출범…총선 예상 의석수는? ③ 황교안 결국 종로 피하나? - 최민희 / 전 더불어민주당 의원 - 김용남 / 전 자유한국당 의원 #시사법정 ① 대법원, ‘국정농단’ 차은택·장시호 파기환송 ② 병무청, '성매매알선' 승리에 입영 통지 - 신유진 / 변호사

#각설하G0! ① 신종 코로나, “가짜뉴스 부적절” VS “중국인 입국금지” ② 민주당 원종건 후폭풍…김의겸 · 정봉주는? ③ 바른미래 탈당한 안철수, 신당 창당하나? - 김남국 / 전 더불어민주당 법률위 부위원장 - 김용남 / 전 자유한국당 의원 #시사법정 ① 김기춘 오늘 대법 선고…직권남용 기준 나오나? ② 법원 "이상호, 故김광석 아내에 1억 배상" - 양지열 / 변호사

#각설하G0! ① 검찰 중간간부 인사…중앙지검 차장검사 전원 교체 ② 이낙연, 종로 출마·공동 선대위원장 확정 ③ 황교안 영수회담 제안…명절 앞둔 정치권 풍경 - 최민희 / 전 더불어민주당 의원 - 김용남 / 전 자유한국당 의원 #시사법정 ① 법원 “김경수 킹크랩 시연 봤다” 잠정 판단 ② '가세연'의 마구잡이식 폭로, 문제없나? - 신유진 / 변호사

#각설하G0! ① 이낙연 민주당 복귀…황교안의 선택은? ② 1호 공약은 “무료 와이파이” vs “재정 건전화” ③ 한국당, 공천관리위원장에 김형오 전 국회의장 임명 ④ 유승민 빼고 보수통합?…안철수 19일 귀국 - 최민희 / 전 더불어민주당 의원 - 김용남 / 전 자유한국당 의원 #시사법정 ① '억울한 옥살이' 이춘재 8차 사건 재심 결정 ② 98세 노인, 아들과 법정 싸움 벌인 이유는? - 신유진 / 변호사

#각설하G0! ① 추미애 검찰 인사 단행…윤석열 패싱 현실화? ② 친박계 반발에 한국-새보수 통합 논의 급제동 ③ ‘결정적 한방’ 없었던 정세균 청문회 ④ 靑 출신 대거 출마…여야 인재영입 평가는? - 최민희 / 전 더불어민주당 의원 - 김용남 / 전 자유한국당 의원 #시사법정 ① '서지현 인사보복' 안태근 사건 ‘파기환송‘ ② 檢, MB 항소심서 징역 23년 구형 ③ 제3자 펭수 상표권 출원…EBS "법적 절차 진행" - 신유진 / 변호사

#각설하G0! ① 문 대통령, 의인 7인과 새해맞이 산행 ② 공수처법 통과 이후 달라지는 것들 ③ 추미애 임명‧조국 기소…2020 검찰은? - 최민희 / 전 더불어민주당 의원 - 김용남 / 전 자유한국당 의원 #시사법정 ① '선거개입 의혹' 송병기 구속영장 기각 ② 새 주민등록증, '민식이법'…새해 달라지는 것들 - 신유진 / 변호사

#각설하G0! ① 성탄절에도 필리버스터…선거법 표결 D-Day는? ② 조국 오늘 ‘운명의 날’…구속 갈림길 - 최민희 / 전 더불어민주당 의원 - 김용남 / 전 자유한국당 의원 #시사법정 ① 조국 오늘 영장심사…구속 여부 가를 쟁점은? ② 헌재, 한·일 위안부합의 위헌 여부 내일 결론 - 신유진 / 변호사

#각설하G0! ① 석패율제 거부한 민주당, 원포인트 국회 제안 ② “잠이 오냐” 군기잡던 황교안, 공개회의에서 ‘꾸벅’ ③ 무주공산 종로…이낙연·황교안 등판설 - 최민희 / 전 더불어민주당 의원 - 김용남 / 전 자유한국당 의원 #시사법정 ① '골프, 12·12 오찬' 전두환 재판 불출석 논란 ② ‘노조 와해’ 폭로 6년 만…삼성 임원들 실형 선고 - 신유진 / 변호사

#각설하G0! ① 예산안 통과…황교안 “나를 밟고 가라” ② 재판부, 정경심 공소장 변경 불허 ③ 숨진 수사관 휴대전화 놓고 검경 갈등 - 최민희 / 전 더불어민주당 의원 - 김용남 / 전 자유한국당 의원 #시사법정 ① 노소영, 최태원 SK회장에 이혼 맞소송 ② 검찰, 화성연쇄살인 8차 사건 직접 수사 - 신유진 / 변호사

#각설하G0! ① 신임 법무부장관에 추미애 ② ‘나경원 불신임’ 후폭풍 어디까지? ③ 패스트트랙 ‘4+1 공조’ 본격화…향후 전망은? ④ 청와대 경고 다음날 압수수색한 검찰, 왜? - 최민희 / 전 더불어민주당 의원 - 김용남 / 전 자유한국당 의원 #시사법정 ① 상임위 문턱 넘은 '데이터3법'…남은 관문은? ② 7번째 극단 선택…경마장서 무슨 일이? - 신유진 / 변호사

