Podcasts about Interphase

TECH CLUBBERS PODCAST W/ SILENT G Over the past years, Silent G has managed to establish himself in Berlin's electronic music scene. As a DJ, he is a regular behind some of the city's most renowned decks. And with gigs in Vienna and Mongolia, his music has also taken him abroad. In the booth, his main goal is to find the right connection to the dance floor. Depending on the vibe, his sets range from dubby Techno to old school Tribal cuts, funky Hardgroove or driving Techno tools. But he also feels comfortable in more break-heavy spheres, such as Drum'n'Bass and Jungle. Besides DJing, his activities in the local scene include running the event series ZEMENT, and curating the Drum'n'Bass and Jungle night STALACTITES at Paloma. Silent G is also part of the Interphase crew, hosting notorious genre-bending off-location parties. As a producer, he has track releases on the Irish label Space Records and on Nonchalance Music. Another one is scheduled for January 2025 on the Toronto-based imprint Oneiros Records. Follow SILENT G here: Instagram: https://www.instagram.com/silentg_berlin Soundcloud: https://soundcloud.com/silentg-music

Link to bioRxiv paper: http://biorxiv.org/cgi/content/short/2023.07.03.547485v1?rss=1 Authors: Allshire, R., Median-Pritchard, B., Spanos, C., Rappsilber, J., Jeyaprakash, A. A., London, N. Abstract: CENP-A chromatin specifies mammalian centromere identity, and its chaperone HJURP replenishes CENP-A when recruited by the Mis18 complex (Mis18C) via M18BP/KNL2 to CENP-C at kinetochores during interphase. However, the Mis18C recruitment mechanism remains unresolved in species lacking M18BP1, such as fission yeast. Fission yeast centromeres cluster at G2 spindle pole bodies (SPBs) when CENP-ACnp1 is replenished and where Mis18C also localizes. We show that SPBs play an unexpected role in concentrating Mis18C near centromeres through the recruitment of Mis18 by direct binding to the major SPB LInker of Nucleoskeleton and Cytoskeleton (LINC) complex component Sad1. Mis18 recruitment by Sad1 is important for CENP-ACnp1 chromatin establishment and acts in parallel with a CENP-C-mediated Mis18C recruitment pathway to maintain centromeric CENP-ACnp1, but is independent of Sad1-mediated centromere clustering. SPBs therefore provide a non-chromosomal scaffold for both Mis18C recruitment and centromere clustering during G2. This centromere-independent Mis18-SPB recruitment provides a mechanism that governs de novo CENP-ACnp1 chromatin assembly by the proximity of appropriate sequences to SPBs and highlights how nuclear spatial organization influences centromere identity. Copy rights belong to original authors. Visit the link for more info Podcast created by Paper Player, LLC

Link to bioRxiv paper: http://biorxiv.org/cgi/content/short/2022.11.06.515321v1?rss=1 Authors: Oda, H., Sato, Y., Kawashima, S. A., Fujiwara, Y., Mate, P., Wu, E., Vastenhouw, N. L., Kanai, M., Kimura, H. Abstract: In the cytoplasm, filamentous actin (F-actin) plays a critical role in cell regulation, including cell migration, stress fiber formation, and cytokinesis. Recent studies have shown that actin filaments that form in the nucleus are associated with diverse functions. Here, using live imaging of an F-actin-specific probe, superfolder GFP-tagged utrophin (UtrCH-sfGFP), we demonstrated the dynamics of nuclear actin in zebrafish (Danio rerio) embryos. In early zebrafish embryos up to around the high stage, UtrCH-sfGFP increasingly accumulated in nuclei during the interphase and reached a peak during the prophase. After nuclear envelope breakdown (NEBD), patches of UtrCH-sfGFP remained in the vicinity of condensing chromosomes during the prometaphase to metaphase. When zygotic transcription was inhibited by injecting -amanitin, the nuclear accumulation of UtrCH-sfGFP was still observed at the sphere and dome stages, suggesting that zygotic transcription may induce a decrease in nuclear F-actin. The accumulation of F-actin in nuclei may contribute to proper mitotic progression of large cells with rapid cell cycles in zebrafish early embryos, by assisting in NEBD, chromosome congression, and/or spindle assembly. Copy rights belong to original authors. Visit the link for more info Podcast created by Paper Player, LLC

“The Star Gazer” Following the events of Season 1, Picard returns to his life in France with the flame of love unexpectedly flickering in the vineyard. But when his connection to the Romulan Laris comes up against internal defenses, and Starfleet presents him with a mystery, Picard sets out on a journey of personal exploration that leads him to encounters with old friends. In this episode of The Artificial Tango, our rebranded dedicated Star Trek: Picard podcast that carries forward the flame from The Line, hosts C Bryan Jones and Matthew Rushing discuss the start of Season 2, digging into Picard as a character, his connection to Laris, how Seven on Nine serves as a mirror to Picard, how the story sets up a hopeful Federation worth saving, and how the characters transition from the story of Season 1. We also speculate on where the upcoming adventure in Los Angeles might take us. Note: This episode is being published as a joint episode of Interphase, our Star Trek Universe podcast, and The Artificial Tango, with Interphase being used to introduce the name change and transition of The Line. Hosts C Bryan Jones and Matthew Rushing Production C Bryan Jones (Editor and Producer) Matthew Rushing (Executive Producer) Cornelia Reutner (Associate Producer)

“The Star Gazer” Following the events of Season 1, Picard returns to his life in France with the flame of love unexpectedly flickering in the vineyard. But when his connection to the Romulan Laris comes up against internal defenses, and Starfleet presents him with a mystery, Picard sets out on a journey of personal exploration that leads him to encounters with old friends. In this episode of The Artificial Tango, our rebranded dedicated Star Trek: Picard podcast that carries forward the flame from The Line, hosts C Bryan Jones and Matthew Rushing discuss the start of Season 2, digging into Picard as a character, his connection to Laris, how Seven on Nine serves as a mirror to Picard, how the story sets up a hopeful Federation worth saving, and how the characters transition from the story of Season 1. We also speculate on where the upcoming adventure in Los Angeles might take us. Note: This episode is being published as a joint episode of Interphase, our Star Trek Universe podcast, and The Artificial Tango, with Interphase being used to introduce the name change and transition of The Line. Hosts C Bryan Jones and Matthew Rushing Production C Bryan Jones (Editor and Producer) Matthew Rushing (Executive Producer) Cornelia Reutner (Associate Producer)

“The Star Gazer” Following the events of Season 1, Picard returns to his life in France with the flame of love unexpectedly flickering in the vineyard. But when his connection to the Romulan Laris comes up against internal defenses, and Starfleet presents him with a mystery, Picard sets out on a journey of personal exploration that leads him to encounters with old friends. In this episode of The Artificial Tango, our rebranded dedicated Star Trek: Picard podcast that carries forward the flame from The Line, hosts C Bryan Jones and Matthew Rushing discuss the start of Season 2, digging into Picard as a character, his connection to Laris, how Seven on Nine serves as a mirror to Picard, how the story sets up a hopeful Federation worth saving, and how the characters transition from the story of Season 1. We also speculate on where the upcoming adventure in Los Angeles might take us. Note: This episode is being published as a joint episode of Interphase, our Star Trek Universe podcast, and The Artificial Tango, with Interphase being used to introduce the name change and transition of The Line. Hosts C Bryan Jones and Matthew Rushing Production C Bryan Jones (Editor and Producer) Matthew Rushing (Executive Producer) Cornelia Reutner (Associate Producer)

“The Star Gazer” Following the events of Season 1, Picard returns to his life in France with the flame of love unexpectedly flickering in the vineyard. But when his connection to the Romulan Laris comes up against internal defenses, and Starfleet presents him with a mystery, Picard sets out on a journey of personal exploration that leads him to encounters with old friends. In this episode of The Artificial Tango, our rebranded dedicated Star Trek: Picard podcast that carries forward the flame from The Line, hosts C Bryan Jones and Matthew Rushing discuss the start of Season 2, digging into Picard as a character, his connection to Laris, how Seven on Nine serves as a mirror to Picard, how the story sets up a hopeful Federation worth saving, and how the characters transition from the story of Season 1. We also speculate on where the upcoming adventure in Los Angeles might take us. Note: This episode is being published as a joint episode of Interphase, our Star Trek Universe podcast, and The Artificial Tango, with Interphase being used to introduce the name change and transition of The Line. Hosts C Bryan Jones and Matthew Rushing Production C Bryan Jones (Editor and Producer) Matthew Rushing (Executive Producer) Cornelia Reutner (Associate Producer)

Unser Kurs auf der Webseite: https://www.selbstorientiert.org/shop/Zellzyklus-&-Cycline-Kurs-mit-Karteikarten-&-Texten-p432106000 Unser Kurs auf Amazon: Unser YouTube-Video: https://youtu.be/X_N6pwvL0Wo --- Send in a voice message: https://anchor.fm/selbstorientiert/message

Josh and crew blaze through three seasons in The Quiet Year and introduce even more horrors into the world. All while Josh fights for his dream of a city... The post Interphase 07: We Built This City appeared first on Geekspective.

