Podcasts about inducible

Contributor: Jorge Chalit-Hernandez, OMS3 Educational Pearls: Serotonin syndrome occurs most commonly due to the combination of monoamine oxidase inhibition with concomitant serotonergic medications like SSRIs Examples of unexpected monoamine oxidase inhibitors Linezolid - a last-line antibiotic reserved for patients with true anaphylaxis to penicillins and cephalosporins Methylene blue - not mentioned in the podcast due to its uncommon usage for methemoglobinemia Other medications that can interact with SSRIs to cause serotonin syndrome Dextromethorphan - primarily an anti-tussive at sigma opioid receptors that also has serotonin reuptake inhibition Clinical presentation of serotonin syndrome Altered mental status Autonomic dysregulation leading to hypertension (most common), hypotension, and tachycardia Hyperthermia Neuromuscular hyperactivity - tremors, myoclonus, and hyperreflexia Hunter Criteria (high sensitivity and specificity for serotonin syndrome): Spontaneous clonus Inducible clonus + agitation or diaphoresis Ocular clonus + agitation or diaphoresis Tremor + hyperreflexia Hypertonia, temperature > 38º C, and ocular or inducible clonus Management of serotonin syndrome Primarily supportive - benzodiazepines can help treat hypertension, agitation, and hyperthermia. Patients often require repeated and higher dosing of benzodiazepines Avoid antipyretics to treat hyperthermia since the elevated temperature is due to sustained muscle contraction and not central temperature dysregulation In refractory patients, cyproheptadine (a 5HT2 antagonist) may be used as a second-line treatment Patients with temperatures > 41.1º C or 106º F require medically induced paralysis and intubation to control their temperature References Boyer EW, Shannon M. The serotonin syndrome [published correction appears in N Engl J Med. 2007 Jun 7;356(23):2437] [published correction appears in N Engl J Med. 2009 Oct 22;361(17):1714]. N Engl J Med. 2005;352(11):1112-1120. doi:10.1056/NEJMra041867 Dunkley EJ, Isbister GK, Sibbritt D, Dawson AH, Whyte IM. The Hunter Serotonin Toxicity Criteria: simple and accurate diagnostic decision rules for serotonin toxicity. QJM. 2003;96(9):635-642. doi:10.1093/qjmed/hcg109 Ramsay RR, Dunford C, Gillman PK. Methylene blue and serotonin toxicity: inhibition of monoamine oxidase A (MAO A) confirms a theoretical prediction. Br J Pharmacol. 2007;152(6):946-951. doi:10.1038/sj.bjp.0707430 Schwartz AR, Pizon AF, Brooks DE. Dextromethorphan-induced serotonin syndrome. Clin Toxicol (Phila). 2008;46(8):771-773. doi:10.1080/15563650701668625 Thomas CR, Rosenberg M, Blythe V, Meyer WJ 3rd. Serotonin syndrome and linezolid. J Am Acad Child Adolesc Psychiatry. 2004;43(7):790. doi:10.1097/01.chi.0000128830.13997.aa Summarized & Edited by Jorge Chalit, OMS3 Donate: https://emergencymedicalminute.org/donate/

Group Singing-Related Technique Workshops for People with Inducible Laryngeal Obstruction: A Feasibility Study

