Podcasts about Metaphase

Connect to John Gilroy on LinkedIn   https://www.linkedin.com/in/john-gilroy/ Want to listen to other episodes? www.Federaltechpodcast.com Brian Funk from Metaphase summarizes his company in one sentence, “It's all about meeting – meeting the mission with the technology.” What makes his company unique is the focus on drawing down costs in an efficient way. That is a great concept and has worked for Metaphase since its founding in 2013, but today we are uncertainly living in a world of policy. The question to ask, how does Metaphase operate in a world where the next six months are almost impossible to predict. Brian Funk's response is that they support over twenty agencies, it has given them a range of experience so they can select from a wide range of solutions. One example he gives includes a rapid response to a DHS RFI. Instead of sketching a possible solution, Metaphase delivered a fully functional application. That in and of itself, is a demonstration of being able to rapidly adapt to unpredictable situations. Funk also discusses the need for guardrails in AI usage and the potential for AI to enhance both efficiency and security in federal IT.

Drekker Brewing Metaphase is a Double New England IPA by style. Metaphase is brewed with oats and spelt for a fluffy, pillowy texture then double dry hopped with a pile of Citra & Nelson Sauvin. This killer hop combo leads to some luscious notes of orange, pineapple, papaya, & white grape with a beautifully dry finish. This craft beer is 8.3% ABV. In this craft beer review, we will take a look at the color, smell, and taste.

Link to bioRxiv paper: http://biorxiv.org/cgi/content/short/2023.07.31.551204v1?rss=1 Authors: Chen, J. K., Liu, T., Cai, S., Ruan, W., Ng, C. T., Shi, J., Surana, U., Gan, L. Abstract: The structure of chromatin at the nucleosome level inside cells is mysterious. Here we present in situ cryo-ET analyses of chromatin in both G1 and metaphase RPE-1 cells. G1 nucleosomes are concentrated in globular chromatin domains and metaphase nucleosomes are concentrated in the chromatids. Classification analysis reveals that canonical mononucleosomes, ordered stacked dinucleosomes, and mononucleosomes with a disordered gyre-proximal density are abundant in both cell-cycle states. Class averages that have more than two stacked nucleosomes or that have side-by-side dinucleosomes are not detected, suggesting that groups of more than two nucleosomes are heterogeneous. Large multi-megadalton structures are abundant in G1 nucleoplasm, but not found in G1 chromatin domains and metaphase chromatin. The macromolecular phenotypes studied here represent a starting point for the comparative analysis of condensation in normal and unhealthy human cells, in other cell-cycle states, other organisms, and in vitro chromatin assemblies. 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/2023.02.26.530101v1?rss=1 Authors: Zhang, C., Wang, Y., Sun, M., Xin, G., Yang, B., Jiang, Q. Abstract: Precise chromosome congression and segregation require proper assembly of a steady-state metaphase spindle, which is dynamic and maintained by continuous microtubule flux. NuSAP is a microtubule-stabilizing and -bundling protein that promotes chromosome-dependent spindle assembly. However, its function in spindle dynamics remains unclear. Here, we demonstrate that NuSAP regulates the dynamics and length control of the metaphase spindle. Mechanistically, NuSAP facilitates kinetochore capture and spindle assembly via promoting Eg5 binding with microtubules. It also prevents excessive microtubule depolymerization through interacting with Kif2A and reduces its spindle-pole localization. NuSAP is phosphorylated by Aurora A at Ser-240 during mitosis, and this phosphorylation promotes its interaction with Kif2A on the spindle body and reduces its localization to the spindle poles, thus maintaining the proper spindle microtubule flux. NuSAP knockout resulted in shorter spindle formation with faster microtubule flux and chromosome misalignment. Taken together, we uncover that NuSAP participates in spindle assembly, dynamics, and metaphase spindle length control via affecting microtubule flux and Kif2A localization. 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/2022.11.07.515476v1?rss=1 Authors: Soler, N., Chesneau, L., Bouvrais, H., Pastezeur, S., Le Marrec, L., Pecreaux, J. Abstract: The microtubule array, assembled into the mitotic spindle, polymerises from the centrosomes and the chromosomes in many organisms. Their plus ends alternate between growing and shrinking. This dynamic instability plays a key role in pulling on the kinetochores to check the spindle assembly and correct the errors in chromosome attachments. In addition, the minus ends at centrosomes can undergo depolymerisation coordinated with the polymerisation of the plus ends at the kinetochores. Such a mechanism, among others, creates treadmilling, id est a net poleward movement of microtubules called poleward flux. This flux is involved in many roles, chromosome congression in prometaphase, chromosome misattachment detection and correction, spindle length maintenance in metaphase, and synchronous segregation of sister chromatids in anaphase. Interestingly, no poleward flux was measured in the Caenorhabditis elegans single-cell embryo, despite it is equipped with all homologous proteins involved in this mechanism in other organisms. To investigate this peculiarity, we labelled the microtubules and photobleached them in a rectangular region. Surprisingly, we observed that both edges of the bleached zone (fronts) move inwards, closing the dark area. However, the middle of the bleached zone does not move clearly, confirming the absence of a global poleward flow. The dynamics of the microtubules emanating from the centrosomes combined with the diffraction due to microscopy imaging account for the apparent movement of the front on the centrosome side. Therefore, we suggest no flux of the centrosome-anchored (spindle) microtubules. In contrast, on the chromosome side, we observed a front moving poleward, faster than the one on the other side, and dependent on proteins ensuring the attachment and growth of microtubules at kinetochores, NDC-80, CLS-2CLASP, and ZYG-9XMAP215. Besides, we found that the depletion of the depolymerase KLP-7MCAK does not impair this poleward recovery. Finally, the faster recovery is restricted to the spindle region close to the chromosomes. Therefore, we suggest that the kinetochore microtubules undergo a poleward flux, moving with respect to spindle microtubules. Because the kinetochore microtubules are shorter than the half-spindle, this flux is localised close to the chromosomes. Furthermore, it may not rely on treadmilling as KLP-7MCAK is dispensable. This spatially restricted flux found in the nematode may be related to the slow elongation of the spindle during metaphase and may buffer the strong pulling forces exerted by the cortical force generators at the spindle poles. 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 red Solo Cup, Gatorade sport bottle, Harry's Razor and many more common household products were designed in St. Louis. Metaphase Design Group founder and CEO Bryce Rutter's 30-year-old company has quietly been working with some of the most recognizable companies in the world, and he joined the show to share the stories behind his company's success.

