Podcasts about ATPase

In this episode of the Epigenetics Podcast, we talked with Carl Wu from John's Hopkins University about his work on nucleosome remodeling, histone variants, and the role of single-molecule imaging in gene regulation. Our discussion starts with Carl Wu sharing his first significant milestones, a paper in "Cell" and the serendipitous discovery of DNA hypersensitive sites, which transformed our understanding of chromatin accessibility and its implications for gene regulation. As we delve into Dr. Wu's specific areas of research, he elaborates on the biochemistry of nucleosome remodeling and the intricate role of chromatin remodeling enzymes like NURF. We discuss how these enzymes employ ATP hydrolysis to reposition nucleosomes, making DNA accessible for transcription. He then explains the collaborative relationship between chromatin remodelers and transcription factors, showcasing the fascinating interplay that governs gene expression and regulatory mechanisms. The conversation takes a deeper turn as we explore Carl Wu's groundbreaking studies on histone variants, particularly H2AZ. He elucidates the role of SWR1 in facilitating the exchange between H2A and H2AZ in nucleosome arrays. The high-resolution structural insights garnered from recent studies reveal how the enzyme mediates histone eviction and insertion with remarkable precision, providing a clearer picture of chromatin dynamics at a molecular level.   References Wu, C., Bingham, P. M., Livak, K. J., Holmgren, R., & Elgin, S. C. (1979). The chromatin structure of specific genes: I. Evidence for higher order domains of defined DNA sequence. Cell, 16(4), 797–806. https://doi.org/10.1016/0092-8674(79)90095-3 Wu, C., Wong, Y. C., & Elgin, S. C. (1979). The chromatin structure of specific genes: II. Disruption of chromatin structure during gene activity. Cell, 16(4), 807–814. https://doi.org/10.1016/0092-8674(79)90096-5 Wu C. (1980). The 5' ends of Drosophila heat shock genes in chromatin are hypersensitive to DNase I. Nature, 286(5776), 854–860. https://doi.org/10.1038/286854a0 Wu, C., Wilson, S., Walker, B., Dawid, I., Paisley, T., Zimarino, V., & Ueda, H. (1987). Purification and properties of Drosophila heat shock activator protein. Science (New York, N.Y.), 238(4831), 1247–1253. https://doi.org/10.1126/science.3685975 Mizuguchi, G., Shen, X., Landry, J., Wu, W. H., Sen, S., & Wu, C. (2004). ATP-driven exchange of histone H2AZ variant catalyzed by SWR1 chromatin remodeling complex. Science (New York, N.Y.), 303(5656), 343–348. https://doi.org/10.1126/science.1090701 Kim, J. M., Visanpattanasin, P., Jou, V., Liu, S., Tang, X., Zheng, Q., Li, K. Y., Snedeker, J., Lavis, L. D., Lionnet, T., & Wu, C. (2021). Single-molecule imaging of chromatin remodelers reveals role of ATPase in promoting fast kinetics of target search and dissociation from chromatin. eLife, 10, e69387. https://doi.org/10.7554/eLife.69387   Related Episodes Multiple challenges of ATAC-Seq, Points to Consider (Yuan Xue) Pioneer Transcription Factors and Their Influence on Chromatin Structure (Ken Zaret) ATAC-Seq, scATAC-Seq and Chromatin Dynamics in Single-Cells (Jason Buenrostro)   Contact Epigenetics Podcast on X Epigenetics Podcast on Instagram Epigenetics Podcast on Mastodon Epigenetics Podcast on Bluesky Epigenetics Podcast on Threads Active Motif on X Active Motif on LinkedIn Email: podcast@activemotif.com

In this episode, we discuss the mitochondria. You know mitochondria as the "powerhouse" of the cell, but this biological process is biophysics meeting biochemistry. We discuss glycolysis, the TCA cycle (or Citric Acid cycle or Krebs cycle), and Oxidative Phosphorylation. This discussion goes deep inside the atomic molecular level with electrons and protons. Hang in there with the discussion and topic of Autism and understanding of the generational link to a loss of electrons. Jack Kruse https://www.patreon.com/DrJackKruse/postsGlycolysis https://www.ncbi.nlm.nih.gov/books/NBK470170/#:~:text=Glycolysis%20is%20a%20central%20metabolic,use%20in%20other%20metabolic%20pathways.Citric Acid cycle https://en.wikipedia.org/wiki/Citric_acid_cycleOxidative Phosphorylation https://en.wikipedia.org/wiki/Oxidative_phosphorylation0:00 Intro; Light, water, magnetism; The "powerhouse" and understanding of healthy living organisms and different cell types; Photosynthesis and Cell Respiration 4:11 Prokaryotes, Eukaryotes, and Cytochrome C Oxidase (CCO)6:53 Mitochondria and Cellular Respiration 1) Glycolysis, 2) Citric Acid cycle, and 3) Oxidative Phosphorylation (OXPHOS) 8:25 Glycolysis10:03 Citric Acid cycle12:02 OXPHOS13:51 Cytochrome I15:07 Cytochrome II16:08 Cytochrome III17:10 Cytochrome IV and creating water19:56 Not all water is equal/same; Aging; Light and Melanin and Rates of Autism29:20 Cytochrome V and ATPase; Chromophores 33:56 Evolution and losing electrons; Autism and modern health complications37:50 Environmental signals due DNA sequencing; Autism research and Genetic studies40:23 Reviews/Ratings and contact infoX: https://twitter.com/rps47586Hopp: https://www.hopp.bio/fromthespectrumemail: info.fromthespectrum@gmail.com

When you get deep enough into the natural health world, you learn about heavy hydrogen or deuterium. It is a natural contaminant that builds up in our body over time and stalls the nano motor in our mitochondria called the ATPase.  Victor Sagalovsky of Litewater joins us once again to talk about the health benefits of depleting deuterium in the body, not only for disease prevention but also to radically extend human lifespan. He talks about the cancer connection with deuterium, how the Russians discovered it, how your body will intelligently shuttle deuterium to less important tissues, on average how many molecules of HDO are in a glass of water, and how deuterium affects our reflex time. He also talks about human photosynthesis and why he loves melanin, we get into a friendly debate about whole food vs isolated supplements, he shares his theory on what the Ark of the Convenant was, and I ask listener questions including his thoughts on hydrogen infused water, sunlight to deplete deuterium, and the ideal PPM of deuterium in our drinking water. www.drinklitewater.com www.beevitamin.com www.protiumgenerator.com My website: www.matt-blackburn.com Mitolife products: www.mitolife.co Music by George Henner: https://georgehenner.bandcamp.com    

BUFFALO, NY- August 15, 2024 – A new #research paper was #published in Oncotarget's Volume 15 on August 14, 2024, entitled, “A nanobody against the V-ATPase c subunit inhibits metastasis of 4T1-12B breast tumor cells to lung in mice.” The vacuolar H+-ATPase (V-ATPase) is an ATP-dependent proton pump that functions to control the pH of intracellular compartments as well as to transport protons across the plasma membrane of various cell types, including cancer cells. Researchers Zhen Li, Mohammed A. Alshagawi, Rebecca A. Oot, Mariam K. Alamoudi, Kevin Su, Wenhui Li, Michael P. Collins, Stephan Wilkens, and Michael Forgac from Tufts University School of Medicine; Tufts University; Dana Farber Cancer Institute, Harvard Medical School; University of Minnesota School of Medicine; Prince Sattam Bin Abdulaziz University; Korro Bio; SUNY Upstate Medical University; and Foghorn Therapeutics, have previously shown that selective inhibition of plasma membrane V-ATPases in breast tumor cells inhibits the invasion of these cells in vitro. They have now developed a nanobody directed against an extracellular epitope of the mouse V-ATPase c subunit. “We show that treatment of 4T1-12B mouse breast cancer cells with this nanobody inhibits V-ATPase-dependent acidification of the media and invasion of these cells in vitro.” The research team further found that injecting this nanobody into mice implanted with 4T1-12B cells orthotopically in the mammary fat pad inhibited the metastasis of tumor cells to the lungs. “In conclusion, our results indicate that a nanobody directed against an extracellular epitope expressed on the surface of the V-ATPase is able to inhibit activity of cell surface V-ATPases in 4T1-12B breast cancer cells, inhibit in vitro invasion of these cells and inhibit metastasis of these cells to lung following their implantation in the mammary fat pad of mice.” DOI - https://doi.org/10.18632/oncotarget.28638 Correspondence to - Michael Forgac - michael.forgac@tufts.edu Video short - https://www.youtube.com/watch?v=4eLGqvSfAgg Sign up for free Altmetric alerts about this article - https://oncotarget.altmetric.com/details/email_updates?id=10.18632%2Foncotarget.28638 Subscribe for free publication alerts from Oncotarget - https://www.oncotarget.com/subscribe/ Keywords - cancer, vacuolar ATPase, breast cancer, invasion, tumor metastasis, tumor growth About Oncotarget Oncotarget (a primarily oncology-focused, peer-reviewed, open access journal) aims to maximize research impact through insightful peer-review; eliminate borders between specialties by linking different fields of oncology, cancer research and biomedical sciences; and foster application of basic and clinical science. Oncotarget is indexed and archived by PubMed/Medline, PubMed Central, Scopus, EMBASE, META (Chan Zuckerberg Initiative) (2018-2022), and Dimensions (Digital Science). To learn more about Oncotarget, please visit https://www.oncotarget.com and connect with us: Facebook - https://www.facebook.com/Oncotarget/ X - https://twitter.com/oncotarget Instagram - https://www.instagram.com/oncotargetjrnl/ YouTube - https://www.youtube.com/@OncotargetJournal LinkedIn - https://www.linkedin.com/company/oncotarget Pinterest - https://www.pinterest.com/oncotarget/ Reddit - https://www.reddit.com/user/Oncotarget/ Spotify - https://open.spotify.com/show/0gRwT6BqYWJzxzmjPJwtVh MEDIA@IMPACTJOURNALS.COM

In this episode, the Sarah discusses why she does not recommend collagen powders personally. She emphasizes that this is her personal opinion and encourages listeners to make their own decisions. The main reasons she does not recommend collagen powders are the presence of heavy metals and the high deuterium content. She explains that deuterium accumulates in collagen and can disrupt mitochondrial function, leading to health issues. She suggests natural ways to increase collagen production, such as red light therapy and consuming collagenous fatty meats and wild seafood. The speaker also emphasizes the importance of informed consent and individual decision-making.Natural ways to increase collagen production include red light therapy and consuming collagenous fatty meats and wild seafood.Individuals should make their own decisions and prioritize informed consent. Links mentioned: Free deuterium guide - https://www.sarahkleinerwellness.com/opt-in-deuterium Free Protein guide - https://www.sarahkleinerwellness.com/opt-in-protein Mother's Day Sale - https://www.sarahkleinerwellness.com/mothers-day-sale Mini red light therapy course - https://www.sarahkleinerwellness.com/offers/ezR259xA Leptin Master plan practitioner course - https://www.sarahkleinerwellness.com/leptin-master-plan Red light therapy panel - https://emr-tek.com/discount/sarah20 Studies: Collagen is high in deuterium due to the nature of how deuterium makes collagen molecules stronger and more resistant to wear and tear - https://onlinelibrary.wiley.com/doi/abs/10.1002/bip.21305 The real problem is when deuterium accumulates in the mitochondria, and breaks the ATPase (the "motors" of the energy management system of your body) - https://www.mdpi.com/1422-0067/24/4/3107 A lot of collagen powders also contain heavy metals (another recipe for disaster for your mitochondria) - https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7455464/ Timestamps: 00:00 Introduction and Triggered Reactions to Collagen Powders 02:11 The Issue of Heavy Metals in Collagen Powders 05:15 The Impact of Deuterium on Mitochondrial Function 08:21 Natural Ways to Increase Collagen Production 10:01 The Importance of Informed Consent and Individual Decision-Making This video is not medical advice & as a supporter to you and your health journey - I encourage you to monitor your labs and work with a professional! ________________________________________ Get all my free guides and product recommendations to get started on your journey! https://www.sarahkleinerwellness.com/all-free-resources Sign up for my newsletter to get special offers in the future! -https://www.sarahkleinerwellness.com/contact Free Guide to Building your perfect quantum day (start here) - https://www.sarahkleinerwellness.com/opt-in-9d5f6918-77a8-40d7-bedf-93ca2ec8387f My free product guide with all product recommendations and discount codes: https://www.sarahkleinerwellness.com/resource_redirect/downloads/file-uploads/sites/2147573344/themes/2150788813/downloads/84c82fa-f201-42eb-5466-0524b41f6b18_2024_SKW_Affiliate_Guide_1_.pdf

In today's episode, I'm talking about fluoride. Is fluoride a necessity in mouthwash, toothpaste and our water supply? Here are 5 things to consider before making fluoride a part of your routine: Fluoride inhibits the thyroid stimulating hormone output from the pituitary gland. Without a strong signal to the thyroid to make thyroid hormones, T4 and T3 production decrease. Fluoride is much lighter and more reactive than iodine, so it easily displaces iodine in the body.  Fluoride inhibits multiple antioxidant enzymes and lowers glutathione levels. Fluoride exposure leads to cell membrane degeneration and decreases the lifespan of red blood cells. Fluoride depletes our body of energy. Whether we are talking about the thyroid, glutathione, iron or straight up depleting ATP production by inhibiting the enzyme ATPase.  -------- Get the 30 Day Microbiome Upgrade  Start accumulating health with the Shake the Sugar guide Download the Healthy Meals Made Easy PDF   Ask your lifestyle health questions on social media, tag @drwholeness and use #accumulatehealth. -------- Connect with Dr. Matt online:

Dr. Luis Oliviera is parent to a son with an ultra-rare developmental and epileptic encephalopathy.  He's also a researcher with the Michael J. Fox Foundation.  He created an organization bringing parents and researchers together to better understand his son's disorder, which is part of a  group of disorders caused by rare mutations in v-ATPase genes. Dr. Oliviera was interviewed by Dr. Alina Ivaniuk.Visit the v-ATPase Alliance websiteContact the v-ATPase AllianceWhat is v-ATPase?  The International League Against Epilepsy invites you to explore the ILAE Academy: Interactive, practice based online courses for health care professionals who diagnose and treat epilepsy. Find more information at ilae-academy.org. Support the showSharp Waves episodes are meant for informational purposes only, and not as clinical or medical advice.The International League Against Epilepsy is the world's preeminent association of health professionals and scientists, working toward a world where no person's life is limited by epilepsy. Visit us on Facebook, Twitter, and Instagram.

Dr Lazslo Boros joins me for a deep dive into the role of deuterium in causing metabolic disease and the deposition of visceral fat. We go down to the level of the mitochondrion and learn exactly how foods enriched in deuterium, such as processed carbs and seed oils, contribute to metabolic dysfunction by wrecking the ATPase nanomotors in the inner mitochondrial matrix.Dr Boros former professor of paediatrics at UCLA, research scientist and world expert on deutonomics. Deutonomics refers to the study of deuterium,  heavy isotope of hydrogen, and how it interacts with biological systems.You won't want to miss this one!Join my private MEMBERS Q&A Group (USD20/month)✅ https://www.skool.com/dr-maxs-circadian-resetLEARN how to optimise your Circadian Rhythm ✅ Dr Max's Optimal Circadian Health course

We are truly honored to have Dr. Stephanie Seneff as our guest on the podcast. With over five decades of continuous affiliation with MIT and an impressive academic journey, including four degrees from MIT and extensive research experience in various fields, she brings a wealth of knowledge and expertise to our discussion. Dr. Seneff's more recent focus on the crucial role of nutrition and environmental factors, particularly glyphosate and sulfur, in human health adds an amazing perspective to our conversation. Topics: Understanding Deuterium Explanation of deuterium and its distinct atomic structure. The Significance of Low Deuterium Levels Importance for mitochondrial health. Consequences of elevated deuterium levels. Deuterium and ATPase Pumps Impact on cellular ATPase pumps. Glyphosate's Interference with Depletion Glyphosate's role in hindering deuterium depletion enzymes. Deuterium, Viral Infections, and Immunity How deuterium relates to viral infections and immune responses. Glyphosate, Viral Infections, and Symptoms Effects of glyphosate on deuterium depletion during infections. Manifested symptoms due to these interactions. Preventive and Treatment Strategies Recommendations for maintaining mitochondrial health. High-fat foods, Butter, Fermented foods, And hydrogen water. Thanks for tuning in! Get Chloe's Book Today! "⁠⁠⁠⁠⁠⁠⁠75 Gut-Healing Strategies & Biohacks⁠⁠⁠⁠⁠⁠⁠" If you liked this episode, please leave a rating and review or share it to your stories over on Instagram. If you tag @synthesisofwellness, Chloe would love to personally thank you for listening! Follow Chloe on Instagram ⁠⁠⁠⁠⁠⁠⁠⁠⁠⁠⁠⁠⁠@synthesisofwellness⁠⁠⁠⁠⁠⁠⁠⁠⁠⁠⁠⁠⁠⁠ Follow Chloe on TikTok @chloe_c_porter Visit ⁠⁠⁠⁠⁠⁠⁠⁠⁠⁠⁠⁠⁠⁠synthesisofwellness.com⁠⁠⁠⁠⁠⁠⁠⁠⁠⁠⁠⁠⁠⁠ to purchase products, subscribe to our mailing list, and more! Or visit ⁠⁠⁠⁠⁠⁠⁠⁠⁠⁠⁠⁠⁠⁠linktr.ee/synthesisofwellness⁠⁠⁠⁠⁠⁠⁠⁠⁠⁠⁠⁠⁠⁠ to see all of Chloe's links, schedule a BioPhotonic Scanner consult with Chloe, or support the show! Thanks again for tuning in! --- Support this podcast: https://podcasters.spotify.com/pod/show/chloe-porter6/support