Today, I am blessed to have here with me Dr. John Lieurance. John Lieurance, ND, DC, RMA, BS, DABCN (Board Eligible), is a chiropractic neurologist and naturopath with 25 years of experience in private practice. He is a valuable member of the team at Advanced Rejuvenation, a multidisciplinary clinic specializing in alternative and regenerative medicine, naturopathic medicine, chiropractic functional neurology, Functional Cranial Release (FCR), LumoMed, Lyme disease, mold illness, and various other neurological conditions. Notably, Dr. Lieurance extends his expertise internationally, passionately sharing his knowledge and teaching fellow doctors. In this episode, we explore the metabolic sensitivity of the brain and the vital role of maintaining mitochondrial health, particularly in the inner ear. Dr. Lieurance sheds light on how issues like tinnitus, hearing loss, and hyperacusis are linked to poor mitochondrial function, exacerbated by factors such as jab injuries, vaping effects, and infections like COVID. The conversation then shifts to the battle between the external and internal environments, emphasizing the challenge of managing the body's toxic burden. Dr. Lieurance introduces essential oils as a valuable tool for caring for the body's doorways, particularly the mouth and nasal passages. Tune in as we explore the intricacies of the locus coeruleus, the command center for norepinephrine, and its role in stress modulation.  Dr John Lieurance's LIVE event in Sarasota, Florida: https://www.drjohnlieurance.com/elements-of-vitality Use the coupon code: azadi for 5% off Join Ben Azadi's 90 day heavy metal detox program (15 spots available): https://ketokamp.clickfunnels.com/order-page-a  / / E P I S O D E   S P ON S O R S  Wild Pastures: $20 OFF per Box for Life + Free Shipping for Life + $15 OFF your 1st Box! https://wildpastures.com/promos/save-20-for-life-lf?oid=6&affid=132&source_id=podcast&sub1=ad BonCharge: Blue light Blocking Glasses, Red Light Therapy, Sauna Blankets & More. Visit https://boncharge.com/pages/ketokamp and use the coupon code KETOKAMP for 15% off your order.  Text me the words "Podcast" +1 (786) 364-5002 to be added to my contacts list. [06:50] Doorways and Cranial Release for Holistic Health -       The concept of taking care of doorways is derived from Hindu philosophy, specifically the representation of the God Ganesh, the remover of obstacles. In this context, doorways symbolize obstacles that hinder the expression of vitality and health. Dr. Lieurance draws parallels between this Hindu idea and the practice of cranial release, using balloons to open nasal passages as a means of removing obstacles to better health. -       Dr. Lieurance emphasizes the inherent ability of the body to be self-healing and self-regulating. The vital force within the body, responsible for health, can be disrupted, leading to diseases. -       Dr. Lieurance introduces the concept of cranial release, particularly endo-nasal cranial release using balloons. -       Cranial release is explained as a way to address issues such as chronic nasal congestion, sleep apnea, snoring, TMJ, headaches, and other head-related problems. [12:00] Insights on the Metabolic Sensitivity of the Brain and Inner Ear -       Dr. Lieurance emphasizes the brain as the most metabolically sensitive organ, requiring constant energy. The depletion of this energy source affects sensitive tissues, leading to dysfunction. -       He connects poor mitochondrial function, particularly in the inner ear, to common issues like tinnitus, hearing loss, and hyperacusis. -       These problems, exacerbated by factors such as jab injuries, vaping effects, and infections like COVID, highlight the importance of maintaining mitochondrial health for overall well-being. -       The inner ear is described as the "canary in the coal mine," serving as an early indicator of issues associated with poor mitochondrial function. Dr. Lieurance draws attention to the organ's constant need for energy, even during sleep, and its vulnerability to environmental factors. [18:00] Battling Toxic Burden, Inflammation, and the Vital Role of Essential Oils in Doorway Care -       Dr. Lieurance introduces the concept of a constant battle between the external and internal environments, highlighting the challenge of managing the body's toxic burden. -       This battle leads to inflammation, a key factor in health shifts. -       The emphasis is on strategies to take care of the body's doorways, particularly the mouth and nasal passages. -       Essential oils are presented as a valuable tool for caring for doorways. -       Dr. Lieurance explains the role of biofilm, a sticky gelatinous material that microbes use to shield themselves from the immune system. Without proper care of the doorways, biofilm in the nasal passages can accumulate particles from the air, potentially leading to bloodstream entry. [21:10] Locus Coeruleus, Alzheimer's, and the Intricate Balance of Norepinephrine Modulation -       Dr. Lieurance explains that the locus coeruleus is the command central for norepinephrine, the stress hormone. The activation of this area in the brain is responsible for the transition from sleepiness to alertness and focus in the morning. -       While essential for productivity, some individuals struggle to regulate the firing frequency of the locus coeruleus, leading to prolonged stress responses. -       The accumulation of beta-amyloid plaques associated with Alzheimer's disease is found in the locus coeruleus even before the first stage of Alzheimer's. -       Dr. Lieurance suggests a connection between endotoxins and the development of these plaques in the locus coeruleus. The progression of the plaques then extends to the hypothalamus and the broader brain, impacting functions related to mood, dopamine, movement, and memory. [33:20] The Complex Landscape of Nitric Oxide Forms, Supplements, and the Fine Line Between Health Benefits and Risks -       Dr. Lieurance explains that there are three forms of nitric oxide—endothelial, neuronal, and inducible. -       Inducible nitric oxide is often elevated in response to