The final world/universe building game is under way with The Quiet Year with a full table of players! As is tradition, The Quiet Year goes places no one expects! The post Interphase 06: The Planet Itself appeared first on Geekspective.

The Ground Itself continues with Josh, Amber, and Dave playing out the last section of time. Alliances are forged and broken, love is staged, and Jace is THE WORST! The post Interphase 05: The Ace appeared first on Geekspective.

The world building continues with The Ground Itself! Dave and Amber join Josh as they build the history of a space colony as they observe what happens over the decades at J.O.S.H. The post Interphase 04: I Dream of Lemons appeared first on Geekspective.

The world building continues with Life in the Machine! Josh and Brother John play out pilot and mech in a war that pushes their relation to the limit! The post Interphase 03: Brock and Dragoon appeared first on Geekspective.

Now for the conclusion of Microscope! We know the beginning and the end now to fill in the rest of that cake! This episode is all game! The post Interphase 02: A Poor Cover-up appeared first on Geekspective.

It is the beginning of Tales From Mauxferry Metaphase! But what comes before metaphase? Interphase! Josh and the crew start play off with Microscope by Ben Robbins as they build the history of a new universe! You can catch every episode of Tales From Mauxferry Season Interphase streamed live on the Geekspective Network Twitch channel every other Saturday 6 PM EST! You can find more great content from Geekspective on Twitter @geekspective. This episode was edited by Brother John – he can be reached on Twitter @That_Drawer for all your podcast needs.

It is the beginning of Tales From Mauxferry Metaphase! But what comes before metaphase? Interphase! Josh and the crew start play off with Microscope by Ben Robbins as they build the history of a new universe! The post Interphase 01: Bobo The Monkey appeared first on Geekspective.

Captain Kirk's ghost spooks the crew. Join your interphasic hosts as they praise the writers for examining Spock and McCoy's relationship by removing Kirk from the equation, discuss how this episode is connected to the larger Star Trek universe, and debate the viability of the Tholians' languid plan. Spoiler warning! We dive right into a detail-rich discussion of this episode, so if you haven't had a chance to see it yet – beware! Next week: We slip into a slightly-sideways dimension with Geordi, Ro, and a rascally Romulan in Star Trek: TNG's "The Next Phase" Note that this and all episodes of the It's Got Star Trek podcast contain explicit language and, frankly, an unnecessary amount of offensive content, so the show is intended only for adults and really really cool kids. Check out the Feedspot list of Top 60 Star Trek Podcasts for 2021. Please utilize one of the following options if you have an interest in contacting your hosts: Email us at feedback@itsgotstartrek.com Twitter us @ItsGotStarTrek Instagramaphone us @ItsGotStarTrek Facebookify us @ItsGotStarTrek Watch a static image while listening to the podcast on YouTube Telephone us at 202-456-1414 You can also visit www.itsgotstartrek.com and leave a comment or head on over to the It's Got Everything subreddit to join the discussion. Don't say we didn't give you options here!

My AP Biology Thoughts  Unit 4 Cell Communication and Cell CycleWelcome to My AP Biology Thoughts podcast, my name is Chloe McGregor and I am your host for episode #93 called Unit 4 Cell Communication and Cell Cycle: The Cell Cycle. Today we will be discussing the basics of the cell cycle including how it works and what products are made.  Segment 1: Introduction to the Cell CycleThe cell cycle is split up into 2 different parts, each with their own purpose. The first part of the cell cycle is called interphase. The cell spends a majority of its time in this phase. Interphase itself is made up of 3 different stages. These stages are the G1, S, and G2 phase. The first step in interphase is G1. During this step, the cell spends its time growing in size and gaining a sufficient amount of resources. With this abundance of resources, the cell is able to replicate its DNA and intracellular components during the S phase. The third step is G2 where the cell continues its growth and will stop its progression to the mitotic phase if there are any issues or damaged DNA. The second part of the cell cycle is the mitotic phase which has 2 parts. The first part is mitosis. Mitosis consists of four steps: Prophase, metaphase, anaphase, and telophase. More details about these steps will be discussed later in the episode, but generally they work together in order to separate the replicated DNA. The second part of the mitotic phase is cytokinesis where the cell actually splits into 2 new identical daughter cells. Overall, each step in the cell cycle is important to grow the cell and its resources in order to produce 2 identical daughter cells, each with the same copies of DNA. Another important part of the cell cycle is the G0 phase. Although this isn't the generic path of cell replication , a cell may enter the G0 phase if there is a non sufficient amount of resources and nutrients available to proceed to healthy replication. Cells may also enter G0 if they are adult cells that are not necessarily looking to replicate. For example, a lot of cells in your brain and nervous system stop replicating once you reach adulthood, so an injury in these areas could be extremely difficult, or impossible, to heal. Segment 2: More About the Cell CycleTo go more in depth, let's talk about the different steps in mitosis. Once again, these steps are prophase, metaphase, anaphase, and telophase. The goal of mitosis is to separate the replicated DNA on opposite sides of the cell, with both sides having identical copies of the DNA. Prophase begins once G2 is finished and the cell has grown enough. In prophase, the DNA condenses, and the replicated chromosomes have a more visible shape to them. The replicated chromosomes are called sister chromatids and are linked together at a point called the centromere. The next step, metaphase, is when the replicated chromosomes line up in the center of the cell in a line. Anaphase is next, and this is where the spindle fibers on each side of the cell attach to the centromere region on each sister chromatid. Then, the fibers pull the sister chromatids apart, leaving opposite sides of the cell with identical genetic material. Telophase is the last stage of mitosis where each side of the cell begins to form a nuclear envelope around the new genetic material. The chromosomes also unravel back into chromatin. Once mitosis is complete, the cell membrane scrunches in the middle of the cell causing it to physically separate into two identical daughter cells. This step is called cytokinesis. Another important point to touch on is the checkpoints that occur during the cell cycle. Although there is a separate episode on cell cycle regulation, it is important to understand that the cell reaches checkpoints throughout the entire cell cycle which ensure that it is healthy, and that the DNA is replicating correctly. There are 3 checkpoints. These are near the end of G1, between G2 and the M phase, and one...

In this episode, Jules Higa asks a question about Interphase. This is a question you might see from the Biological and Biochemical Foundations of Living Systems section of the MCAT. This podcast is designed for Pre-medical students preparing to take their Medical College Admissions Test (MCAT). This episode is powered by Premed Consultants, an all-inclusive premed advising program. Whether you are starting out as a freshman or about to start prepping for the MCAT, the premed consultants will help you throughout your entire premed process until you get into medical school. Not only is there a full MCAT program utilizing the most effective study tactics, but they will also help you through the entire admissions process as well. If you're interested in one on one mentorship, go to thepremedconsultants.com and you can schedule a free 30 min strategy session to see if Premed Consultants is the right fit for you. If you have any suggestions, concerns, or question, feel free to e-mail us at mcatflashgo@gmail.com We wish you the best of luck on your educational journey!