Neste episódio do Podcast CRUZAMENTO, André Correia e Daniel Guedelha dialogam com Luís Oliveira, Senior Associate Director Research Programs na The Michael J. Fox Foundation, abordando a missão inspiradora da fundação, o impacto global das doenças neurodegenerativas em saúde e economia, e estratégias eficazes para mitigar riscos na Doença de Parkinson. Luis Miguel Oliveira is a Senior Associate Director of Research Programs at The Michael J. Fox Foundation and the Founder of v-ATPase Alliance for rare diseases due to v-ATPase defects. Luís is an expert in the field of Parkinson's disease research and investment with more than 15 years of experience in the neurologic diseases space. Currently, he holds the position of Senior Associate Director of Research Programs at The Michael J. Fox Foundation, where he leads various efforts for therapeutic and biomarker development in priority genetic targets in Parkinson's disease accounting for over $30 million in executed projects. Additionally, Luis Miguel Oliveira is a founder of the v-ATPase Alliance, an organization aimed at connecting families affected by rare diseases related to v-ATPase with the research and medical communities to further advance therapeutic development. With a strong commitment to advance novel treatments from the bench to bedside, Luis Miguel Oliveira serves as a Review Editor for Frontiers in Neurology, Neuroscience and Psychiatry, and as an Investment Advisor for Portugal Ventures, where he leverages his expertise to guide investment decisions in promising ventures. Recently, Luis Miguel Oliveira had an integral contribution to a landmark study that validated the first biomarker for Parkinson's disease diagnosis. This significant achievement was reported in Lancet Neurology with broader international media coverage and is changing the Parkinson's research and drug development fields by enabling an objective characterization of the disease based on its biological features. Luis Miguel Oliveira's educational background includes a degree in Biochemistry from the Faculty of Sciences of Lisbon University. He further pursued his academic journey with a PhD in Molecular Biophysics from the same institution. During his postdoctoral research experience, he conducted his studies at renowned institutions such as the Max Planck Institute for Biophysical Chemistry in Germany and Columbia University in New York. Other relevant references | Outras referências relevantes: v-ATPase Alliance (website) The Michael J. Fox Foundation (website) Contacts | Contactos: CruzamentoPodcast.com cruzamentopodcast@gmail.com LinkedIN: Cruzamento Twitter: @cruzamentofm Facebook: @podcastcruzamento Instagram: @cruzamentopodcast YouTube: Podcast Cruzamento

Link to bioRxiv paper: http://biorxiv.org/cgi/content/short/2023.08.04.552066v1?rss=1 Authors: Charles-Orszag, A., van Wolferen, M., Lord, S. J., Albers, S.-V., Mullins, D. Abstract: Type IV pili are ancient and widespread filamentous organelles found in most bacterial and archaeal phyla where they support a wide range of functions, including substrate adhesion, DNA uptake, self aggregation, and cell motility. In most bacteria, PilT-family ATPases disassemble adhesion pili, causing them to rapidly retract and produce twitching motility, important for surface colonization. As archaea do not possess homologs of PilT, it was thought that archaeal pili cannot retract. Here, we employ live-cell imaging under native conditions (75{degrees}C and pH 2), together with automated single-cell tracking, high-temperature fluorescence imaging, and genetic manipulation to demonstrate that S. acidocaldarius exhibits bona fide twitching motility, and that this behavior depends specifically on retractable adhesion pili. Our results demonstrate that archaeal adhesion pili are capable of retraction in the absence of a PilT retraction ATPase and suggests that the ancestral type IV pilus machinery in the last universal common ancestor (LUCA) relied on such a bifunctional ATPase for both extension and retraction. 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/2023.07.19.548188v1?rss=1 Authors: Chai, Y., Li, D., Gong, W., Ke, J., Tian, D., Chen, Z., Guo, A., Guo, Z., Li, W., Feng, W., Ou, G. Abstract: KIF1A, a microtubule-based motor protein responsible for axonal transport, is linked to a group of neurological disorders known as KIF1A-associated neurological disorder (KAND). Current therapeutic options for KAND are limited. Here, we introduced the clinically relevant KIF1A(R11Q) variant into the C. elegans homolog UNC-104, resulting in uncoordinated animal behaviors. Through genetic suppressor screens, we identified intragenic mutations in UNC-104's motor domain that rescued synaptic vesicle localization and coordinated movement. We showed that two suppressor mutations partially recovered motor activity in vitro by counteracting the structural defect caused by R11Q at KIF1A's nucleotide-binding pocket. We found that supplementation with fisetin, a plant flavonol, improved KIF1A(R11Q) worms' movement and morphology. Notably, our biochemical and single-molecule assays revealed that fisetin directly restored the ATPase activity and processive movement of human KIF1A(R11Q) without affecting wild-type KIF1A. These findings suggest fisetin as a potential intervention for enhancing KIF1A(R11Q) activity and alleviating associated defects in KAND. 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/2023.07.10.548302v1?rss=1 Authors: Richardson, H. E., Portela, M., Mukherjee, S., Paul, S., La Marca, J. E., Parsons, L. M., Veraksa, A. Abstract: The tumour suppressor, Lethal (2) giant larvae (Lgl), is an evolutionarily conserved protein that was discovered in the vinegar fly, Drosophila, where its depletion results in tissue overgrowth and loss of cell polarity and tissue architecture. Our previous studies have revealed a new role for Lgl in linking cell polarity and tissue growth through regulation of the Notch (proliferation and differentiation) and the Hippo (negative tissue growth control) signalling pathways. Moreover, Lgl regulates vesicle acidification, via the Vacuolar ATPase (V-ATPase), and we showed that Lgl inhibits V-ATPase activity through Vap33 (a Vamp (v-SNARE)-associated protein, involved in endo-lysosomal trafficking) to regulate the Notch pathway. However, how Lgl acts to regulate the Hippo pathway was unclear. In this current study, we show that V-ATPase activity inhibits the Hippo pathway, whereas Vap33 acts to activate Hippo signalling. Using an in vivo affinity-purification approach we found that Vap33 binds to the actin cytoskeletal regulators RtGEF (Pix, a Rho-type guanine nucleotide exchange factor) and Git (G protein-coupled receptor kinase interacting ArfGAP), which also bind to the Hpo protein kinase, and are involved in the activation of the Hippo pathway. Vap33 genetically interacts with RtGEF and Git in Hippo pathway regulation. Additionally, we show that the ADP ribosylation factor Arf79F (Arf1), which is a Hpo interactor, is involved in the inhibition of the Hippo pathway. Altogether our data suggests that Lgl acts via Vap33 to activate the Hippo pathway by a dual mechanism, 1) through interaction with RtGEF/Git/Arf79F, and 2) through interaction and inhibition of the V-ATPase, thereby controlling epithelial tissue growth. 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/2023.06.30.547267v1?rss=1 Authors: Zamal, M. Y., Venkataramana, C., Subramanyam, R. Abstract: In the phototrophic alphaproteobacteria, photosynthesis is performed by pigment-protein complexes, including the light-harvesting complexes known as LH1 and LH2. The photosystem also encompasses carotenoids to assist in well-functioning of photosynthesis. Most photosynthetic bacteria are exposed to various abiotic stresses, and here, the Rhodobacter (R.) alkalitolerans were extracted from the alkaline pond. We report the comparative study of photosynthetic apparatus of R. alkalitolerans in various light intensities in relation to this bacterium's high pH tolerance ability. We found that as the light intensity increased, the stability of photosystem complexes decreased in normal pH (npH pH 6.8{+/-}0.05) conditions, whereas in high pH (hpH pH 8.6{+/-}0.05) acclimation was observed. The content of bacteriochlorophyll a, absorbance spectra, and circular dichroism data shows that the integrity of photosystem complexes is less affected in hpH compared to npH conditions. Sucrose density and LP-BN of photosystem complexes also shows that LH2 is more affected in npH than hpH, whereas RC-LH1 monomer or dimer has shown interplay between monomer and dimer in hpH although the dimer and monomer both increased in npH. Additionally, the phosphatidylcholine (PC) levels have increased in hpH conditions. Moreover, qPCR data showed that the subunit -c of ATPase levels was overexpressed in hpH. Consequently, the P515 measurement shows that more ATP production is required in hpH, which dissipates the protons from the chromatophore lumen. This could be the reason the photosystem protein complex destabilized due to more lumen acidification. To maintain homeostasis in hpH, the antiporter NhaD expressed more than in the npH condition. 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/2023.06.25.546476v1?rss=1 Authors: Xie, Z., Chai, Y., Zhu, Z., Shen, Z., Zhao, Z., Xiao, L., Du, Z., Ou, G., Li, W. Abstract: Asymmetric cell divisions (ACDs) generate two daughter cells with identical genetic information but distinct cell fates through epigenetic mechanisms. However, the process of partitioning different epigenetic information into daughter cells remains unclear. Here, we demonstrate that the nucleosome remodeling and deacetylase (NuRD) complex is asymmetrically segregated into the surviving daughter cell rather than the apoptotic one during ACDs in Caenorhabditis elegans. The absence of NuRD triggers apoptosis via the EGL-1-CED-9-CED-4-CED-3 pathway, while an ectopic gain of NuRD enables apoptotic daughter cells to survive. We identify the vacuolar H+-adenosine triphosphatase (V-ATPase) complex as a crucial regulator of NuRD's asymmetric segregation. V-ATPase interacts with NuRD and is asymmetrically segregated into the surviving daughter cell. Inhibition of V-ATPase disrupts cytosolic pH asymmetry and NuRD asymmetry. We suggest that asymmetric segregation of V-ATPase may cause distinct acidification levels in the two daughter cells, enabling asymmetric epigenetic inheritance that specifies their respective life-versus-death fates. 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/2023.06.21.545956v1?rss=1 Authors: Klickstein, J. A., Johnson, M. A., Antonoudiou, P., Maguire, J., Paulo, J., Gygi, S., Weihl, C. C., Raman, M. Abstract: Mutations in the AAA+ ATPase p97 (also known as valosin containing protein, VCP) cause multisystem proteinopathy 1 (MSP-1) which includes amyotrophic-lateral sclerosis (ALS); however, the pathogenic mechanisms that contribute to motor neuron loss in familial ALS caused by p97 mutations remain obscure. Here, we use two distinct induced pluripotent stem cell models differentiated into spinal motor neurons to investigate how p97 mutations perturb the motor neuron proteome. Using multiplexed quantitative proteomics in these cells, we find that motor neurons harboring the p97 R155H mutation have deficits in lysosome quality control and the selective autophagy of lysosomes (lysophagy). p97 R155H motor neurons are unable to efficiently clear damaged lysosomes and have reduced viability. Additionally, lysosomes in mutant motor neurons have increased pH compared to their wildtype counterparts. The endo-lysosomal damage repair (ELDR) complex is required for clearance of damaged lysosomes and involves UBXD1-p97 interaction which is disrupted in mutant motor neurons. Finally, we report that inhibition of the ATPase activity of p97 R155H using the ATP competitive inhibitor CB-5083 rescues lysophagy defects in mutant motor neurons. These results add to the increasing evidence that endo-lysosomal dysfunction is a key aspect of disease pathogenesis in p97-related disorders. 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/2023.06.21.545957v1?rss=1 Authors: Hernandez-Ono, A., Zhao, Y. P., Murray, J. M., Ostlund, C. A., Lee, M. J., Ginsberg, H. N., Worman, H. J., Shin, J.-Y. Abstract: TorsinA is an atypical ATPase that lacks intrinsic activity unless it is bound to its activators lamina-associated polypeptide 1 (LAP1) in the perinuclear space or luminal domain-like LAP1 (LULL1) throughout the endoplasmic reticulum. However, the interaction of torsinA with LAP1 and LULL1 has not yet been shown to modulate a defined physiological process in mammals in vivo. We previously demonstrated that depletion of torsinA from mouse hepatocytes leads to reduced liver triglyceride secretion and marked steatosis, whereas depletion of LAP1 had more modest similar effects. We now show that depletion of LULL1 alone does not significantly decrease liver triglyceride secretion or cause steatosis. However, simultaneous depletion of both LAP1 and LULL1 from hepatocytes leads to defective liver triglyceride secretion and marked steatosis similar to that observed with depletion of torsinA. Our results demonstrate that torsinA and its activators dynamically regulate a physiological process in mammals in vivo. 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/2023.06.21.545896v1?rss=1 Authors: Wuenkhaus, D., Tang, R., Nyame, K., Laqtom, N. N., Schweizer, M., Rosato, A. S., Kroegsaeter, E. K., Wollnik, C., Abu-Remaileh, M., Grimm, C., Hermey, G., Kuhn, R., Gruber-Schoffnegger, D., Markmann, S. Abstract: Mutations in the lysosomal membrane protein CLN3 cause Juvenile Neuronal Ceroid Lipofuscinosis (JNCL). Activation of the lysosomal ion channel TRPML1 has previously been shown to be beneficial in several neurodegenerative disease models. Here, we tested whether TRPML1 activation rescues disease-associated phenotypes in CLN3-deficient retinal pigment epithelial (ARPE-19 CLN3-KO) cells. ARPE-19 CLN3-KO cells accumulate LAMP1 positive organelles and show lysosomal storage of mitochondrial ATPase subunit C (SubC), globotriaosylceramide (Gb3), and glycerophosphodiesters (GPDs), whereas lysosomal bis(monoacylglycero)phosphate (BMP/LBPA) lipid levels were significantly decreased. Activation of TRPML1 reduced lysosomal storage of Gb3 and SubC but failed to restore BMP levels in CLN3-KO cells. TRPML1-mediated decrease of storage was TFEB-independent, and we identified TRPML1-mediated enhanced lysosomal exocytosis as a likely mechanism for clearing storage including GPDs. Therefore, ARPE-19 CLN3-KO cells represent a human cell model for CLN3 disease showing many of the described core lysosomal deficits, some of which can be improved using TRPML1 agonists. Copy rights belong to original authors. Visit the link for more info Podcast created by Paper Player, LLC

We are talking pigweed. We are talking ATPase in diatoms. We are talking trees. What more do you want?!