Metaphase is upon us! Join us as we play Beam Saber this season. Everything is brand new and is the perfect opportunity to start listening. This season we will follow an up and coming squad known as the Thundermorphs and their rag tag pilots. The post Metaphase 01: The One Way Trip Pt 1. appeared first on Geekspective.

Metaphase is upon us! Josh, Amber, Dave, and Kyle start the newest full season of Tales From Mauxferry now! Join them as they create their pilots and squad for Beam Saber! The post Metaphase 00: Beam Saber Character and Squad Creation appeared first on Geekspective.

This is Draw The Line Radio Show with Jacki-e, presenting the best music from female producers and DJs. Helping me Draw the Line with her guest mix in the second hour, it's Mizzrazz who's from Mannheim in Germany. She's co-founder of Side Effect, a collective for electronic dance music. Tech house and melodic techno are her favourite genres and her sets feature groovy, rhythms and intense basslines. She is currently learning music production and will be releasing her own tracks soon. Links for Mizzrazz:- Soundcloud:- https://soundcloud.com/mizzrazz Facebook:- https://www.facebook.com/mizzrazz In my mix in the first hour I'm playing tracks by Carly Wilford, Erika Krall, Beth Lydi, Miss Melera, Demah and lots more. If you like the tracks we play, please support the artists by buying their music. It's time to say NO to gender imbalance in dance music. It's time to Draw The Line!! Draw The Line Radio Show is produced for radio by Sergio Erridge and is A Darker Wave production. Track list 1st hour mixed by Jacki-E 1. Erika Krall – Hipazon (original mix) Trip & Dream. 2. Miss Melera – Stunning (original mix) Parquet Recordings. 3. Sascha Wieberknecht, Reanna Peris – Detroit Frequency (original mix) Wechselstrom Music. 4. Andrey Pitkin, Christina Novelli – Talking To You (No Hopes remix) Promo Party. 5. Clarity – Girl Get That (original mix) Confession. 6. La Fleur – Eavesdropper (original mix) Power Plant Records. 7. Beth Lydi – Pull Through (original mix) Plump Records. 8. Mai, Mai, Carly Wilford – Together for the Love (original mix) Basement Sounds. 9. Isabelle Gaultier – Ping Pong (original mix) AnalogMusiq promo to be released in September 2021. 10. Erika Krall – Losing Myself (original mix) Trip & Dream. 11. Demah – Love or Lust (Black Chapel remix) DeepDownDirty. 12. Puls'em – Perc ‘n' Ride (original mix) Crackhouse Recordings. 13. Carly Wilford – Generation X ft Mr V (original mix) Toolroom. 14. Mila Dietrich – Human Fate (original mix) Techgnosis Records. 15. Keah, Zephixx – Sky (original mix) Pure Dope Digital. 16. Jesika Jane, NEM3SI$ – The Pusher (Chris Coles and Latex Zebra remix) Onhcet Republik promo. 2nd hour Mizzrazz – An exclusive guest mix for Draw The Line Radio Show. 1. Ackermann - So Hot (original mix) IGNITE! Records. 2. Brothers Black - Annexe (Hiver remix) Bade Records. 3. Heiko Laux - R U Rec (original mix) Kanzleramt Music. 4. Michael Klein - Blk Drp 301 (original mix) BLK DRP. 5. Paride Saraceni - Cold Summer (original mix) Truesoul. 6. Tale Of Us – Anywhere (original mix) Afterlife. 7. Amotik – Chau (original mix) Amotik. 8. Pierre Blanche - Ares (original mix) Davotab Records. 9. The Micronaut - Jet (Marquez III remix) Voltage Musique. 10. Analog Agenda - Cloud by Cloud (Kape Blanc remix) Supdub Records. 11. Steam Shape - Nucleus (original mix) !Organism Rec. 12. Steam Shape – Ocsid (original mix) Metaphase. 13. Luigi Madonna - Unconditional Beauty (original mix) Drumcode. 14. Adam Beyer, Layton Giordani & Green Velvet - Data Point (original mix) Drumcode. 15. Cari Lekebusch - Fire Alarm (original mix) Drumcode

On the podcast today I have Naomi Gillespie a Naturopath, Functional Medicine Practitioner, Remedial Massage Therapist, Acupuncturist, and Founder of Origin Pain Relief Therapy. She has owned and operated clinics and treated 1000's of patients in the Gold Coast, Northern NSW, Western Australia, and Mornington Victoria.She is a keynote speaker on joy, work/life balance, psychological safety, and holistic wellbeing. She has lectured for large corporations, retail chains, government institutions, and the corporate health sector. Naomi has also been working on bodies for over 26 years and has treated over 80 000 people. Yep, that's a lot! She has developed her own pain relief therapy that is quite amazing and corrects pain in minutes within a couple of sessions. It sounds crazy but it's true. world. She is also the founder and creator of Badass Beat the Bulge, the Metaphase Method, author, and director at abundance health.Naomi has tried every diet you can think of, you name it, Yet she did not find any success losing weight. So for the last 20 years, she experimented and researched science around nutrition, exercise, metabolism, and hormones. She has created a program using the best science out there that uses your body to lose weight.Diets don't work because they're one-size-fits-all and mostly designed by men who don't have our awesome (and sometimes crazy) hormones.If you're sick of the time-wasting, crappy cardboard food, exercising until you drop, and wasting money on s*** that doesn't work, then you should listen to our podcast.You can get in touch with Naomi :Website: http://www.badassbeatthebulge.com/Facebook: https://www.facebook.com/naomi.gillespie74/aboutworkand_education