#각설하G0! ① 황교안 단식 8일 만에 병원 이송 ② 나경원, ‘총선 전 북미정상회담 자제’ 요구 ③ 선거 앞둔 김기현 울산시장 수사, 靑 ‘하명’ 의혹 - 최민희 / 전 더불어민주당 의원 - 김용남 / 전 자유한국당 의원 #시사법정 ① 대법원, 박근혜 '국정원 특활비' 사건 파기환송 ② '환경부 블랙리스트' 김은경 전 장관 첫 공판 - 신유진 / 변호사

#각설하G0! ① 여야, 文 대통령 '국민과의 대화'에 엇갈린 평가 ② 황교안, 이틀째 단식투쟁…국회서 '천막 최고위' ③ 물갈이·출마·불출마…총선 준비로 떠들썩한 정치권 - 최민희 / 전 더불어민주당 의원 - 김용남 / 전 자유한국당 의원 #시사법정 ① 윤중천 성범죄 무죄…​​​​​​​김학의도 혐의 벗나? ② 민원인에 “마음에 든다” 연락한 순경 처벌 면해…왜? - 신유진 / 변호사

#각설하G0! ① 나경원 대표 검찰 출석…패스트트랙 향방은? ② 조국 오늘 피의자로 검찰 출석 ③ 더불어민주당‧자유한국당 ‘험지’ 출마 논란 - 최민희 / 전 더불어민주당 의원 - 김용남 / 전 자유한국당 의원 #시사법정 ① '위안부 손해배상' 소송 3년 만에 첫 재판 ② 유승준 '비자 소송' 내일 결론…입국 길 열릴까 - 신유진 / 변호사

#각설하G0! ① 황교안 긴급 기자회견, 어떻게 봤나? ② 강기정 사과에도···줄다리기만 하다 파행된 예결위 - 최민희 / 전 더불어민주당 의원 - 김용남 / 전 자유한국당 의원 #시사법정 ① 법원 "땅콩회항 대한항공, 박창진에 7천만원 배상" ② '순위 조작' 프로듀스X 101 PD 2명 구속 - 신유진 / 변호사

#각설하G0! ① 문 대통령 모친 강한옥 여사 별세…오늘 발인 ② 자유한국당 총선 ‘인재 영입 1호’ 오늘 발표 ③ 이해찬 ‘조국 사태’ 사과…“국민께 송구” ④ “검찰, 조국 내사” 유시민 주장에 여당서도 “근거 빈약” - 김 현 / 전 더불어민주당 의원 - 김용남 / 전 자유한국당 의원 #시사법정 ① '강제징용 배상 판결' 1년…배상은 언제쯤? ② 임신 34주 낙태 중 '아기 울음'…의사 살인 혐의 - 신유진 / 변호사

#각설하G0! ① 정경심 구속영장 발부, 어떻게 보나? ② 자유한국당, 표창장에 공천 가산점 논란 ③ 이낙연, 오늘 오전 아베와 20분 면담 - 최민희 / 전 더불어민주당 의원 - 김용남 / 전 자유한국당 의원 #시사법정 ① 대법 "혼인 중 태어난 자녀, 유전자 달라도 친자식" ② ‘징역 3년’ 신격호 롯데 명예회장 형집행정지 결정 ③ '성폭행 부인' 김준기 전 동부회장 구속영장 신청 - 신유진 / 변호사

#각설하G0! ① 공수처법 ‘여야 평행선’ 좁힐 수 있을까? ② 文, 법무차관 불러 “검찰개혁 시급하다” - 최민희 / 전 더불어민주당 의원 - 김용남 / 전 자유한국당 의원 #시사법정 ① "설리법 만들어야"…악플방지법이란? ② ‘신림동 주거침입 사건’ 강간 미수 무죄 논란 - 신유진 / 변호사

#각설하G0! ① 특수부 폐지 발표 2시간 만에 전격사퇴, 배경은? ② ‘포스트 조국’ 정국 어떻게 되나? - 최민희 / 전 더불어민주당 의원 - 김용남 / 전 자유한국당 의원 #하재근의 문화쌀롱 ① 설리 비보에 연예계 추모 행렬…악플 자성 목소리도 ② 사우디 물들인 BTS…K팝 중동 진출 본격화하나? ③ 싸이월드 홈페이지 다시 열렸다…일부 복구 시작 - 하재근 / 문화평론가

#각설하G0! ① 국감장 곳곳 조국 공방…여상규 발언 논란 확산 ② 취임 한달 맞은 조국, 검찰개혁안 발표 - 최민희 / 전 더불어민주당 의원 - 김용남 / 전 자유한국당 의원 #시사법정 ① 국감서도 잊혀진 '사법개혁' 진행 상황은? ② 집행유예 중 재범자 1만 명 이상…"엄정 조치 필요" - 신유진 / 변호사

#각설하G0! ① 조국 “피의사실 공표 제한, 가족 수사 뒤에 할 것” ② 이주영·심재철, 자유한국당 7명째 릴레이 삭발 ③ 유은혜·김현미, 내년 총선 불출마? - 최민희 / 전 더불어민주당 의원 - 김용남 / 전 자유한국당 의원 #시사법정 ① 하 중사 ‘공상’ 판정 논란…보훈처 “곧 재심의 진행” ② “DLF 불완전판매 아닌 사기”…투자자들 공동 소송 - 신유진 / 변호사

#각설하G0! ① 조국 청문회 내일 열기로…남은 변수는? ② 여야 법사위, 조국 청문회 증인 11명 합의 ③ 조국 딸 생기부 공개한 주광덕 논란 - 최민희 / 전 더불어민주당 의원 - 김용남 / 전 자유한국당 의원 #시사법정 日, 도쿄올림픽 경기장 내 욱일기 허용 - 신유진 / 변호사