This week, please join authors Marc Sabatine and Prakriti Gaba, as well as Associate Editor Amit Khera, as they discuss the article "Association Between Achieved Low-Density Lipoprotein Cholesterol Levels and Long-Term Cardiovascular and Safety Outcomes: An Analysis of FOURIER-OLE." Dr Greg Hundley: Welcome listeners, to this April 18th issue of Circulation on the Run and I am Dr. Greg Hundley, Associate Editor, Director of the Pauley Heart Center at VCU Health in Richmond, Virginia. Dr Peder Myhre: And I'm Dr. Peder Myhre, Social Media Editor from Akershus University Hospital and University of Oslo. Dr Greg Hundley: Peder, today's feature discussion, very interesting. We're going to evaluate the association between what's achieved with LDL cholesterol lowering, and then also long-term cardiovascular and safety outcomes. But before we get to that, how about we grab a cup of coffee and discuss some of the other articles in the issue? Would you like to go first? Dr Peder Myhre: Yes, Greg. I would love to. And the first paper is from the World of Preclinical Science and it comes to us from corresponding author, Jan Magnus Aronsen from University of Oslo in Norway. And perhaps, as you know, Greg, cardiomyocyte contraction and relaxation depend on the activity of the sarcoplasmic reticulum CA+2 ATPase 2, abbreviated SERCA2, and lowered levels or reduced activity of SERCA2, as seen in chronic heart failure, weakens contractile force and delays relaxation and no available therapy involves direct manipulation of the SERCA2 activity. And Greg, phosphodiesterase 3A is proposed to be present in the SERCA2 interactome limit SERCA2 activity and disruption of phosphodiesterase 3A from SERCA2 might thus be a strategy to develop SERCA2 activators. And in this study, the authors investigated and mapped SERCA2 and phosphodiesterase 3A and assessed this in experiments assessing the binding between these two in cardiomyocytes and in vesicles. Dr Greg Hundley: Wow Peder, sounds very interesting. So what did they find and how about the clinical implications of the findings? Dr Peder Myhre: So Greg phosphodiesterase 3A bounded directly to SERCA2 in the cardiomyocyte. So that's the first finding. Second, they demonstrated that SERCA2 phosphodiesterase 3A disruption increased SERCA2 activity independently of the catalytic activity of phosphodiesterase 3A in both normal and failing cardiomyocytes. And third, SERCA2 activity by the optimized SERCA2 phosphodiesterase 3A disruptor peptide OPT F reduced mortality and improved contractility after aortic binding in mice. So the clinical implication is that direct targeting of phosphodiesterase 3A binding to SERCA2 could be a novel approach to increase SERCA2 activity and thus cardiac contractility in patients with heart failure. Dr Greg Hundley: Very nice Peder. What a great new finding in the world of preclinical science. Well my paper is going to delve into the world of clinical science and involves patients with stroke. So Peder in this study led by corresponding author, Dr. Dileep Yavagal from University of Miami Miller School of Medicine performed a survey in 75 countries through the Mission Thrombectomy 2020+ Global Network between November of 2020 and February of 2021 to determine the availability of mechanical thrombectomy for large vessel occlusion in patients with stroke. Now Peder, the primary endpoints were the current annual mechanical thrombectomy availability, the mechanical thrombectomy operator availability and the mechanical thrombectomy center availability. All of these availabilities were defined as the proportion of estimated large vessel occlusion for patients receiving mechanical thrombectomy in a given region annually. Dr Peder Myhre: Okay, Greg, so this is really an access question. So in essence, what is the availability of mechanical thrombectomy worldwide? So what did they find? Dr Greg Hundley: Right, great Peder. So what they found, the authors received 887 responses from 67 countries and low-income countries had 88% lower mechanical thrombectomy availability compared to high-income countries. The global mechanical thrombectomy operator availability was 16.5% of optimal, and the mechanical thrombectomy center availability was only 20.8% optimal. And with these results, the authors indicate that global cooperation and targeted region-specific public health interventions, including all stakeholders involved in stroke care delivery, are really needed to rapidly increase access to this brain-saving and disability-sparing treatment with mechanical thrombectomy really worldwide. Dr Peder Myhre: Oh wow. What a beautiful summary, Greg. Thank you so much. And we also have some other interesting papers in the mailbag today. We have an exchange of letters between Dr. Yang and Dr. O'Donoghue regarding the article “Long Term evolocumab in Patients with Established Atherosclerotic Cardiovascular Disease.” Dr Greg Hundley: Great Peder, and also Professor Perera has a Frontiers article entitled “Unloading the Left Ventricle in Venoarterial ECMO in Who, When and How?” and then finally there's a Research Letter from Professor Verma entitled “Prevalence of Diabetes and Cardiovascular Risk in the Middle East and Africa: The Primary results of the PACT-MEA Study.” Well Peder, how about we jump to that feature discussion? Dr Peder Myhre: Can't wait. Dr Greg Hundley: Welcome listeners to this feature discussion on April 18th and we have with us today Prakriti Gaba and Marc Sabatine from Brigham and Women's Hospital and our own associate editor, Dr. Amit Khera. Welcome everyone. Well Marc, we'll start with you. Can you describe for us some of the background information that really helps constitute the preparation of your study and what was the hypothesis that you wanted to address? Dr. Marc Sabatine: Yeah, thanks Greg and thanks for having us. So we've seen in a variety of epidemiologic cohorts the association between LDL cholesterol and the risk of adverse cardiovascular events like in Framingham Heart Study and UK Biobank. But in those cohorts, in these industrial societies, we don't have the benefit of lots of data in individuals with very low levels of LDL cholesterol and so we had the opportunity with the FOURIER study that was the randomized comparison of evolocumab PCSK9 inhibitor versus placebo to get patients down to extremely low levels of LDL cholesterol and evolocumab. We were able to get individuals down to about 30 mg/dL. And so in addition to all the studies we've done showing the comparison of evolocumab to placebo, we also then had the chance to use FOURIER, and as you'll hear from PK, FOURIER-OLE, the open-label extension, as a cohort to then examine patients' new baseline, if you will, their new achieved LDL cholesterol and then it's association not only with cardiovascular events but safety events. And so the hypothesis is that there would be a relationship with the lower the LDL cholesterol, the lower the risk of cardiovascular events and we wanted to explore how far down that went. And then the second one for safety would be that there wouldn't be any association between low levels of LDL cholesterol and a variety of safety outcomes that rightly or wrongly people have ascribed to low levels of LDL cholesterol. Dr Greg Hundley: Thanks so much, Marc. Well listeners, now we're going to turn to PK, the first author, on this very interesting paper and PK, Marc mentioned to us the FOURIER-OLE study. Maybe describe for us here your study design and then what specifically was your study population? Dr. Prakriti Gaba (PK): Yeah, definitely. Thanks so much for the introduction. So the study population included 27,564 patients with stable atherosclerotic cardiovascular disease and LDL cholesterol levels that were greater than or equal to 70 mg/dL or non-HDL cholesterol greater than or equal to a 100 mg/dL on statin therapy. The patients who then went on to the FOURIER-OLE or the open-label extension part of the trial consisted of about 6,635 patients. And so in this study we essentially evaluated the combination of those populations in 2 separate analyses. We then categorized patients according to 6 pre-specified bins based on their achieved LDL cholesterol levels at designated time points and those ranged from LDL levels of less than 20 mg/dL all the way up to 100 mg/dL. And then we looked at their baseline characteristics and evaluated the cardiovascular and safety outcomes that Dr. Sabatine mentioned earlier. Dr Greg Hundley: Very nice PK. Well we've got a great listening audience today and they're anxious to hear your study results, so can you share those with us please? Dr. Prakriti Gaba (PK): Definitely. So over the course of more than 77,000 patient years of follow-up, we found that there was a monotonic relationship between achieving lower LDL cholesterol levels down to very low levels of less than 20 mg/dL and a lower annualized risk of the primary efficacy endpoint, which was a composite of 5 individual endpoints. We also found that there was a similar relationship observed between lower LDL achieved, LDL cholesterol levels and a lower annualized risk of the secondary efficacy endpoint, and then when we looked at safety, there were actually no clear monotonic trends between lower achieved LDL levels and the risk of any of the 8 adverse events and these included things like serious adverse events, hemorrhagic stroke or muscle related events. Dr Greg Hundley: Very nice PK and I'm sure our listeners are wanting to know, did you find any discrepancy in your results based on either age or gender? Dr. Prakriti Gaba (PK): That's an excellent question, and we did look at age and gender throughout. I think across the board the results were pretty consistent, but additional subanalyses will further address this question. Dr Greg Hundley: Very good. Well listeners now we're going to turn to one of our associate editors, Dr. Amit Khera, who has helped move this article through the process of evaluation with the editorial team. Amit, you have many papers come across your desk, what attracted you to this paper and then how do we put this study's results really in the context of other studies that have sought to dramatically lower LDL cholesterol? Dr. Amit Khera: Well first thanks a lot Greg for allowing me to participate today. I want to congratulate Drs. Gaba and Sabatine on a fantastic paper and the minute I saw it, and you know can tell when you've done this for a while what's a great paper, and this one certainly is and we work closely with them to try to make it better and enhance the analyses and as a group, I think we achieved that. I was fortunate to write an accompanying editorial that you'll see. So I got to take a pretty deep dive in this paper and I want to just talk about sort of what's important here, why is this important, and I think as Dr. Gaba mentioned, there's two sides to this. There's the efficacy side where you talk about LDL lowering and getting to very low levels. Now mind you, they got to, what I call, ultra low levels, even explored for a down to a median of 7 mg/dL, so really, really low. And first I think what our listeners need to know when we look at guidelines, these numbers of 70 or 55, these are completely arbitrary and they're based on what was observed in clinical trials, what was achieved in high intensity versus moderate intensity statins in IMPROVE-IT. There's no biology behind that, and I think what this study does is reminds us there is no biology behind how low we need to go. This group previously published their shorter-term data approximately 2 years with this construct of lower is better and I think that's fine, but people worry, particularly on the safety side about extension, and we'll get to that in a minute, so where this fits is it gives us even more reassurance that lower is better, reminds us there's no biologic basis of that even down to very low levels. And so what does that mean? I think that comes back to guidelines. We have some discrepancy between European, ACC, Multisociety Guidelines that are around 55 and so from a guideline perspective, I think we'll see a little bit more enthusiasm about lower cut points or lower thresholds. And from a clinical standpoint, as a clinician it reminds me that when I see someone that's very high risk, there's no magic to achieving a number that if the risk is high, we need to be quite intensive and get their LDL down as low as possible and as safely as possible. I do want to also acknowledge, there's not, to your point about context, the IMPROVE-IT study also showed very low levels show additional efficacy and there's also a lot of other data, genetics and ecologic data supporting this. So this is... we look at Bayesian analysis that this is consistent that we're seeing across different platforms. I do want to talk about safety too, Greg. That's really important because honestly this is when it comes to patient level, the safety part of it. We as clinicians may have comfort with very low levels, but the safety is important. I also want to, just from a steady design, this is post-hoc, so those that achieve very low levels are different. You can see that in their table 1, but these investigators did lots of things. They did pretty extensive multi-variable analysis, they looked at time-dependent LDLs, they looked at it multiple different ways, but as mentioned, there really did not seem to be a safety signal. And this is where time matters. Safety in two years, interesting, but safety 5 to 8 years really offers us much more reassurance. So I think that's where this really comes in about that safety piece with the extended analysis. So again, I think from a guideline perspective, from a clinical perspective, there's so many implications from this paper and I really hope people take the time to take a deep dive and also put it in context, like you said, to the other literature where this is not standalone, but it's corroborating what we're seeing. Dr Greg Hundley: Very nice, amit. Marc, I want to come back to you, just two quick questions thinking about the preparation of your study. One, did you sample cognition? One thing we hear about frequently in dramatic lowering of LDL cholesterol are questions around cognition, particularly in the elderly? Dr. Marc Sabatine: Yeah, it's a great question Greg. So first of all, in the FOURIER study itself, there was an embedded study called EBBINGHAUS that Bob Giugliano from our group led that actually did formal neurocognitive testing in individuals using basically a iPad-like test. We also collected the usual neurocognitive adverse events as part of safety collecting. So 2 ways, the general asking about any adverse events and then the specific neurocognitive testing. We had previously reported out the results of EBBINGHAUS that there wasn't any relationship between evolocumab and the low LDL cholesterol and the risk of any neurocognitive AEs. We just were able to recently do this OLE analysis over time for the major adverse cardiovascular events and for the general safety events including cognition, so all that looked good. As PK indicated, we're now digging into the EBBINGHAUS formal neurocognitive testing, which was also extended out. So stay tuned for those results. Dr Greg Hundley: Very nice. And then eligibility, maybe just walk us through that really quickly. Patients that are going to be randomized to this form of therapy, were they already on high-dose statins? Who exactly did we randomize in this trial? Dr. Marc Sabatine: Yeah, so at the get-go, as PK indicated, these are patients with atherosclerotic cardiovascular disease, so they had a prior MI, prior stroke, symptomatic PAD. They were to be on an optimized lipid-lowering regimen, optimized statin therapy, so for close to 70%, that was a high-intensity statin. We had a small percentage on ezetimibe, but that's because we hadn't yet published the results of the IMPROVE-IT trial that Amit mentioned when we were enrolling in FOURIER, but it was a well-treated population on statin therapy. So these results would apply to your typical patient with ASCVD who's on a good statin regimen. Dr Greg Hundley: Very nice. Well, listeners now we're going to go back to both of our authors and investigators, as well as Dr. Khera. PK we'll start with you. What do you see as the next study to really be performed in this sphere of research? Dr. Prakriti Gaba (PK): I think that's an excellent question. I think with the data presented here now we know that the lower the LDL, the lower the risk of adverse cardiovascular events and that having a low LDL is safe in the long term. I think moving forward, as Dr. Khera mentioned, there needs to be a shift of these recommendations into the guidelines. So I think additional studies confirming these findings is what we need, but we do have the evidence available. Dr Greg Hundley: Very nice. And Marc? Dr. Marc Sabatine: Yeah, and I agree with PK of course. I think there's a couple things that we want to look at. We had looked in the parent FOURIER trial and found some groups who were higher risk, who seemed to have a bigger benefit early on, and those by and large were people who had a lot of athero. But as Amit indicated that the parent FOURIER trial was relatively short at about in two and a quarter years median follow up, and so now we have the benefit of an additional half decade of follow up in a subset of people and so now we're starting to look through and see the subgroups where we saw some differential benefit and this was a paper we published in circulation soon after we published the primary results of FOURIER. Now we have the ability to go through and look at those same subgroups and see what happens now with an additional 5 years. And so that'll be quite interesting, I think, to see how those groups play out now over time. I think as Amit indicated, time is critical. We know the benefit of lipid lowering really tends to grow with time. We saw that in FOURIER, we saw that in FOURIER-OLE and then as Amit indicated, I think for safety also it's now very reassuring, being able to go out to not two and a half years, but 5, 6, 7, 8 years of safety follow up. Dr Greg Hundley: Very nice. Well, listeners we're next going to turn to Dr. Khera. I'm going to put him on the spot a little bit. I don't know if many of you know he's a cardiologist with expertise in primary prevention. So here we've really focused today, I think, on a very unique set of results in secondary prevention. Amit, as you think about studies to be performed in the future, is there a role for really lowering LDL cholesterol as a primary prevention target? Dr. Amit Khera: The short answer is absolutely. I think, to be fair, you can't necessarily directly extrapolate these results 'cause it's a secondary prevention population, but I think if we step back for a second, is there any reason I think this wouldn't work in primary prevention, there's not, and I think there's tons of genetic data, tons of other long-term data that suggests that lower is better than primary prevention. I think the challenge, as you know, is just from primary prevention is it's just about the number that you need to treat and primary prevention is pretty profound in terms of to lower events. So this is where the trade-off comes. I think even in their study, we do have to appreciate while lower is better, when you have very low levels and you're going to even lower, let's say when you go from in secondary prevention from 50 to 40, as much as that sounds valuable, that delta's pretty small and then the absolute risk reduction is still going to be pretty small for those individuals and that's only magnified in primary prevention. So the short answer is I have no reason to believe that lower is better is not applicable in primary prevention, but I do know that the cost and what entails to get there, you don't get as much return on investment. I do want to say one last thing though. We're talking about lower is better, and I know these investigators know this well, but it's not only just how low but how long and I think that's where primary prevention about to go to clinic and I play the long game for primary prevention that we know we've magnified these benefits over the long term and even a little bit early in life can pay off long dividends. So that's how I look at it. Dr Greg Hundley: Very nice. Well, listeners we want to thank Dr. Prakriti Gaba, Dr. Marc Sabatine, both from Brigham and Women's Hospital and also our own associate editor, Dr. Amit Khera from University of Texas Southwestern Medical Center for bringing us this study involving patients with arteriosclerotic cardiovascular disease indicating that long-term achievement of lower LDL cholesterol levels down to values less than 20 mg/dL was associated with a lower risk of cardiovascular outcomes and not and not an increase in the risk of significant safety related events. Well, on behalf of Peder, Carolyn and myself, we want to wish you a great week and we will catch you next week on the run. Dr. Greg Hundley: This program is copyright of the American Heart Association 2023. The opinions expressed by speakers in this podcast are their own and not necessarily those of the editors or of the American Heart Association. For more, please visit ahajournals.org.

For almost 30 years, the protein KRAS has been the "undruggable target" of cancer research.  Gene mutations in KRAS are responsible for 33% of all human cancers, including pancreatic, lung, and colon cancer.A new molecule identified by Dr. Bhairavi Tolani, an assistant professor at the University of California, San Francisco  upends conventional concepts of directly targeting the protein KRAS as a treatment strategy. Instead she suggests an alternative avenue for treatment of these deadliest cancers.Read her original research in Nature Biotechnology here: https://doi.org/10.1038/s41587-022-01386-z