Do you have trouble with weight loss or keeping it off? We get to a certain age and it just seems that much harder. Even though we do everything right, everything just seems to go wrong. No matter what we do we just don’t seem to get the results we're looking for. Don’t you find it interesting that the weight loss industry is a multi-billion dollar industry? There are so many diets and exercise plans out there and they should work - but they don’t. All we hear is trust the process and you’ll be fine. It's very frustrating and quite depressing.  Most people just give up and eat all the food and drink all the wine. That makes us feel like failures on so many levels. Wouldn’t it be great to do a plan that works to use your hormones and metabolism as your superpowers instead of your burden? To truly understand how you as an individual lose weight.  Nutritionists are actually trained in doing the wrong thing for women. Most food plans are done for women by men (weird right?) or done by people who have never had a weight problem. Hence the birth of Badass Beat The Bulge System. Naomi had enough and so had her clients. Luckily research is her favorite and she found loopholes in the way exercise and diet plans are done. She developed the metaphase method. It works - at last - without excess training or starving.   Learn more about Naomi and Badass Beat the Bulge click here. 

This podcast reviews the results of the whole genome sequencing study by Samur and colleagues that identified a genomic signature associated with superior survival in patients with newly diagnosed multiple myeloma. Disclosures: SAH has served on advisory boards or as a consultant for Adaptive Biotechnologies, Amgen, Celgene, Genentech, GSK, Oncopeptides, Sorrento; Takeda; has received research funding from Oncopeptides.     This JCO Podcast provides observations and commentary on the JCO article “Genome-Wide Somatic Alterations in Multiple Myeloma Reveals a Superior Outcome Group” by Samur et al. My name is Sarah Holstein, and I am an Associate Professor at the University of Nebraska Medical Center in Omaha, Nebraska in the United States. My oncologic specialty is plasma cell dyscrasias. I do not have any relationships to disclose related to these studies. The clinical heterogeneity of myeloma has long been appreciated as it is clear there is a broad range of disease behavior, with some patients having indolent disease and others having very aggressive disease. As a result, there has been significant interest in developing risk stratification systems that classify patients into different risk groups, thus providing some information about prognosis and potentially inform treatment decisions. Historically, staging systems were based on factors related to tumor burden. However, it is increasingly evident that the underlying disease biology is a key modulator of risk. Our ability to detect disturbances in the myeloma genome has changed dramatically over time. Metaphase karyotyping represents our lowest “power of magnification”. These studies led to the recognition that in general, hyperdiploidy involving odd-numbered chromosomes is associated with lower-risk disease while high-risk disease can involve translocations of chromosome 14, monosomy 13 and monosomy 17. Use of fluorescent in-situ hybridization (FISH) allowed for a higher power of magnification and identification of more subtle chromosomal abnormalities. Next, gene expression profiling studies utilizing small panels of genes, enabled classification of patients into different risk categories, although there was little concordance between the panels used in the various studies. The advent of deep whole genome sequencing technology has facilitated a much more “high-powered” view of the myeloma genome. In the article that accompanies this podcast, diagnostic bone marrow specimens were obtained from 183 patients enrolled in the IFM/DFCI 2009 study. This phase 3 study enrolled newly diagnosed transplant-eligible patients (up to age 65). All patients received three cycles of lenalidomide, bortezomib, dexamethasone (RVD) induction, underwent stem cell collection and then received consolidation with either 5 cycles of RVD or a single autologous stem cell transplant followed by 2 cycles of RVD. Of note, in the French portion of this study, all patients subsequently received one year of lenalidomide maintenance, while in the US portion, all patients received lenalidomide until progression. The French portion has already been published and showed a 14 month PFS benefit for the transplant arm compared to the non-transplant arm. Results from the US portion have not yet been released, but I would speculate that the PFS and OS for both arms will be superior to the French counterparts, given the existing data supporting the benefit of prolonged lenalidomide maintenance. Deep whole genome sequencing, with a median tumor depth of 75X, was used to interrogate the myeloma genome. Mutational signatures were based on identified single nucleotide variants, small insertions and deletions. The genomic scar score (GSS) was calculated based on allele-specific copy number alterations. A GSS of 5 or less was the cut-off for inclusion in the low GSS group. The goals of the study were to describe mutational loads and processes in order to establish genomically-defined subgroups, gain insight into patterns of evolution from clonal to subclonal mutations, and correlate these findings with clinical outcomes and more traditional risk factors. There were several key findings. First, mutational load varied amongst myeloma subgroups, with hyperdiploid myeloma having the lowest mutational load and t(14;16) having the highest mutational load. Second, analysis of mutational patterns led to identification of five separate mutational processes that contributed to eight mutational signatures. These five processes included: 1) an age-related process, 2) an AID/APOBEC process, 3) somatic hypermutation, 4) DNA repair, and 5) processes with unidentified etiology including the clock-like signature. Samur et al., found that these various processes contributed to different myeloma subgroups in different ways. For example, the age-related process was high in hyperdiploid myeloma, the APOBEC-related process was high in t(14;16) myeloma, the clock-like signature was high in t(4;14) myeloma and the DNA repair process was high in del(17p) and t(11;14) myeloma. Furthermore, analysis of these mutational patterns from a clonal vs subclonal perspective enabled insight into mutational development patterns of different subgroups of myeloma. Next, the GSS was correlated with mutational signature and clinical outcome data. The authors found that the frequency of a low GSS was higher in t(11;14) myeloma and lower in del17p, gain1q21, del1p and del13 subgroups. Patients in the low GSS group had a trend towards a longer median PFS and a statistically significant longer 4-year OS rate than other patients. In particular, patients with both low GSS and gain of chromosome 9 had a superior outcome compared to all other subgroups, with an OS probability of 100%. Patients with either low GSS and no gain(9) or with high GSS and gain(9) had intermediate outcome while those with high GSS and t(4;14), gain(1q) or no gain(9) had the worst OS. Although the numbers are quite small, an interesting finding was that for patients in the low-risk group (low GSS + gain(9)), there was a significantly superior PFS in favor of those patients in the transplant arm compared to the non-transplant arm. Overall, no statistically significant differences were found between the four subgroups and factors such as ISS stage, response, or achievement of MRD negativity.  The numbers in each subgroup were quite small though. Finally, the authors demonstrated that the low GSS could separate hyperdiploid myeloma into low-risk and high-risk subgroups. Overall, these studies are interesting because they provide insight into the mutational processes that drive different subgroups of myeloma and offer a potential method by which to differentiate low-risk hyperdiploid myeloma from high-risk hyperdiploid myeloma. The finding that there was a difference in PFS for patients who underwent transplant vs no transplant in the low-risk group (low GSS, gain(9)) is very intriguing. There has been much discussion centered around whether low-risk patients really “need” to go through a stem cell transplant since in general their outcomes are good. The present study, aside from highlighting the fact that our traditional methods of identifying low-risk disease are likely inadequate, raises the hypothesis that patients with low-risk disease may benefit even more from transplant than other risk groups. However, it is noted that this subgroup consisted of only 28 patients. In addition, it is not clear from the manuscript whether patients were in the US vs French portion of the study, thus differences in study design (i.e., lenalidomide maintenance duration) could impact PFS findings. Clearly, this type of whole genome sequencing analysis will need to be applied to additional prospective studies in order to validate the novel risk stratification system. For now, these results are not practice-changing. However, they provide a potential glimpse into the future, where whole genome sequencing analysis is performed as readily as FISH analysis, and where enrichment strategies using genomic markers are used to design clinical trials. Aside from studies evaluating the use of venetoclax in t(11;14)-positive myeloma and other studies focused on high-risk disease that encompass a variety of high-risk chromosomal abnormalities, the field of myeloma has not yet moved into an era of precision medicine. Whether whole genome sequencing can finally usher us into that era remains to be determined. While this plasma cell-centric analysis is certainly revealing, it is likely that in order to maximally target myeloma, the genomics analysis must be coupled with an equally in-depth understanding of the host’s immune system. This concludes this JCO Podcast. Thank you for listening.