The cell cycle is more than just memorizing the position of chromosomes on a slide, it can help us understand growth and development, lead us to treatments for cancer, or repair damaged organs and limbs. I go over the major steps of the cell cycle including interphase and the mitotic phase. Education level - high school to non-majors

Serialization in Star Trek. When it comes to discussion of serialization in Star Trek, Deep Space Nine is the go-to series—at least prior to Discovery and Picard. The Original Series was famously episodic, and The Next Generation, for the most part, followed that same pattern. DS9 itself barely moved away from the model in its first season, but then things started to change. By the time Voyager came along, the winds of serialization were blowing. But how do these two Next Generation spinoffs compare when it comes to story and character threads that span the seasons? In this episode of Interphase, host C Bryan Jones is joined by Chris Chaplin to discuss the history of serialization in Star Trek—especially comparing Deep Space Nine and Voyager—through plot threads, character arcs, interaction with various species, and more. We also break down the narrative structure of the two series season by season. Lastly, we ask the question, “Is Star Trek episodic by nature?” Host C Bryan Jones Guest Chris Chaplin Production C Bryan Jones (Editor and Producer) Matthew Rushing (Executive Producer) Renee Roberts (Associate Producer)

Serialization in Star Trek. When it comes to discussion of serialization in Star Trek, Deep Space Nine is the go-to series—at least prior to Discovery and Picard. The Original Series was famously episodic, and The Next Generation, for the most part, followed that same pattern. DS9 itself barely moved away from the model in its first season, but then things started to change. By the time Voyager came along, the winds of serialization were blowing. But how do these two Next Generation spinoffs compare when it comes to story and character threads that span the seasons? In this episode of Interphase, host C Bryan Jones is joined by Chris Chaplin to discuss the history of serialization in Star Trek—especially comparing Deep Space Nine and Voyager—through plot threads, character arcs, interaction with various species, and more. We also break down the narrative structure of the two series season by season. Lastly, we ask the question, “Is Star Trek episodic by nature?” Host C Bryan Jones Guest Chris Chaplin Production C Bryan Jones (Editor and Producer) Matthew Rushing (Executive Producer) Renee Roberts (Associate Producer)

Watching TOS for the first time, in 2020. For many of us, the five-year mission of Kirk, Spock, McCoy, and the crew of the USS Enterprise is what hooked us on Star Trek and made us lifelong fans. But there are many who have never seen The Original Series, and still others who are just now finding Star Trek itself for the first time. What is it like to watch those episodes from the 1960s with modern eyes and sensibilities, without decades of seeing the mythos be built layer by layer, and the approach to storytelling evolve with the times? Can the expectations of today be set aside to allow the messages of Star Trek to shine through the veneer of yesterday? In this episode of Interphase, host C Bryan Jones is joined by Michael Pfeffer, who has just finished his first-ever viewing of Star Trek: The Original Series—an experience that was, in fact, his first contact with the Star Trek television franchise in general. How did he connect with the style, themes, storytelling, and attitudes of 1960s TV? Let’s find out. Chapters Intro (00:00:00) Connecting with Star Trek (00:02:42) Starting with TOS (00:05:48) Target Audience (00:09:12) More About Characters (00:16:01) Science, Technology, and Cultures (00:30:52) What Is Star Trek About? (00:38:34) Klingons and Romulans as Analogue (00:42:45) Temporal Insensitivity (00:48:06) Favorite Episodes (00:53:36) Final Thoughts (01:00:09) Closing (01:04:40) Host C Bryan Jones Guest Michael Pfeffer Production C Bryan Jones (Editor and Producer) Matthew Rushing (Executive Producer) Renee Roberts (Associate Producer)

Watching TOS for the first time, in 2020. For many of us, the five-year mission of Kirk, Spock, McCoy, and the crew of the USS Enterprise is what hooked us on Star Trek and made us lifelong fans. But there are many who have never seen The Original Series, and still others who are just now finding Star Trek itself for the first time. What is it like to watch those episodes from the 1960s with modern eyes and sensibilities, without decades of seeing the mythos be built layer by layer, and the approach to storytelling evolve with the times? Can the expectations of today be set aside to allow the messages of Star Trek to shine through the veneer of yesterday? In this episode of Interphase, host C Bryan Jones is joined by Michael Pfeffer, who has just finished his first-ever viewing of Star Trek: The Original Series—an experience that was, in fact, his first contact with the Star Trek television franchise in general. How did he connect with the style, themes, storytelling, and attitudes of 1960s TV? Let’s find out. Chapters Intro (00:00:00) Connecting with Star Trek (00:02:42) Starting with TOS (00:05:48) Target Audience (00:09:12) More About Characters (00:16:01) Science, Technology, and Cultures (00:30:52) What Is Star Trek About? (00:38:34) Klingons and Romulans as Analogue (00:42:45) Temporal Insensitivity (00:48:06) Favorite Episodes (00:53:36) Final Thoughts (01:00:09) Closing (01:04:40) Host C Bryan Jones Guest Michael Pfeffer Production C Bryan Jones (Editor and Producer) Matthew Rushing (Executive Producer) Renee Roberts (Associate Producer)

Lower Decks pre-launch speculation. Starfleet is preparing to become animated once again in the new CBS All Access series Star Trek: Lower Decks. Mike McMahan, writer and producer of the Cartoon Network animated comedy series Rick and Morty and creator of the Hulu animated series Solar Opposites, is introducing us to the people who really make a starship run. Apart from two 2019 installments of Short Treks—“Ephraim and Dot” and “The Girl Who Made the Stars”—Lower Decks marks the first “cartoon” incarnation of the franchise since Star Trek: The Animated Series (1973–74) and follows the support crew serving on one of Starfleet’s least important ships, the USS Cerritos, in the year 2380. In this premiere episode of Interphase, Trek.fm’s new Star Trek Universe podcast, host C Bryan Jones is joined by Hayley Stoddart, Zachary Fruhling, and Chris Chaplin to discuss our expectations, whether Star Trek is the right platform for comedy, our thoughts on the crew and the ship, and whether a fractured fandom can—or should—be reunited. Host C Bryan Jones Guests Hayley Stoddart, Zachary Fruhling, and Chris Chaplin Production C Bryan Jones (Editor and Producer) Matthew Rushing (Executive Producer) Renee Roberts (Associate Producer)

Lower Decks pre-launch speculation. Starfleet is preparing to become animated once again in the new CBS All Access series Star Trek: Lower Decks. Mike McMahan, writer and producer of the Cartoon Network animated comedy series Rick and Morty and creator of the Hulu animated series Solar Opposites, is introducing us to the people who really make a starship run. Apart from two 2019 installments of Short Treks—“Ephraim and Dot” and “The Girl Who Made the Stars”—Lower Decks marks the first “cartoon” incarnation of the franchise since Star Trek: The Animated Series (1973–74) and follows the support crew serving on one of Starfleet’s least important ships, the USS Cerritos, in the year 2380. In this premiere episode of Interphase, Trek.fm’s new Star Trek Universe podcast, host C Bryan Jones is joined by Hayley Stoddart, Zachary Fruhling, and Chris Chaplin to discuss our expectations, whether Star Trek is the right platform for comedy, our thoughts on the crew and the ship, and whether a fractured fandom can—or should—be reunited. Host C Bryan Jones Guests Hayley Stoddart, Zachary Fruhling, and Chris Chaplin Production C Bryan Jones (Editor and Producer) Matthew Rushing (Executive Producer) Renee Roberts (Associate Producer)

It's the conclusion of series 3, Eleanor, Nick and Steve are back in the shed to bicker about corona, dog corneas and the final postdoc soap box! A HUGE thank you to @Elamanor for her wonderful help with the pod, we'll miss you! If you like TheScienceShed, the best thing you can do is share it on social media, and to follow us all on twitter, and please leave us a rating or review on iTunes! twitter @SteveTheChemist, @theevanslab, @Elamanor & @TheScienceShed

This week, we’re diving into a science paper that was hard to miss. It’s all about creating artificial photosynthesis using spinach chloroplasts. We also talk about our favourite genome duplication events and news in the world of science.

Folge 032 - Mitose | Die Teilung des Zellkerns | Genetik Teil 4 Show Notes: Bitte unterstützt den Biologie Passion Podcast finanziell ➤ paypal.me/biologiepassionpdcst Hier gehts zum zugehörigen Blogartikel auf meiner Webseite. Wenn dir die Podcastfolge gefallen hat, würde mich eine kurze Bewertung auf iTunes freuen. Trag dich in meinen Newsletter ein, wenn du über neue Podcastfolgen informiert werden willst. Vielen Dank fürs Zuhören!