Link to bioRxiv paper: http://biorxiv.org/cgi/content/short/2023.04.03.535492v1?rss=1 Authors: Manandhar, B., Pandzic, E., Deshpande, N., Chen, S.-Y., Wasinger, V., Kockx, M., Glaros, E. N., Ong, K. L., Thomas, S. R., Wilkins, M., Whan, R., Cochran, B. J., Rye, K.-A. Abstract: Objective: High cholesterol levels in pancreatic {beta}-cells cause oxidative stress and decrease insulin secretion. {beta}-cells can internalize apolipoprotein (apo) A-I, which increases insulin secretion. This study asks whether internalization of apoA-I improves {beta}-cell insulin secretion by reducing oxidative stress. Approach: Ins-1E cells were cholesterol-loaded by incubation with cholesterol-methyl-{beta}-cyclodextrin. Insulin secretion in the presence of 2.8 or 25 mM glucose was quantified by radioimmunoassay. Internalization of fluorescently labelled apoA-I by {beta}-cells was monitored by flow cytometry. The effects of apoA-I internalization on {beta}-cell gene expression was evaluated by RNA sequencing. ApoA-I binding partners on the {beta}-cell surface were identified by mass spectrometry. Mitochondrial oxidative stress was quantified in {beta}-cells and isolated islets with MitoSOX and confocal microscopy. Results: An F1-ATPase {beta}-subunit on the {beta}-cell surface was identified as the main apoA-I binding partner. {beta}-cell internalization of apoA-I was time-, concentration-, temperature-, cholesterol- and F1-ATPase {beta}-subunit-dependent. {beta}-cells with internalized apoA-I (apoA-I+ cells) had higher cholesterol and cell surface F1-ATPase {beta}-subunit levels than {beta}-cells without internalized apoA-I (apoA-I- cells). The internalized apoA-I co-localized with mitochondria and was associated with reduced oxidative stress and increased insulin secretion. The ATPase inhibitory factor 1, IF1, attenuated apoA-I internalization and increased oxidative stress in Ins-1E {beta}-cells and isolated mouse islets. Differentially expressed genes in apoA-I+ and apoA-I- Ins-1E cells were related to protein synthesis, the unfolded protein response, insulin secretion and mitochondrial function. Conclusions: These results establish that {beta}-cells are functionally heterogeneous and apoA-I restores insulin secretion in ?-cells with elevated cholesterol levels by improving mitochondrial redox balance. 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/2023.03.31.534207v1?rss=1 Authors: van Veen, S., Kourti, A., Ausloos, E., Van Asselberghs, J., Van den Haute, C., Baekelandt, V., Eggermont, J., Vangheluwe, P. Abstract: Polyamine homeostasis is disturbed in several human diseases, including cancer, which is hallmarked by increased intracellular polyamine levels and an upregulated polyamine transport system (PTS). So far, the polyamine transporters contributing to the elevated levels of polyamines in cancer cells have not yet been described, despite the fact that polyamine transport inhibitors are considered for cancer therapy. Here, we tested whether upregulation of candidate polyamine transporters of the P5B-transport ATPase family is responsible for the increased PTS in the well-studied breast cancer cell line MCF7 compared to the non-tumorigenic epithelial breast cell line MCF10A. We found that MCF7 cells present elevated expression of a previously uncharacterized P5B-ATPase ATP13A4, which is responsible for the elevated polyamine uptake activity. Furthermore, MCF7 cells are more sensitive to polyamine cytotoxicity, as demonstrated by cell viability, cell death and clonogenic assays. Importantly, overexpression of ATP13A4 WT in MCF10A cells induces a MCF7 polyamine phenotype, with significantly higher uptake of BODIPY-labelled polyamines and increased sensitivity to polyamine toxicity. In conclusion, we establish ATP13A4 as a new polyamine transporter in the human PTS and show that ATP13A4 may play a major role in the increased polyamine uptake of breast cancer cells. ATP13A4 therefore emerges as a candidate therapeutic target for anticancer drugs that block the PTS. 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/2023.03.07.531620v1?rss=1 Authors: Zaver, S. A. A., Sarkar, M. K., Egolf, S., Zou, J., Tiwaa, A., Capell, B. C., Gudjonsson, J. E., Simpson, C. L. Abstract: Mutation of the ATP2A2 gene encoding sarco-endoplasmic reticulum calcium ATPase 2 (SERCA2) was linked to Darier disease more than two decades ago; however, there remain no targeted therapies for this disorder causing recurrent skin blistering and infections. Since Atp2a2 knockout mice do not phenocopy its pathology, we established a human tissue model of Darier disease to elucidate its pathogenesis and identify potential therapies. Leveraging CRISPR/Cas9, we generated human keratinocytes lacking SERCA2, which replicated features of Darier disease, including weakened intercellular adhesion and defective differentiation in organotypic epidermis. To identify pathogenic drivers downstream of SERCA2 depletion, we performed RNA sequencing and proteomic analysis. SERCA2-deficient keratinocytes lacked desmosomal and cytoskeletal proteins required for epidermal integrity and exhibited excess MAP kinase signaling, which modulates keratinocyte adhesion and differentiation. Immunostaining patient biopsies substantiated these findings with lesions showing keratin deficiency, cadherin mis-localization, and ERK hyper-phosphorylation. Dampening ERK activity with MEK inhibitors rescued adhesive protein expression and restored keratinocyte sheet integrity despite SERCA2 depletion or chemical inhibition. In sum, coupling multi-omic analysis with human organotypic epidermis as a pre-clinical model, we found that SERCA2 haploinsufficiency disrupts critical adhesive components in keratinocytes via ERK signaling and identified MEK inhibition as a treatment strategy for Darier disease. 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/2023.02.26.530105v1?rss=1 Authors: Bi, P. Y., Killackey, S. A., Schweizer, L., Arnoult, D., Philpott, D. J., Girardin, S. E. Abstract: Mitochondrial stress inducers, such as the proton ionophore carbonyl cyanide m-chlorophenyl hydrazone (CCCP) and the ATPase pump inhibitor oligomycin, trigger the DELE1-HRI branch of the integrated stress response (ISR) pathway. Previous studies performed using epitope-tagged forms of DELE1 showed that these stresses induced the accumulation of a cleaved form of DELE1, DELE1-S, which stimulates HRI. Here, we report that mitochondrial protein import stress (MPIS) is an overarching stress that triggers the DELE1-HRI pathway, and that endogenous DELE1 could be cleaved into two forms, DELE1-S and DELE1-VS, the latter accumulating only upon non-depolarizing MPIS. We further showed that DELE1 specifically senses MPIS triggered by the inhibition of the TIM23 complex at the inner mitochondrial membrane (IMM). While MPIS can also cause mitophagy induction through engagement of the NLRX1-RRBP1 pathway, we observed that DELE1-HRI and NLRX1-RRBP1 signaling were engaged independently upon MPIS. Surprisingly, our results suggest that in our cellular model the mitochondrial protease OMA1 was dispensable for DELE1 cleavage upon MPIS. Instead, we identified a key role for another mitochondrial protease, HtrA2, in mediating the cleavage of DELE1 into DELE1-S and DELE1-VS. Our data further suggest that DELE1 is likely cleaved into DELE1-S by HtrA2 in the cytosol, while the DELE1-VS form might be generated during halted translocation of the protein into mitochondria. Together, this study identifies MPIS as the overarching stress detected by DELE1 and identifies HtrA2 as a critical protease involved in DELE1 processing. 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/2023.02.24.529955v1?rss=1 Authors: Hasan, S., Fernandopulle, M. S., Humble, S. W., Frankenfield, A. M., Li, H., Prestil, R., Johnson, K. R., Ryan, B. J., Wade-Martins, R., Ward, M. E., Hao, L. Abstract: Progranulin (PGRN) is a lysosomal protein implicated in various neurodegenerative diseases. Over 70 mutations discovered in the GRN gene all result in reduced expression of PGRN protein. However, the detailed molecular function of PGRN within lysosomes and the impact of PGRN deficiency on lysosomal biology remain unclear. Here we leveraged multifaceted proteomic techniques to comprehensively characterize how PGRN deficiency changes the molecular and functional landscape of neuronal lysosomes. Using lysosome proximity labeling and immuno-purification of intact lysosomes, we characterized lysosome compositions and interactomes in both human induced pluripotent stem cell (iPSC)-derived glutamatergic neurons (i3Neurons) and mouse brains. Using dynamic stable isotope labeling by amino acids in cell culture (dSILAC) proteomics, we measured global protein half-lives in i3Neurons for the first time and characterized the impact of progranulin deficiency on neuronal proteostasis. Together, this study indicated that PGRN loss impairs the lysosome degradative capacity with increased levels of v-ATPase subunits on the lysosome membrane, increased catabolic enzymes within the lysosome, elevated lysosomal pH, and pronounced alterations in neuron protein turnover. Collectively, these results suggested PGRN as a critical regulator of lysosomal pH and degradative capacity, which in turn influences global proteostasis in neurons. The multi-modal techniques developed here also provided useful data resources and tools to study the highly dynamic lysosome biology in neurons. 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/2023.02.22.529417v1?rss=1 Authors: Hino, C., Chan, G., Jordaan, G., Chang, S. S., Saunders, J., Bashir, M. T., Hansen, J. E., Gera, J., Weisbart, R. H., Nishimura, R. N. Abstract: Heat shock proteins (HSPs), especially Hsp70 (HSPA1), have been associated with cellular protection from various cellular stresses including heat, hypoxia-ischemia, neurodegeneration, toxins, and trauma. Endogenous HSPs are often synthesized in direct response to these stresses but in many situations are inadequate in protecting cells. The present study addresses the transduction of Hsp70 into cells providing protection from acute oxidative stress by H2O2. The recombinant Fv-Hsp70 protein and two mutant Fv-Hsp70 proteins minus the ATPase domain, and minus the ATPase and terminal lid domains were tested at 0.5 and 1.0 uM concentrations after two different concentrations of H2O2 treatment. All three recombinant proteins protected SH-SY5Y cells from acute H2O2 toxicity. This data indicated that the protein binding domain was responsible for cellular protection. In addition, experiments pretreating cells with inhibitors of antioxidant proteins catalase and gamma-glutamylcysteine synthase (GGCS) before H2O2 resulted in cell death despite treatment with Fv-Hsp70, implying that both enzymes were protected from acute oxidative stress after treatment with Fv-Hsp70. This study demonstrates that Fv-Hsp70 is protective in our experiments primarily by the protein-binding domain. The Hsp70 terminal lid domain was also not necessary for protection. Cellular protection was protective via the antioxidant proteins catalase and GGCS. 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/2023.02.06.527293v1?rss=1 Authors: Wang, B., Martini-Stoica, H., Qi, C., Lu, T.-C., Wang, S., Xiong, W., Qi, Y., Xu, Y., Sardiello, M., Li, H., Zheng, H. Abstract: Transcription factor EB (TFEB) mediates gene expression through binding to the Coordinated Lysosome Expression And Regulation (CLEAR) sequence. TFEB targets include subunits of the vacuolar ATPase (v-ATPase) essential for lysosome acidification. Single nucleus RNA-sequencing (snRNA-seq) of wild-type and PS19 (Tau) transgenic mice identified three unique microglia subclusters in Tau mice that were associated with heightened lysosome and immune pathway genes. To explore the lysosome-immune relationship, we specifically disrupted the TFEB-v-ATPase signaling by creating a knock-in mouse line in which the CLEAR sequence of one of the v-ATPase subunits, Atp6v1h, was mutated. We show that the CLEAR mutant exhibited a muted response to TFEB, resulting in impaired lysosomal acidification and activity. Crossing the CLEAR mutant with Tau mice led to higher tau pathology but diminished microglia response. These microglia were enriched in a subcluster low in mTOR and HIF-1 pathways and was locked in a homeostatic state. Our studies demonstrate a physiological function of TFEB-v-ATPase signaling in maintaining lysosomal homoeostasis and a critical role of the lysosome in mounting a microglia and immune response in tauopathy and Alzheimer disease. 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/2023.02.03.526715v1?rss=1 Authors: Elezaby, A., Lin, A. J., Vijayan, V., Pokhrel, S., Bechara, L. R., Ostberg, N. P., Queliconi, B. B., Campos, J. C., Ferreira, J. C., Haileselassie, B., Mochly-Rosen, D. Abstract: Cardiac troponin I (cTnI) is a sarcomeric protein critical to myocyte contraction. Unexpectedly, we found that some cTnI localized to the mitochondrial matrix in the heart, inhibited mitochondrial functions when stably expressed in non-cardiac cells and increased opening of the mitochondrial permeability transition pore under oxidative stress. Direct, specific, and saturable binding of cTnI to ATP synthase was demonstrated in vitro, using immune-captured ATP synthase, and in cells using proximity ligation assay. cTnI binding doubled F1F0 ATPase activity, whereas skeletal troponin I and several human mutant cTnI variants associated with familial hypertrophic cardiomyopathy did not. A rationally-designed ten amino acid peptide, P888, inhibited cTnI binding to ATP synthase and cTnI-induced increase in ATPase activity in vitro and reduced cardiac injury following transient ischemia in vivo. We therefore suggest that mitochondria-associated cTnI may inhibit cardiac ATP synthase under basal conditions; pharmacological agents that release this inactivating effect of cTnI and thus preventing ATP hydrolysis during cardiac ischemia may increase the reservoir of functional mitochondria to reduce cardiac injury. 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/2023.01.31.526435v1?rss=1 Authors: LEVEQUE, C., MAULET, Y., WANG, Q., RAME, M., MOCHIDA, S., SANGIARDI, M., YOUSSOUF, F., IBORRA, C., SEAGAR, M., VITALE, N., EL FAR, O. Abstract: V-ATPase is an important factor in synaptic vesicle acidification and is implicated in synaptic transmission. Rotation of the extra-membranous V1 sector drives proton transfer through the membrane-embedded multi-subunit V0 sector of the V-ATPase. Intra-vesicular protons are then used to drive neurotransmitter uptake by synaptic vesicles. V0a and V0c, two membrane subunits of the V0 sector have been shown to interact with SNARE proteins and their photo-inactivation rapidly impairs synaptic transmission. V0d, a soluble subunit of the V0 sector strongly interacts with its membrane embedded subunits and is crucial for the canonic proton transfer activity of the V-ATPase. Our investigations show that the loop 1.2 of V0c interacts with complexin, a major partner of the SNARE machinery and that V0d1 binding to V0c inhibits this interaction, as well as V0c association with SNARE complex. Injection of recombinant V0d1 in rat superior cervical ganglion neurons rapidly reduced neurotransmission. In chromaffin cells, V0d1 overexpression and V0c silencing modified in a comparable manner several parameters of unitary exocytotic events. Our data suggest that V0c subunit promotes exocytosis via interactions with complexin and SNAREs and that this activity can be antagonized by exogenous V0d. 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/2023.01.25.525394v1?rss=1 Authors: Tillotson, R., Yan, K., Ruston, J., de Young, T., Cordova, A., Turcotte-Cardin, V., Yee, Y., Taylor, C., Visuvanathan, S., Babbs, C., Ivakine, E. A., Sled, J. G., Nieman, B. J., Picketts, D. J., Justice, M. J. Abstract: ATRX is a chromatin remodelling ATPase that is involved in transcriptional regulation, DNA damage repair and heterochromatin maintenance. It has been widely studied for its role in ALT-positive cancers, but its role in neurological function remains elusive. Hypomorphic mutations in the X-linked ATRX gene cause a rare form of intellectual disability combined with alpha-thalassemia called ATR-X syndrome in hemizygous males. Patients also have facial dysmorphism, microcephaly, musculoskeletal defects and genital abnormalities. Since complete deletion of ATRX in mice results in early embryonic lethality, the field has largely relied on conditional knockout models to assess the role of ATRX in multiple tissues. Given that null alleles are not found in patients, a more patient-relevant model was needed. Here, we have produced and characterised the first patient mutation knock-in model of ATR-X syndrome, carrying the most common patient mutation, R246C. This is one of a cluster of missense mutations located in the chromatin interaction domain that disrupts its function. The knock-in mice recapitulate several aspects of the patient disorder, including craniofacial defects, microcephaly and impaired neurological function. They provide a powerful model for understanding the molecular mechanisms underlying ATR-X syndrome and for testing potential therapeutic strategies. 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/2022.12.16.520763v1?rss=1 Authors: Raman, M., Ahlstedt, B. A., Ganji, R., Mukkavalli, S., Paolo, J., Gygi, S. P. Abstract: Endoplasmic reticulum (ER) protein homeostasis (proteostasis) is essential to facilitate proper folding and maturation of proteins in the secretory pathway. Loss of ER proteostasis due to cell stress or mutations in ER proteins can lead to the accumulation of misfolded or aberrant proteins in the ER and triggers the unfolded protein response (UPR). In this study we find that the p97 adaptor UBXN1 is an important negative regulator of the UPR. Loss of UBXN1 significantly sensitizes cells to ER stress and activates canonical UPR signaling pathways. This in turn leads to widespread upregulation of the ER stress transcriptional program. Using comparative, quantitative proteomics we show that deletion of UBXN1 results in a significant enrichment of proteins belonging to ER-quality control processes including those involved in protein folding and import. Notably, we find that loss of UBXN1 does not perturb p97-dependent ER associated degradation (ERAD). Our studies indicate that loss of UBXN1 increases translation in both resting and ER-stressed cells. Surprisingly, this process is independent of p97 function. Taken together, our studies have identified a new role for UBXN1 in repressing translation and maintaining ER proteostasis in a p97 independent manner. 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/2022.11.29.518367v1?rss=1 Authors: Alonso-Gil, D., Cuadrado, A., Gimenez-Llorente, D., Rodriguez-Corsino, M., Losada, A. Abstract: Cohesin organizes the genome through the formation of chromatin loops. NIPBL activates cohesin's ATPase and is essential for loop extrusion, but its requirement for cohesin loading is currently unclear. Here we have examined the effect of reducing NIPBL levels on the behavior of the two cohesin variants carrying STAG1 or STAG2 by combining a flow cytometry assay to measure chromatin-bound cohesin with analyses of its genome-wide distribution and genome contacts. We show that NIPBL depletion results in increased cohesin-STAG1 on chromatin that further accumulates at CTCF positions while cohesin-STAG2 diminishes genome-wide. Our data support a model in which NIPBL is not required for initial association of cohesin with chromatin but it is for loop extrusion, which in turn facilitates stabilization of cohesin-STAG2 at CTCF positions after being loaded elsewhere. In contrast, cohesin-STAG1 is loaded and stabilized at CTCF sites even under low NIPBL levels, but genome folding is severely impaired. 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/2022.11.14.516427v1?rss=1 Authors: Li, T. Y., Wang, Q., Gao, A. W., Li, X., Mottis, A., Shong, M., Auwerx, J. Abstract: Lysosomes are central platforms for not only the degradation of macromolecules but also the integration of multiple signaling pathways. However, whether and how lysosomes mediate the mitochondrial stress response (MSR) remain largely unknown. Here, we demonstrate that lysosomal acidification via the vacuolar H+-ATPase (v-ATPase) is essential for the transcriptional activation of the mitochondrial unfolded protein response (UPRmt). Mitochondrial stress stimulates v-ATPase-mediated lysosomal activation of the mechanistic target of rapamycin complex 1 (mTORC1), which then directly phosphorylates the MSR transcription factor, activating transcription factor 4 (ATF4). Disruption of mTORC1-dependent ATF4 phosphorylation blocks the UPRmt, but not other similar stress responses, such as the UPRER. Finally, ATF4 phosphorylation downstream of the v-ATPase/mTORC1 signaling is indispensable for sustaining mitochondrial redox homeostasis and protecting cells from reactive oxygen species (ROS)-associated cell death upon mitochondrial stress. Thus, v-ATPase/mTORC1-mediated ATF4 phosphorylation via lysosomes links mitochondrial stress to UPRmt activation and mitochondrial function resilience. 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/2022.11.04.515188v1?rss=1 Authors: Bidaud-Meynard, A., Nicolle, O., Bourdais, A., Duclos, M., Saleh, J., Ruemmele, F., Farin, H. F., Delacour, D., Moshous, D., Michaux, G. Abstract: Intestinal microvillus atrophy is a major cause of enteropathies such as idiopathic or congenital diarrhea that are often associated with severe morbidity. It can be caused by genetic disorders, inflammatory diseases, toxins or pathogens. In particular, Microvillus inclusion disease (MVID) is characterized by a chronic intractable diarrhea and a severe microvillus atrophy. It is triggered by mutations in MYO5B, STX3, MUNC18.2 or UNC45A which alter epithelial polarity by affecting apical trafficking in intestinal epithelial cells. Furthermore, we recently established that the depletion of the V0 sector of the V-ATPase complex induces an MVID-like phenotype in C. elegans. In this study we investigated the function of the V0-ATPase complex in mouse intestinal organoids. We found that its depletion also triggers a very severe microvillus atrophy in this model. Furthermore, we established that the polarity of intestinal cells is affected in a patient carrying mutations in TCIRG1 which encodes a V0-ATPase subunit. However, V0-ATPase depletion does not recapitulate other MVID-specific phenotypes such as subapical vesicle accumulation and Rab11+ endosomes mislocalization. Finally, we found that the apical localization of the V0-ATPase is disrupted in MVID patients. Altogether these results suggest a role for the V0-ATPase in microvillus atrophy which might be independent from apical trafficking. 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/2022.11.01.514550v1?rss=1 Authors: Nguyen, L. D., Wei, Z., Silva, M. C., Barberan-Soler, S., Rabinovsky, R., Muratore, C. R., Stricker, J. M. S., Hortman, C., Young-Pearse, T. L., Haggarty, S. J., Krichevsky, A. M. Abstract: MicroRNAs (miRNAs) are short RNAs that regulate fundamental biological processes. miR-132, a key miRNA with established functions in Tau homeostasis and neuroprotection, is consistently downregulated in Alzheimer's disease (AD) and other tauopathies. miR-132 overexpression rescues neurodegenerative phenotypes in several AD models. To complement research on miRNA-mimicking oligonucleotides targeting the central nervous system, we developed a high-throughput-screen coupled high-throughput-sequencing (HTS-HTS) in human induced pluripotent stem cell (iPSC)-derived neurons to identify small molecule inducers of miR-132. We discovered that cardiac glycosides, which are canonical sodium-potassium ATPase inhibitors, selectively upregulated miR-132 in the sub-M range. Coordinately, cardiac glycoside treatment downregulated total and phosphorylated Tau in rodent and human neurons and protected against toxicity by glutamate, N-methyl-D-aspartate, rotenone, and A{beta} oligomers. In conclusion, we identified small-molecule drugs that upregulated the neuroprotective miR-132 and ameliorated neurodegenerative phenotypes. Our dataset also represents a comprehensive resource for discovering small molecules that modulate specific miRNAs for therapeutic purposes. 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/2022.08.25.505358v1?rss=1 Authors: Becchi, S., Chieng, B., Bradfield, L., Capallan, R., Leung, B., Balleine, B. W. Abstract: The loss of neurons in parafascicular thalamus (Pf) and of their inputs to dorsomedial striatum (DMS) are associated with Lewy body disease (LBD) and Parkinsons disease dementia (PDD) and have been linked to the effects of neuroinflammation. In rats, these inputs regulate the function of striatal cholinergic interneurons (CINs) that are necessary for the flexible encoding of the action-outcome (AO) associations for goal-directed action. We found that these inputs modify the encoding, not retrieval, of new AO associations and cause burst-pause firing of CINs in the DMS during AO remapping. These adaptive effects were abolished by neuroinflammation in the Pf, resulting in the loss of goal-directed control when the rats were required to update AO associations after a change in contingency. We found that the neuronal and behavioral deficits induced by inflammation in the Pf were rescued by administration of selegiline, a MAO-B inhibitor that we found also enhances ATPase activity in CINs, suggesting a potential treatment for cognitive deficits associated with inflammation affecting the function of midline thalamic nuclei and related structures. Copy rights belong to original authors. Visit the link for more info Podcast created by PaperPlayer