#각설하G0! ① 국회 정개특위, ‘선거법 개정안’ 통과 ② 일정 합의 후 더 거세진 공방…조국 청문회 어떻게 되나? ③ 자유한국당 연찬회, 어떤 말 오갔나? - 최민희 / 전 더불어민주당 의원 - 김용남 / 전 자유한국당 의원 #시사법정 ① KT 전 사장 "김성태, 딸 채용 직접 청탁" ② 대법원, 오늘 '국정농단' 최종심 선고 - 김영주 / 변호사

#각설하G0! ① 靑 “청문회서 가리자”…조국 검증 ‘정면 돌파’ ② 김문수 “박근혜 저주 받을 것” vs 김무성 “연사 잘못 선택” ③ 한일 외교장관 회동…‘지소미아’ 연장 여부는? - 최민희 / 전 더불어민주당 의원 - 김용남 / 전 자유한국당 의원 #시사법정 ① ‘가슴 보형물’ 시술 받고 희귀암…2천 명 “집단 소송” ② 페이스북 VS 방통위 1심 판결…'역차별' 해소되나 - 신유진 / 변호사

#각설하G0! ① 문 대통령 광복절 경축사에 담긴 의미는? ② 황교안 “文정권 실패…국정대전환 위해 싸울 것” - 최민희 / 전 더불어민주당 의원 - 김용남 / 전 자유한국당 의원 #시사본부 시사법정 ① '세월호 조작' 김기춘, 1심 집행유예…김장수·김관진 '무죄' ② 김구·유관순…판결문 속 독립운동가 - 조수진 / 변호사

#각설하G0! ① 강제징용 배상 1+1 안에 피해자 동의 없었다? ② “과거 한국으로 매춘 관광”…아베 보좌관의 망언 ③ “文 대통령 친일파” 공세에 노영민 “정론관 가라” ④ 유승민에 ‘러브콜’ 보낸 나경원 - 최민희 / 전 더불어민주당 의원 - 김용남 / 전 자유한국당 의원 #시사법정 교단 재판국 "명성교회 부자세습은 무효" - 신유진 / 변호사

#각설하G0! ① 합참은 미사일이라는데 北 “신형 방사포 시험사격” ② 황교안 “내 머릿속에 친박 비박 없어” ③ 양정철 민주연구원 “日보복 총선서 긍정” 보고서 논란 - 김봉준 / 전 청와대 인사비서관 - 김용남 / 전 자유한국당 의원 #시사본부 시사법정 ① “징용배상 판결 땐 보복”…日, 朴정부에 '내정간섭‘? ② 은행 직원의 사기행각…은행 책임은 어디까지? - 도진기 / 변호사

#각설하G0! ① 정치권, 러·일 도발 규탄…‘안보 위기’ 공방 ② 26개월 만에 靑 떠나는 조국…법무부장관 ‘예약’? - 최민희 / 전 의원 - 김희정 / 전 의원 #시사본부 시사법정 ① 법원, ‘사법농단’ 양승태 직권 보석 결정 ② 참사 8년 만에 결론난 가습제살균제 사건 - 오상원 / 변호사

#각설하G0! ① 한일 갈등에 정치권 친일 공방 가열 ② 오늘 文-5당 대표 회동, 어떤 대화 오갈까? ③ MB “정두언 만나려 했는데 안타깝다” - 최민희 / 전 더불어민주당 의원 - 조해진 / 전 새누리당 의원 #노변의 시사법정 ① ‘삼바’ 김태한 ‘분식회계 혐의’로 첫 구속영장 ② '합의 성관계'도 처벌…개정 아·청법 내용은? - 노영희 / 변호사

#각설하G0! ① 마이크 켜진 줄 몰랐던 윤석열 “윤대진 보호하려고…” ② 이낙연 “김상조, 너무 많은 말 하고 있다” 비판 ③ 최경환, 대법 징역 5년 확정…의원직 상실 - 현근택 / 더불어민주당 법률위 부위원장 - 김용남 / 전 자유한국당 의원 #노변의 시사법정 ① 대법원 "유승준 입국금지 결정 지나치다" ② '성폭행' 혐의 강지환 이르면 오늘 영장 여부 결정 - 노영희 / 변호사

#각설하G0! ① 이채익 “경찰청장 외압 안 느꼈다”…여야 4당 비판 ② 심상정 버티기 돌입…민주-정의, ‘해고’ 앙금 풀릴까? ③ “조건없는 일정 합의” vs “北 어선 국정조사” - 최민희 / 전 더불어민주당 의원 - 김용남 / 전 자유한국당 의원 #노변의 시사법정 ① 日 경제 보복, WTO 제소 검토…승소 가능성은? ② MB, 뇌물 혐의 51억 추가…1심보다 양형 세질까? - 노영희 / 변호사

#각설하G0! ① 조국 입각설에 야당 “선전포고” 반발 ② 국회, 내일 본회의서 상임·예결위원장 선출 추진 ③ “울고 싶다” 자유한국당 ‘엉덩이춤’ 논란 - 최민희 / 전 더불어민주당 의원 - 김용남 / 전 자유한국당 의원 #노변의 시사법정 ① 송중기·송혜교 이혼조정 돌입…추후 절차는? ② 김명환 민주노총 위원장 구속적부심…석방 가능성은? ③ '채용비리' 권성동 무죄…김성태는? - 노영희 / 변호사