Folge 031 - Der Ablauf des Zellzyklus | Genetik Teil 3 Show Notes: Bitte unterstützt den Biologie Passion Podcast finanziell ➤ paypal.me/biologiepassionpdcst Hier gehts zum zugehörigen Blogartikel auf meiner Webseite. Wenn dir die Podcastfolge gefallen hat, würde mich eine kurze Bewertung auf iTunes freuen. Trag dich in meinen Newsletter ein, wenn du über neue Podcastfolgen informiert werden willst. Vielen Dank fürs Zuhören!

THIS WEEK:  Dr. Noah Snyder gives us an inspiring peek into Interphase Materials, a company that specializes in the many places where technology interfaces with biology, which makes for a wide-ranging set of applications.  During this week's show, Noah speaks with Scot MacTaggart about how Interphase improves outcomes in medicine, and improves the efficiency of both Navy ships and HVAC systems. Be sure to listen for Noah's advice about customer discovery, the pitfalls of startup fashion, and the quest for true purpose he thinks is key to his happiness as well as his success.

Ask the Mentors Episode Topic: What are you finding is working best for your clients the bezwecken Progonb-l4x Phytob-l4x or the Pro-adapt and est-adapt?Client Case: Helping female client balance their blood sugar if they are Type 1 Diabetic. Client has tried Paleo, keto, vegan. Experiencess really low life threatening blood sugar drops, but they are medically induced.  Is there any correlation to IgA Immunodeficiency detected with blood work and Sec IgA levels on the GI Map?Would Secretory IgA on GI Map be low if a person was taking immunosuppressant drugs?Decreasing Sec IgA, what are some strategies outside of these topics to increase Sec IgA?Amazon A-P and Amazon A-F, how to take, with or without food?Client Case: Female client who is severely constipated, doing the GI Map but wants some relief ahead of time while she is on the Interphase plus. Suggestions beside magnesium citrateDoes P5P and B6 have similar dosing? Dosing: 100 mg/day of Pyridoxine hydrochloride or 25+ mg/day of P-5-P  

It’s an “Ask The Mentors” episode! Today's topics: 1. Dutch Test:  Taking a deeper look into Testosterone, Estrogen and Cortisol 2. 205 Comparison Worksheet and the FDN optimum range 3. Boosting testosterone in women 4. 401H and E.Coli results 5. Hysterectomy: How to prepare physically and mentally 6. Working on gut specific challenges with clients that are breastfeeding 7. Biofilm Busters: Biocidin vs. Interphase  8. Parasite Cleanse: Deciding on which binders to use

Nabeel Hyatt is a venture partner at Spark Capital where he invests in entrepreneurs using that rare combination of design and technology to transform markets. He is currently on the board of Spark Capital’s investments in Cruise (acquired by General Motors), Fig, Harmonix, Postmates, Proletariat, and Thalmic Labs. Nabeel was previously cofounder and CEO of Conduit Labs, which was sold to Zynga in 2010, where he then became General Manager up through the IPO. Prior to that he was variously cofounder, head of product, and CEO at a variety of hardware and software companies including MIT Media Lab spin-out Ambient Devices, Teamtalk (BSkyB), and Interphase.    In Today’s Episode You Will Learn: 1.) How did Nabeel make his way into the world of VC? 2.) What are the biggest lessons Nabeel can apply from his time in the trenches to being a VC at Spark? How has his investment strategy and decision making changed over time? 3.) What were the biggest takeaways for Nabeel as an observer and investor in Oculus? 4.) What is the story behind Spark's investment in Cruise (recently acquired by GM)? How did Nabeel come to meet Kyle and the team? What was the product like? How did it evolve? 5.) What does Cruise’s acquisition mean for the autonomous car industry? What are the inherent challenges for the industry as a whole? How will they be overcome and what timeline are you placing on the industry to come into fruition? Items Mentioned In Today’s Episode:  Nabeel’s Fave Blog: AVC Nabeel’s Most Recent Investment: Fig  As always you can follow The Twenty Minute VC, Harry and Nabeel on Twitter here! If you would like to see a more colourful side to Harry with many a mojito session, you can follow him on Instagram here!   The Twenty Minute VC is brought to you by Leesa, the Warby Parker or TOMS shoes of the mattress industry. Lees have done away with the terrible mattress showroom buying experience by creating a luxury premium foam mattress that is order completely online and ships for free to your doorstep. The 10 inch mattress comes in all sizes and is engineered with 3 unique foam layers for a universal, adaptive feel, including 2 inches of memory foam and 2 inches of a really cool latex foam called Avena, design to keep you cool. All Leesa mattresses are 100% US or UK made and for every 10 mattresses they sell, they donate one to a shelter. Go to Leesa.com/VC and enter the promo code VC75 to get $75 off!  

Interphase centrosomes flare up Centrosomes undergo dramatic changes in size and structure during the rapid cell cycles of early Drosophila embryos. Lerit et al. reveal that a scaffold formed by the proteins centrosomin and PLP is required to maintain the activity of interphase centrosomes, which is essential for nuclear spacing and proper chromosome segregation. This biosights episode presents the paper by Lerit et al. from the July 6th, 2015, issue of The Journal of Cell Biology and includes an interview with two of the paper's authors, Dorothy Lerit and Nasser Rusan (National Heart, Lung, and Blood Institute, Bethesda, MD). Produced by Caitlin Sedwick and Ben Short. See the associated paper in JCB for details on the funding provided to support this original research. Subscribe to biosights via iTunes or RSS View biosights archive The Rockefeller University Press biosights@rockefeller.edu

Hi to all our listeners from all corners of the globe, we’ve got another bumper pack of fresh and interesting diverse music to fill you lugholes and in this edition we have introduced a couple of electronic prog rock tracks and yes there are some vocals, do I hear groans?, but in a more spacey, psychedelic way. On a sad note, we’ve lost another legend in the music world, Daevid Allen, the eccentric Australian-born guitarist, singer and poet who was an original member of the psychedelic rock group Soft Machine and went on to form Gong, an aggressively avant-gard band with Allen’s whimsical, mystical, sometimes impenetrable and often very funny lyrics, whose influence far exceeded its relatively modest success, recently passed away in Australia with cancer, (ones life is something to treasure, memories last forever) Lets get straight into the music we love to hear, with two tracks from Mac Of Bionight stunning new double album Memories Sequences Podcast playlist No62 Mac of BIOnight ‘Memories’ 1.56 (album Memories vol 01) *** http://www.macvibes.com/ Mac of BIOnight ’Soft Chaos’ 9.48 (album Memories vol 02) Moonbooter ‘Cosmonaut Leonow’ 18.37 (Live 4 CD set) http://www.mellowjet.com/ Moonbooter ‘Epic Moments’ 25.50 “ “ “ “ Mystical Light ‘Secrets Of Taklamakan’ 34.18 (album Beyond The Horizon) http://www.mysticallight.de/ Mystical Light’ ‘Eternal Peace’ 41.33 “ “ “ “ “ Digitonal ‘The Beating Of Her Heart’ 48.35 www.justmusic.com Forest Fang ‘Burnt Offerings’ 53.00 (album Letters To The Farthest Star) www.spottedpeccary.com Digitonal ‘Eighteen’ 64.12 (album Beautiful Broken) Michael Bruckner ‘Vagueness Part 1’ 68.17 (album Fog Music 35) https://michaelbrueckner.bandcamp.com/music *** Howard Givens & Craig Padilla ’Tide Of The Opposition Moon’ 78.10 ( album Life Flows Water) www.spottedpeccary.com Annemarie Borg ’Nacre’ 86.03 (album Iridescent Perceptions) http://www.annemarieborg.com Annemarie Borg ‘Requiem’ 93.00 Though Guild ‘The Ebbing Universe’ 98.12 (album Third Voyage) http://www.hrresonance.blogspot.com/ Thought Guild ‘Moon Blossom Meditation’ 105.16 “ “ “ Sine Amplitude ‘Arpworld’ 109.00 http://www.tma-music.com Sonic Federation ‘Arrival At The Village’ 115.54 (album The Village ) https://sonicfederation.bandcamp.com/ Numina & Zero Ohms ‘Secrets of the Treasure House of Stars’ 126.24 (album Broken Stars Thorough Brilliant Clouds) www.spottedpeccary.com Spacemegalithe ‘Live in Paris 21 February 2015’ 140.34 http://mowgli9.wix.com/spacemegalithe *** Ars & Sylphides ‘Detatch’ 147.05 (album Interphase 3) http://interphase.bandcamp.com/album/interphase-3-2015 Ars Sonor, M.Nomized, Mystified & Sylphides ‘Cellular Fugue’ 152.15 (album Interphase 3) Sound Awakener ‘Learning To Drown’ 153.43 (album September Traveler) http://soundawakener.weebly.com/ Mark Jenkins ‘RA8543’ 158.32 (album Change Of Cosmic Address) http://www.markjenkinsmusic.com Chris Jenkins ‘Ancient Chronicles Of Silent Souls’ 162.09 (album Change Of Cosmic Address) http://chrisjenkinse-music.bandcamp.com Edit ***