In this episode of the Brawn Body Health and Fitness Podcast, Dan is joined by Brunde Broady to discuss Calcium ATPase, the little-known enzyme at the root of your overall cellular health. Brunde's quest began as a mother, searching for answers to her son Knute's health challenges. Her knowledge evolved through countless hours of painstaking research. Brunde's life mission proceeds as an author, leading her to write The Calcium Connection, which empowers you to take charge of your health and understand how Calcium ATPase affects the very balance of your well-being. Brunde is also the director of Neurodon, a biopharmaceutical company which develops therapeutics for significant diseases like diabetes and Alzheimer's, Parkinson's, and Huntington's disease. Neurodon recently received a $2 million grant from the National Institute on Aging at the National Institutes of Health to further essential Alzheimer's research. Brunde's goal is to share her discovery and knowledge with you, so you can make educated decisions regarding your health, lifestyle, and of course, Calcium ATPase. For more on Brunde Broady, you can check out her website at https://www.brundebroady.com/ For more on Neurodon, you can check out their website here https://neurodon.net/ To purchase Brunde's Book on Amazon, click here: https://www.amazon.com/Calcium-Connection-Little-Known-Enzyme-Cellular/dp/1510763910?msclkid=50ca155bcf3211eca8ffddc6fd6b47a8 To keep up to date with everything we are currently doing on the podcast, be sure to subscribe and follow @brawnbody on social media! This episode is brought to you by CTM band recovery products - the EXACT soft tissue recovery technology used by Dan. CTM Band was founded by Dr. Kyle Bowling, a sports medicine practitioner who treats professional athletes (and was a guest on the Brawn Body Podcast!). You can check out their website here: https://ctm.band/collections/ctm-band ... while you're there, be sure to use the coupon code "BRAWN10" for 10% off! This episode is also brought to you by Ice shaker! Use our affiliate link to help support future podcast episodes and development by clicking here: https://www.iceshaker.com?sca_ref=1520881.zOJLysQzKe Make sure you SHARE this episode with a friend who could benefit from the information we shared! Check out everything we do, including blog posts, fitness programs, and more by clicking here: https://linktr.ee/brawnbodytraining Liked this episode? Leave a 5-star review on your favorite podcast platform! --- This episode is sponsored by · Anchor: The easiest way to make a podcast. https://anchor.fm/app --- Send in a voice message: https://anchor.fm/daniel-braun/message Support this podcast: https://anchor.fm/daniel-braun/support

On Episode 15 of the Stroke Alert Podcast, host Dr. Negar Asdaghi highlights two articles from the April 2022 issue of Stroke: “Kawasaki Disease May Increase the Risk of Subsequent Cerebrovascular Disease” and “Effect of Moderate and Severe Persistent Hyperglycemia on Outcomes in Patients With Intracerebral Hemorrhage.” She also interviews Dr. François Gros-Louis about his article “Moyamoya Disease Susceptibility Gene RNF213 Regulates Endothelial Barrier Function.” Dr. Negar Asdaghi:                        1) How would you counsel the parent of a child who has just recovered from Kawasaki disease on their child's future risk of having a stroke? 2) Should we or should we not treat stress hyperglycemia in the setting of acute intracerebral hemorrhage? 3) What is the CRISPR-Cas9 gene editing technology? And why, if you haven't heard of it already, you most definitely should be listening to this podcast? We're back here with the April issue of the Stroke Alert Podcast, and this is the latest in Stroke. Stay with us. Dr. Negar Asdaghi:                        Welcome back to another extremely informative Stroke Alert Podcast. My name is Negar Asdaghi. I'm an Associate Professor of Neurology at the University of Miami Miller School of Medicine, and your host for the monthly Stroke Alert Podcast. The April 2022 issue of Stroke contains a range of really exciting papers and topics. In the paper titled "Vascular Response to Spreading Depolarization Predicts Stroke Outcome," we have a really interesting in vivo mouse model of ischemic stroke, looking at the spreading patterns of cortical depolarization and the subsequent vascular response to this by way of hyperemia. The researchers from Zurich University, led by Dr. Binder and colleagues, walk us through how the patterns of hyperemia can actually predict the severity of subsequent ischemic injury. Dr. Negar Asdaghi:                        In a separate paper in this issue of the journal, we're reminded of how the classic NIH Stroke Scale can underestimate the severity of neurological symptoms and outcomes in patients with posterior circulation infarcts. In the paper led by Dr. Alemseged and colleagues, the investigators from the Royal Melbourne Hospital in Australia evaluate the prognostic accuracy of the Posterior NIH Stroke Scale, which is the modified version of the classic NIH Stroke Scale, in predicting the outcomes of patients with posterior circulation infarcts. Dr. Negar Asdaghi:                        I encourage you to review these papers in addition to listening to our podcast today. Later in the podcast, I have the great pleasure of interviewing Dr. François Gros-Louis from Laval University in Quebec to discuss the latest in gene editing technology and how this technology has helped his team unravel the biological function of RNF213 susceptibility gene in Moyamoya disease. But first with these two articles. Dr. Negar Asdaghi:                        Kawasaki disease, which was first described in 1976, is an acute febrile illness predominantly affecting children younger than five years of age. In addition to fever, other clinical signs of the disease include rash, bilateral conjunctival injection, cervical lymphadenopathy, swelling of the hands and feet, and irritation and inflammation of the mouth, lips, and throat. Now, for those of us like me who are adult neurologists, here is a quick review of the pathophysiology of Kawasaki disease. Dr. Negar Asdaghi:                        This is a medium vessel vasculopathy, most significantly affecting the coronary arteries, a vasculopathy that is characterized by three linked pathological processes, necrotizing arteritis, subacute to chronic vasculitis, and luminal myofibroblastic proliferation. So, simply put, these processes can lead to stenotic lesions in various vascular beds, which are affected by this disease. Dr. Negar Asdaghi:                        And as we mentioned earlier, the most recognized vascular blood vessels affected by Kawasaki disease are the coronary arteries, which can lead to myocardial ischemia, infarction, and sudden death in these cases. However, involvement of other vascular beds, including cerebral vessels, are also increasingly reported as part of Kawasaki disease. Dr. Negar Asdaghi:                        So, in the current issue of the journal, Dr. Chien-Heng Lin from the Division of Pediatric Pulmonology at China Medical University Children's Hospital in Taiwan and colleagues studied the subsequent risk of cerebrovascular events in patients with Kawasaki disease. Using the National Health Insurance Research Database of Taiwan, they collected data on 8467 children with Kawasaki disease from 2000 to 2012. And for each child with Kawasaki, data was also collected on four randomly selected non-Kawasaki disease children who were matched with the Kawasaki cohort for sex, urbanization level of residence, and parental occupation. Dr. Negar Asdaghi:                        So, that gave them a sample size of over 33,000 children for their non-Kawasaki cohort. And then they compared the risk of subsequent stroke in children between the two cohorts. The study period for any given patient would end when the said patient was either diagnosed with a cerebrovascular disease or withdrew from the research database. Dr. Negar Asdaghi:                        So, in terms of their demographics, 61% of patients in the Kawasaki group were boys; 88% of the Kawasaki cohort were younger than five years of age. So, here are the findings. Number one, the incident rate of subsequent cerebrovascular disease was 14.7 per hundred thousand person years in the Kawasaki cohort versus only 4.6 per hundred thousand person years in the non-Kawasaki cohort. That's greater than a threefold higher incidence rate of cerebrovascular disorders for children who had Kawasaki disease before. Dr. Negar Asdaghi:                        This finding was independent of other potential confounders, which they adjusted for in their multivariate analysis. Now, the length of follow up was a median of 9.8 years for the entire cohort. And on the issue of time, they found two important associations. The first finding was that when the follow-up time was stratified by five-year periods, Kawasaki disease cohort patients showed a significantly higher risk of developing a stroke within the first five years after being diagnosed. Dr. Negar Asdaghi:                        And the second important association was that when they looked at the age at the time of diagnosis of Kawasaki, children who were younger than five years at the time of diagnosis were at a significantly higher risk of having a future stroke as compared to those who were older than five at the time of diagnosis. Dr. Negar Asdaghi:                        So, simply put, the risk of subsequent stroke was higher in children who acquired the disease at a younger age, and that risk was higher in the first few years after the diagnosis of Kawasaki disease. The authors discuss a number of putative mechanisms to link Kawasaki with stroke. The most important being a cardiac source of embolism that we already alluded to, but other etiologies, including medium vessel cerebral vasculitis, or hypercoagulability in the setting of increased systemic inflammation, and even Kawasaki disease-associated aneurysmal rupture to cause hemorrhagic forms of stroke, are discussed in the paper and should be considered in the correct setting in children with a prior history of this disease. Dr. Negar Asdaghi:                        So, what we learned from this large population-based pediatric study is that Kawasaki disease does indeed increase the risk of subsequent cerebrovascular disorders, and its influence is stronger in children who are diagnosed with this condition under the age of five, and the time period during which the risk of stroke is the highest is within the first five years after the diagnosis. Dr. Negar Asdaghi:                        In the setting of spontaneous intracerebral hemorrhage, or ICH, much research has focused on the association between hypertension and blood pressure-lowering therapies and hematoma expansion and functional outcomes, but a lot less attention relatively has been given to the impact of hyperglycemia and ICH-related outcomes. Dr. Negar Asdaghi:                        The current guidelines state that serum glucose should be monitored and both hypo- and hyperglycemia should be avoided in the setting of ICH. The older studies have given us inconsistent results as to whether or not hyperglycemia can increase the risk of ICH-related mortality. More recent studies have suggested that perhaps persistent hyperglycemia is indeed a predictor of poor neurological outcomes in ICH, but these results come from smaller single-center studies, which require further confirmation. And this confirmation is exactly what Dr. Adnan Qureshi from Zeenat Qureshi Stroke Institute and the Department of Neurology at University of Missouri and colleagues aim to give us in their study titled "Effect of Moderate and Severe Persistent Hyperglycemia on Outcomes in Patients With Intracerebral Hemorrhage." Dr. Negar Asdaghi:                        So, they use data from the ATACH-2 study, and a quick reminder that ATACH-2 was a multicenter randomized control trial of a thousand patients with acute spontaneous intracerebral hemorrhage enrolled within four and a half hours from symptom onset, and patients were randomized to either the intensive blood pressure control treatment arm to maintain their systolic blood pressure goal of 110 to 139 millimeter of mercury versus standard treatment arm, which was keeping their systolic blood pressure above 140, between 140 to 179 millimeter of mercury, in the first 24 hours after randomization. Dr. Negar Asdaghi:                        You will recall that enrollment of ATACH-2 was stopped early because of futility after pre-specified interim analysis. The main results of the trial was published in 2016 in New England Journal of Medicine, and the primary results did not show a lower rate of death or disability in patients assigned to the intensive treatment group. Dr. Negar Asdaghi:                        So, in the current paper, in this current issue of the journal, the authors looked at the glycemic status of the patients enrolled in the trial. As part of the trial, patients had a complete chemistry panel at baseline, 24, 48, and 72 hours from onset. So, they used the glucose measurement from this panel and defined moderate hyperglycemia as serum glucose level of over 140 and under 180 and severe hyperglycemia as serum glucose levels of equal or greater than 180. Dr. Negar Asdaghi:                        Now, persistent hyperglycemia was if two consecutive serum glucose levels were in the moderate or severe categories. And, very simply, they looked at the effects of hyperglycemia on ICH outcomes. And importantly, they evaluated whether hyperglycemia modified the effects of intensive blood pressure reduction on outcomes of ICH. So, of the thousand participants in ATACH-2, 11% had persistent moderate hyperglycemia, and 17% had severe persistent hyperglycemia. Those in the hyperglycemic group were more likely to be diabetic, not surprisingly, more likely to have a history of hypertension and dyslipidemia as compared to the normal glycemic patients. Dr. Negar Asdaghi:                        And here are the results. Number one, serious adverse events were higher in the hyperglycemic groups, whether we're talking about the moderate or the severe hyperglycemic groups. This is despite the fact that the rate of hematoma expansion and perihematomal edema was not different based on the hyperglycemic status. However, the hyperglycemic patients were more likely to have serious adverse events, which were operationally defined as complications that were not expected to have occurred from the study intervention, in this case, the intensive hypertensive therapy, and resulted in either death or prolonged hospitalization or persistent or significant disabilities. Now, serious renal adverse events, which are, of course, expected as a complication for aggressive blood pressure therapy, were also significantly higher in the hyperglycemic category. Dr. Negar Asdaghi:                        Now, their next important finding was that overall, both moderate and severe hyperglycemia was associated with higher odds of 90 days disability or death post-ICH adjusting for typical variables that could predict these outcomes, such as GCS score, hematoma volume, presence or absence of intraventricular hemorrhage, amongst other factors that they accounted for. Dr. Negar Asdaghi:                        Now, number three, this is perhaps the most important finding of the study. Among patients without a preexisting history of diabetes, both moderate and severe hyperglycemia increased the risk of death and disability at 90 days after adjusting for all the potential confounders, but hyperglycemia was not associated with these poor outcomes in those with a prior history of diabetes. I'm going to pause here to let this information sink in. Let's go over them again, stress hyperglycemia in non-diabetics was associated with poor ICH outcomes, but high sugars in diabetics did not predict the same poor outcomes. And finally, they looked at the possible interactions between the glycemic status and the ATACH-2 intervention, which as we alluded to earlier, which was intensive versus standard blood pressure therapy, and it turns out that the intensive systolic blood pressure reduction was indeed associated with a lower rate of hematoma expansion only in patients with normal glycemia, but not in those with moderate or severe hyperglycemia. Dr. Negar Asdaghi:                        So, this is again food for thought. Simply put, if the sugars are not well controlled, it appears that intensive blood pressure control would not lower the rate of hematoma expansion. Blood pressure lowering works when the sugar levels are controlled. So, overall, here are the two simple messages of this study. Number one, hyperglycemia in the acute setting of intracerebral hemorrhage is associated with poor outcomes or death only in those with stress hyperglycemia, meaning in those who have high sugar levels, but are not diabetic. Dr. Negar Asdaghi:                        Number two, there seems to be an important interaction between the acute glycemic status of the patients and how intensive blood pressure control can prevent hematoma expansion, in that intensive BP control is only effective in prevention of hematoma expansion if the sugar levels are normal. So, a lot of thought-provoking and hypothesis-generating findings, and definitely more to come on this topic. Dr. Negar Asdaghi:                        Moyamoya disease, or MMD, is an idiopathic disorder characterized by progressive stenosis of the supraclinoid internal carotid artery and its main branches in subsequent formation of a network of abnormal lenticulostriate collaterals. First described in Japan, the term "Moyamoya" is a Japanese expression for the puff of smoke and describes the characteristic appearance of the tangled and abnormal collateral vessels that are seen in angiography in various stages of the Moyamoya disease. Dr. Negar Asdaghi:                        Epidemiological studies have revealed several risk factors associated with Moyamoya disease, including Asian ethnicity, female gender, and a family history of the disorder. Given that 15% of MMD patients have a family history of this disease, it's not surprising that genetic factors are suspected to underlie its pathogenesis. Now, a polymorphism in the ring finger protein 213, or RNF213, gene on chromosome 17 has been identified as the strongest genetic susceptibility factor for Moyamoya disease specifically in the East Asian population. Dr. Negar Asdaghi:                        But despite the many advances in understanding the pathophysiology of MMD, as well as advances in animal models and genetic studies, to date, none of the animal models of RNF213 have quite replicated the vascular abnormalities that are typically seen in human Moyamoya disease. Dr. Negar Asdaghi:                        The scientists feel