stress and infections. While supplements are commonly used to increase nitric oxide, it's important to be aware that these supplements typically target the inducible form. -       Elevation of nitric oxide can lead to vasodilation, which can have clinical benefits such as enhanced athletic performance, improved sexual function, and lower blood pressure. -       However, chronic elevation, particularly inducible nitric oxide, is associated with chronic infections and may be linked to conditions like depression. [36:40] The Clinical and Personal Applications of Methylene Blue -       Dr. Lieurance has been actively using methylene blue clinically and personally. He notes its efficacy in treating chronic infections and highlights its remarkable impact on individuals with COVID, leading to dramatic changes within a day. -       Methylene blue, known for absorbing red light and reflecting blue, has an affinity for mitochondria. Originally a dye for clothing, it was later found beneficial for staining tissues and microbes. -       Dr. Lieurance underscores the dye's ability to highlight differences in tissues based on their mitochondrial content and its historical role in studying microbes for drug development, particularly in malaria research. -       Methylene blue's affinity for mitochondria makes it an exciting component in photobiomodulation. AND MUCH MORE!  Resources from this episode: ●      Website: https://www.drjohnlieurance.com ●      Elements of Vitality (Code “AZADI”): https://tickets.drjohnlieurance.com/events/advancedrejuvenation/1023550 ●      Advanced Rejuvenation: https://www.advancedrejuvenation.us ●      MitoZen: https://www.mitozen.club/ketokamp (use code ketokamp) ●      Centner Academy: https://centneracademy.com/ ●      Follow Dr. John Lieurance ●      Facebook: https://www.facebook.com/askdrjohn/ ●      Instagram: https://www.instagram.com/drjohnlieurance  Join Ben Azadi's 90 day heavy metal detox program (15 spots available): https://ketokamp.clickfunnels.com/order-page-a  Keto Kamp Podcast review on Medium, thank you Tim: https://timebl.substack.com/p/tims-grateful-8-deb / / E P I S O D E   S P ON S O R S  Wild Pastures: $20 OFF per Box for Life + Free Shipping for Life + $15 OFF your 1st Box! https://wildpastures.com/promos/save-20-for-life-lf?oid=6&affid=132&source_id=podcast&sub1=ad BonCharge: Blue light Blocking Glasses, Red Light Therapy, Sauna Blankets & More. Visit https://boncharge.com/pages/ketokamp and use the coupon code KETOKAMP for 15% off your order.  Text me the words "Podcast" +1 (786) 364-5002 to be added to my contacts list. // F O L L O W ▸ instagram | @thebenazadi | http://bit.ly/2B1NXKW ▸ facebook | /thebenazadi | http://bit.ly/2BVvvW6 ▸ twitter | @thebenazadi http://bit.ly/2USE0so ▸clubhouse | @thebenazadi Disclaimer: This podcast is for information purposes only. Statements and views expressed on this podcast are not medical advice. This podcast including Ben Azadi disclaim responsibility from any possible adverse effects from the use of information contained herein. Opinions of guests are their own, and this podcast does not accept responsibility of statements made by guests. This podcast does not make any representations or warranties about guests qualifications or credibility. Individuals on this podcast may have a direct or non-direct interest in products or services referred to herein. If you think you have a medical problem, consult a licensed physician.

Have you ever worked with an athlete or a young patient who fears exercise because it can trigger what feels like an asthma attack? Or maybe you've helped someone who feels like they can't breathe when they smell a strong perfume or have acid reflux. Only none of this is asthma…it's a form of airway-induced obstructions.  In this podcast episode, Sara Davis, an SLP and the founder of Vox Fit discusses exercise-induced laryngeal obstruction (formerly known as vocal fold dysfunction/paradoxical vocal fold movement disorder) and inducible airway obstruction due to other triggers. She opens up about the impact of high stress and pressure on individuals with upper airway disorders and the importance of behavioral therapy in managing symptoms.  Learn about the use of scopes for biofeedback, techniques for managing airway conditions, and the role of education and empowerment in helping patients understand and manage their symptoms. Understand the importance of tailoring breathing techniques to specific sports and activities and the need for individualized treatment. You'll walk away from this episode with a greater understanding of airway-induced obstructions, desensitization training, and techniques for addressing exercise-induced laryngeal obstruction.  Get the show notes and references: https://syppodcast.com/299 Timestamps: The conference on upper airway disorders and conditions [00:01:09] Establishment of the Global Initiative for Inducible Laryngeal Obstruction (GLO) [00:02:22] Inducible laryngeal obstruction (ILO) and exercise-induced laryngeal obstruction (EILO) [00:05:01] The fear cycle and high stress [00:06:55] Importance of the first visit and education [00:08:07] Differentiating upper airway disorders from asthma [00:11:26] The role of scope for biofeedback teaching [00:13:31] Understanding the larynx as a gatekeeper [00:14:50] Teaching breathing techniques and gaining control over the airway [00:18:08] Establishing Triggers and Sensory Awareness [00:20:31] Managing Cough and Throat Clearing [00:21:36] Breathing Techniques for Vocal Fold Opening [00:22:43] Modifying breathing techniques for sports [00:27:53] Assessing exercise-induced condition [00:30:55] Desensitization training for triggers [00:33:32] Reducing sensory receptors in the larynx [00:35:35] Ellerby techniques for exercise-induced laryngeal obstruction [00:36:58] The importance of fullness and depth of breath [00:40:09] Breathing techniques during high-intensity exercise [00:41:17] The benefits of respiratory muscle trainers and nasal breathing [00:42:09] The post 299 – Airway Induced Obstructions: What are they, and why should Med SLPs know? appeared first on Swallow Your Pride Podcast.