Mit Gästen plaudern wir in dieser Episode über Sci-Fi, Fantasy und Co. im Spannungsfeld zwischen Utopie und Dystopie. Autoren und eine Auswahl ihrer Bücher, die in dieser Episode erwähnt wurden: Vonda N. McIntyre: Superluminal (1983) (Heyne) Starfarers: (bastei Lübbe) Starfarers (1989) Kontakt (1991) Metaphase (1992) Nautilus (1994) Star Trek – The Original Series Die erste Mission (1986) Der Entropie-Effekt (1981) Romanfassungen von Star Trek Filmen: (Heyne) Star Trek II: Der Zorn des Khan. Star Trek III: Auf der Suche nach Mister Spock. Star Trek IV: Zurück in die Gegenwart Star Wars The Crystal Star (1994) Hanns Kneifel, Raumpatrouille Orion (Heftserie Terra Astra 1971 - 1985) Ray Bradbury, Die Mars Chroniken (1950) (Diogenes) Alan Dean Foster, Alien: Drei Romane in einem Band (Heyne) Die Homanx-Reihe (Reihe in 14 Bänden) (Heyne) Mark Lawrence, Waffenschwestern – Reihe in 3 Bänden bis jetzt auf Deutsch (Fischer) The Broken Empire – Reihe auf Deutsch: Prinz der Dunkelheit, König der Dunkelheit und Kaiser der Dunkelheit (Heyne) Jim Butcher, Die dunklen Fälle des Harry Dresden (Reihe in 15 Bänden) (Feder und Schwert Verlag) Codex Alera – Reihe in 6 Bänden (Blanvalet TB Verlag) Robert Jordan, Das Rad der Zeit – Reihe in 15 Bänden (Piper) Andy Weir, Der Marsianer (Heyne) Artemis (Heyne) Book Lover's Companion auf twitter: https://twitter.com/book_companion Zuhörerpost an: bookcompanioncontact@gmail.com Music: English Country Garden by Aaron Kenny Video Link: https://youtu.be/mDcADD4oS5E --- Send in a voice message: https://anchor.fm/ez-fiction/message

Mit Gästen plaudern wir in dieser Episode über Sci-Fi, Fantasy und Co. im Spannungsfeld zwischen Utopie und Dystopie. Autoren und eine Auswahl ihrer Bücher, die in dieser Episode erwähnt wurden: Vonda N. McIntyre: Superluminal (1983) (Heyne) Starfarers: (bastei Lübbe) Starfarers (1989) Kontakt (1991) Metaphase (1992) Nautilus (1994) Star Trek – The Original Series Die erste Mission (1986) Der Entropie-Effekt (1981) Romanfassungen von Star Trek Filmen: (Heyne) Star Trek II: Der Zorn des Khan. Star Trek III: Auf der Suche nach Mister Spock. Star Trek IV: Zurück in die Gegenwart Star Wars The Crystal Star (1994) Hanns Kneifel, Raumpatrouille Orion (Heftserie Terra Astra 1971 - 1985) Ray Bradbury, Die Mars Chroniken (1950) (Diogenes) Alan Dean Foster, Alien: Drei Romane in einem Band (Heyne) Die Homanx-Reihe (Reihe in 14 Bänden) (Heyne) Mark Lawrence, Waffenschwestern – Reihe in 3 Bänden bis jetzt auf Deutsch (Fischer) The Broken Empire – Reihe auf Deutsch: Prinz der Dunkelheit, König der Dunkelheit und Kaiser der Dunkelheit (Heyne) Jim Butcher, Die dunklen Fälle des Harry Dresden (Reihe in 15 Bänden) (Feder und Schwert Verlag) Codex Alera – Reihe in 6 Bänden (Blanvalet TB Verlag) Robert Jordan, Das Rad der Zeit – Reihe in 15 Bänden (Piper) Andy Weir, Der Marsianer (Heyne) Artemis (Heyne) Book Lover's Companion auf twitter: https://twitter.com/book_companion Zuhörerpost an: bookcompanioncontact@gmail.com Music: English Country Garden by Aaron Kenny Video Link: https://youtu.be/mDcADD4oS5E --- Send in a voice message: https://anchor.fm/ez-fiction/message

Topic: Autism Representation in Comics Guest: Megan Psychology: Multicultural Representation and Diversity Description: This podcast features Megan's story and experience of living with autism. She is an artist who has a love for manga and openly discusses how she longs for representation in manga and comics. Afterward hearing her story, this podcast touches on two comics: Superb and Metaphase that have main superhero characters living with down syndrome. Reference: Lion Forge Comics, Superb 2017-2019 Superb Vol 1. Life After the Fallout Metaphase by Alterna comics 2015

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!