Willkommen in der Welt der Biologie! Mein Name ist Alia Korth und heute geht es um Mitose. Als Mitose bezeichnet man den Vorgang der Zellteilung bei Zellen, die einen Zellkern besitzen. Aus einer Mutterzelle werden also zwei Tochterzellen. Man teilt die Mitose in die folgenden 5 Phasen ein: In der Interphase werden die Chromosomen, also die DNS, verdoppelt. Während der Prophase ziehen sich die Chromatidfäden zusammen, es entstehen Paare, die von einem sich bildenden Zentromer zusammen gehalten werden. Anschließend wandern die Zentriolen zu den Polen, also den gegenüberliegenden Seiten des Zellkerns. Nun löst sich die Wand des Zellkerns auf. Im Anschluss daran kommt die Metaphase, in welcher sich die Chromosomen an der Äquatorialebene, der Mitte des Zellkerns, ausrichten. Es bilden sich Spindelfasern, welche zu den Zentromeren wandern. In dem Moment, in dem die Chromatiden der Chromosome auseinander gezogen werden, beginnt die Anaphase. Darauf folgend, in der Telophase, bildet sich die Zellwand des Zellkerns wieder, die Zentriolen bauen sich ab und die Äquatorialebene zieht sich zusammen. Es entstehen zwei Tochterzellen. Die eigentliche Mitose ist nun abgeschlossen, nun wachsen die Tochterzellen. Dies nennt man Zytokinese. Für die Phasen der Mitose gibt es verschiedene Merksprüche. Ich finde den Spruch “Ich pauke Mitose alle Tage.” am einfachsten, die Anfangsbuchstaben der Worte sind die Anfangsbuchstaben der einzelnen Phasen. Wenn euch die vielen Fachbegriffe, die wir in dieser Folge nicht alle ausführlich erklären konnten, teilweise noch nicht klar sind, dann schaut doch einfach in unserem Glossar auf www.in2minuten.com nach. Dort haben wir alle unklaren Begriffe noch mal kurz erklärt. Wenn ihr noch Fragen oder Anregungen habt, dann schreibt mir einfach eine E-Mail an biologie@in2minuten.com. Weitere “in 2 Minuten” Podcasts findet ihr auch im Internet unter www.in2minuten.com. Vielen Dank für’s Zuhören und bis zum nächsten Mal!

http://darienps.org/teachers/otterspoor/podcasts/cellcycle2.mp3Interphase and Mitosis

Verschiedene Arbeiten der letzten Jahre konnten zeigen, dass sich in vielen, verschiedenen Zelltypen genreiches, transkriptionell aktives und früh replizierendes Chromatin bevorzugt im Inneren der Zellkerne aufhält, während das genarme, transkriptionell inaktive und spät replizierende Chromatin vorrangig an der Zellkernperipherie zu finden ist. Dennoch ist bislang noch nicht wirklich verstanden, welche der Chromatineigenschaften, wie die lokale Gendichte, die Expression oder die Replikationszeit, tatsächlich einen ausschlaggebenden Einfluss auf die räumliche Anordnung im Zellkern haben und welche dieser Eigenschaften nur aufgrund ihrer Korrelation mit diesen „dominanten“ Merkmalen eine spezifische Verteilung im Interphasekern aufweisen. Um dieses Problem zu untersuchen, stellten wir Pools aus BAC Klonen von HSA 11, 12, 18 und 19 für R-und G-Banden-spezifische Regionen, genreiche bzw. genarme Segmente sowie für hoch bzw. niedrig exprimierte Gene zusammen. Mit Hilfe der multicolor 3D-FISH Technik, Bildverarbeitung und computergestützter, quantitativer Auswertungen wurde die Lage dieser BAC Pools im Zellkern sowie ihre Anordnung bezüglich ihrer Chromosomenterritorien analysiert. Sowohl in den humanen Lymphozyten wie in den humanen Fibroblasten fanden wir den R-Banden Pool, den genreichen Pool sowie den Pool, der die hoch exprimierten Gene enthielt, weiter im Zellkerninneren als ihre jeweils korrespondierenden Pools (G-Banden, genarmer Pool, bzw. niedrig exprimierte Gene). Für jeden BAC Pool wurde mittels sorgfältiger Datenbankrecherche die mittlere lokale Gendichte, der mittlere GC Gehalt, die Replikationszeit sowie das mittlere Expressionsniveau bestimmt. Anschließend wurde eine Korrelationsanalyse dieser Parameter mit der berechneten mittleren, relativen Position der Pools im Zellkern durchgeführt. Die höchste Korrelation ergab sich für die Gendichte, während wir zeigen konnten, dass das Expressionsniveau, die Zuordnung zu einer R- oder G.Bande, sowie das Replikationstiming offensichtlich so gut wie keinen Einfluss auf die radiale Anordnung des Chromatins im Zellkern hat. Diese radiale Positionierung der verschiedenen Pools spiegelte sich auch in ihrer Anordnung bezüglich der Chromosomenterritorien wieder. Diese zeigen eine polare Anordnung in Bezug auf den Zellkern: Genreiche Segmente waren zum Mittelpunkt des Zellkerns hin orientiert, während die genarmen Segmente in der Hälfte des CTs zu finden waren, die sich in Richtung der Peripherie erstreckte. Etwas weniger deutlich ausgeprägt wurde diese Anordnung auch für die R-/G-Banden Pools sowie für die von der transkriptionellen Aktivität abhängigen Pools beobachtet. Dies spricht für eine deutliche strukturelle Transformation bei der Umwandlung der Metaphasenchromosomen zu den CTs der Interphase, die Territorien haben eine hohe Plastizität. Wir konnten bestätigen, dass die extrem genreiche und hoch transkriptionell aktive Region 11p15.5 oft weit aus ihrem CT herausragt. Ein ähnliches Verhalten konnte jedoch nicht für die ebenfalls sehr genreichen und transkriptionell aktiven Segmente des Chromosoms 12 beobachtet werden, was gegen die Annahme spricht, das dieses Phänomen des „looping outs“ ein typische Anordnung für Chromatinabschnitte mit solchen extremen Eigenschaften ist. Wir konnten ebenfalls keine Unterschiede für die Verteilung der BAC Pools der Chromosomen 12, 18 und 19 bezüglich der Oberfläche der CTs finden. R- und G-Banden, genreiche und genarme Segmente sowie hoch und niedrig exprimierte Gene scheinen gleichmäßig im gesamten Territorium verteilt zu sein. Die äußere, das CT einschließende Oberfläche scheint entgegen der Erwartung offensichtlich kein wichtiger Reaktionsort für besonders genreiche bzw. hoch exprimierte Sequenzen zu sein.

The article reviews the existing methods of multicolor FISH on nuclear targets, first of all, interphase chromosomes. FISH proper and image acquisition are considered as two related components of a single process. We discuss (1) M-FISH (combinatorial labeling + deconvolution + widefield microscopy); (2) multicolor labeling + SIM (structured illumination microscopy); (3) the standard approach to multicolor FISH + CLSM (confocal laser scanning microscopy; one fluorochrome - one color channel); (4) combinatorial labeling + CLSM; (5) non-combinatorial labeling + CLSM + linear unmixing. Two related issues, deconvolution of images acquired with CLSM and correction of data for chromatic Z-shift, are also discussed. All methods are illustrated with practical examples. Finally, several rules of thumb helping to choose an optimal labeling + microscopy combination for the planned experiment are suggested. Copyright (c) 2006 S. Karger AG, Basel.