Link to bioRxiv paper: http://biorxiv.org/cgi/content/short/2023.04.17.536878v1?rss=1 Authors: Bedolla, A. M., McKinsey, G., Ware, K., Santander, N., Arnold, T., Luo, Y. Abstract: The recent proliferation of new Cre and CreER recombinase lines provides researchers with a diverse toolkit to study microglial gene function. To determine how best to apply these lines in studies of microglial gene function, a thorough and detailed comparison of their properties is needed. Here, we examined four different microglial CreER lines (Cx3cr1CreER(Litt), Cx3cr1CreER(Jung), P2ry12CreER, Tmem119CreER), focusing on (1) recombination specificity; (2) leakiness - degree of non-tamoxifen recombination in microglia and other cells; (3) efficiency of tamoxifen-induced recombination; (4) extra-neural recombination -the degree of recombination in cells outside the CNS, particularly myelo/monocyte lineages (5) off-target effects in the context of neonatal brain development. We identify important caveats and strengths for these lines which will provide broad significance for researchers interested in performing conditional gene deletion in microglia. We also provide data emphasizing the potential of these lines for injury models that result in the recruitment of splenic immune cells. 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/2023.03.26.534268v1?rss=1 Authors: Li, S., Wang, Y., Stoel, M. v. d., Zhou, X., Madhusudan, S., Kanerva, K., Nguyen, V. D., Eskici, N., Olkkonen, V. M., Zhou, Y., Raivio, T., Ikonen, E. Abstract: Auxin-inducible degron (AID) technology is powerful for chemogenetic control of proteolysis. However, generation of human cell lines to deplete endogenous proteins with AID remains challenging. Typically, homozygous degron-tagging efficiency is low and overexpression of an auxin receptor requires additional engineering steps. Here, we establish a one-step genome editing procedure with high-efficiency homozygous tagging and auxin receptor expression. We demonstrate its application in 5 human cell lines, including embryonic stem (ES) cells. The method allowed isolation of AID single-cell clones in 10 days for 11 target proteins with greater than 80% average homozygous degron-tagging efficiency in A431 cells, and greater than 50% efficiency for 5 targets in H9 ES cells. The tagged endogenous proteins were inducibly degraded in all cell lines, including ES cells and ES-cell derived neurons, with robust expected functional readouts. This method facilitates the application of AID for studying endogenous protein functions in human cells, especially in stem cells. 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/2023.03.25.534211v1?rss=1 Authors: Mariano, N. C., Rusin, S. F., Nasa, I., Kettenbach, A. N. Abstract: Protein phosphorylation is an essential regulatory mechanism that controls most cellular processes, including cell cycle progression, cell division, and response to extracellular stimuli, among many others, and is deregulated in many diseases. Protein phosphorylation is coordinated by the opposing activities of protein kinases and protein phosphatases. In eukaryotic cells, most serine/threonine phosphorylation sites are dephosphorylated by members of the Phosphoprotein Phosphatase (PPP) family. However, we only know for a few phosphorylation sites which specific PPP dephosphorylates them. Although natural compounds such as calyculin A and okadaic acid inhibit PPPs at low nanomolar concentrations, no selective chemical PPP inhibitors exist. Here, we demonstrate the utility of endogenous tagging of genomic loci with an auxin-inducible degron (AID) as a strategy to investigate specific PPP signaling. Using Protein Phosphatase 6 (PP6) as an example, we demonstrate how rapidly inducible protein degradation can be employed to identify dephosphorylation SITES and elucidate PP6 biology. Using genome editing, we introduce AID-tags into each allele of the PP6 catalytic subunit (PP6c) in DLD-1 cells expressing the auxin receptor Tir1. Upon rapid auxin-induced degradation of PP6c, we perform quantitative mass spectrometry-based proteomics and phosphoproteomics to identify PP6 substrates in mitosis. PP6 is an essential enzyme with conserved roles in mitosis and growth signaling. Consistently, we identify candidate PP6c-dependent phosphorylation sites on proteins implicated in coordinating the mitotic cell cycle, cytoskeleton, gene expression, and mitogen-activated protein kinase (MAPK) and Hippo signaling. Finally, we demonstrate that PP6c opposes the activation of large tumor suppressor 1 (LATS1) by dephosphorylating Threonine 35 (T35) on Mps One Binder (MOB1), thereby blocking the interaction of MOB1 and LATS1. Our analyses highlight the utility of combining genome engineering, inducible degradation, and multiplexed phosphoproteomics to investigate signaling by individual PPPs on a global level, which is currently limited by the lack of tools for specific interrogation. 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/2023.03.15.531679v1?rss=1 Authors: Schyrr, F., Alonso-Calleja, A., Vijaykumar, A., Gebhard, S., Sarkis, R., Lopes, S. F., Oggier, A., De Leval, L., Nombela-Arrieta, C., Naveiras, O. Abstract: Hematopoietic Stem and Progenitor Cells (HSPCs) reside in the hematopoietic niche, a structure that regulates the balance of cellular quiescence, self-renewal and commitment in a demand-adapted manner. The bone marrow (BM) hematopoietic niche is formed by several cellular players, mainly endothelial cells, osteoblasts, adipocytes, and stromal cells. While the BM niche forms a complex structure, evidence exists for simpler, albeit functional, extramedullary hematopoietic niches. However, the composition of what constitutes the simplest unit of an HSPC supportive microenvironment remains largely unknown. Here, we show that the adult adrenal gland can be transformed into a hematopoietic supportive environment. Upon splenectomy and hormonal stimulation, the adult adrenal gland can be induced to recruit and host HSPC function, including serial transplantation. Furthermore, the adrenal stroma contains a CXCL12+ population, reminiscent of BM CXCL12-Abundant Reticular (CAR) cells. Mirroring this, we found CXCL12+ cells in patient samples obtained from a local cohort of myelolipoma, a benign adrenal tumor composed of adipose and hematopoietic tissue that constitutes the most common site of extramedullary hematopoiesis specific to the adult. We present our model as a novel tool to increase our understanding of the physiology of hematopoietic support and to facilitate the development of a boneless niche model. 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/2023.02.02.526918v1?rss=1 Authors: Singh, G., Lakhotia, S. C. Abstract: Oncogenic cells recruit diverse cellular survival machineries, including the highly conserved heat shock proteins (Hsps), to counter stressful conditions during tumour progression. Despite important roles of Hsps in several cancers, poor understanding of their regulation leaves major gaps in identifying mechanisms of cellular stress responses exploited by cancer cells. Following our earlier report of stress inducible Hsp70 expression only in a few cells in polarity defective tumorous clones, we now show that Hsp70 is expressed only in neoplastic tumours. Hsp70s expression at 72h after clone induction is mostly limited to a few lgl- ykiOE cells exhibiting mesenchymal features in hypoxic zone closer to tracheae, although all tumorous cells express hsp70 transcripts. Down-regulation of the hsp70a but not hsp70b cluster transcripts substantially suppresses growth of lgl- ykiOE clones without affecting their early establishment. However, over-expression of Hsp70 or Hsp70-cochaperone DnaJ suppress lgl- ykiOE clones growth at early stage. This spatially and temporally regulated expression of Hsp70 in lgl- ykiOE clones is independent of HSF but requires dFOXO and JNK signalling, while a nearly similar pattern of Hsp70 expression in lgl- RasV12 clones requires HSF, rather than dFOXO. Such context dependent Hsp70 regulation provides novel insight into stress regulatory machinery in cancer cells. 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/2023.01.30.526325v1?rss=1 Authors: El, K., Bauer, B. M., Chen, Y.-C., Jeong, J.-W., Fueger, P. T. Abstract: Type 1 Diabetes (T1D) is caused by autoimmune-mediated beta cell destruction. Following beta cell injury, the pancreas attempts to launch a cellular repair and regenerative program, yet it fails to completely restore functional beta cell mass. One component of this regenerative program is epidermal growth factor receptor (EGFR) signaling. However, upon irreparable beta cell damage, EGFR signaling is dampened, disrupting attempts to restore functional beta cell mass and maintain normoglycemia. We previously demonstrated that the negative feedback inhibitor of EGFR, Mitogen-inducible gene 6 (Mig6), is induced by the pro-inflammatory cytokines central to the autoimmune-mediated beta cell destruction. We also established that pro-inflammatory cytokines suppress EGFR activation, and siRNA-mediated suppression of Mig6 restores EGFR signaling. Thus, we hypothesized that pro-inflammatory cytokines induce nitric oxide production and that in turn induced Mig6, disrupting EGFR repair mechanisms. We determined that NO induces Mig6, attenuating EGFR signaling, and NO synthase inhibition blocks the cytokine-mediated induction of Mig6, thereby restoring cytokine-impaired EGFR signaling. To that end, we treated mice lacking pancreatic Mig6 and control mice with a streptozotocin (STZ) to induce beta cell death and diabetes in a way that mimics the onset and progression of T1D. Whereas STZ-treated control mice became hyperglycemic and had reduced beta cell mass, STZ-treated Mig6 pancreas-specific knock out (PKO) mice remained euglycemic and glucose tolerant due to preserved beta cell mass. The restoration of beta cell mass in PKO mice was accompanied by enhanced beta cell proliferation. Thus, our work suggests that Mig6 is a promising target to preserve beta cell mass before overt T1D. Copy rights belong to original authors. Visit the link for more info Podcast created by Paper Player, LLC