#각설하G0! ① 오늘 6월 임시국회 개회…자유한국당 돌아오나? ② 황교안 “외국인에 똑같은 임금 불공정” 발언 논란 ③ 해상판 ‘노크 귀순’…정치권 공방 - 최민희 / 전 더불어민주당 의원 - 김용남 / 전 자유한국당 의원 #노변의 시사법정 ① 손혜원 발목 잡은 '보안자료'…비밀 여부가 관건 ② “글씨체가 달라요”…양승태 재판의 ‘시간 끌기’ - 노영희 / 변호사

#각설하G0! ① 김정은, 조문단 대신 조의문 보낸 이유는? ② 문 대통령 해외순방이 해외유람? 靑 “정정요청” ③ 청와대의 연이은 정치권 압박, 속내는? - 최민희 / 전 더불어민주당 의원 - 김용남 / 전 자유한국당 의원 #노변의 시사법정 ① '밀수 혐의' 조현아·이명희 1심 집행유예 ② 홍상수 이혼 소송, 14일 선고…'파탄주의' 판결 가능성은? - 노영희 / 변호사

#각설하G0! ① 청와대 VS 자유한국당, 회동 형식 논란 ② 오늘 황교안 취임 100일, 성과와 과제는? ③ 전국 도는 양정철…‘총선 행보’ 촉각 - 최민희 / 전 더불어민주당 의원 - 김용남 / 전 자유한국당 의원 #시사본부 이슈-2 국립현충원 ‘친일파’ 묘소, 해결 방법은? - 심용환 / 역사학자 (성공회대 외래교수)

#시사본부 이슈-3 "구명조끼 미착용 해왔다"…유람선 패키지, 문제 없었나? - A 씨 / 헝가리 현지 전직 가이드 #각설하G0! ① 민주당 “강효상 윤리위에 제소” VS 한국당 “총선 공작” ② 나경원 “서훈-양정철 회동 청와대가 감찰해야” - 최민희 / 전 더불어민주당 의원 - 김용남 / 전 자유한국당 의원 #노변의 시사법정 ① 김용균 없는 김용균법 개정 촉구 ② “여성 환자 정신착취” 김현철 처벌 가능성은? - 노영희 / 변호사

#각설하G0! ① 이해찬 VS황교안 ‘독재자의 후예’ 공방 ② 국회 정상화 협상 다시 교착…자유한국당 복귀 명분은? ③ 노무현 전 대통령 서거 10주기 추도식 - 권혁기 / 전 청와대 춘추관장 - 김용남 / 전 자유한국당 의원 #시사본부 현장연결 노 전 대통령 10주기 추도식…정치권 대거 참석 - 최민희 / 전 더불어민주당 의원 #노변의 시사법정 ① 윤중천, '김학의 사건'으로 6년 만에 재구속 ② 막바지 치닫는 'KT 부정 채용 수사'…김성태도 곧 소환? - 노영희 / 변호사

ADVANCE has spent the better part of a decade bringing his trademark and sound to events throughout the country. ADVANCE prides himself on versatility in his sets and throwing the memorable events in his home market - Minneapolis, MN.   While the Twin Cities may not be known as an entertainment hot bed, the young veteran continues to use his and forward thinking to help shape a scene traditionally found in larger regions. He can routinely be found at landmark venues such as SPADE, First Ave and many more.   ADVANCE has never been one to rest his laurels and formed "3Way Marketing Group" with his partners in 2010 to help harness his influence around the city. G0 95.3 Radio, Red Bull and L'Oreal now tap him to provide soundtrack for events and on-air mixes. In addition to him sharing the stage with elite DJ’s and artists such as R3HAB, Sander Van Doorn, Lil Wayne, and J. Cole.   Follow ADVANCE facebook.com/officialdjadvance instagram.com/djadvance twitter.com/djadvance soundcloud.com/theofficialdjadvance   Tracklist Mask Off (Advance Remix) - Future 7 Rings (Fraze Remix) - Ariana Grande MO Bamba (XaeboR Remix) - Sheck Wes Spend It (Advance Remix) - Dae Dae Hustlin (Chizzle & Rever Deep Remix) - Rick Ross 2012 - Party Favor Sicko Mode (Advance Remix) - Travis Scott Non Stop (Awoltalk Remix) - Drake Feel Free (Blvk Sheep Remix) - RL Grime Pony (Alamaki & Uki Dancehall Remix) - Ginuwine Police In Helicopter -  (Benny L Remix) - John Holt Going Bad (Spryte Remix) - Meek Mill My Cherie Amour (Lemi Vice x Action Jackson) - Stevie Wonder I Want You Back - Jackson 5 Lets Go Crazy - Prince Mo Bounce (Deadly Zoo Remix) - Iggy Azalia Wow (Dj Scene Bootleg) - Post Malone Thotiana (Knock2 Remix) - Blueface Wild Thoughts (Bees Knees Remix) – Rihanna  