Tue, 7 Oct 2003 12:00:00 +0100 https://edoc.ub.uni-muenchen.de/1658/ https://edoc.ub.uni-muenchen.de/1658/1/Maierhofer_Christine.pdf Maierhofer, Christine ddc:570, ddc:500, Fa

In der Arbeit wurde die Methodik einer Kombination aus einer Immunzytochemischen Färbung und Fluoreszenz-in-situ-Hybridisierung (FISH) mit Sonden für 6p, 8p und 8q an Interpahsezellen etabliert. Anschließend erfolgte die Analyse von Tumorzelllinien und Primärkulturen, die aus Knochenmarkaspiraten von Patienten mit Prostatakarzinom gewonnen worden waren. Sowohl in den Zelllinien wie auch in den Primärkulturen der disseminierten Prostatakarzinomzellen wurden für alle drei untersuchten Chromosomenlokalisationen numerische Aberrationen entdeckt. In fast allen Tumorpräparaten zeigte sich eine verschieden große Zellpopulation mit normalem Hybridisierungsmuster (je zwei Signale für 6p, 8q und 8p) und mindestens ein Zellklon mit pathologischem Muster. Ein konsistentes Muster für 6p25, 8q24.3 und 8p23 Aberrationen, das in allen untersuchten Karzinomen vorhanden oder vorherrschend gewesen wäre, ließ sich nicht definieren. Dominierend war ein gemischtes Bild verschiedener Kombinationen von Aberrationen und eine ausgeprägte Heterogenität vorhanden. Ebenso wie an den in der Literatur beschriebenen Zellen aus Primärtumoren waren auch in den disseminierten Zellen 8p-Verluste und zum Teil 8q-Zugewinne und 6p-Zugewinne zu finden. Die Zugewinne dieser Chromosomenregionen waren überwiegend in den Zelllinien nachweisbar. Hervorzuheben ist der Verlust von 6p in sieben der elf Primärkulturen, eine Aberration, die bisher nur in einem einzigen Fall eines primären Prostatakarzinom-Gewebe beschrieben wurde.

Um die Anordnung von Chromosomen in Zellkernen der Interphase zu untersuchen, wurde ein Verfahren aus der Computergeometrie adaptiert. Dieser Ansatz basiert auf der Zerlegung von dreidimensionalen Bildvolumen mithilfe des Voronoi-Diagramms in konvexe Polyeder. Die graphenorientierte, geometrische Struktur dieses Verfahrens ermöglicht sowohl eine schnelle Extraktion von Objekten im Bildraum als auch die Berechnung morphologischer Parameter wie Volumina, Oberflächen und Rundheitsfaktoren. In diesem Beitrag wird exemplarisch die dreidimensionale Morphologie von XChromosomen in weiblichen Interphasezellkernen mithilfe dieser drei Parameter untersucht. Um diese Zellkerne mit lichtoptischen Methoden zu untersuchen, wurden die Territorien der X-Chromosomen mit einem molekularcytogenetischen Verfahren fluoreszierend dargestellt. Zur Unterscheidung des aktiven und inaktiven X-Chromosoms wurde das Barr-Körperchen zusätzlich markiert und mithilfe eines Epifluoreszenzmikroskops, ausgerüstet mit einer CCD-Kamera, aufgenommen. Anschließend wurden 1 2 - 2 5 äquidistante, lichtoptische Schnitte der X-Chromosomenterritorien mit einem konfokalen Laser Scanning Mikroskop (CLSM) aufgenommen. Diese lichtoptischen Schnitte wurden mithilfe des Voronoi-Verfahrens segmentiert und analysiert. Methoden aus der Computergraphik wurden zur Visualisierung der Ergebnisse eingesetzt. Es konnte gezeigt werden, daß mithilfe des Voronoi-Verfahrens Chromosomen- Territorien anhand der morphologischen Parameter zuverlässig beschrieben werden können.

We report on multicolor fluorescence in situ hybridization protocols for the simultaneous visualization of deletion-prone regions for carrier detection of Duchenne/ Becker (DMD/BMD) muscular dystrophy. Cosmid and yeast artificial chromosome (YAC) clones specific for preferentially deleted subregions of the dystrophin gene were labeled differentially and detected with three different fluorochromes using digital imaging microscopy. This approach allows for an assessment of the carrier status of female relatives even in families where no index patient is available. Cosmid and YAC clones, and different probe-generation protocols are compared with respect to their feasibility for carrier detection. The use of histone-depleted interphase nuclei (Halo-preparations) for deletion mapping is demonstrated and shown to have a resolution power of 5 kb.

Fri, 1 Jan 1993 12:00:00 +0100 https://epub.ub.uni-muenchen.de/9386/1/9386.pdf Ward, D. C.; Cremer, Thomas; Fischer, C.; Lipp, Martin; Lengauer, Christoph; Ried, Thomas

A human yeast artificial chromosome (YAC) library was screened by polymerase chain reaction with oligonucleotide primers defined for DNA sequences of the BCR gene and the protooncogenes c-raf-1, c-fms, and c-erB-2. Alu-PCR-generated human DNA sequences were obtained from the respective YAC clones and used for fluorescence in situ hybridization experiments under suppression conditions. After chromosomal in situ suppression hybridization to GTG-banded human prometaphase chromosomes, seven of nine initially isolated YAC clones yielded strong signals exclusively in the chromosome bands containing the respective genes. Two clones yielded additional signals on other chromosomes and were excluded from further tests. The band-specific YACs were successfully applied to visualize specific structural chromosome aberrations in peripheral blood cells from patients with myelodysplasia exhibiting del(5)(q13q34), chronic myeloid leukemia and acute lymphocytic leukemia with t(9;22)(q34;q11), acute promyelocytic leukemia (M3) with t(15;17)(q22;q21), and in a cell line established from a proband with the constitutional translocation t(3;8)(p14.2;q24). In addition to the analysis of metaphase spreads, we demonstrate the particular usefulness of these YAC clones in combination with whole chromosome painting to analyze specific chromosome aberrations directly in the interphase nucleus.

Triple fluorescence in situ hybridization with a plasmid DNA library from sorted human chromosomes 8 in combination with bacteriophage clones flanking the breakpoint in 8q24 of the Burkitt lymphoma cell line Jl was used for the specific delineation of this breakpoint in individual tumor cells. With this approach, tumor-specific breakpoints in translocation chromosomes can be detected at all stages of the cell cycle with high specificity.

Tue, 1 Jan 1991 12:00:00 +0100 https://epub.ub.uni-muenchen.de/9352/1/9352.pdf Cremer, Christoph; Stelzer, Ernst; Wienberg, Johannes; Jauch, Anna; Kharboush, I.; Remm, B.; Cremer, Thomas

Mon, 1 Jan 1990 12:00:00 +0100 https://epub.ub.uni-muenchen.de/9401/1/9401.pdf Cremer, Thomas; Jauch, Anna; Emmerich, Patricia; Walt, H. dd

Six patients, five with acute myeloid leukemia (AML) and one with a myelodysplastic syndrome (MDS), were found to have monosomy 7 by conventional cytogenetics at diagnosis. Repetitive DNA sequences from the heterochromatic region of human chromosomes 1 and 7 were used as probes for in situ hybridization experiments on interphase cells of these patients. A double hybridization protocol was used to reveal the particular chromosomes as distinct spots or clusters of signals within interphase nuclei. The chromosome 1 sequence served as an internal control. Simultaneous detection of the sequences showed the presence of two normal number 1 chromosomes and a missing 7 chromosome from individual cells. While cytogenetic preparations showed only -7 metaphases in 3 AML and 1 MDS patients, in situ hybridization of interphase cells showed many normal cells as well as the presence of -7 in fully mature granulocytes. One AML patient studied in remission showed only normal metaphases yet had 9% interphase cells with a missing 7 and relapsed within 3 months. We conclude that examination of interphase cells by in situ hybridization provides clinically useful data since every cell including mature granulocytes can be examined, the lineage of a cell can be determined, and efficacy of differentiation therapy can be evaluated.