In this session, first recorded at the 2022 IO360° Summit, Dr Semenza will talk about how his work is impacting cancer immunotherapy. Dr Semenza's groundbreaking discovery of hypoxia-inducible factors paves the way for the development of drugs that could kill cancer cells by cutting off the oxygen supply tumors need to grow and improve the response to immunotherapies. Key areas addressed include: Regions of intratumoral hypoxia are a common feature of advanced cancersHypoxia-inducible factors increase the expression of multiple proteins that mediate immune evasionHIF inhibitors stimulate anti-tumor immunity and improve the response to immune checkpoint blockade Gregg Semenza, MD, PhD, Director, Vascular Program, Institute for Cell Engineering and Professor of Genetic Medicine, Johns Hopkins University School of Medicine Learn more about the IO360° Summit at www.io360summit.com

Link to bioRxiv paper: http://biorxiv.org/cgi/content/short/2023.01.09.523268v1?rss=1 Authors: Faust, T. E., Feinberg, P. A., O'Connor, C., Kawaguchi, R., Chan, A., Strasburger, H., Masuda, T., Amann, L., Knobeloch, K.-P. E., Prinz, M., Schaefer, A., Schafer, D. P. Abstract: Cre/LoxP technology has revolutionized genetic studies and allowed for spatial and temporal control of gene expression in specific cell types. The field of microglial biology has particularly benefited from this technology as microglia have historically been difficult to transduce with virus or electroporation methods for gene delivery. Here, we interrogate four of the most widely available microglial inducible Cre lines. We demonstrate varying degrees of recombination efficiency and spontaneous recombination, depending on the Cre line and loxP distance. We also establish best practice guidelines and protocols to measure recombination efficiency in microglia, which could be extended to other cell types. There is increasing evidence that microglia are key regulators of neural circuit structure and function. Microglia are also major drivers of a broad range of neurological diseases. Thus, reliable manipulation of their function in vivo is of utmost importance. Identifying caveats and benefits of all tools and implementing the most rigorous protocols are crucial to the growth of the field of microglial biology and the development of microglia-based therapeutics. 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/2023.01.08.523142v1?rss=1 Authors: Collins, A., Scott, R., Wilson, C. L., Abbate, G., Ecclestone, G. B., Biddles, D., Oakley, F., Mann, J., Mann, D. A., Kenneth, N. S. Abstract: Liver fibrosis is the excessive accumulation of extracellular matrix proteins that occurs in most types of chronic liver diseases. Fibrosis is associated with the activation of hepatic stellate cells (HSCs) which transdifferentiate into a myofibroblast like phenotype that is contractile, proliferative and profibrogenic. Hypoxia-inducible factor 1 (HIF1), an oxygen-sensitive transcription factor, is elevated during HSC activation and promotes the expression of profibrotic mediator HIF target genes. HIF activation during HSC activation can by either due to localised decreases in oxygen levels, or through oxygen-independent mechanisms that are not completely understood. Here we describe a role for the deubiquitinase UCHL1 in regulating HIF levels and activity during HSC activation and liver fibrosis. Increased HIF1 expression correlated with induction of UCHL1 mRNA and protein with HSC activation. Genetic deletion or chemical inhibition of UCHL1 impaired HIF activity through reduction of HIF1 levels. UCHL1 specifically cleaves the degradative ubiquitin chains from HIF1 leading to increased HIF1 levels, even in sufficiently oxygenated cells. Furthermore, our mechanistic studies have shown that UCHL1 elevates HIF activity through specific cleavage of degradative ubiquitin chains, elevates levels of pro-fibrotic gene expression and increases proliferation rates. These results demonstrate how small molecule inhibitors of DUBs can modulate the activity of HIF transcription factors in liver disease. Furthermore, inhibition of HIF activity via modulation of the ubiquitin-proteasomal degradation pathway may represent a therapeutic opportunity with other HIF-related pathologies. 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/2022.12.20.521030v1?rss=1 Authors: Sanyal, S., Kouznetsova, A., Bjorkegren, C. Abstract: Targeted protein degradation systems developed for eukaryotes employ cytoplasmic machineries to perform proteolysis. This has prevented mitochondria-specific analysis of genome maintaining proteins that localize to both mitochondria and nucleus. Here, we present an inducible mitochondria-specific protein degradation system in Saccharomyces cerevisiae based on the Mesoplasma florum Lon (mf-Lon) protease and its corresponding ssrA tag (called PDT). We show that mitochondrially targeted mf-Lon protease efficiently and selectively degrades a PDT- tagged reporter protein localized to the mitochondrial matrix. The degradation can be induced by depleting adenine from the medium and tuned by altering the promoter strength of the MF-LON gene. Finally, we demonstrate that mf-Lon degrades endogenous, dually localized proteins inside mitochondria. In summary, our system is an efficient tool for analysis of intricate mitochondria-nuclear crosstalk essential for proper mitochondrial function. 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/2022.11.14.516419v1?rss=1 Authors: Sankaranarayanan, I., Tavares-Ferreira, D., Mwrigi, J. M., Mejia, G. L., Burton, M. D., Price, T. J. Abstract: Chemotherapy-induced peripheral neuropathy (CIPN) is a primary dose-limiting side effect caused by antineoplastic agents, such as paclitaxel. This causes damage to peripheral nerves and the dorsal root ganglia (DRG). Currently, there are no effective treatments for CIPN, which can lead to long-term morbidity in cancer patients and survivors. Neuro-immune interactions occur in CIPN and have been implicated both in the development and progression of the disease and disease resolution. We investigated the potential role of Inducible co-stimulatory molecule (ICOS) in the resolution of CIPN pain-like behaviors in mice. ICOS is an immune checkpoint molecule that is expressed on the surface of activated T cells and promotes proliferation and differentiation of T cells. We found that intrathecal administration of ICOS agonist antibody (ICOSaa) alleviates mechanical hypersensitivity caused by paclitaxel and facilitates the resolution of mechanical sensitivity in female mice. Administration of ICOSaa reduced astrocyte-gliosis in the spinal cord and satellite cell gliosis in the DRG of mice previously treated with paclitaxel. Mechanistically, ICOSaa intrathecal treatment promoted pain resolution by increasing interleukin 10 (IL-10) expression in the dorsal root ganglion. In line with these observations, blocking IL-10 receptor (IL-10R) activity occluded the effects of ICOSaa treatment on CIPN behavior in female mice. Suggesting a broader activity in neuropathic pain, ICOSaa also partially resolved mechanical hypersensitivity in the spared nerve injury (SNI) model. Our findings support a model wherein ICOSaa administration induces IL-10 expression to facilitate neuropathic pain relief in female mice. ICOSaa treatment is in clinical development for solid tumors and given our observation of T cells in the human DRG, ICOSaa therapy could be developed for combination chemotherapy - CIPN clinical trials. 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/2022.10.27.514052v1?rss=1 Authors: Whitlock, J. M., de Castro, L. F., Collins, M. T., Chernomordik, L. V., Boyce, A. M. Abstract: Metabolic bone diseases are a collection of disorders resulting in diminished skeletal integrity and changes in bone mass due to perturbations in the life-long process of bone remodeling. Perturbations in the number, size and nuclear multiplicity of osteoclasts underpin the development of diverse metabolic bone diseases that impact greater than 13% of adults over age 50 world-wide. Each metabolic bone disease (e.g., osteoporosis, Pagets disease, fibrous dysplasia (FD), osteopetrosis) presents with unique phenotypes, rises from distinct etiologies and progresses with disparate severities, but all are underpinned by a breakdown in osteoclast formation/function. These perturbations of osteoclast formation/function either stem from or cause dysfunctional osteoclast-osteoblast coordination. Unfortunately, a mechanistic understanding of osteoclast-osteoblast coordination and communication is lacking and represents a major barrier to understanding the biology underpinning bone remodeling and the development of effective treatments targeting this process. Here we have developed an inducible ex vivo culture model that models osteoclast-osteoblast coordination in the bone remodeling compartment. Doxycycline addition to cultures activates GsR201C expression and RANKL release from osteoprogenitors, which elicits the differentiation and fusion of neighboring preosteoclasts. In turn, multinucleated osteoclast formation promotes the proliferation of osteoprogenitors, accompanied by the robust release of RANK+ extracellular vesicles, all within ~4 days. This system recapitulates many aspects of the complex osteoclast-osteoblast coordination required for the function of the bone remodeling compartment in both health and diseases underpinned by excessive osteoclast formation. Moreover, based on the ease of isolation, culture, reproducibility and the general adaptability of these cultures to a variety of assays, we expect that this new model will expedite the investigation of osteoclast-osteoblast coordination and osteoclast fusion in bone remodeling and offer a powerful tool for evaluating signaling cascades and novel therapeutic interventions in osteoclast-linked skeletal 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/2022.09.19.508525v1?rss=1 Authors: Choi, E. Y., Franco, D., Stapf, C. A., Gordin, M., Chow, A., Cover, K. K., Chandra, R., Lobo, M. K. Abstract: Substance use disorder is a debilitating chronic disease and a leading cause of disability around the world. The nucleus accumbens (NAc) is a major brain hub that mediates reward behavior. Studies demonstrate exposure to cocaine is associated with molecular and functional imbalance in two NAc medium spiny neuron subtypes (MSNs), dopamine receptor 1 and 2 enriched D1-MSNs and D2-MSNs. Our previous reports showed that repeated cocaine exposure induced transcription factor early growth response 3 (Egr3) mRNA in NAc D1-MSNs, while reducing it in D2-MSNs. Here, we report our findings of repeated cocaine exposure inducing cell subtype specific bidirectional expression of the Egr3 corepressor NGFI-A-binding protein 2 (Nab2). Using CRISPR activation and interference (CRISPRa and CRISPRi) tools combined with Nab2 or Egr3 targeted sgRNAs, we mimicked these bidirectional changes in Neuro2a cells. Furthermore, we investigated D1-MSN and D2-MSN subtype specific expressional changes of histone lysine demethylases Kdm1a, Kdm6a and Kdm5c in NAc after repeated cocaine exposure. Since Kdm1a showed bidirectional expression patterns in D1-MSNs and D2-MSNs, like Egr3, we developed a light inducible Opto-CRISPR-KDM1a system. We were able to downregulate Egr3 and Nab2 transcripts and cause bidirectional expression changes in D1-MSNs and D2-MSNs similar to cocaine exposure in Neuro2A cells. In contrast, our Opto-CRISPR-p300 activation system induced the Egr3 and Nab2 transcripts and caused bidirectional transcription regulations in D1-MSNs and D2-MSNs. Our study sheds light on the expression patterns of Nab2 and Egr3 in specific NAc MSN subtypes in cocaine action and uses CRISPR tools to further mimic these expression patterns. Copy rights belong to original authors. Visit the link for more info Podcast created by PaperPlayer