that this is related to how little is known about the exact biological function of RNF213 gene and the protein it encodes. So, in the current issue of the journal, in the study titled "Moyamoya Disease Susceptibility Gene RNF213 Regulates Endothelial Barrier Function," Dr. François Gros-Louis from CHU de Québec Research Center at Laval University in Québec and colleagues aim to study the biological functions of RNF213 using a novel genome editing technology by the name of CRISPR-Cas9 technology. Dr. Negar Asdaghi:                        Joining me now is Dr. Gros-Louis himself to discuss the findings of this paper. Dr. Gros-Louis is a Professor of Neurosciences at the Department of Surgery at Laval University. He holds the Canada Research Chair in Brain Disease Modeling and is the Director of the Induced Pluripotent Stem Cell Platform research in Québec. Dr. Negar Asdaghi:                        Good morning, François. Welcome to our podcast. And thank you so much for joining us. Dr. François Gros-Louis:               My pleasure. Dr. Negar Asdaghi:                        François, you have to promise to hold my clinician's hand through this interview as obviously these are some foreign subjects for us, but very excited to learn from your study and learn from you on the association between RNF213 and the pathophysiology of what happens in Moyamoya disease. Now, before we talk about your paper, can you please talk to us about some basic concepts? What is the RNF213 protein? Dr. François Gros-Louis:               Yes. The RNF213 gene is thought to be involved in mediating protein, protein interactions. The protein also contains a domain which is associated with an ATPase activity. This gene is a susceptibility gene for Moyamoya disease, as you mentioned in the introduction, vascular disorder of intracranial arteries. It's encoded in ubiquitously expressed protein. The protein is found to be expressed throughout the cytocell with the partial association in the intracellular membrane and cytoskeleton. Its expression varies according to the tested tissue type or location or cellular types. Dr. François Gros-Louis:               Although the function of RNF213 protein is unknown, studies suggest that it plays a role in the proper development of blood vessels, cell proliferation, and inflammation. Recently, RNF213 has been reported to be associated with angiogenesis. However, little is known about its endogenous function or its pathogenic role in Moyamoya disease. Our results are in line with these results and indicate that RNF213 could also be a key regulator of cerebral endothelial integrity, whose disruption could be an early pathological mechanism leading to Moyamoya disease. Dr. Negar Asdaghi:                        So, just to continue on this, there's quite a bit of research already done on association of the RNF213 gene, that's located, as we noted earlier, on chromosome 17, and basically susceptibility of development of Moyamoya disease. Can you give our listeners a brief overview of this genetic connections and what was known from past research? Dr. François Gros-Louis:               Yeah, there is a couple polymorphism describing this gene, the most frequent, the most prevalent genetic study have identified the variant R4810K, meaning for arginine is replaced by another amino acid at the position of 4810 within the protein. It's a large protein and a large gene and a susceptible gene and a risk factor for developing Moyamoya disease. Dr. François Gros-Louis:               So, people bearing this variant have a higher chance to develop the disease. This is a loss of function variant, also called inactivating mutation, meaning that the mutated gene product have less or no function. So, this variant is found in heterozygous, meaning one copy, or two copy homozygous in Moyamoya disease patients. While patient bearing homozygous mutation develop a more severe disease with earlier age of onset and worse prognosis, patients bearing heterozygous mutation can also develop the disease. Dr. François Gros-Louis:               So, strong evidence suggests that the carrying rate of RNF213 R4810K mutant is closely related and give a higher chance to develop the disease. Interestingly, also with colleagues, we found that there are other variants within this genes leading to what we think is a gain of function mutation have been associated also with other cerebrovascular disease, such as intracranial aneurysms. Dr. Negar Asdaghi:                        So, François, this is very interesting. Let me recap what you mentioned so I know that I understood it. So, this is an interesting gene, this RNF213, and basically evidence shows that mutations in the RNF213, whether it's loss of function or gain of function, both can result in variety of cerebrovascular disorders. And interestingly, the phenotype of the disease when it comes to loss of function of this gene is actually correlated with whether a person is a carrier, homozygous carrier of this gene, loss of function, or heterozygous carrier of the gene. Dr. Negar Asdaghi:                        So, very interesting information for clinicians who treat patients with Moyamoya disease, specifically those who have a family history of Moyamoya disease, so perhaps a higher chance of carrying a genetic susceptibility gene. Now, we want to get to the paper that you published in this issue of the journal, but I think before we talk about your paper, we also have to have a basic understanding of this CRISPR-Cas9 technology, which is the new genome editing technology that you use in your experiments. Can you please give us a little bit of an overview of this technology? Dr. François Gros-Louis:               Yes. CRISPR-Cas9 gene editing is genetic engineering technique in molecular biology by which the genomes of living organisms may be modified. This technology allows genetic material to be added, removed, or altered at particular location in the genome. Several approaches to genome editing have been developed. Recent one is known as CRISPR-Cas9. So, the CRISPR-Cas9 system has generated a lot of excitement in the scientific community because it is faster, cheaper, and more accurate, and also more efficient than other existing genome editing methods. It's clearly revolutionizing the field in research. Dr. Negar Asdaghi:                        So, it's very exciting. It's truly a new chapter in gene targeting research and editing research. So, now we're ready to hear about your study. And I guess the first part of the study was just to look at how various cells in vitro that you used had expressed RNF213. Can you please tell us about the first part of your experiments? Dr. François Gros-Louis:               Yeah. We first wanted to know where the protein is expressed or where the protein is more highly expressed. So, we found by doing immunofluorescence analysis that the RNF213, so we confirmed that it's ubiquitously expressed in the cytoplasm of different cellular types. So, we found that significant difference also in the expression of RNF213 protein levels in several endothelial cells, where we found it's been highly expressed when compared to other endothelial cells isolated from different other body location, meaning outside of the CNS. So, it's highly expressed also when compared to smooth muscle cells or fibroblasts. Dr. Negar Asdaghi:                        Okay. So, just again, to recap for our listeners, this is, this RNF213 protein, is ubiquitously expressed in many different cell types, but you did find a significantly higher expression rates in endothelial cells, specifically those endothelial cells that were derived from cerebrovasculature. So, that's the first exciting part of the experiments that you showed in the study. Now, using the CRISPR-Cas9 technology, you and your team were able to successfully create an in vitro RNF213 knockout model. Can you please tell us about these models and also the main findings of your study? Dr. François Gros-Louis:               Yeah, so taken together, the results we presented in the article indicate that RNF213 could be a key regulator of cerebral, endothelial and tight junctions integrity whose disruption could be an early pathological mechanisms leading to Moyamoya disease. So, we established for the first time an easily reproducible and stable in vitro 3D model generated using the CRISPR-Cas9 gene editing technology. Dr. François Gros-Louis:               This advanced 3D culture approach has emerged as an excellent system to recapitulate histopathological feature reminiscent to disease pathogenesis. So, 3D cell culture approach is different from standard 2D culture, where cells are cultured, monolayered into a Petri dish. And we have results showing that the 3D cell culture system better mimic the in vivo conditions in terms of cell to cell and cell to matrix interaction and lead to histopathological phenotypic feature can be observed in cell culture, in a 3D fashion. Quite interestingly, alongside of providing the first evidence for the role of RNF213, the maintenance of endothelial barrier and the potential implication of this gene in the expression of maturation of tight junctions. So, we define a novel role for PECAM-1 as well in barrier impairment as a part of the disease pathogenic mechanisms. Dr. Negar Asdaghi:                        Okay. And now this is really interesting. So, I wanted to, again, recap some of the important points that you raised here. First of all, your in vitro models are different than the classic in vitro models, where 2D cells were basically grown in a Petri dish. You are trying to, more and more, replicating what happens, for instance, in blood vessels, where you have endothelial cells overlying mesenchymal cells underneath them, so tunica intima and then tunica media, and so you have 3D cells, where various types of cells are overlying each other in a more in vivo representation of what happens in blood vessels, which is truly interesting. Dr. Negar Asdaghi:                        And what you found was, in sort of summary, was that these knockout endothelial cells ended up having abnormal tight junctions and abnormal connectivity, which basically would lead in an in vivo model to abnormal leaky blood brain barrier, if this were truly in the in vivo model. Does that summarize the findings of the paper? Dr. François Gros-Louis:               Yes, perfectly. Dr. Negar Asdaghi:                        Perfect. And so I want to also give us a chance to talk about the important pro-inflammatory aspects of these knockout cells. You did find that a number of cytokines were expressed in excess in those RNF213 deficient cells. Can you please elaborate on those findings? Dr. François Gros-Louis:               So, to further investigate whether inflammation plays an important role in RNF213-associated Moyamoya disease development, we indeed performed experiments to study pro-inflammatory cytokines and analyze the immune secretome profiles of cerebral RNF213 deficient endothelial cells. So, then the cells can secrete different cytokines or different other proteins. So, by analyzing the secretome, we found an end secretion of a few pro-inflammatory cytokines indicating that inflammation may also play a central role in the initiation of the immune response in the pathogenesis of the disease. Dr. Negar Asdaghi:                        So, this is exciting, François. For years, we thought about the pathophysiology of Moyamoya disease as a disorder involving large vessels. And perhaps the initial thought was that it starts with excessive proliferation of smooth muscles within the middle layer of the cerebral blood vessels, in tunica media, and then perhaps subsequently there will be other abnormalities, including the intimal hyperplasia that is classically seen in Moyamoya. Dr. Negar Asdaghi:                        Your study seems to propose a shift in that pathophysiological paradigm, where the problem seems to start from endothelial cells, so inside of the blood vessels and the tunica intima, and then gradually would go out to the middle layers, and, of course, proposes the hyperinflammatory state in the Moyamoya disease as well. So truly interesting. Do you think that that is the new or rather a paradigm shift for pathophysiology of MMD? Dr. François Gros-Louis:               That's a great question. Our results certainly demonstrated that endothelial cells are involving in the disease pathogenesis in Moyamoya disease, but it doesn't exclude the possibility that other cell types might also be involved in the disease pathogenesis. We know, like you mentioned, that a blood vessel is formed by two different cell layers, tunica intima, media, and adventitia, containing, respectively, endothelial cells, smooth muscle cells, and fibroblasts. So which cells are to be blamed in Moyamoya disease is a question of many ongoing results studies over the years. Dr. François Gros-Louis:               So, using tissue-engineered approach to reconstruct small caliber blood vessels, as we developed in my lab, in combination with patient-derived stem cells, in which adult cells isolated from a patient of any individuals can be reprogrammed into stem cells and re-differentiated into different cell types in occurrence, smooth muscle, fibroblasts, or endothelial cells. We would like to generate blood vessels in which each of the different cellular layers will harbor or not, or a combination with RNF213 mutants. So, this will hopefully help us to elucidate this question. Dr. Negar Asdaghi:                        That's perfect. So, François, before we end the interview, I wanted to ask two more questions. So, what should be our top two takeaway messages from your study? Dr. François Gros-Louis:               We believe that the innovative transdisciplinary approach to generate, for the first time, as we describe in the article, an in vitro 3D model recapitulating important diseases features. So, this model could become a unique tool in precision medicine to study Moyamoya disease or other RNF213-associated pathologies. So, our study provides, for the first time, role of RNF213 in the maintenance of blood-brain barrier and the potential implication of RNF213 in the expression and maturation of tight junctions. Taken together, our data define a novel role for PECAM-1 in the blood-brain barrier impairment in Moyamoya disease. Dr. François Gros-Louis:               So, better characterization of each, also this regulated inflammatory molecules, we found taken separately could reveal a crucial information and help elaborate a more precise approach. Hence, this pro-inflammatory signature could be used as a circulatory biomarker for the follow-up of Moyamoya disease patients and to manage an appropriate treatment, according to the pathology progression. Dr. Negar Asdaghi:                        François, this is great. And last, I want to digress a little bit and ask you about the future of gene editing. I think it's important to end our interview with a little bit of a discussion regarding the future of CRISPR-Cas9 technology. In subatomic quantum physics, people talk about the God particles. And I feel that the CRISPR-Cas9 technology is, in a way, like playing God, if you agree. What do you think is the future for gene editing, and how do you see that helping us in terms of treatment of genetic causes of cerebrovascular disorders? Dr. François Gros-Louis:               Yes, gene editing is, like I said, revolutionizing, of course, experimental therapies for genetic disorder and generated excitement for new and improved gene therapies. We can think that it will be possible to correct any gene mutations associated with a disease to reestablish the normal or natural gene function and help treating the targeted diseases. But also, to me, the future of genome editing also resides in optimizing next generation disease models. The use of genome editing, in particular, the CRISPR-Cas9 technology, has extended to potential in generating new personalized model for a number of disorder, not only including Moyamoya disease or other cerebrovascular diseases, but also diseases like Alzheimer's, ALS, or Parkinson's disease, for which obtaining patient sample is difficult. Dr. François Gros-Louis:               No one wants to give up a bit of his brain. So modeling it, this disease, in vitro will be really helpful in combination also gene editing with the stem cells, induced pluripotent stem cells technology, will allow the generation of better model to mimic human disease and reflects the genetic drivers that govern specific pathology. So, the synergy between IPS cell-based model system and gene editing will play a pivotal role in the root of precision medicine and clinical translation in the future. Dr. Negar Asdaghi:                        Dr. François Gros-Louis, it was a pleasure learning from you. And we look forward to the endless possibilities brought by the future of genome editing technology. Dr. François Gros-Louis:               It was a pleasure discussing with you. Dr. Negar Asdaghi:                        Thank you for joining us. Dr. Negar Asdaghi:                        And this concludes our podcast for the April 2022 issue of Stroke. Please be sure to check out this month's table of contents for the full list of publications, including a series of Focused Updates on the topic of blood pressure management in stroke, organized by Dr. Else Sandset. I would also like to draw your attention to two scientific statements from the American Heart Association, which appear in print in the April issue. The first one is titled "Identifying Best Practices to Improve Evaluation and Management of In-Hospital Stroke," and the second one is on the effect of marijuana use on brain health. Dr. Negar Asdaghi:                        And now, to end our podcast, last month, in honor of the 2022 Olympic Games, and to celebrate those with determination to survive and push despite the most difficult of circumstances, we ended our podcast with the story of a refugee Olympic athlete. Dr. Negar Asdaghi:                        Sadly, since our last podcast, the world has seen even darker days of war, mass immigration, displacement, and human suffering. At times like this, we're reminded that although not all of us can help everyone, but at least each of us can do something to help someone, and the comfort in knowing that what we do in the field of medicine, from daily patient care to the scientific work leading to the next medical breakthrough, every action is a step forward in reducing the suffering of another person. And what better way to do this than staying alert with Stroke Alert. Dr. Negar Asdaghi:         This program is copyright of the American Heart Association, 2022. The opinions expressed by speakers in this podcast are their own and not necessarily those of the editors or of the American Heart Association. For more, visit AHAjournals.org.