In this free flowing episode we begin our look at further Reznor-Ross collaborations. For the first of two installments we will be covering the major theatrical releases that aren't named The Social Network. That would be Girl With The Dragon Tattoo, Gone Girl, Patriots Day, and Bird Box. Join us for discussions of the movies, how the music works in the film as well as on it's own, and dissect a few stand out tracks. Warning: earmuffs are required during the Birdbox discussion. Links:https://thefilmstage.com/features/interview-trent-reznor-discusses-the-girl-with-the-dragon-tattoo/https://www.stereogum.com/1709852/watch-trent-reznor-discuss-his-gone-girl-score-on-cbs-this-morning/video/https://www.youtube.com/watch?v=xeS8MNPocBw (GWTDT trailer)https://www.youtube.com/watch?v=2-_-1nJf8Vg (gone girl trailer)https://www.youtube.com/watch?v=OAtkXofiAzk (patriots day trailer) https://www.youtube.com/watch?v=INJ2bPFy108 (bird box trailer)What the dumbass monster from birdbox looks like https://www.express.co.uk/entertainment/films/1069204/Bird-Box-Netflix-monster-look-like-blindfold-suicide-alien-Sandra-Bullock-horror-filmHumans are idiots https://www.youtube.com/watch?v=of6dh59l-G0 

Emptying Out The MailbagOn this eighteenth episode, Em & Alan reply to all the listener feedback they've received since the last time they went over listener feedback, which was at least 3 episodes ago. This gives them a chance to revisit Marah in the Mainsail, The Prince of Egypt, and Audiofeed.They also talk about the value of community, and reiterate the fact that ALL ARE WELCOME in the Dorkness to Light community.Click on the player below to listen to the episode:Right-click to download the episode.You may also subscribe to the podcast through iTunes or the RSS Feed.Link: Faith and FandomLink: The Redeemed Otaku PodcastLink: The Nomad PodcastWe would love to hear from you about this podcast episode, or the podcast in general. Send e-mail feedback to dorknesstolight@gmail.com You can follow the network on twitter @DorknessToLight or Alan @ProfessorAlan

This week in Steins;Gate 0 Biweekly the gang talks about S;G0 episodes 3 and 4 and touch upon deep philosophical topics such as trying to have sex with your phone, the female casts' ever expanding chest sizes, and the purest form of humanity: Alexis Leskinen.

Le 151e épisode de "Deux minutes avant de dormir"

In the past view decades the therapy of malignant diseases has improved significantly. However especially metastatic diseases and relapses still require the development of new therapeutic modalities. Most chemotherapeutic agents induce apoptosis. A better understanding of apoptosis signaling is needed in order to combine cytostatic drugs in a more efficient way and to develop new targeted therapies. Conventional cytotoxic drugs and new therapies target p53, a key regulator of apoptosis. p53 is one of the most intensively studied target proteins but the heterogeneity and diversity of the function of p53 remains unclear. We investigated the importance of p53 in the intrinsic and extrinsic apoptosis signaling, via a variety of molecular biological approaches (Wachter et al., 2013). Apoptosis is induced intrinsically by the release of pro-apoptotic proteins from the inter-membrane space of the mitochondrion (eg. doxorubicin) or via activation of death receptors on the surface of the cell (eg. TRAIL, Tumor necrosis factor Related Apoptosis Inducing Ligand). As described in the literature, wildtype p53 leaded to an increase in intrinsic cell death, whereas different p53 mutations reduced apoptosis. In our work we discovered that the influence of p53 status on extrinsic cell death induction was much more multifaceted. The presence of p53 in tumor cell lines and xenograft tumor cells resulted in augmented, unaffected or decreased cell death. With great interest we discovered that the replacement of wildtype p53 by mutant p53 did not disturb the extrinsic apoptosis inducing capacity. As a next step we studied the influence of cell cycle arrest induced by p53 activation on TRAIL apoptosis sensitivity (Ehrhardt/Wachter et al., 2013). Cell cycle arrest was induced in tumor cell lines and patient samples in G0, G1 or G2 with cytostatic drugs, phase-specific inhibitors or RNA interference against cyclinB and E. Molecular or biochemical arrest at any point of the cell cycle augmented the rate of apoptosis by TRAIL. Consequently, when cell cycle arrest was deactivated by adding caffeine, the antitumor activity of TRAIL was decreased. The extrinsic apoptosis by TRAIL was increased in senescent tumor cells. This is especially important for the therapy of minimal residual disease and for the prevention of relapse caused by resting tumor cells. Additionally, we studied the impact of p53 activation on the rate of apoptosis induced by combinations of cytostatic drugs (Ehrhardt, Schrembs, Moritz, Wachter et al., 2011). A single drug is not very effective in cancer therapy therefore malignant diseases are usually treated with a combination of cytostatic agents. Co-administration of anthracyclines and vinca alkaloids on the same day is a standard of care for hematopoietic malignancies. If both drugs are applied simultaneously, doxorubicin inhibits apoptosis induction of vincristine. While it is well understood that doxorubicin activates p53 and induces cell-cycle arrest, we observed that cell-cycle arrest disabled inactivation of anti-apoptotic Bcl-2-family members by vincristine. Therefore, it follows that vincristine was unable to activate downstream apoptosis signaling. Cycling cells are required for vincristine to kill cancer cells efficiently. In summary we have shown the heterogeneity of p53 in extrinsic and intrinsic apoptosis signaling. The impact of the p53 status of tumor cells on extrinsic apoptosis signaling should be considered, especially in the context of therapeutic approaches that aim to restore p53 function to enable cell death. p53 can act pro- and anti-apoptotic, depending on the individual patient tumor cell and on drug and signal interactions. Finally, this suggests that patients benefit from an individualized therapeutic regimen. Further investigation is required in order to better understand the therapeutic potential of these complex interactions.