Top 500, Superman 1, Flash 50, Amazing Spider-Man 1, X-23 1, Farm Hand, Outpost Zero, Incredibles II: Crisis in Mid-Life, She Could Fly, It Came Out On A Wednesday, Metaphase, Ruinworld, Little Girl, Wild’s End GN 3, Black Hops, Relay 1, Die! Die! Die!, Incredibles II movie review,  Goosebumps 2 trailer, Doctor Who teaser, Batman: Arkham Clayface, Batgirl: Stephanie Brown Vol 1 TP, News, More!   Comics Details: Superman 1 by Brian Michael Bendis, Ivan Reis, Joe Prado, Alex Sinclair Flash 50 by Joshua Williamson, Howard Porter, Hi-Fi Amazing Spider-Man 1 by Nick Spencer, Ryan Ottley, Cliff Rathburn, Laura Martin, Humberto Ramos, Victor Olazaba, Edgar Delgado X-23 by Mariko Tamaki, Juan Cabal, Nolan Woodard She Could Fly 1 by Christopher Cantwell, Martin Morazzo, Miroslav Mrva Incredibles II: Crisis in Mid-Life and Other Stories 1 by Christos Gage, Landry Walker, Gurihiru, J Bone, Andrea Greppi, Roberta Zanotta, Dan Jackson, Angela Capolupo Farmhand 1 by Rob Guillory, Taylor Wells Outpost Zero 1 by Sean Kelley McKeever, Alexandre Tefenkgi, Jean-Francois Beulieu It Came Out On A Wednesday 1 by Peter Simeti, Eastin DeVerna, Jeremy Massie, Terry Mayo, Ben Slabak, Troy Vevasis, Scott Wilson, Salo Farias, Aleksander Jovic, Ken Knudtsen, Javi Laparra, Michael Oppenheimer, Eli Powell Metaphase 1 by Kelly Williams, Chip Reece Ruinworld 1 by Derek Laufman Little Girl 1 by Pat Shand, Olivia Pelaez and Devil’s Due Comics Wild’s End: The Journey’s End by Dan Abnett, INJ Culbard Black Hops U.S.A. G.I. 1 by Mark Pellegrini, Timothy Lim, Dave Dorman Relay 1 by Zac Thompson, Donny Cates, Andy Clarke, Dan Brown Die! Die! Die! 1 by Robert Kirkman, Scott Gimple, Chris Burnham, Nathan Fairbairn   Comics Countdown, 19 Jul 2018: Wild’s End: Journey’s End by Dan Abnett, INJ Culbard Outpost Zero 1 by Sean Kelley McKeever, Alexandre Tefenkgi, Jean-Francois Beulieu Flash 50 by Joshua Williamson, Howard Porter, Hi-Fi Black Science 37 by Rick Remender, Matteo Scalera, Moreno DiNisio Mech Cadet Yu 10 by Greg Pak, Takeshi Miyazawa, Triona Farrell Bloodshot Salvation 11 by Jeff Lemire, Doug Braithwaite, Jordie Bellaire Farmhand 1 by Rob Guillory, Taylor Wells Nancy Drew 2 by Kelly Thompson, Jenn St-Onge, Triona Farrell Oblivion Song 5 by Robert Kirkman, Lorenzo De Felici, Annalisa Leoni Detective Comics 984 by Bryan Hill, Miguel Mendonca, Diana Egea, Adriano Lucas  

This episode contains new school releases at old school 145 BPM tempo, featuring Kamikaze Space Programme, Backrill, DJ D Redd, as well as a new release from Ankara, Turkey based producer Emir Hazir. This show aired originally 27 March 2018 11 PM Eastern Time US/28 March 2018 0400 GMT/UTC on Fnoob Techno Radio, fnoobtechno.com. 00:00 Fnoob Techno Radio, Station ident/intro 00:40 Ceephax Acid Crew, Alien Beacon (Acid Fourniture EP, self-released on Bandcamp) 05:28 Kamikaze Space Programme, Half Life A (Void Coefficient EP, Mord MORD048) 10:34 T-Dok, Pure Blood (Closing Mouths EP, Gynoid Audio GYNOIDD167) 13:25 Cannibal Cooking Club, Corupt (DJ Ze MigL Remix) (Disorder EP, Riot Radio Records) 20:09 Reynard D. Fox, Mic Break 21:52 T-Dok, Pure Blood (Again! LOL) 23:57 Daeron, Finite Incantatem (Finite Incantatem EP, Archivio 01) 29:45 Emir Hazir, "Taraba" (Obstacle EP, Fnoob Digital 18) 35:29 Reynard D. Fox, Mic Break 36:53 Klankman, "Polyacid" (World of Phenomena EP, Mord MORD039) 41:48 Karkz, Klamer & Køzløv, "Metaphase" (The Techno's Children TTC003) 47:04 Backrill, "Subtle Niffle Gate" (Subtle Niffle Gate EP, Bandcamp self-release) 52:16 DJ D-Redd "Waste" (Self-released on Soundcloud) 57:14 Reynard D. Fox, Mic Break 58:18 Joey Beltram, "Game Form" (Robert Armani Remix)(Tresor 33)

Chip Reece and his son, Ollie wanted to bring some diversity to the comic book/ graphic novel scene. Ollie, born with Down Syndrome, and his dad wanted to create a new superhero and highlight their story. Together they created "Metaphase," a graphic novel that brings some insights & understanding of Down's to the audience. I HIGHLY encourage you to order Metaphase: https://www.amazon.com/Metaphase-Chip-Reece/dp/1934985384/ref=la_B078P3G8D5_1_1?s=books&ie=UTF8&qid=1515525857&sr=1-1 You can follow Chip on Twitter: @ChipReece

Today we have Chip Reece in the Haven! Comic creator of Metaphase. Beautiful project full of heart and originality.