This podcast is an Interview with Brunde Broady who has become an expert in a little know enzyme called Calcium ATPase. She provides valuable information on how important ATPase is to your health and longevity. It reinforces the importance of our fucus on doing things that help us at a cellular level.

This week, join author Tristram Bahnson and Associate Editor Changsheng Ma as they discuss the article "Association Between Age and Outcomes of Catheter Ablation Versus Medical Therapy for Atrial Fibrillation: Results from the CABANA Trial." Dr. Carolyn Lam: Welcome to Circulation On The Run, your weekly podcast summary and backstage pass to the journal and its editors. We're your co-hosts. I'm Dr. Carolyn Lam, Associate Editor from the National Heart Center and Duke National University of Singapore. Dr. Greg Hundley: And I'm Dr. Greg Hundley, Associate Editor, Director of the Poly Heart Center at VCU Health in Richmond, Virginia. Dr. Carolyn Lam: Guess what, Greg? For today's feature paper, we are going to be looking at a very interesting analysis from the CABANA trial, this time, looking at the association between age and outcomes of catheter ablation versus medical therapy for atrial fibrillation. Cool, huh? Okay, but first, let's go through some other important papers in today's issue. Why don't I let you go first? Dr. Greg Hundley: Well, Carolyn, my first paper pertains to the cost effectiveness of coronary artery bypass surgery, and it comes to us from the STICH trial. Dr. Carolyn Lam: Ah, very important question, but please remind us what the STICH trial is again. Dr. Greg Hundley: Right, Carolyn. So the Surgical Treatment for Ischemic Heart Failure trial, or STICH demonstrated that coronary artery bypass grafting reduced all-cause mortality rates out to 10 years compared with medical therapy alone in patients with ischemic cardiomyopathy and reduced left ventricular function, defined as an ejection fraction of less than or equal to 35%. Now in this study, the authors led by Dr. Derek Chew at University of Calgary examined the economic implications of these results using a decision-analytic patient-level simulation model to estimate the lifetime costs and benefits of CABG versus medical therapy alone, using patient-level resource use and clinical data collected from the STICH trial. Dr. Carolyn Lam: Again, really important study. And what did they find? Dr. Greg Hundley: Right, Carolyn. So first, using their patient-level simulation model incorporating resource use and clinical data collected from the STICH trial, they found that coronary artery bypass grafting was estimated to cost $63,989 per quality-adjusted life year gain compared to medical therapy alone. Second, in STICH eligible patients with left ventricular ejection fraction of less than 35% in coronary artery disease amenable to CABG, routine use of CABG increased the quality-adjusted life expectancy compared to medical therapy alone for an increased cost within current benchmarks for good value in healthcare within the United States. Then finally, Carolyn, together with the improved clinical outcomes seen in the 10 year extended follow-up of STICH, the findings in this study provide additional economic support for the use of coronary artery bypass grafting in patients with ischemic cardiomyopathy eligible for STICH. Dr. Carolyn Lam: Wow, thanks Greg. Well, this next study contributes to the understanding of the effect of lifestyle and genetic risk on the lifetime risk of coronary heart disease. Interesting? Well, listen up. This is from Dr. deVries from UT Health Science Center at Houston and colleagues who aimed to quantify remaining lifetime risk and years free of coronary heart disease according to polygenic risk and the AHA's Life's Simple 7 guidelines in the population base cohort of ARIC. As a reminder, the Life's Simple 7 by the AHA consists of smoking status, body weight, total cholesterol, blood glucose, blood pressure, physical activity, and diet. Dr. Greg Hundley: Ah, Carolyn. So genes versus lifestyle. So what did they find? Dr. Carolyn Lam: Participants with high polygenic risk may offset their lifetime risk of coronary heart disease by up to 50% through managing their health according to the Life's Simple 7's recommendations, depending on ancestry. Individuals with high polygenic risk scores and ideal Life's Simple 7 scores had 4.5 to 20 more coronary heart disease free years than individuals with high polygenic risk scores, but low Life's Simple 7 scores and again, depending on ancestry. Appropriate management of lifestyle and clinical risk factors of coronary heart disease play larger roles in the overall lifetime risk of coronary heart disease than presently available genetic information. Thus, communicating the effects of Life's Simple 7 measures and polygenic risk on coronary heart disease in terms of absolute risk may have important implications for education, policy, and environmental changes, which can benefit not only high risk individuals, but the whole population. Dr. Greg Hundley: Wow, Carolyn, really informative study and so nicely summarized. So Carolyn, my next paper comes to us from the world of preclinical science and it's from Professor Yan from Shanghai, Ruijin University School of Medicine. So Carolyn, previous studies have suggested that mitochondrial dysfunction plays critical roles in the progression of heart failure. However, the underlying mechanisms often remain unclear. Now since kinases have been reported to modulate mitochondrial function team investigated the effects of dual specificity tyrosine regulate kinase one B on mitochondrial, bio energetics, cardiac hypertrophy, and heart failure. Dr. Carolyn Lam: Wow. Okay. So what did they find Greg? Dr. Greg Hundley: Right, Carolyn. So this team found that Dual Specificity Tyrosine-Regulated Kinase 1B, our DYRK1B expression was clearly up regulated in failing human myocardium as well as in hypertrophic mirroring hearts and cardiac specific DYRK1B over expression resulted in cardiac dysfunction, accompanied by a decline in the left ventricular ejection fraction, as well as the fraction shortening. And it increased left ventricular myocardial fibrosis. Carolyn in striking contrast to DYRK1B over expression, the deletion of DYRK1B mitigated tack-induced cardiac hypertrophy and heart failure. In addition, the authors found that DYRK1B was positively associated with impaired mitochondrial bio-energetics by directly binding with stat three to increase its phosphorylation and nuclear accumulation. Thereby ultimately contributing toward the down regulation of PG C one alpha. Now, furthermore, the inhibition of DYRK1B or stat three activity using specific inhibitors was able to restore cardiac performance by rejuvenating mitochondrial bio-energetics. Dr. Carolyn Lam: Cool, Greg. So could you give us a take home? Dr. Greg Hundley: Right. So in summary then, Carolyn, taken together, the findings of this study provide new insights into the previously unrecognized role of DYRK1 beta in mitochondrial bio-energetics and the progression of cardiac hypertrophy in heart failure. Dr. Carolyn Lam: Fantastic. Thanks, Greg. Well, other papers in today's issue include an exchange of letters between Doctors Nie and Wollert on the article myeloid derived growth factor protects against pressure overload induced heart failure by preserving sarcoplasmic reticulum calcium, ATPase expression in cardiomyocytes. There's an AHA update [AHA Advocacy Page] paper by Dr. Churchwell on improving heart health through value-based payment. An ECG Challenge by Dr. Murphy on a “Curious ECG Morphology of a Cardiac Device.” An On My Mind paper by Dr. Figtree on “Sublingual Nitrates for Patients as a Default in the Post ACS Discharge Pack. Is the Time for a Rethink?” Dr. Greg Hundley: Right? Carolyn. Boy, this issue is really packed with great articles. There's a Perspective piece from Professor Stewart entitled “Myocardial Edema Provides A Link Between Pulmonary Arterial Hypertension and Pericardial Effusion.” There's a wonderful Frontiers in medicine piece from Professor Kandzari entitled “A Clinical Trial Design Principles and Outcomes Definitions for Device-Based Therapies for Hypertension: A Consensus Document from the Hypertension Academic Research Consortium.” And then finally, Carolyn, there's a Research Letter from Professor Wold entitled “E-Cigarette Aerosol Reduces Left Ventricular Function in Adolescent Mice. Well, Carolyn, how about we get onto those results from the CABANA trial?” Dr. Carolyn Lam: Let's go, Greg. Dr. Greg Hundley: Well, listeners, we are now here for our feature discussion and we have with us today, Dr. Tristram Bahnson from Duke University and one of our own Associate Editors, Dr. Changsheng Ma from Beijing. Welcome gentlemen. Tristram, we will start with you first. Could you describe for us some of the background pertaining to this particular research study and what was the hypothesis that you wanted to address? Dr. Tristram Bahnson: Sure. Being an active electrophysiologist, a challenge we've had over the years is to try to figure out for whom catheter ablation would be a preferred therapy. I've had the privilege of being part of the CABANA study team over the last several years. As listeners might recall, the CABANA trial was a very large trial looking specifically at hard endpoints, including mortality, to try to determine whether or not catheter ablation provides significant benefits to patient. Apart from what we already knew over the years, which is the catheter ablation was more effective than drug therapy to reduce AFib recurrences. That study, the CABANA proper study was published in 2019. Dr. Tristram Bahnson: In the course of that study, pre-specified subgroup analyses were done initially reporting unadjusted outcomes for important clinically relevant subgroups. We found in that initial study that patients with heart failure, minorities, and patients of young age in particular appeared to do better with catheter ablation than with drug therapy. So with that as background, the CABANA study team embarked to focus on each of those subgroups and the heart failure paper was published in 2021, the minorities paper also in 2021 and the subject of our discussion now, the relationship between age and outcome in the CABANA study cohort is a subject of study today. Dr. Greg Hundley: Describe just quickly Tristram the hypothesis you wanted to test here and then in order to test that hypothesis, what was the study population that you included and what was your study design? Dr. Tristram Bahnson: So the focus was on the relationship between age and outcome in CABANA, and this was pre-specified substudy of the CABANA population. So it's probably worthwhile going over who got into the CABANA trial and to remind folks the CABANA trial enrolled 2,204 patients across 126 sites at 10 countries and randomized them one to one to a treatment strategy of either catheter ablation or drug therapy for simple traumatic atrial fibrillation that in the judgment of the treating physicians warranted therapy, patients had to have had at least two episodes of PAF or one episode of persistent AFib documented by ECG or ambulatory recordings within the six months prior to enrollment and they hadn't have failed more than one anuric drug. In other words, they would have to have been reasonable candidates for drug therapy, should they be so randomized. Dr. Tristram Bahnson: In addition, patients that were less than 65 years of age, had to have some additional factors that would increase the likelihood that outcome events would occur. They had to have a CHADSVASC score greater than one. That was not required of the older subjects follow up was 48 and a half months for the population at large, with the interportal range of follow up between 30 and 62 months. The patients had regular follow up every three months for the first year and then six months thereafter. In addition, 1,240 patients received a recording device that allowed them to provide either prescribed episodic recordings or recordings for when they were symptomatic and they also provided 96 hour holters every six months throughout the duration of the trial. Dr. Tristram Bahnson: So that's the population that we were working with. The study design, as I said, focused on trying to tease out the relationship between age and outcomes and the primary outcomes of the CABANA trial included the primary outcome, which was a composite. It included all cause mortality, disabling, stroke, serious bleeding or cardiac arrest, and the key secondary endpoints that were looked at included mortality and cardiovascular hospitalization and AF recurrence. Dr. Greg Hundley: Very nice. Describe for us your results. Dr. Tristram Bahnson: So we actually took a deeper dive into the subgroup of age, and we did a couple things that we thought would be valuable. One was to consider age as a continuous variable because after all, it's pretty arbitrary to bin people into age groups. I think the initial analysis did so with the CABANA proper publication in 2019 to correspond with the break points that we use for CHADSVASC scoring, but we elected to consider age as a continuous variable and we also elected to do adjusted Cox proportional hazard models to account for the various clinical factors that of course varied with age, such as their CHADSVASC score, the occurrence of structural heart disease, like valvular heart disease or coronary disease, the proportion of women, which typically increases with age and did so in this population. The key endpoints that we examined were the CABANA endpoints, including the primary composite endpoint of total mortality, mortality, or CB hospitalization and AF recurrence. Dr. Tristram Bahnson: So at the end of the day, we had 766 patients who were less than 65, 1,130 that were between 65 and 74 and 308 that were greater than 75. Mind you, CABANA admitted patients with any kind of AFib. As a matter of fact, more than half of the study population had persistent or longstanding persistent atrial fibrillation, which is not typical of many studies that have been published, looking at the relative benefits of catheter ablation. We had an unexpected finding that was hinted at, at the initial CABANA study and that was the benefit of catheter ablation was greatest in the younger patients and the benefits of catheter ablation relative to drug therapy seemed to decrease with advancing age at enrollment, which was the age criterion that we based the analysis this on and that this effect was primarily driven by changes in mortality. Dr. Tristram Bahnson: For the composite endpoint in CABANA, which was total mortality, serious stroke, serious bleeding and cardiac arrest, we saw that the adjusted hazard ratio increased average of 27% for every decade in advancing age, where the age was defined as that at enrollment, and for the total mortality endpoint, the adjusted hazard ratio increased an average of 46% for every 10 year increment in age at enrollment. For all age groups, catheter ablation was superior to drug therapy, a relative to a reduction in AFib consistent with many other studies. The benefit was a reduction in the adjusted hazard ratio of about 50%. So catheter ablation was agnostic to age in terms of the benefit of reducing AFib, but was not agnostic to age with result to these mortality inclusive endpoints. We did notice that there was a trend towards a relative benefit of drug therapy for the oldest age group, but we interpreted that result with caution for a variety of reasons. The oldest age group was least well represented and comprised less than 10% of the CABANA population and less than half of the next best well represented age group, which was the less than 65's. Dr. Tristram Bahnson: In looking carefully at the data, we could find no plausible explanation for why the older age group might do better with drug therapy. Again, it was not significant by an intention to treat analysis, but there was a trend towards drug therapy getting better with the oldest age group. We noticed that there was no excess mortality in the old age group within six months of treatment, so it didn't seem like it was related to some adverse procedural effect. We saw no evidence of more advanced forms of AFib in the oldest age group, because they had as good AFib suppression as others, and had the same distribution of paroxysmal versus persistent forms of AFib as the other age groups. There was no difference in crossover after all, if more patients in the old age group crossed over from drug to ablation therapy, who might expect that to be a confounder. Dr. Tristram Bahnson: We did see something that was very unusual and unexpected, which is that the mortality of the oldest age group treated with drugs was actually less than their mortality in catheter ablation, which is the issue at hand, but also less than the other age groups, which was unexpected and even less than all but the youngest age group treated with catheter ablation. So we can't explain this finding. It was not statistically significant. At the end of the day, we don't believe that elderly patients who have drug refractory AFib that is symptomatic should be denied ablation. Dr. Greg Hundley: Well, thank you so much, Tristram, for these very intriguing results. Changsheng, you have many papers that come across your desk. What drew you to this particular paper? Dr. Changsheng Ma: Yes. Dr. Bunch and colleagues should be commanded for the understand and taking important subgroup analysis of CABANA study. There has also been interest in whether the risk and the benefit of ablation may be modulated by patient age. The current analysis suggests that the related benefit of ablation was characterized for those less than 65 years of age are a tiny bit by the increasing age. It is important to emphasize that the current analysis result should not be interpreted to suggest that the cancer ablation has less value in idly patients. As a casual ablation must treated before recurrence across all age groups. Dr. Changsheng Ma: The current analysis is assuming we should know age related increase in safety constant in patients and taking ablation therapy. So we must be cautious not to over incorporate the result of the sub-group analysis, especially in the context of CABANA trial, treating in the permanent effect of ITT analysis. So I think it can be a possible that reach age related gradings in the relatively treatment benefits of the ablation is finding a challenge. Secondly, the CABANA trial was not a oral subgroup analysis. So the variation of treatment effect across the different age group were in the further resource. That's my opinion. Dr. Greg Hundley: Thank you very much. Well, gentlemen, what do you see is the next study that needs to be performed in this sphere of research and Tristram, we'll start with you. Dr. Tristram Bahnson: Well, clearly the clinical task at hand, for those of us who treat patients is to advise patients about relative benefits of therapy when there are choices at hand. And in the case of atrial fibrillation, the fundamental choice obviously is whether or not to pursue catheter ablation or to pursue medical therapy, either for rhythm or rate control. An important part of that decision making is to understand which patients would derive the most benefit from one versus the other therapy. And that need is perhaps the genesis of why we embarked on these subgroup analysis, which admittedly need to be interpreted with caution are not powered to give definitive results, but can certainly help guide future research. So we have noted in the CABANA trial that heart failure patients might do better and that's consistent with other studies looking specifically at heart failure with reduced ejection fraction. So we're contemplating additional studies to help tease that population out since in CABANA, in particular, our heart failure population was mostly those with a preserved ejection fraction and clinical heart failure. Dr. Tristram Bahnson: With regard to age, I think it'll be important to do studies to try to understand what factors resulted in the young patients apparently doing better with ablation. Again, this is hypothesis generating in terms of our result with this paper. So it'd be very interesting to find out whether there are some subsets of patients with younger ages or patients who have the relevant characteristics of the young age patients who would derive particular benefit from catheter ablation. This would obviously require a variety of approaches, including prospective randomized studies and carefully done population studies. So this issue about which patients really derive a significant mortality benefit it from catheter ablation is an important one that has not yet been teased out completely. Dr. Greg Hundley: Thank you. And Changsheng, do you have anything to add? Dr. Changsheng Ma: Yes. I think two streams say it's a very important topic for, you know, who have more and more, the older patients. So we need to answer the question, how about the real influence of age on the outcomes of the atrial fibrillation patients with ablation. So in future, we should consider randomized trial, but I think it's very difficult. So maybe we have to wait more and more, you know, other study to have a trend, how about the outcome for all the patients. It becomes too difficult for a new randomizedtrial. Dr. Greg Hundley: Very nice. Well listeners, we want to thank Dr. Tristram Bahnson from Duke University and Dr. Changsheng Ma from Beijing for bringing us the results from this substudy of the CABANA trial indicating that the mortality related benefits of catheter ablation for atrial fibrillation appeared to decrease for every 10 year increment in age, above the age of 65 years. Well, on behalf of Carolyn and myself, we want to wish you a great week and we will catch you next week on the run. Dr. Greg Hundley: This program is copyright of the American heart association, 2022. The opinions expressed by speakers in this podcast are their own and not necessarily those of the editors or of the American Heart Association for please visit ahajournals.org.

Every drop of water that you drink is not always H2O; sometimes it is D2O. There are three isotopes of hydrogen: protium, deuterium and tritium. One glass of water has on average 6 drops of D2O (deuterium), which doesn't sound like a lot, but over several years it adds up and impairs the ATPase of the mitochondria. Robert Slovak and Victor Sagalovsky are the founders of a company called Litewater. They offer deuterium depleted water that is the lowest parts per million of any available, which makes it the most affordable to use for a deuterium depletion strategy. They describe the exact mechanisms how deuterium impairs mitochondrial energy production, quantum tunneling, and decreases oxygen utilization. They answer questions from listeners including: thoughts on primary water, kangen water, freezing water at home to make your own DDW and the synergy between DDW and molecular hydrogen tablets. Order Litewater: https://www.drinklitewater.com Use the discount code BLACKBURN to save 10% on your first order My website: www.matt-blackburn.com Mitolife products: www.mitolife.co Music by George Henner: https://georgehenner.bandcamp.com  

I wanted to record this episode to clear up any confusion with regards to fat loss and weight loss (the number on the scales). As these are two very different things demonstrated by the episode photo. On the right taken on the day this was recorded and the other several years back. Would it surprise you if I said I weighed the same in both pictures? Remember fat and muscle weigh the same but muscle takes up less room. Hence the ability to alter your physique.  Strong is definitely the new skinny, so steer clear of those scales or better still hide or ditch them and maybe think about your goals and how you can achieve a healthy balance on your transformation journey. So here are 12 helpful reasons as to why you're not seeing the right number on the scales and what you should focus on instead. Hydration We are made up of around 60% water and this decreases as you get older or if you gain more fat. So it's important to stay hydrated. However drinking fizzy water or diet soda drinks can cause gas and bloating! Carbs Are the main fuel source for your body and brain which are converted into glucose and stored as glucogen in your liver and muscles. Did you know that glycogen molecules hold a fair amount of water - 1 gram of glycogen holds just under 3 grams of water (2.7g) If you are eating more carbs than your body is utilising, you muscles will look fuller.  Water is stored inside the muscle cells Intracellular (potassium based) (inside the cells) holds 2/3rds of the water Extracellular (sodium based) holds 1/3rd With 75% of this held in the interstitial space, means only 8% of total body water is held in the plasma (not the same as blood) Regulated by sodium potassium ATPASE enzyme. Sodium Eating a meal high in salt will cause your body to retain water. If you are regularly consuming high salt foods in excess of what your body needs, this can contribute to weight gain. Aim to keep potassium and magnesium rich foods in your diet to minimise the bloat Potassium rich examples - Such as Sweet Potatoes, Spinach, Beetroots, Bananas and Oranges Magnesium rich examples - Whole Grains, Nuts, Avocados, legumes, Seeds, Tofu, Dark Chocolate Post Workout weight gain Weight bearing exercises that put extra stress on the muscles can cause short term inflammation causing temporary water weight retention Menstrual Cycle /Time of the Month Common to feel bloated and with the added hormonal cravings to eat more…this is not the time to jump on the scales Therefore if using be sure to measure week 1 to week to week 1, week 2 to week 2 etc Sleep Lack of sleep can increase your hormone gherkin (released in your stomach) telling your brain you are hungry Being sleep deprived then means you have less leptin (released by fat cells) telling your brain that you are full This can lead to eating foods higher in sugar and fat and most likely calories. You've gained Muscle Muscle weighs the same as fat ut just takes up less room. So you could have a complete body transformation and only lose a few pounds in the process as your body now has more lean muscle and less fat on board. Muscles are 75% water in comparison to fat at just 10% How to define the process without using the scales Clothes - are they fitting better? Photos- A camera never lies! Resting Heart Rate. A healthy RHR is between 60-100 beats per min Are you lifting heavier weights You have more energy , getting better quality sleep Hopefully you found this episode informative and interesting, DM if you have any questions.  I'd be super grateful if you would rate/review on iTumes/Spotify to help the show grow