Link to bioRxiv paper: http://biorxiv.org/cgi/content/short/2022.08.30.503668v1?rss=1 Authors: Licht-Murava, A., Meadows, S. M., Palaguachi, F., Song, S. C., Bram, Y., Zhou, C., Jackvony, S., Schwartz, R. E., Froemke, R. C., Orr, A. L., Orr, A. G. Abstract: TDP-43 pathology is prevalent in dementia but the cell type-specific effects of TDP-43 are not clear and therapeutic strategies to alleviate TDP-43-linked cognitive decline are lacking. We found that patients with Alzheimer's disease (AD) or frontotemporal dementia (FTD) have aberrant TDP-43 accumulation in hippocampal astrocytes. In mouse models, induction of widespread or hippocampus-targeted accumulation in astrocytic TDP-43 caused progressive memory loss and localized changes in antiviral gene expression. These changes were cell-autonomous and correlated with impaired astrocytic defense against infectious viruses. Among the changes, astrocytes had elevated levels of interferon-inducible chemokines and neurons had elevated levels of the corresponding chemokine receptor CXCR3 in presynaptic terminals. CXCR3 stimulation altered presynaptic function and promoted neuronal hyperexcitability, akin to the effects of astrocytic TDP-43, and blockade of CXCR3 reduced this activity. Ablation of CXCR3 also prevented TDP-43-linked memory loss. Thus, astrocytic TDP-43 dysfunction contributes to cognitive impairment through aberrant chemokine-mediated astrocytic-neuronal interactions. Copy rights belong to original authors. Visit the link for more info Podcast created by PaperPlayer

Commentary by Dr. Valentin Fuster

Nov 21The papers behind the pod:1. A non-human primate in vitro functional assay for the early evaluation of TB vaccine candidates. npj Vaccines, January 2021. https://doi.org/10.1038/s41541-020-00263-72. Inducible dominant negative ErbB2 rat spermatogonial line for generation of transgenic rat model and dissecting ERBB2 tyrosine kinase mediated pathways. Experimental Oncology, June 2019. https://doi.org/10.32471/exp-oncology.2312-8852.vol-41-no-2.130263. Characterization of drug responses of mini patient-derived xenografts in mice for predicting cancer patient clinical therapeutic response. Cancer Communications, September 2018. https://doi.org/10.1186/s40880-018-0329-5Visit the AAALAC website to learn more about this year's Global 3Rs Award winners: https://www.aaalac.org/awards/global-3rs-winners/It's the third Thursday of November, and you're listening to 3 Minute 3Rs, your monthly recap of efforts to replace, reduce and refine the use of animals in research. This month, we're celebrating our fourth birthday, and we're also celebrating this year's Global AAALAC/IQ Award winners. Three papers were recognized this year, by scientists in Europe, North America and the Pacific Rim. First let's hear about the European winner: Dr Rachel Tanner at the University of Oxford.Tuberculosis is a serious global health problem with rising cases and deaths, in part, due to the current vaccine being insufficient. To develop a new vaccine, non-human primates are necessary relevant animal model, but efforts to refine their use are needed. Recently, a team has developed an in vitro assay to refine and expedite early tuberculosis vaccine testing. This assay uses non-human primate blood or cell samples to measure the ability to control mycobacterial replication. Importantly, it allows vaccine candidates to be tested without infecting non-human primates with tuberculosis directly. Rather, blood samples can be taken before and after vaccination and tested in vitro to examine efficacy. This work is a promising 3Rs tool to facilitate early testing of new vaccine candidates.To learn more, read the full paper online. Next, let's move to North America and Dr Andrew Syvyk from Texas A&M University, who won the prize for work on a simpler way to generate transgenic animal models. Such models can be tailored to study specific genes, for example by introducing a gene construct into a zygote or embryonic stem cell, but this is a long process that requires numerous steps and manipulations, and therefore a lot of animals.In his winning paper, Syvyk describes a simpler approach using immature sperm cells, or spermatogonia. The model involves manipulating spermatogonial stem cells from a donor male, which are then transplanted into recipient males. These can be used to breed transgenic animals directly, eliminating mosaicism and reducing the number of animals required.The paper discusses using this approach to study ERBB2, a protein involved in multiple biological processes and forms of cancer. As well as being used to generate transgenic animals, the cells can be used for in vitro research, further reducing reliance on in vivo models for investigating molecular pathways.Want to learn more about the model? Find the paper in Experimental Oncology via the link in the description.And finally, the winner... See acast.com/privacy for privacy and opt-out information.

Go online to PeerView.com/UNQ860 to view the activity, download slides and practice aids, and complete the post-test to earn credit. In this activity, a nephrology expert discusses the latest evidence for hypoxia-inducible factor prolyl-hydroxylase (HIF-PH) inhibitors for the treatment of anemia associated with chronic kidney disease (CKD). Upon completion of this activity, participants will be able to: Describe the epidemiology, pathology, and burden of anemia associated with CKD, Discuss key limitations associated with the current model of treatment for anemia associated with CKD, including in patients with NDD-CKD and DD-CKD, Explain the rationale for and potential implications of targeting HIF stabilization with HIF-PH inhibitors as a treatment strategy for anemia in CKD.

Go online to PeerView.com/UNQ860 to view the activity, download slides and practice aids, and complete the post-test to earn credit. In this activity, a nephrology expert discusses the latest evidence for hypoxia-inducible factor prolyl-hydroxylase (HIF-PH) inhibitors for the treatment of anemia associated with chronic kidney disease (CKD). Upon completion of this activity, participants will be able to: Describe the epidemiology, pathology, and burden of anemia associated with CKD, Discuss key limitations associated with the current model of treatment for anemia associated with CKD, including in patients with NDD-CKD and DD-CKD, Explain the rationale for and potential implications of targeting HIF stabilization with HIF-PH inhibitors as a treatment strategy for anemia in CKD.

Go online to PeerView.com/UNQ860 to view the activity, download slides and practice aids, and complete the post-test to earn credit. In this activity, a nephrology expert discusses the latest evidence for hypoxia-inducible factor prolyl-hydroxylase (HIF-PH) inhibitors for the treatment of anemia associated with chronic kidney disease (CKD). Upon completion of this activity, participants will be able to: Describe the epidemiology, pathology, and burden of anemia associated with CKD, Discuss key limitations associated with the current model of treatment for anemia associated with CKD, including in patients with NDD-CKD and DD-CKD, Explain the rationale for and potential implications of targeting HIF stabilization with HIF-PH inhibitors as a treatment strategy for anemia in CKD.

Go online to PeerView.com/UNQ860 to view the activity, download slides and practice aids, and complete the post-test to earn credit. In this activity, a nephrology expert discusses the latest evidence for hypoxia-inducible factor prolyl-hydroxylase (HIF-PH) inhibitors for the treatment of anemia associated with chronic kidney disease (CKD). Upon completion of this activity, participants will be able to: Describe the epidemiology, pathology, and burden of anemia associated with CKD, Discuss key limitations associated with the current model of treatment for anemia associated with CKD, including in patients with NDD-CKD and DD-CKD, Explain the rationale for and potential implications of targeting HIF stabilization with HIF-PH inhibitors as a treatment strategy for anemia in CKD.