Top 300 September sales, Clone Conspiracy 1, Solo 1, Mosaic 1, Great Lakes Avengers 1, Reborn 1, Lost Boys 1, Warlords of Appalachia 1, Star Wars Classified, Iceman, Justice League vs. Power Rangers, Alterna: Chair and Metaphase. Details: Clone Conspiracy 1 by Dan Slott, Jim Cheung, Ron Frenz, John Dell; Solo 1 by Gerry Duggan, Geoffrey Thorne, Paco Diaz, Israel Silva; Mosaic 1 by Geoffrey Thorne, Khary Randolph, Emilio Lopez; Great Lakes Avengers 1 by Zac Gorman, Will Robson, Tamra Bonvillain; Reborn 1 by Mark Millar, Greg Capullo, Jonathan Glapion, Fco Plascencia; Lost Boys 1 by Tim Seeley, Scott Godlewski, Patricia Mulvihill; Warlords of Appalachia 1 by Phillip Johnson, Jonas Scharf, Doug Garbark; Chair by Peter Simeti, Kevin Christensen; Metaphase by Chip Reece, Kelly Williams 12 October Comics Countdown: 10. Wrath of the Eternal Warrior 12 by Robert Venditti, Robert Gill, Michael Spicer 9. Amazing Spider-Man: Clone Conspiracy 1 by Dan Slott, Jim Cheung, Ron Frenz, John Dell 8. Black Monday Murders 3 by Jonathan Hickman, Tomm Coker, Michael Garland 7. Supergirl 2 by Steve Orlando, Brian Ching, Michael Atiyeh 6. All-Star Batman 3 by Scott Snyder, John Romita Jr., Danny Miki, Dean White, Declan Shalvey, Jordie Bellaire 5. Deadpool 20 by Gerry Duggan, Matteo Lolli, Guru eFX 4. Deathstroke 4 by Christopher Priest, Joe Bennett, Mark Morales, Jeromy Cox 3. Black Science 25 by Rick Remender, Matteo Scalera, Moreno Dinisio 2. Kill or Be Killed 3 by Ed Brubaker, Sean Phillips, Bettie Breitweiser 1. American Vampire Anthology 2 by Scott Snyder, Rafael Albuquerque, Joelle Jones, Christopher Mitten, Marguerite Bennett, Mirka Andolfo, Clay Chapman, Richard Isanove, Steve Orlando, Artyom Trakhanov, Elliot Kalan, Andrea Mutti, Shawn Aldridge, Szymon Kudranski, Kieron Gillen, Leila Del Duca, Renato Guedes, Afua Richardson

Reviews: Black Bat #1, Movement #1, Suicide Risk #1, Ten Grand #1, Iron Man 3 Jimmy jets through a quickie solo show before heading off to TCAF (Toronto Comics Arts Festival) this weekend. He gives his thoughts on the recent Free Comic Book Day, meeting Natalie Dormer and interviewing Neil Gaiman about his upcoming Doctor Who episode. No news this week but a brief bit about his plans for TCAF. As always, listener feedback, Top 3 and more! Leave your iTunes comments! 5 stars and nothing but love! Thanks for listening!

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!

DUBSTEP.FM ARCHIVE - 2010-11-13 - Dank Dealz With Special Guest Metaphase

Der Erfolg einer Gravidität hängt von einem komplexen Zusammenspiel molekularer Faktoren von Gameten, Embryonen und Feten mit ihrer maternalen Umgebung ab. Eine intakte embryo-maternale Kommunikation ist die Voraussetzung für die Entstehung und Aufrechterhaltung der Trächtigkeit. Von den molekularen Signalen, die an dieser Interaktion beteiligt sind, sind bis heute nur wenige bekannt. Für ein besseres Verständnis dieser molekularen Faktoren beim Rind wurden auf maternaler Seite embryo-induzierte Veränderungen des Endometriums am 18. Tag der Trächtigkeit auf Proteomebene untersucht. Diese so genannte Periimplantationsphase wurde anhand monozygoter Zwillinge analysiert. Jeweils einer der beiden Zwillinge erhielt einen Transfer zweier in vitro produzierter Blastozysten, der korrespondierende Zwilling bekam einen Scheintransfer und diente als nicht trächtige Kontrolle. Endometriumproben dreier Zwillingspaare wurden mit der zweidimensionalen Differenz-Gelelektrophorese (2D-DIGE) in Kombination mit der Minimalmarkierung untersucht. Um als biologisch relevant angesehen zu werden, mussten die Protein-Spots die folgenden strengen Kriterien erfüllen: Abundanzfaktor ≥ 2; Spots in allen Gelen vorhanden; Student’s t-Test p ≤ 0,01. Vier abundanzveränderte Protein-Spots konnten detektiert und mittels PMF und MALDI-TOF/TOF Analyse identifiziert werden: Die Isocitrate Dehydrogenase 1 (NADP+, soluble) spielt eine Rolle bei der Vorbereitung des Endometriums auf die Implantation. Acyl-CoA-binding Protein ist ein wichtiges Element des Steroidmetabolismus. Rho GDP Dissociation Inhibitor beta interagiert mit Proteinen der Rho-Familie, wobei Rho A an der Implantation beteiligt ist. 20 α Hydroxysteroid-Dehydrogenase ist am Progesteron-Metabolismus beteiligt. Die vier trächtigkeitsrelevanten Proteine lieferten interessante Kandidaten für weiterführende Untersuchungen. Die erfolgreiche Entstehung der Trächtigkeit basiert jedoch nicht nur auf der Vorbereitung der maternalen Umgebung, ebenso essentiell ist die präzise Entwicklung männlicher und weiblicher Gameten. Eine korrekte Eizellreifung stellt die Grundlage für eine erfolgreiche Befruchtung und die darauf folgende Embryonalentwicklung dar. Da in vivo eine Analyse der Oozytenmaturation nicht durchführbar ist, wurde auf der embryonalen Seite die In-vitro-Maturation (IVM) boviner Oozyten auf Proteomebene untersucht. Die IVM ist beim Rind eine routinemäßig durchgeführte Prozedur und der erste und limitierende Schritt für assistierte Reproduktionstechniken (ART) wie die In-vitro-Produktion (IVP) von Embryonen. Die Eizellreifung ist definiert als die Phase von der Vollendung der ersten Reifeteilung bis zur Metaphase der Meiose II. Oozyten wurden aus Ovarien präpariert und in vitro maturiert. Auf Proteomebene wurden ungereifte mit in vitro gereiften bovinen Oozyten verglichen. Aufgrund der Limitierung der Probenmenge (10 – 20 Oozyten pro Ovar = 0,9 – 1,8 µg Protein) wurde eine 2D-DIGE-basierte Analyse mit der ultra-sensitiven Sättigungsmarkierung durchgeführt. So konnten die quantitativen Analysen mit nur 0,5 µg Gesamtprotein pro 2D-Gel durchgeführt werden. Sechs unabhängige biologische Replikate von ungereiften bzw. gereiften Oozyten wurden analysiert. Die Auswertung der Analyse nach den oben genannten Kriterien ergab die Detektion von 38 abundanzveränderten Proteinen, von denen zehn mittels nano-LC-MS/MS Analyse eindeutig identifiziert werden konnten: Dihydrolipoamide S-Succinyltransferase und 6-Phosphogluconolactonase sind Enzyme des Energiemetabolismus. Translationally-controlled Tumor Protein könnte bei der Wiederaufnahme der Meiose eine Rolle spielen. Clusterin interagiert mit Komponenten des Komplementsystems und könnte eine Schutzfunktion vor Komplement-vermittelten Abbauprozessen haben. Peroxiredoxin-3 und Glutathion S-Transferase Mu5 sind in das Redox-System involviert. Durch den 2D-Gel-basierten Ansatz war es möglich, drei Formen der Glutathion S-Transferase zu detektieren. Elongation Factor 1 gamma und das Protein 14-3-3 ε sind in den Regulationsmechanismus des Maturation Promoting Factor (MPF) involviert. MPF ist ein Schlüsselenzym der Zellzykluskontrolle. Einige dieser abundanzveränderten Proteine, die während der IVM boviner Oozyten detektiert wurden, könnten wichtige Hinweise zur Optimierung bestehender in vitro Kultursysteme geben. Die abundanzveränderten Proteine, die in den hier durchgeführten 2D-basierten Analysen detektierten werden konnten, sind interessante Kandidaten für weitere, tiefer gehende Analysen im Zusammenhang mit der embryo-maternalen Kommunikation.