Cellular senescence appears to be a phenomenon fundamentally ingrained within the aging process and linked to age-related diseases. Characterized broadly by permanent cessation of the cell cycle, cellular senescence may not be as permanent as once thought. Researchers from Incheon National University and Korea University conducted a new study exploring analogs of oxazoloquinoline and their potential to alleviate cellular senescence. Their trending research paper was published as the cover of Aging (Aging-US) Volume 14, Issue 2, and entitled, “Targeting regulation of ATP synthase 5 alpha/beta dimerization alleviates senescence.” Full blog - https://www.impactjournals.com/journals/blog/aging/trending-with-impact-therapeutic-strategy-improves-cell-senescence/ DOI - https://doi.org/10.18632/aging.203858 Correspondence to - Youngjoo Byun - yjbyun1@korea.ac.kr and Joon Tae Park - joontae.park@inu.ac.kr Sign up for free Altmetric alerts about this article - https://oncotarget.altmetric.com/details/email_updates?id=10.18632%2Foncotarget.203858 Keywords - aging, senescence amelioration, KB1541, ATPase synthase 5, OXPHOS About Aging-US Launched in 2009, Aging-US publishes papers of general interest and biological significance in all fields of aging research and age-related diseases, including cancer—and now, with a special focus on COVID-19 vulnerability as an age-dependent syndrome. Topics in Aging-US go beyond traditional gerontology, including, but not limited to, cellular and molecular biology, human age-related diseases, pathology in model organisms, signal transduction pathways (e.g., p53, sirtuins, and PI-3K/AKT/mTOR, among others), and approaches to modulating these signaling pathways. Please visit our website at http://www.Aging-US.com​​ and connect with us: SoundCloud - https://soundcloud.com/Aging-Us Facebook - https://www.facebook.com/AgingUS/ Twitter - https://twitter.com/AgingJrnl Instagram - https://www.instagram.com/agingjrnl/ YouTube - https://www.youtube.com/agingus​ LinkedIn - https://www.linkedin.com/company/aging/ Pinterest - https://www.pinterest.com/AgingUS/ Aging-US is published by Impact Journals, LLC: http://www.ImpactJournals.com​​ Media Contact 18009220957 MEDIA@IMPACTJOURNALS.COM

Aging-US published a Special Collection on Eye Disease which included "Role of Citicoline in an in vitro AMD model" which reported that citicoline is the exogenous form of the nootropic, Cytidine 5'-diphosphate-choline that exerts its neuroprotective effects in the brain as well as in the eye. Citicoline alleviates apoptotic effects as evidenced by diminished AnnexinV/PI and Caspase-3/7 staining, downregulation of apoptosis genes, enhanced cell viability, and reduced oxidative stress in AMD RPE cybrid cells. However, further studies are required to establish the merit of citrusoline as a cytoprotective molecule in AMD and to decipher the molecular underpinnings of its mechanism of action. Dr. M. Cristina Kenney from The University of California Irvine said, "Citicoline is the international nonproprietary name given to the exogenous pharmacological form of Cytidine 5'-diphosphate-choline (CDP-Choline, CDPCho), a naturally occurring endogenous nucleotide compound that is water-soluble and has a molecular weight of 488.32 g/mol" Citicoline maintains neuronal membrane integrity, influences neurotransmitter levels, increases norepinephrine and dopamine levels in the central nervous system, restores the activity of membrane sodium/potassium ATPase and mitochondrial ATPase, and enhances brain function. Owing to these mechanisms, it has been successfully used as a neuroprotective agent to prevent neuronal aging and improve memory and learning in vitro. It has been extensively used in preclinical studies and clinical trials for neurodegenerative diseases including Parkinson's disease and glaucoma. This in vitro study supports our hypothesis as Citicoline conferred significant protection against apoptotic cell death that was in-part mediated by damaged mtDNA from AMD patients. The Kenney Research Team concluded in their Aging-US Research Output, "although further studies with Citicoline/ AMD RPE cybrid cells are underway, these results present novel findings that identify Citicoline to be a potential protector that attenuates apoptotic cell death in AMD. Citicoline is available as an over-the-counter dietary supplement in the U.S. and offers the advantage of easy access that shortens considerably the transition from lab bench to clinic." Full Text - https://www.aging-us.com/article/103164/text Correspondence to: M. Cristina Kenney email: mkenney@uci.edu Keywords: Citicoline, age-related macular degeneration (AMD), neuroprotection, RPE, mitochondria About Aging-US Launched in 2009, Aging-US publishes papers of general interest and biological significance in all fields of aging research as well as topics beyond traditional gerontology, including, but not limited to, cellular and molecular biology, human age-related diseases, pathology in model organisms, cancer, signal transduction pathways (e.g., p53, sirtuins, and PI-3K/AKT/mTOR among others), and approaches to modulating these signaling pathways. To learn more about Aging-US, please visit http://www.Aging-US.com or connect with @AgingJrnl Aging-US is published by Impact Journals, LLC please visit http://www.ImpactJournals.com or connect with @ImpactJrnls Media Contact 18009220957x105 MEDIA@IMPACTJOURNALS.COM

How do β-adrenergic receptor subtypes regulate immune function in the heart? In this unique episode of The AJP-Heart and Circ Podcast, Consulting Editor Dr. Kristine DeLeon-Pennell (Medical University of South Carolina) interviews two authors about their two articles published recently in AJP-Heart and Circulatory Physiology. Dr. Laurel Grisanti (University of Missouri, Columbia) discussed her study (Tanner et al.) on the important role for β2-adrenergic receptor expression on immune cells in the development of heart failure in response to chronic catecholamine elevation. Using a chronic isoproterenol infusion model of heart failure, Dr. Grisanti and co-authors concluded that the immune cell expression of β2-adrenergic receptor is an important contributor to the detrimental responses seen with chronic elevations in catecholamine. The macrophage populations lacking β2-adrenergic receptor largely retained their reparative phenotype and failed to illicit pro-inflammatory macrophage recruitment. Dr. Petra Eder-Negrin (University Hospital, Würzburg) discussed her work (Cellini et al.) in context of Dr. Grisanti's study, underscoring the mechanistic link between sodium potassium -ATPase and β-adrenergic stimulation in the post-MI heart. Eder-Negrin and co-authors found that sodium potassium ATPase alpha 2 overexpressing cardiomyocytes are a crucial adaption, providing an important functional reserve for the heart to cope with chronic stress more efficiently. Sodium potassium ATPase alpha 2 overexpression could be an alteration to attenuate heart failure. How do these research studies connect? Listen now.   Miles A. Tanner, Charles A. Maitz, and Laurel A. Grisanti Immune cell β2-adrenergic receptors contribute to the development of heart failure Am J Physiol Heart Circ Physiol, published September 15, 2021. DOI: 10.1152/ajpheart.00243.2021   Antonella Cellini, Dorina Höfler, Paula A. Arias-Loza, Sandra Bandleon, Tanja Langsenlehner, Michael Kohlhaas, Christoph Maack, Wolfgang R. Bauer, and Petra Eder-Negrin The α2-isoform of the Na+/K+-ATPase protects against pathological remodeling and β-adrenergic desensitization after myocardial infarction Am J Physiol Heart Circ Physiol, published September 15, 2021. DOI: 10.1152/ajpheart.00808.2020

https://magtein.com/products/magtein-orginal-formula enter code DREW Today's show is packed with value. From a track athlete and wrestler, to a community pharmacist, and now a cardiovascular research scientist, doctor of pharmacy, and author of at least 5 life-changing books, Dr. James Dinicolantonio is here to help blow the lid off of nutrition myths and legends. 0:57 "A lot of my patients were suffering from symptoms when their doctor put them on a diuretic to lower their blood pressure, or told them to reduce their salt intake. They felt terrible." Dr. James has hundreds of positive reviews on his books The Mineral Fix, The Salt Fix, The Longevity Solution, and others. Many of these reviews are very personal stories of people sharing how their lives have been changed and improved after following his research and advice. 2:17 "Being more omnivorous optimizes all minerals, better than being a vegan or a carnivore." The average American is not only missing out on the quantity of vegetables they need daily, but they're also not eating enough variety of fruits and vegetables. Our diet does affect our pH balance in the body. Yes, the body does regulate pH on its own, but our bicarbonate levels can go down if we eat high acidic foods without offsetting them with some type of base. 3:28 "Two different waters that are pretty high in bicarbonate and minerals, one is Gerolsteiner, and the other is San Pellegrino." If you have a diet higher in animal foods, your body might struggle a bit to maintain the proper acid levels. So fruits and vegetables or at least bicarbonate mineral water are essential for helping the body balance itself. "5:09 "In order for a neuron to fire and for muscles to move, sodium has to go into the cell, potassium gets kicked out and it's called sodium-potassium ATPase, and it requires ATP and magnesium to do that. So we're alive because we're these electrical, basically these walking bags of salt, fluids, and electrolytes." Both physical performance and cognitive abilities depend heavily on minerals and electrolytes. There's so much information and electrical transfer that happens throughout our body. None of our cells would be proper conductors for that electrical transfer without minerals. Magnesium is required for over 600 enzymatic functions in the body, to produce ATP, to create protein and DNA and RNA. They're necessary for hormones and neurotransmitters. So life really doesn't exist without magnesium. Most people aren't even getting the recommended daily allowance of magnesium, let alone the right VARIETY of magnesiums. Further Resources: The Salt Fix: Why the Experts Got It All Wrong--and How Eating More Might Save Your Life - Dr. James DiNicolantonio The Mineral Fix: How to Optimize Your Mineral Intake for Energy, Longevity, Immunity, Sleep and More - Dr. James DiNicolantonio http://drjamesdinic.com/ https://www.instagram.com/drjamesdinic Follow Drew: @Twitter https://twitter.com/drewcanole @Instagram https://www.instagram.com/drewcanole/ @Facebook https://www.facebook.com/people/Drew-Canole/100044261357988/ Visit www.mysoulcbd.com/drew and get 15% off your next order!

Researchers from Virginia Commonwealth University, Translational Genomics Research Institute, and the Banner Alzheimer's Institute took part in a research study experimenting with combinations of therapeutic agents which they believe may improve neurodegenerative disorders. In 2021, their paper was published in Aging's Volume 13, Issue 13, and entitled, “Inhibition of heat shock proteins increases autophagosome formation, and reduces the expression of APP, Tau, SOD1 G93A and TDP-43.” “In this paper we examined using isogenic colon cancer cells [with] several existing drugs that function by increasing autophagy and degrading misfolded proteins.” “Aberrant expression of chaperone proteins is found in many human pathologies including cancer, in virology and in AD, ALS and HC.” In this study, researchers tested drugs that have been used preclinically and clinically in several anticancer studies. The drugs used were: AR12, an antiviral chaperone ATPase inhibitor; Neratinib, a tyrosine kinase inhibitor; a combination of AR12 and Neratinib; Fingolimod, an immunosuppressive sphingosine l-phosphate receptor modulator; MMF, monomethyl fumarate; and a combination of Fingolimod and MMF. The cells they tested these drug combinations on in vitro included Vero cells (African Green Monkey kidney cells), isogenic HCT116 colon cancer cells (genetically manipulated colon cancer cells), and GBM6 cells (glioblastoma cancer stem cells). They also used plasmids, antibodies, and siRNAs. Researchers acknowledged that the use of non-neuronal cells may be a limitation of this study. “Our present studies were performed in non-neuronal cells and as a caveat, it is possible that our data in HCT116 and Vero cells will not be reflective of the same processes in neuronal cells.” Despite this caveat, results from their research were promising. Some combinations of these drugs were capable of knocking down many disease specific proteins that form toxic aggregates inside cells and in extracellular environments via autophagy. Full blog - https://www.impactjournals.com/journals/blog/aging/trending-with-impact-new-drug-combinations-inhibit-stress-proteins/ Sign up for free Altmetric alerts about this article - https://oncotarget.altmetric.com/details/email_updates?id=10.18632%2Foncotarget.203297 DOI - https://doi.org/10.18632/aging.203297 Full text - https://www.aging-us.com/article/203297/text Correspondence to: Paul Dent email: paul.dent@vcuhealth.org Keywords: Alzheimer's, chaperone, GRP78, autophagy, neratinib About Aging-US Launched in 2009, Aging-US publishes papers of general interest and biological significance in all fields of aging research and age-related diseases, including cancer—and now, with a special focus on COVID-19 vulnerability as an age-dependent syndrome. Topics in Aging-US go beyond traditional gerontology, including, but not limited to, cellular and molecular biology, human age-related diseases, pathology in model organisms, signal transduction pathways (e.g., p53, sirtuins, and PI-3K/AKT/mTOR, among others), and approaches to modulating these signaling pathways. Please visit our website at http://www.Aging-US.com​​ or connect with us on: Twitter - https://twitter.com/AgingJrnl Facebook - https://www.facebook.com/AgingUS/ SoundCloud - https://soundcloud.com/aging-us​ YouTube - https://www.youtube.com/agingus​ LinkedIn - https://www.linkedin.com/company/aging​ Aging-US is published by Impact Journals, LLC please visit http://www.ImpactJournals.com​​ or connect with @ImpactJrnls Media Contact 18009220957 MEDIA@IMPACTJOURNALS.COM

Find the Memorizing Pharmacology book here: https://adbl.co/3wAZEmN The body system we continue to cover is gastrointestinal and omeprazole, esomeprazole, lansoprazole, pantoprazole are all proton pump inhibitors PPIs.  TonyPharmD YouTube Channel here: https://www.youtube.com/c/tonypharmd Suffixes Omeprazole (Prilosec) with the -prazole suffix, p-r-a-z-o-l-e suffix is a true proton pump inhibitor, abbreviated PPI. We want to watch out for aripiprazole (Abilify) and brexpiprazole (Rexulti) which are antipsychotics, not PPIs but have the -piprazole ending, p-i-p-r-a-z-o-l-e. Also, some drug cards say the ending is -azole, but that is not an actual suffix, that is a chemical group, using that ending might have you confuse antifungals like fluconazole (Diflucan) for PPIs, so again, the PPI suffix is -prazole. You will notice that omeprazole (Prilosec) and esomeprazole (Nexium) are very similar and it's that omeprazole contains two molecules, a left and right mirror image and esomeprazole only contains the left-handed image. In Latin, left is sinister, so the “es, e-s” represents that only left-handed side. Why does that matter? That left-handed molecule is the active molecule. Mechanism of Action (MOA) PPIs or “prazoles” work by blocking your stomach's parietal cells which normally release hydrogen ions contributing to the stomachs' acidity. This, without the proton pump inhibitor, could lead to heartburn or possible GI ulceration. The proton pump inhibitor blocks the hydrogen/potassium ATPase pump preventing protons from going in the stomach. This raises the pH, making it more basic, and removes the excessive acid. Indications We then use proton pump inhibitors to manage heartburn, gastroesophageal reflux disease (GERD), peptic ulcer disease, and Barrett's esophagus. Barrett's esophagus is a condition where the acid reflux damages the esophagus causes it to redden. Many times patients who are on chronic NSAIDs or anticoagulants have a higher GI bleed risk and a proton pump inhibitor is for prophylaxis rather than active treatment. Dosing Traditional dosing is to give the PPI 30 to 60 minutes before breakfast. A concern comes when the medication does not seem to work, but it is not the medication, rather, the patient is taking with or even after breakfast. Make sure you know which is which. Also, H2 blockers work a bit more quickly, so the patient might expect a similar timetable with a PPI, let them know that it will take a bit longer. Clinical Considerations Acute use for a few weeks, especially with over-the-counter lengths of time, usually 2 weeks, tends to cause few side effects. Long term, however, we have concerns of B-12 deficiency, increased fracture risk, C. Diff, an opportunistic infection. Again, B-12 deficiency comes because the now less acidic stomach does not do as good a job at absorbing B-12. Before we start this section, here's a reminder contrasting enzyme inhibition and enzyme induction. A drug that inhibits and enzyme blocks the enzyme somewhat increasing drug levels making the patient toxic. A drug that induces and enzyme, makes the enzyme work better reducing drug levels and making the patient subtherapeutic. CYP2C19 inhibition can happen with citalopram (Celexa) and escitalopram (Lexapro), so in this case the antidepressant drug levels can go up leading to QTc prolongation. That's why we have dosing maximums on citalopram of 20 milligrams daily with someone on omeprazole. CYP2C19 induction with omeprazole and clopidogrel (Plavix) is one class example as clopidogrel is a pro-drug and by inducing the enzyme to break down more clopidogrel, the enzyme lowers clopidogrel levels. A pro-drug is one that is not quite the drug yet, the liver may have to metabolize it into a drug. Clopidogrel itself is an antiplatelet drug, so reducing the effectiveness of an antiplatelet drug while trying to prevent myocardial infarction (heart attacks) and strokes.   Note, prescribers can use cilostazol (Pletal) for intermittent claudication, a problem with blood flow in the legs where they might be in pain for short distances and the drug allows them to walk further is also a concern. Using lansoprazole or a similar PPI might create a favorable effect. Some drugs need an acidic environment for absorption like iron supplements and lowering the acidity runs counter to the best situation for iron. Adding ascorbic acid, vitamin C can help. Cefuroxime (Ceftin) is a second-generation cephalosporin antibiotic with good gram-positive coverage, but one might change to another antibiotic if they see omeprazole in the chart. Mesalamine (Pentasa) for ulcerative colitis and itraconazole (Sporanox) and antifungal both both benefit from an acidic stomach.    

Esomeprazole is available over the counter and by prescription. The commonly known brand name is Nexium. Esomeprazole is a substituted benzimidazole proton pump inhibitor (PPI). It comes in a delayed release capsule and as delayed release granule packets. When treating GERD and other hypersecretory conditions a typical dosing range is between 20-40 mg po qd-bid. Esomeprazole is also used as part of a multi-drug regimen for treating H. pylori infection. The mechanism of action behind esomeprazole is through the suppression of acid secretion by inhibiting hydrogen-potassium ATPase. Esomeprazole is the S-enantiomer of omeprazole (Prilosec). Since esomeprazole affects stomach acid it may alter how well other drugs work. For example, absorption of drugs may decrease which may lead to a decrease in efficacy of those drugs. Esomeprazole is best when taken 1 hour before a meal and best if taken before the first meal of the day. If needed, the capsule can be opened. Go to DrugCardsDaily.com for my episode show notes which will contain a drug summary, quiz, and a link to FREE drug card sheets. SUBSCRIBE on Spotify or Apple Podcasts or search for us on your favorite place to listen to podcasts. I will go over the Top 100-200 Drugs as well as throwing in some recently released drugs that peak my interest. Also, if you'd like to say hello, suggest a drug, or leave any constructive feedback on the show I'd really appreciate it! Leave a voice message at anchor.fm/drugcardsdaily or message us through twitter @drugcardsdaily --- Send in a voice message: https://anchor.fm/drugcardsdaily/message