Chromosomal in situ suppression (CISS)-hybridization of biotinylated phage DNA-library inserts from sorted human chromosomes was used to decorate chromosomes 1 and 7 specifically from pter to qter and to detect structural aberrations of these chromosomes in irradiated human peripheral lymphocytes. In addition, probe pUC1.77 was used to mark the Iq12 subregion in normal and aberrant chromosomes 1. Low LET radiation (60Co--rays; 1.17 and 1.33 MeV) of lymphocyte cultures was performed with various doses (D = 0, 2, 4, 8 Gy) 5 h after stimulation with phytohaemagglutinin. Irradiated cells were cultivated for an additional 67 h before Colcemid arrested metaphase spreads were obtained. Aberrations of the specifically stained chromosomes, such as deletions, dicentrics, and rings, were readily scored after in situ hybridization with either the 1q12 specific probe or DNA-library inserts. By the latter approach, translocations of the specifically stained chromosomes could also be reliably assessed. A linear increase of the percentage of specifically stained aberrant chromosomes was observed when plotted as a function of the square of the dose D. A particular advantage of this new approach is provided by the possibility to delineate numerical and structural chromosome aberrations directly in interphase nuclei. These results indicate that cytogenetic monitoring of ionizing radiation may be considerably facilitated by CISS-hybridization.

DNA libraries from sorted human gonosomes were used selectively to stain the X and Y chromosomes in normal and aberrant cultured human cells by chromosomal in situ suppression (CISS-) hybridization. The entire X chromosome was stained in metaphase spreads. Interphase chromosome domains of both the active and inactive X were clearly delineated. CISS-hybridization of the Y chromosome resulted in the specific decoration of the euchromatic part (Ypter-q11), whereas the heterochromatic part (Yq12) remained unlabeled. The stained part of the Y chromosome formed a compact domain in interphase nuclei. This approach was applied to amniotic fluid cells containing a ring chromosome of unknown origin (47,XY; +r). The ring chromosome was not stained by library probes from the gonosomes, thereby suggesting its autosomal origin. The sensitivity of CISS-hybridization was demonstrated by the detection of small translocations and fragments in human lymphocyte metaphase spreads after irradiation with 60Co-gamma-rays. Lymphocyte cultures from two XX-males were investigated by CISS-hybridization with Y-library probes. In both cases, metaphase spreads demonstrated a translocation of Yp-material to the short arm of an X chromosome. The translocated Y-material could also be demonstrated directly in interphase nuclei. CISS-hybridization of autosomes 7 and 13 was used for prenatal diagnosis in a case with a known balanced translocation t(7;13) in the father. The same translocation was observed in amniotic fluid cells from the fetus. Specific staining of the chromosomes involved in such translocations will be particularly important, in the future, in cases that cannot be solved reliably by conventional chromosome banding alone.

In situ hybridization of human chromosome 18 and X-specific alphoid DNA-probes was performed in combination with three dimensional (3D) and two dimensional (2D) image analysis to study the interphase distribution of the centric heterochromatin (18c and Xc) of these chromosomes in cultured human cells. 3D analyses of 18c targets using confocal laser scanning microscopy indicated a nonrandom disposition in 73 amniotic fluid cell nuclei. The shape of these nuclei resembled rather flat cylinders or ellipsoids targets were preferentially arranged in a domain around the nuclear center, but close to or associated with the nuclear envelope. Within this domain, however, positionings of the two targets occurred independently from each other, i.e., the two targets were observed with similar frequencies at the same (upper or lower) side of the nuclear envelope as those on opposite sides. This result strongly argues against any permanent homologous association of 18c. A 2D analytical approach was used for the rapid evaluation of 18c positions in over 4000 interphase nuclei from normal male and female individuals, as well as individuals with trisomy 18 and Bloom's syndrome. In addition to epithelially derived amniotic fluid cells, investigated cell types included in vitro cultivated fibroblastoid cells established from fetal lung tissue and skin-derived fibroblasts. In agreement with the above 3D observations 18c targets were found significantly closer (P < 0.01) to the center of the 2D nuclear image (CNI) and to each other in all these cultures compared to a random distribution derived from corresponding ellipsoid or cylinder model nuclei. For comparison, a chromosome X-specific alphoid DNA probe was used to investigate the 2D distribution of chromosome X centric heterochromatin in the same cell types. Two dimensional Xc-Xc and Xc-CNI distances fit a random distribution in diploid normal and Bloom's syndrome nuclei, as well as in nuclei with trisomy X. The different distributions of 18c and Xc targets were confirmed by the simultaneous staining of these targets in different colors within individual nuclei using a double in situ hybridization approach.

Double in situ hybridization with mercurated and biotinylated chromosome specific DNA probes in combination with digital image analysis provides a new approach to compare the distribution of homologous and nonhomologous chromosome targets within individual interphase nuclei. Here we have used two DNA probes representing tandemly repeated sequences specific for the constitutive heterochromatin of the human chromosomes 1 and 15, respectively, and studied the relative arrangements of these chromosome targets in interphase nuclei of human lymphocytes, amniotic fluid cells, and fibroblasts, cultivated in vitro. We have developed a 2D-image analysis approach which allows the rapid evaluation of large numbers of interphase nuclei. Models to test for a random versus nonrandom distribution of chromosome segments are discussed taking into account the three-dimensional origin of the evaluated 2D-distribution. In all three human diploid cell types the measurements of target-target and target-center distances in the 2D-nuclear image revealed that the labeled segments of the two chromosomes 15 were distributed both significantly closer to each other and closer to the center of the nuclear image than the labeled chromosome 1 segments. This result can be explained by the association of nucleolus organizer regions on the short arm of chromosome 15 with nucleoli located more centrally in these nuclei and does not provide evidence for a homologous association per se. In contrast, evaluation of the interphase positioning of the two chromosome 1 segments fits the random expectation in amniotic fluid and fibroblast cells, while in experiments using lymphocytes a slight excess of larger distances between these homologous targets was occasionally observed. 2D-distances between the labeled chromosome 1 and 15 segments showed a large variability in their relative positioning. In conclusion our data do not support the idea of a strict and permanent association of these homologous and nonhomologous targets in the cell types studied so far.

Sun, 1 Jan 1989 12:00:00 +0100 https://epub.ub.uni-muenchen.de/9331/1/9331.pdf Walt, H.; Cremer, Thomas; Hofmann, M. C.; Jauch, Anna; Emmerich, Patricia

Sun, 1 Jan 1989 12:00:00 +0100 https://epub.ub.uni-muenchen.de/9333/1/9333.pdf Bannwart, F.; Hofmann, M. C.; Cremer, Thomas; Emmerich, Patricia; Walt, H.

A method of in situ hybridization for visualizing individual human chromosomes from pter to qter, both in metaphase spreads and interphase nuclei, is reported. DNA inserts from a single chromosomal library are labeled with biotin and partially preannealed with a titrated amount of total human genomic DNA prior to hybridization with cellular or chromosomal preparations. The cross-hybridization of repetitive sequences to nontargeted chromosomes can be markedly suppressed under appropriate preannealing conditions. The remaining single-stranded DNA is hybridized to specimens of interest and detected with fluorescent or enzymelabeled avidin conjugates following post-hybridization washes. DNA inserts from recombinant libraries for chromosomes 1, 4, 7, 8, 13, 14, 18, 20, 21, 22, and X were assessed for their ability to decorate specifically their cognate chromosome; most libraries proved to be highly specific. Quantitative densitometric analyses indicated that the ratio of specific to nonspecific hybridization signal under optimal preannealing conditions was at least 8:1. Interphase nuclei showed a cohesive territorial organization of chromosomal domains, and laserscanning confocal fluorescence microscopy was used to aid the 3-D visualization of these domains. This method should be useful for both karyotypic studies and for the analysis of chromosome topography in interphase cells.

Chromosome aberrations in two glioma cell lines were analyzed using biotinylated DNA library probes that specifically decorate chromosomes 1, 4, 7, 18 and 22 from pter to qter. Numerical changes, deletions and rearrangements of these chromosomes were radily visualized in metaphase spreads, as well as in early prophase and interphase nuclei. Complete chromosomes, deleted chromosomes and segments of translocated chromosomes were rapidly delineated in very complex karyotypes. Simultaneous hybridizations with additional subregional probes were used to further define aberrant chromosomes. Digital image analysis was used to quantitate the total complement of specific chromosomal DNAs in individual metaphase and interphase cells of each cell line. In spite of the fact that both glioma lines have been passaged in vitro for many years, an under-representation of chromosome 22 and an over-representation of chromosome 7 (specifically 7p) were observed. These observations agree with previous studies on gliomas. In addition, sequences of chromosome 4 were also found to be under-represented, especially in TC 593. These analyses indicate the power of these methods for pinpointing chromosome segments that are altered in specific types of tumors.