Zusammenfassung Die Protease Separase trägt zur Regulation mitotischer und meiotischer Vorgänge entscheidend bei. Ihre klassische Funktion ist die Induktion der Schwesterchromosomen-trennung durch Spaltung des Cohesin-Proteinkomplexes, der die Schwesterchromatiden von der S-Phase bis zur Mitose gepaart hält. Separase wird am Ende der Metaphase durch Ubiquitin-abhängigen Abbau ihres Inhibitors Securin aktiviert. Ein zweiter Separase-Inhibitionsmechanismus ist die Hemmung durch Cyclin B1/Cdk1 („Cyclin Dependent Kinase 1“). Dafür ist Separase-Phosphorylierung durch Cdk1 notwendig (Stemmann et al., 2001). In vielen Modellorganismen hat Separase Funktionen, die über die Anaphase-Induktion hinausgehen. So trägt sie in S. cerevisiae beispielsweise zur Cdk1-Inaktivierung beim Meiose I-Meiose II-Übergang bei. Diese Separase-Funktion benötigt die proteolytische Separase-Aktivität nicht, ist jedoch abhängig vom Securin-Abbau. Für andere Funktionen der Separase hingegen könnte die Separase-abhängige Spaltung noch nicht identifizierter Substrate notwendig sein. In der vorliegenden Arbeit wird deshalb die Etablierung der IVEC-Methode („In Vitro Expression Cloning“) zur Identifizierung neuer Separase-Substrate vorgestellt. Mittels IVEC wurde - basierend auf der proteolytischen Separase-Aktivität - aus einer menschlichen cDNA-Bibliothek das In-vitro-Separase-Substrat GASP isoliert. Des Weiteren wurde die Separase-Hemmung durch Cyclin B1/Cdk1 näher untersucht. In der vorliegenden Arbeit konnte gezeigt werden, dass die Phosphorylierung von Separase durch Cyclin B1/Cdk1 für ihre Inhibition zwar notwendig, aber nicht hinreichend ist. Nach Phosphorylierung der Separase assoziiert die Kinase stabil mit der Protease, und erst diese Komplexbildung führt letztendlich zur Inhibition der proteolytischen Separase-Aktivität. Cyclin B1/Cdk1 ist also ein nicht-katalytisch wirkender Separase-Inhibitor. Die zeitlich korrekte Separase-Aktivierung ist für die fehlerlose Chromosomentrennung essentiell. Da Zellen ohne Securin ihre Chromosomen jedoch akkurat und zum richtigen Zeitpunkt trennen, muss es alternative Separase-Inhibitionsmechanismen geben. Die Separase-Hemmung durch Cyclin B1/Cdk1-Bindung könnte dieser gesuchte Securin-unabhängige Mechanismus sein, da der Separase-Cyclin B1/Cdk1-Komplex in Zellen bereits vor der Anaphase nachgewiesen werden kann und Cyclin B1 - wie Securin - am Ende der Metaphase Ubiquitin-vermittelt abgebaut wird. Securin und Cyclin B1/Cdk1 können nicht gleichzeitig an Separase binden. Die beiden Inhibitoren sind also Komponenten parallel und nicht konvergent wirkender Regulationsmechanismen. Die Phosphorylierung von Separase an Serin 1126 ist für ihre Cyclin B1/Cdk1-abhängige Inhibition essentiell (Stemmann et al., 2001). Daneben konnte in der hier vorgestellten Arbeit eine zweite Domäne in Separase identifiziert werden, die ebenfalls sowohl für die Inhibition der proteolytischen Separase-Aktivität als auch für die Komplexbildung mit Cyclin B1/Cdk1 nötig ist. Da diese zweite Cyclin B1/Cdk1-Bindungsdeterminante Sequenzhomologie zu dem Cdc6-Protein aufweist, wurde sie CLD („Cdc6 Like Domain“) genannt. Cdc6 ist ein konserviertes Protein, das in S. cerevisiae Cdk1-Inhibitionsaktivität besitzt. Dazu bindet es abhängig von der Phosphorylierung seines Aminoterminus direkt an B-Typ-Cycline, die sich im Komplex mit ihren Cdks befinden (Mimura et al., 2004). Durch Phosphatase-behandlung und Mutationsanalyse konnte bewiesen werden, dass die Interaktion zwischen Separase und Cyclin B1/Cdk1 auch von Phosphorylierung der Protease innerhalb ihrer CLD abhängt. Dies legt nahe, dass die Separase-CLD wie der Cdc6-Aminoterminus direkte Kontakte mit der Cyclin-Untereinheit der Kinase ausbildet. Serin 1126-Phosphorylierung ist dagegen indirekt an der Kinase-Bindung beteiligt. Denn erstens wird sie nach der Etablierung des Komplexes für seinen Erhalt nicht mehr benötigt (Holland et al., 2006), und zweitens ist sie für die Wechselwirkung zwischen CLD-enthaltenden Separasefragmenten und der Kinase abkömmlich. Ein zunächst favorisiertes Bindungsmodell, bei dem die Polo-Kinase an phosphoryliertes Serin 1126 bindet, um danach die Bindung von Cyclin B1 durch Phosphorylierung der CLD zu vermitteln, konnte ausgeschlossen werden. Stattdessen bewirkt die Phosphorylierung von Serin 1126 wohl eine Konformationänderung der CLD, die dadurch in die Lage versetzt wird, starke Wechselwirkungen mit der Cyclin B1-Untereinheit der Kinase einzugehen. Überraschenderweise ist im Separase-Cyclin B1/Cdk1-Komplex auch die Kinase inaktiv. Diese unerwartete Separase-Funktion als Cdk1-Inhibitor ist in Oozyten der Maus für den Übergang von der Meiose I in die Meiose II von entscheidender Bedeutung. Denn die Inhibition der Separase-Cyclin B1/Cdk1-Komplexbildung durch Mikroinjektion entsprechender Antikörper in Maus-Oozyten verhindert den Ausstoß des ersten Polkörpers, d.h., die Eizellen können den Meiose I-Meiose II-Übergang nicht vollziehen. In diesen Oozyten sinkt die Cdk1-Aktivität am Ende der Meiose I nicht wie bei Kontroll-Oozyten ab. Diese persistente Cdk1-Aktivität ist der Grund für den verhinderten Übergang von Meiose I nach -II, da künstliche Cdk1-Inhibition in Anwesenheit des inhibitorischen Antikörpers den Polkörperausstoß wiederherstellt. In mitotischen Zellen steigt der unter endogenen Bedingungen mit Separase assoziierte Anteil von Cyclin B1/Cdk1 in der Anaphase - d.h. nach dem Abbau seines Bindungskompetitors Securin - an. Übertragen auf die Meiose bedeutet das, dass Securin-Abbau die Induktion der Anaphase mit der Separase-abhängigen Cdk1-Inaktivierung koppelt.