Transcript: The Sun is a typical main sequence star by which astronomers mean that the Suns properties lie in the middle of the range of stellar properties on the main sequence. It’s intermediate in mass, in size, in temperature, and luminosity compared to the most and least massive main sequence stars. The Sun has a spectral type G2 which gives it a photosphere temperature of fifty-seven hundred Kelvin. Spectral types O, B, A, F, G, K, and M are subdivided on a decimal scale running from O1 to O8 to O9 to B0, B1, etcetera. The Sun as a G2 star is a little cooler than a G0 star. The Sun is typical in most properties, but most stars are less massive than the Sun and the very highest mass main sequence stars are extremely rare.

The apoptotic mode of cell death (programmed cell death) is an active and defined process that plays an important role in the development of multicellular organisms and in the regulation and maintenance of cell populations in tissues under physiologic and pathologic conditions. Since dysregulation of apoptosis is associated with the progress of many diseases, induction of apoptosis is an interesting pharmacological target for the therapy of many diseases. Our study shows that the novel semisynthetic pentacyclic triterpenoid C-KβBA has a profound antiproliferative effect on different tumor cell lines and that it induces apoptosis of tumor cells in vitro and vivo. Previous studies have shown that the mammalian target of rapamycin (mTOR) is a key regulator for many cells activities, and that the perturbation of this signaling pathway is implicated in many diseases and metabolic disorders. Accordingly targeting the mTOR signaling pathway seems to be a promising therapeutic approach for the treatment of various diseases and metabolic disorders. We have shown in our study that C-KβBA downregulates the mTOR signaling pathway in vitro and vivo, which leads to the induction of cell cycle arrest in the G0 cell cycle phase as well as downregulation of the protein translation machinery. Together, the presented data strongly suggest that C-KβBA could serve as a lead compound for the development of novel treatment modalities for diseases and disorders associated with defects in the apoptosis machinery, or the mTOR signaling pathway, or both.

Recent work has underlined the importance of animal models in discovery and characterisation of molecular mechanisms determining radiosensitivity and radioresistance. Enhanced sensitivity of LEC rats to ionizing radiation in terms of the acute radiation syndrome was investigated in the present work on the cellular level and compared to that of LE rats. To understand the molecular basis for the increased radiation sensitivity a series of studies were performed, which included the classical clonogenic survival assay, investigation of double strand break repair by means of PFGE and gH2AX evaluation, comet assay for evaluation of repair of single strand breaks and alkaline labile sites, and analysis of cell cycle progression of asynchronous fibroblast population. Survival assay, PFGE, and H2AX analysis were performed in a standardised experimental system - confluent fibroblasts, synchronized in G1 phase of cell cycle, and comet assays were performed in G0 lymphocytes. The data suggests a mild radiosensitization of LEC fibroblasts compared to LE. The results of studies using the selected model did not reflect the degree of animal sensitivity on the molecular level, since values of dose modifying factor (DMF) were much lower in fibroblasts (DMF2 = 1.32) compared to that of animal sensitivity (DMF = 2.36 for bone marrow syndrome (LD50/30) and DMF = 1.95 of intestinal death (LD50/7)). The investigation of DNA repair and cell cycle did not reveal a significant defect in the studied pathways in synchronized fibroblasts and cell cycle progression was not different from wild type cells. The presented data contradict the published LEC cellular phenotype. Of the possible candidate genes, which are located in the radiosensitivity locus, several were further analysed. Among those, Gata-2 appeared to be the most promising of the positional and functional candidates. However, no mutation in the coding sequence could be identified and mRNA expression levels were similar between control and LEC cells. The presented data suggests that radiosensitivity of LEC rats might be attributed to a mechanism specific for certain target tissue, like bone marrow, or enhanced in cell cycle stages other than G0/G1.