Go online to PeerView.com/UNQ860 to view the activity, download slides and practice aids, and complete the post-test to earn credit. In this activity, a nephrology expert discusses the latest evidence for hypoxia-inducible factor prolyl-hydroxylase (HIF-PH) inhibitors for the treatment of anemia associated with chronic kidney disease (CKD). Upon completion of this activity, participants will be able to: Describe the epidemiology, pathology, and burden of anemia associated with CKD, Discuss key limitations associated with the current model of treatment for anemia associated with CKD, including in patients with NDD-CKD and DD-CKD, Explain the rationale for and potential implications of targeting HIF stabilization with HIF-PH inhibitors as a treatment strategy for anemia in CKD.

Go online to PeerView.com/UNQ860 to view the activity, download slides and practice aids, and complete the post-test to earn credit. In this activity, a nephrology expert discusses the latest evidence for hypoxia-inducible factor prolyl-hydroxylase (HIF-PH) inhibitors for the treatment of anemia associated with chronic kidney disease (CKD). Upon completion of this activity, participants will be able to: Describe the epidemiology, pathology, and burden of anemia associated with CKD, Discuss key limitations associated with the current model of treatment for anemia associated with CKD, including in patients with NDD-CKD and DD-CKD, Explain the rationale for and potential implications of targeting HIF stabilization with HIF-PH inhibitors as a treatment strategy for anemia in CKD.

Go online to PeerView.com/UNQ860 to view the activity, download slides and practice aids, and complete the post-test to earn credit. In this activity, a nephrology expert discusses the latest evidence for hypoxia-inducible factor prolyl-hydroxylase (HIF-PH) inhibitors for the treatment of anemia associated with chronic kidney disease (CKD). Upon completion of this activity, participants will be able to: Describe the epidemiology, pathology, and burden of anemia associated with CKD, Discuss key limitations associated with the current model of treatment for anemia associated with CKD, including in patients with NDD-CKD and DD-CKD, Explain the rationale for and potential implications of targeting HIF stabilization with HIF-PH inhibitors as a treatment strategy for anemia in CKD.

Go online to PeerView.com/UNQ860 to view the activity, download slides and practice aids, and complete the post-test to earn credit. In this activity, a nephrology expert discusses the latest evidence for hypoxia-inducible factor prolyl-hydroxylase (HIF-PH) inhibitors for the treatment of anemia associated with chronic kidney disease (CKD). Upon completion of this activity, participants will be able to: Describe the epidemiology, pathology, and burden of anemia associated with CKD, Discuss key limitations associated with the current model of treatment for anemia associated with CKD, including in patients with NDD-CKD and DD-CKD, Explain the rationale for and potential implications of targeting HIF stabilization with HIF-PH inhibitors as a treatment strategy for anemia in CKD.

Go online to PeerView.com/UNQ860 to view the activity, download slides and practice aids, and complete the post-test to earn credit. In this activity, a nephrology expert discusses the latest evidence for hypoxia-inducible factor prolyl-hydroxylase (HIF-PH) inhibitors for the treatment of anemia associated with chronic kidney disease (CKD). Upon completion of this activity, participants will be able to: Describe the epidemiology, pathology, and burden of anemia associated with CKD, Discuss key limitations associated with the current model of treatment for anemia associated with CKD, including in patients with NDD-CKD and DD-CKD, Explain the rationale for and potential implications of targeting HIF stabilization with HIF-PH inhibitors as a treatment strategy for anemia in CKD.

Go online to PeerView.com/UNQ860 to view the activity, download slides and practice aids, and complete the post-test to earn credit. In this activity, a nephrology expert discusses the latest evidence for hypoxia-inducible factor prolyl-hydroxylase (HIF-PH) inhibitors for the treatment of anemia associated with chronic kidney disease (CKD). Upon completion of this activity, participants will be able to: Describe the epidemiology, pathology, and burden of anemia associated with CKD, Discuss key limitations associated with the current model of treatment for anemia associated with CKD, including in patients with NDD-CKD and DD-CKD, Explain the rationale for and potential implications of targeting HIF stabilization with HIF-PH inhibitors as a treatment strategy for anemia in CKD.

Go online to PeerView.com/UNQ860 to view the activity, download slides and practice aids, and complete the post-test to earn credit. In this activity, a nephrology expert discusses the latest evidence for hypoxia-inducible factor prolyl-hydroxylase (HIF-PH) inhibitors for the treatment of anemia associated with chronic kidney disease (CKD). Upon completion of this activity, participants will be able to: Describe the epidemiology, pathology, and burden of anemia associated with CKD, Discuss key limitations associated with the current model of treatment for anemia associated with CKD, including in patients with NDD-CKD and DD-CKD, Explain the rationale for and potential implications of targeting HIF stabilization with HIF-PH inhibitors as a treatment strategy for anemia in CKD.

Go online to PeerView.com/UNQ860 to view the activity, download slides and practice aids, and complete the post-test to earn credit. In this activity, a nephrology expert discusses the latest evidence for hypoxia-inducible factor prolyl-hydroxylase (HIF-PH) inhibitors for the treatment of anemia associated with chronic kidney disease (CKD). Upon completion of this activity, participants will be able to: Describe the epidemiology, pathology, and burden of anemia associated with CKD, Discuss key limitations associated with the current model of treatment for anemia associated with CKD, including in patients with NDD-CKD and DD-CKD, Explain the rationale for and potential implications of targeting HIF stabilization with HIF-PH inhibitors as a treatment strategy for anemia in CKD.

Go online to PeerView.com/UNQ860 to view the activity, download slides and practice aids, and complete the post-test to earn credit. In this activity, a nephrology expert discusses the latest evidence for hypoxia-inducible factor prolyl-hydroxylase (HIF-PH) inhibitors for the treatment of anemia associated with chronic kidney disease (CKD). Upon completion of this activity, participants will be able to: Describe the epidemiology, pathology, and burden of anemia associated with CKD, Discuss key limitations associated with the current model of treatment for anemia associated with CKD, including in patients with NDD-CKD and DD-CKD, Explain the rationale for and potential implications of targeting HIF stabilization with HIF-PH inhibitors as a treatment strategy for anemia in CKD.

Go online to PeerView.com/UNQ860 to view the activity, download slides and practice aids, and complete the post-test to earn credit. In this activity, a nephrology expert discusses the latest evidence for hypoxia-inducible factor prolyl-hydroxylase (HIF-PH) inhibitors for the treatment of anemia associated with chronic kidney disease (CKD). Upon completion of this activity, participants will be able to: Describe the epidemiology, pathology, and burden of anemia associated with CKD, Discuss key limitations associated with the current model of treatment for anemia associated with CKD, including in patients with NDD-CKD and DD-CKD, Explain the rationale for and potential implications of targeting HIF stabilization with HIF-PH inhibitors as a treatment strategy for anemia in CKD.

Dr. Alireza Mansouri interviews Dr. Matthew Dankner about his recent paper "Invasive growth associated with Cold-Inducible RNA-Binding Protein expression drives recurrence of surgically resected brain metastases" published online in Neuro-Oncology in January, 2021.