Um eine Parthenogenese zu verhindern, arretieren reife Oozyten von Wirbeltieren in der Metaphase der Meiose II. Diese biochemische Aktivität wurde 1971 als Zytostatischer Faktor (engl. Cytostatic Factor; CSF) beschrieben. Einzelne wichtige Komponenten wurden im Laufe der Zeit identifiziert, aber deren Zusammenspiel noch nicht aufgeklärt. Eine wichtige Rolle spielt dabei der Anaphase Fördernder Komplex (engl.Anaphase promoting complex/Cyclosome;APC/C), eine Ubiquitin-Ligase welche Zellzyklus regulierende Proteine dem Abbau zuführt und somit den Beginn der Anaphase ermöglicht. Der APC/C ist in reifen Oozyten inaktiv und wird nach der Befruchtung aktiviert, so dass der Arrest aufgehoben wird. Des Weiteren sind für den Eintritt in die Anaphase II die Aktivitäten zweier Kinasen nötig. Erstens erfolgt während der Befruchtung ein Anstieg der Konzentration des intrazellulären Calciums, dies führt zur Aktivierung der Calmodulin-abhängigen-kinase-II (CaMKII). Allerdings waren die Substrate dieser Kinase bis jetzt unbekannt. Zweitens ist die Polo-like-kinase-1 (Plk1) essentiell für die Aufhebung des Metaphase II - Arrests. In Xenopus Eiextrakt konnte gezeigt werden, dass die Aktivität der Xenopus Plk1 (Plx1) essentiell für den Eintritt in die Anaphase ist. Kürzlich wurde ein Inhibitor des APC/C in einem Yeast-Two-Hybrid-Screen mit inaktiver Plx1 als bait gefunden – Xenopus-Emi1-verwandtes-Protein-1 (engl. Xenopus-Emi1-related protein-1; XErp1). Die Depletion dieses Proteins in Xenopus-Ei-Extrakt führt zu einem verfrühten Eintritt in die Anaphase. Im Rahmen meiner Doktorarbeit konnte gezeigt werden, dass CaMKII und Plx1 kooperieren, um XErp1 nach der Befruchtung zu inaktivieren, indem sie XErp1 für den Abbau markieren. Auch das humane Protein wurde kloniert und es wurde damit begonnen Versuche in Säugetierzelllinien durchzuführen. Erste Hinweise lassen darauf schließen, dass das humane Protein in gleicher Weise reguliert wird wie XErp1.

Fri, 22 Oct 2004 12:00:00 +0100 https://edoc.ub.uni-muenchen.de/2794/ https://edoc.ub.uni-muenchen.de/2794/1/Schroeder_Reiter_Elizabeth.pdf Schroeder-Reiter, Elizabeth ddc:570, ddc:500, Fak

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.

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.

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.

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.

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.

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.

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