Repeated DNAs from the constitutive heterochromatin of human chromosomes 1 and 18 were used as probes in nonradioactive in situ hybridization experiments to define specific numerical and structural chromosome aberrations in three human glioma cell lines and one neuroblastoma cell line. The number of spots detected in interphase nuclei of these tumor cell lines and in normal diploid nuclei correlated well with metaphase counts of chromosomes specifically labeled by in situ hybridization. Rapid and reliable assessments of aneuploid chromosome numbers in tumor lines in double hybridization experiments were achieved, and rare cells with bizarre phenotype and chromosome constitution could be evaluated in a given tumor cell population. Even with suboptimal or rare chromosome spreads specific chromosome aberrations were delineated. As more extensive probe sets become available this approach will become increasingly powerful for uncovering various genetic alterations and their progression in tumor cells.

Thu, 1 Jan 1987 12:00:00 +0100 https://epub.ub.uni-muenchen.de/9324/1/9324.pdf Cremer, Christoph; Schlegel, W.; Emmerich, Patricia; Jauch, Anna; Hausmann, Michael; Cremer, Thomas; Loos, Peter

The localization of chromosome 18 in human interphase nuclei is demonstrated by use of radioactive and nonradioactive in situ hybridization techniques with a DNA clone designated L1.84. This clone represents a distinct subpopulation of the repetitive human alphoid DNA family, located in the centric region of chromosome 18. Under stringent hybridization conditions hybridization of L1.84 is restricted to chromosome 18 and reflects the number of these chromosomes present in the nuclei, namely, two in normal diploid human cells and three in nuclei from cells with trisomy 18. Under conditions of low stringency, cross-hybridization with other subpopulations of the alphoid DNA family occurs in the centromeric regions of the whole chromosome complement, and numerous hybridization sites are detected over interphase nuclei. Detection of chromosome-specific target DNAs by non-radioactive in situ hybridization with appropriate DNA probes cloned from individual chromosomal subregions presents a rapid means of identifying directly numerical or even structural chromosome aberrations in the interphase nucleus. Present limitations and future applications of interphase cytogenetics are discussed.

In spite of Carl Rabl's (1885) and Theodor Boveri's (1909) early hypothesis that chromosomes occupy discrete territories or domains within the interphase nucleus, evidence in favor pf this hypothesis has been limited and indirect so far in higher plants and animals. The alternative possibility that the chromatin fiber of single chromosomes might be extended throughout the major part of even the whole interphase nucleus has been considered for many years. In the latter case, chromosomes would only exist as discrete chromatin bodies during mitosis but not during interphase. Both possibilities are compatible with Boveri's well established paradigm of chromosome individuality. Here we show that an active human X chromosome contained as the only human chromosome in a Chinese hamster x man hybrid cell line can be visualized both in metaphse plates and in interphase nuclei after in situ hybridization with either 3H- or biotin-labeled human genomic DNA. We demonstrate that this chromosome is organized as a distinct chromatin body throughout interphase. In addition, evidence for the territorial organization of human chromosomes is also presented for another hybrid cell line containing several autosomes and the human X chromosome. These findings are discussed in the context of our present knowledge of the organization and topography of interphase chromosomes. General applications of a strategy aimed at specific staining of individual chromosomes in experimental and clinical cytogenetics are briefly considered.

Two cloned repetitive DNA probes, pXBR and CY1, which bind preferentially to specific regions of the human X and Y chromosome, respectively, were used to study the distribution of the sex chromosomes in human lymphocyte nuclei by in situ hybridization experiments. Our data indicate a large variability of the distances between the sex chromosomes in male and female interphase nuclei. However, the mean distance observed between the X and Y chromosome was significantly smaller than the mean distance observed between the two X-chromosomes. The distribution of distances determined experimentally is compared with three model distributions of distances, and the question of a non-random distribution of sex chromosomes is discussed. Mathematical details of these model distributions are provided in an Appendix to this paper. In the case of a human translocation chromosome (XqterXp22.2::Yq11Y qter) contained in the Chinese hamster x human hybrid cell line 445 x 393, the binding sites of pXBR and CY1 were found close to each other in most interphase nuclei. These data demonstrate the potential use of chromosome-specific repetitive DNA probes to study the problem of interphase chromosome topography.

Sun, 1 Jan 1984 12:00:00 +0100 https://epub.ub.uni-muenchen.de/9318/1/9318.pdf Cremer, Christoph; Nakanishi, K.; Baumann, Hella; Cremer, Thomas

In 1885 Carl Rabl published his theory on the internal structure of the interphase nucleus. We have tested two predictions of this theory in fibroblasts grown in vitro from a female Chinese hamster, namely (1) the Rabl-orientation of interphase chromosomes and (2) the stability of the chromosome arrangement established in telophase throughout the subsequent interphase. Tests were carried out by premature chromosome condensation (PCC) and laser-UV-microirradiation of the interphase nucleus. Rabl-orientation of chromosomes was observed in G1 PCCs and G2 PCCs. The cell nucleus was microirradiated in G1 at one or two sites and pulse-labelled with 3H-thymidine for 2h. Cells were processed for autoradiography either immediately thereafter or after an additional growth period of 10 to 60h. Autoradiographs show unscheduled DNA synthesis (UDS) in the microirradiated nuclear part(s). The distribution of labelled chromatin was evaluated in autoradiographs from 1035 cells after microirradiation of a single nuclear site and from 253 cells after microirradiation of two sites. After 30 to 60h postincubation the labelled regions still appeared coherent although the average size of the labelled nuclear area fr increased from 14.2% (0h) to 26.5% (60h). The relative distance dr, i.e. the distance between two microirradiated sites divided by the diameter of the whole nucleus, showed a slight decrease with increasing incubation time. Nine metaphase figures were evaluated for UDS-label after microirradiation of the nuclear edge in G1. An average of 4.3 chromosomes per cell were labelled. Several chromosomes showed joint labelling of both distal chromosome arms including the telomeres, while the centromeric region was free from label. This label pattern is interpreted as the result of a V-shaped orientation of these particular chromosomes in the interphase nucleus with their telomeric regions close to each other at the nuclear edge. Our data support the tested predictions of the Rabl-model. Small time-dependent changes of the nuclear space occupied by single chromosomes and of their relative positions in the interphase nucleus seem possible, while the territorial organization of interphase chromosomes and their arrangement in general is maintained during interphase. The present limitations of the methods used for this study are discussed.

Unsynchronized cells of an essentially diploid strain of female Chinese hamster cells derived from lung tissue (CHL) were laser-UV-microirradiated (=257 nm) in the nucleus either at its central part or at its periphery. After 7–9 h postincubation with 0.5 mM caffeine, chromosome preparations were made in situ. Twenty-one and 29 metaphase spreads, respectively, with partial chromosome shattering (PCS) obtained after micro-irradiation at these two nuclear sites, were Q-banded and analyzed in detail. A positive correlation was observed between the frequency of damage of chromosomes and both their DNA content and length at metaphase. No significant difference was observed between the frequencies of damage obtained for individual chromosomes at either site of microirradiation. The frequency of joint damage of homologous chromosomes was low as compared to nonhomologous ones. Considerable variation was noted in different cells in the combinations of jointly shattered chromosomes. Evidence which justifies an interpretation of these data in terms of an interphase arrangement of chromosome territories is discussed. Our data strongly argue against somatic pairing as a regular event, and suggest a considerable variability of chromosome positions in different nuclei. However, present data do not exclude the possibility of certain non-random chromosomal arrangements in CHL-nuclei. The interphase chromosome distribution revealed by these experiments is compared with centromere-centromere, centromere-center and angle analyses of metaphase spreads and the relationship between interphase and metaphase arrangements of chromosomes is discussed.

Laser UV microirradiation of Chinese hamster interphase cells combined with caffeine post-treatment produced different patterns of chromosome damage in mitosis following irradiation of a small area of the nucleus that may be classified in three categories: I) intact metaphase figures, II) chromosome damage confined to a small area of the metaphase spread, III) mitotic figures with damage on all chromosomes. Category III might be the consequence of a non-localized distortion of nuclear metabolism. By contrast, category II may reflect localized DNA damage induced by microirradiation, which could not be efficiently repaired due to the effect of caffeine. If this interpretation is right, in metaphase figures of category II chromosome damage should occur only at the irradiation site. The effect might then be used to investigate neighbourhood relationships of individual chromosomes in the interphase nucleus.