Im Rahmen der vorliegenden Doktorarbeit wurde zum ersten Mal die Keimbahn eines Käfers erfolgreich genetisch transformiert. Der von uns zu diesem Zweck in Zusammenarbeit mit Ernst Wimmer entwickelte Transformationsmarker 3xP3-EGFP hat inzwischen sein Potential als spezies-unabhängiges Markergen auch in weiteren Invertebraten-Spezies unter Beweis gestellt und damit das Spektrum transformierbarer Taxa beträchtlich erweitert. In Tribolium konnten Transformationsereignisse mit 3xP3-EGFP für drei verschiedene Transposons - Hermes, Minos und piggyBac - erzielt werden. Die Effizienz betrug dabei 1,4% (Hermes), 11,4% (Minos) bzw. 56% (piggyBac) der fertilen G0 und gehört damit zu den höchsten Werten, die in der Literatur für Insekten berichtet wurden. Bei Minos konnte die Effizienz durch die Verwendung von Transposase mRNA statt einem DNA Helper-Plasmid weiter auf 32,4% gesteigert werden. Für piggyBac und Minos wurde ferner in Zusammenarbeit mit anderen Labors gezeigt, daß es sich bei den meisten Transposoninsertionen um unabhängige Einzelintegrationen handelt, die auf verschiedene Chromosomen verteilt sind und stabil weitervererbt werden. Die Größe zusätzlich transferierter Fremd-DNA kann dabei bei piggyBac mindestens bis zu 9,5 kb betragen. Schließlich konnte noch ein piggyBac Element durch Helperinjektion mit einer Rate von 28,1% remobilisiert werden. Zusammen mit der Anfälligkeit für enhancer trap Effekte können daher mit diesem System alle relevanten Transposon-basierenden Techniken zur funktionellen Genomanalyse angewandt werden. Als erste praktische Anwendung wurden D. melanogaster Sequenzen für anteriore und posteriore mRNA-Lokalisierung (bicoid-3’UTR und oskar-3’UTR), sowie ein bicoid-abhängiger Minimalpromotor in Tribolium eingeführt. Allerdings konnten durch diese Ansätze keine Komponenten oder Mechanismen eines ggf. konservierten maternalen Systems nachgewiesen werden. Ein Konstrukt mit 5,2 kb der upstream Sequenzen von Tc’hunchback mit lacZ als Reportergen war hingegen in der Lage, das endogene hunchback-Muster größtenteils nachzubilden. Das frühere Ergebnis von Christian Wolff mit Tc’hunchback in Drosophila, wonach dieses Fragment alle wesentlichen regulatorischen Elemente enthält, konnte daher in transgenen Käfern bestätigt werden. Zusätzlich zu dem als sehr riskant eingestuften Transformations-Projekt wurde parallel ein weiteres Projekt durchgeführt, die Analyse der homöotischen Mutanten wurm und überlänge. In beiden Mutanten ist vor allem die Identität der posterioren Segmente ab A9 verändert. In wurm sind die Segmente A9-A11 nach A8 transformiert und die telsonalen Anhänge Urogomphi und Pygopodien fehlen. In überlänge ist nur A9 wie A8 ausgebildet und demzufolge nicht mit dem Telson fusioniert. Es fehlen nur die Urogomphi. überlänge bildet zusätzlich ein ektopisches Stigma im ersten thorakalen Segment. Es wurde gezeigt, daß es sich bei den betroffenen Genen um zwei verschiedene Loci handelt, die beide nicht im homöotischen Komplex liegen. Obwohl der Phänotyp von wurm weitgehend der RNAi-Phänokopie von Abdominal-B entspricht, konnte also keiner dieser beiden Loci einem bekannten Hox-Gen zugeordnet werden. Als mögliches Kandidatengen für diese Loci wurde daher das Tribolium-Homolog des regionsspezifischen homöotischen Gens spalt kloniert. Die Expression von spalt entspricht weitgehend der von Dm’spalt, mit einer anterioren und einer posterioren Domäne, einer dorsalen Expression an den seitlichen Rändern des Keimstreifs, sowie einem komplexen Muster im Nervensystem. Mit Hilfe der kürzlich entwickelten Technik der parentalen RNAi wurde die Funktion dieses Gens untersucht. In sal–– Phänokopien finden sich, wie in Drosophila, anteriore und posteriore Veränderungen von Segmentidentitäten. So wird das abdominale Segment A9 in Richtung anteriore abdominale Segmente transformiert. Dadurch tritt ein zusätzliches Stigma auf und die Pygopodien gehen verloren, das Segment fusioniert aber weiterhin mit dem Telson. Im Gegensatz zu Drosophila wird aber anterior nicht das Labium verändert, sondern die Identität der Maxille wird partiell in Richtung Mandibel transformiert: statt dem Enditen der Maxille wird ein mandibel-ähnlicher Zahn gebildet. Damit kommt offenbar auch spalt nicht als Locus in Frage, der in wurm oder überlänge seine Funktion verloren hat. Möglicherweise spielen diese beiden Loci eine Rolle als den HOX-Genen übergeordnete regulatorische Gene, oder als Co-Faktor von Abd-B. Damit sind wurm und überlänge als interessante (und aus Drosophila nicht bekannte) Spieler im homöotischen System der Insekten identifiziert, was weitere Untersuchungen als sehr lohnend erscheinen läßt. Vor allem aber hat dieses Teilprojekt die Evolution des spalt-Gens erhellt, das in weniger abgeleiteten Insekten offenbar eine essentielle Rolle bei der Spezifizierung von Mandibel versus Maxille spielt. Diese Funktion ist in Drosophila vermutlich im Zuge der Reduktion der Mandibel verloren gegangen.

Der eukaryontische Zellzyklus wird durch die Aktivität verschiedener Cyclinabhängiger Kinasen (CDKs) reguliert. Die zelluläre Menge des CDK-Inhibitorproteins p27Kip1 spielt eine entscheidende Rolle beim Übergang der Zelle von der G1- zur SPhase. Die Menge von p27Kip1 steigt während der G0- oder der G1-Phase an und nimmt zu Beginn der S-Phase rasch wieder ab. Die Bindung von p27Kip1 an die CDKKomplexe der G1-Phase inaktiviert diese und verhindert dadurch die Initiation der SPhase. Eine verminderte Menge von p27Kip1 am G1/S-Phaseübergang findet man dagegen häufig in verschiedenen Tumorgeweben. Die geringere zelluläre Menge des Inhibitors ist dabei mit einer hohen Patientensterblichkeit und einem aggressiven Verlauf der Erkrankung verbunden. Die zelluläre Aktivität und Menge von p27Kip1 wird entscheidend durch Proteine reguliert, die mit p27Kip1 interagieren. In dieser Arbeit wurden deshalb mit Hilfe von rekombinantem p27Kip1 Interaktionspartner des Inhibitors in HeLa-Zellextrakt identifiziert. Es konnte gezeigt werden, daß p27Kip1 an die CDK-Proteine und an Grb2 bindet. Grb2 ist ein Adapterprotein der Signaltransduktion. Die Interaktion zwischen p27Kip1 und Grb2 könnte damit, nach Stimulation der Zelle durch verschiedene Mitogene, die Signalweitergabe mit der Zellzyklusmaschinerie verbinden. Die zu dieser Interaktion notwendige Domäne in p27Kip1 konnte in weiteren Analysen auf eine acht Aminosäuren lange Prolin-reiche Region eingegrenzt werden. Auf der anderen Seite interagiert Grb2 vornehmlich über seine C-terminale SH3-Domäne mit p27Kip1. Die beiden mit p27Kip1 nah verwandten Inhibitorproteine p21Cip1 und p57Kip2 interagieren dagegen nicht mit Grb2. In einer erweiterten Analyse wurden 41 verschiedene rekombinante SH3-Domänen auf eine Interaktion mit p27Kip1 hin getestet. Es konnte gezeigt werden, daß p27Kip1 nur mit der C-terminalen SH3-Domäne von Grf40/Mona und der SH3-Domäne der Tyrosinkinase Lyn wechselwirkt. Die Interaktion der Tyrosinkinase Lyn in vivo führte zur Hypothese, daß p27Kip1 durch Lyn phosphoryliert werden könnte. Im zweiten Teil dieser Arbeit wurde deshalb die Tyrosinphosphorylierung von p27Kip1 untersucht. In Phosphoaminosäureanalysen mit metabolisch markierten Zellen konnte gezeigt werden, daß p27K i p 1 in vivo an Tyrosinresten phosphoryliert wird. Diese Zusammenfassung 13 Phosphorylierung konnte durch rekombinant hergestellte Tyrosinkinasen und verschiedene Tyrosin/Phenylalanin-Austausche in p27Kip1 auf Tyrosin 88 und 89 eingegrenzt werden. Nach Kristallstrukturdaten des trimeren Komplexes aus p27Kip1, CDK2 und Cyclin A kommt der Tyrosinrest 88 von p27Kip1 in der ATP-Bindetasche der Kinase zu liegen und blockiert diese. Es wurde deshalb untersucht, inwieweit eine Phosphorylierung von p27Kip1 an Tyrosin 88 oder 89 Einfluß auf die Aktivität des Inhibitors hat. Die Tyrosinphosphorylierung von p27Kip1 verhindert nicht die Bindung an den CDK-Komplex. Allerdings konnte mit in vitro-phosphoryliertem p27Kip1 gezeigt werden, daß eine Tyrosinphosphorylierung zu einer etwa 40%-igen Reduktion der Aktivität des Inhibitors führt. Diese Ergebnis konnte in vivo bestätigt werden. Interessanterweise verstärkt die Tyrosinphosphorylierung des Inhibitors die Phosphorylierung von p27Kip1 an Threonin 187 durch den gebundenen CDK-Komplex. Die Phosphorylierung von p27Kip1 an Threonin 187 ist in der Zelle ein initiales Signal zum Abbau von p27Kip1 durch das 26S-Proteasom. Das so markierte p27Kip1 wird von einem E3-Ligase-Komplex erkannt und ubiquitiniert. Es wurde deshalb untersucht, welchen Einfluß die Tyrosinphosphorylierung auf den Abbau von p27Kip1 besitzt. In Halbwertszeitbestimmungen mit einer SH3-bindedefizienten Form von p27Kip1 und einer Tyrosin/Phenylalanin-Austauschform von p27Kip1 konnte zeigten werden, daß beide Formen, im Vergleich zu unverändertem p27Kip1, eine höhere Stabilität aufweisen. Die Interaktion von p27Kip1 mit der Tyrosinkinase Lyn und die Inaktivierung des Inhibitors durch die Phosphorylierung von Tyrosinresten zeigt eine Möglichkeit auf wie p27Kip1, in Abhängigkeit von mitogenen Stimuli, reguliert werden kann. Die in dieser Arbeit gefundene Interaktion von Grb2, Grf40 und Lyn mit p27Kip1 verbindet damit die Signaltransduktion mit der Zellzykluskontrolle.

In human diploid fibroblasts (HDFs) the cell nucleus is oval in shape, quite large in xy-diameter (10-20µm), but flat in z-direction (5µm). In these nuclei chromosome territories typically lie side by side or slightly above each other. The question whether these arrangements are ordered or variable has yielded conflicting answers. We hybridised an improved 7-fluorochrome MFISH probe set on 3D-preserved cell nuclei, fixed with buffered 4% paraformaldehyde. A LEICA wide field microscope with an 8-filter-wheel and an automated z-step motor was used for imaging of the 7 fluorochromes plus DAPI. Multicolor images from nuclei were taken as serial sections in z-direction. After deconvolution, a specifically developed Program goldFISH Saracoglu K. et al. 2001 was used to classify the images according to the labelling scheme. The classification algorithm corresponds to the procedure previously used for metaphase spreads, now adopted to 3-D studies of chromosome territory arrangements in the cell nucleus. The analysis of 30 G0-fibroblast nuclei and 30 prometaphase rosettes revealed a pronounced variability of chromosome territory neighbourhoods, as described by Allison D. C. et al. 1999, but in contrast to Nagele R. et al. 1995. However we noted a distinct radial order: small chromosomes were located close to the centre while large chromosomes were positioned towards the nuclear rim. This non-random radial positioning could also be observed in prometaphase rosettes