Link to bioRxiv paper: http://biorxiv.org/cgi/content/short/2020.09.22.308692v1?rss=1 Authors: Zocher, S., Overall, R. W., Berdugo-Vega, G., Rund, N., Karasinsky, A., Adusumilli, V., Steinhauer, C., Scheibenstock, S., Haendler, K., Schultze, J. L., Calegari, F., Kempermann, G. Abstract: Dynamic DNA methylation controls gene-regulatory networks underlying cell fate specification. How DNA methylation patterns change during adult hippocampal neurogenesis and their relevance for adult neural stem cell differentiation and related brain function has, however, remained unknown. Here, we show that neurogenesis-associated de novo DNA methylation is critical for maturation and functional integration of adult-born hippocampal neurons. Cell stage-specific bisulfite sequencing revealed a pronounced gain of DNA methylation at neuronal enhancers, gene bodies and binding sites of pro-neuronal transcription factors during adult neurogenesis, which mostly correlated with transcriptional up-regulation of the associated loci. Inducible deletion of both de novo DNA methyltransferases Dnmt3a and Dnmt3b in adult neural stem cells specifically impaired dendritic outgrowth and synaptogenesis of new-born neurons, resulting in reduced hippocampal excitability and specific deficits in hippocampus-dependent learning and memory. Our results highlight that, during adult neurogenesis, remodeling of neuronal methylomes is fundamental for proper hippocampal function. Copy rights belong to original authors. Visit the link for more info

Link to bioRxiv paper: http://biorxiv.org/cgi/content/short/2020.09.15.297259v1?rss=1 Authors: Riemslagh, F. W., van der Toorn, E., Verhagen, R. F. M., Maas, A., Bosman, L. W., Hukema, R. K., Willemsen, R. Abstract: The hexanucleotide G4C2 repeat expansion in the first intron of the C9ORF72 gene explains the majority of frontotemporal dementia (FTD) and amyotrophic lateral sclerosis (ALS) cases. Numerous studies have indicated the toxicity of dipeptide repeats (DPRs) which are produced via repeat-associated non-AUG (RAN) translation from the repeat expansion and accumulate in the brain of C9FTD/ALS patients. Mouse models expressing the human C9ORF72 repeat and/or DPRs show variable pathological, functional and behavioral characteristics of FTD and ALS. Here, we report a new Tet-on inducible mouse model that expresses 36x pure G4C2 repeats with 100bp upstream and downstream human flanking regions. Brain specific expression causes the formation of sporadic sense DPRs aggregates upon 6 months dox induction but no apparent neurodegeneration. Expression in the rest of the body evokes abundant sense DPRs in multiple organs, leading to weight loss, neuromuscular junction disruption, myopathy and a locomotor phenotype within the time frame of four weeks. We did not observe any RNA foci or pTDP-43 pathology. Accumulation of DPRs and the myopathy phenotype could be prevented when 36x G4C2 repeat expression was stopped after 1 week. After 2 weeks of expression, the phenotype could not be reversed, even though DPR levels were reduced. In conclusion, expression of 36x pure G4C2 repeats including 100bp human flanking regions is sufficient for RAN translation of sense DPRs and evokes a functional locomotor phenotype. Our inducible mouse model highlights the importance of early diagnosis and treatment for C9FTD/ALS patients. Copy rights belong to original authors. Visit the link for more info

Inducible pluripotent stem cells (iPSCs) are cultured cells with the ability to become any cell type present in the species from which it came. Jeanne Loring (Professsor Emeritus at Scripps Research and the Chief Scientific Officer at Aspen Biosciences) is an expert in this area. In this episode, she described how iPSCs are made and what they can be used for. Spoiler alert: skin cells and lots of important stuff. One example is to create dopamine neurons that may be used as a treatment for Parkinson's disease.  Outside of human health, they may help to save endangered species. The Northern White Rhino is functionally extinct as there are only two females left. The good news is that back in the 1970s, when the era of gene manipulation was just beginning, someone at the San Diego Wild Animal Park had the foresight to take skin cells from animals and deep freeze them in liquid nitrogen. The Northern White Rhino's future is in those frozen samples. It's a fascinating and exciting story.

Ben Valsler introduces the oxygen sensors that help life react to changing conditions, and were key to the 2019 Nobel prize in physiology or medicine

Naked Jane owner Kadri Gedelec has a lot to say about CBD and the unique process his company Naked Jane uses to distinguish themselves above and beyond the plethora of competitors who have jumped on the CBD oil trend bandwagon. Kadri recognizes not all CBD is created equal and uses a seven step process to create only the worlds highest quality of CBD oil. Make sure you check out why Naked Jane's process is unique and why their benefits outweigh their competition, even if that competition now offers their CBD in hamburger form. Naked Jane CBD oilEverything You Wanted to Know About CBD Oilby Jim GoetzColts Neck, NJAllow me to clarify that we are not poteads, stoners or what you could consider, "druggies". We never were.I (Jim Goetz), personally grew up a "normal" kid, playing with my blocks, He-Man and Teenage Mutant Ninja Turtles and ran around outside until my mom had to come find me and drag me in for dinner.When I met Chantea in college, I was a clean cut college baseball player with a huge substance abuse problem. I ate copious amounts of pecan pie, took large amounts of creatine (until I burped poweder), and drank large amounts of protein shake and water. I guess other than the pecan pie (something I had never been exposed to living in the Northeast all my life up until that point), these so called "problems" were typical of any jock and most likely still are. Until recently, the closest thing I have come to what would be considered a "substance" is utilizing nootropics for school, work and training such as nicotine and adrafinil along with melatonin and L-theanine. My mushrooms come in the form of chaga and lions mane. Of course the nootropics made by Alexander and Stefan of Nooflux are awesome! If you have not heard this podcast check out the show notes and take a gander after you are done here. You may already know or have done it yourself but many now are experimenting with edible tetrahydrocannabinol (THC) for exercise performance, and also experimenting with vaporizing indica-rich strains of marijuana for creativity, relaxation and sleep. In these show notes and podcast, we delve into a derivative of the cannabis plant family that is now being used as an infusion into a fast food burger. It has some pretty massive payoffs for balancing your endocrine system, relieving anxiety, modulating chronic stress, shutting down inflammation and chronic pain, decreasing blood sugar, decreasing appetite and lowering abdominal obesity.Sit back, grab your favorite nootropic and learn about a form of cannabis with zero psychoactive effects of weed but gives you all of the good stuff you want. Cannabis 101When you hear someone discuss the topic of marijuana, typically they are referring to THC. This is the part of the plant (known as a hemp plant), that induces a state of euphoria. Some may enjoy this state while others find it annoying. Some however, believe it makes certain movies and cartoons, such as the entire Adult Swim, more than simply mildly palpable.The THC content is what the majority of recreational marijuana users desire. Back in the 1960's, bontanists were able to increase the amount of THC in marijuana (weed) from 3% to 5% to a whopping 28%, which is the amount typically found in the plant today. Back in the 60's, individuals would find a mild euphoric state as compared to today, which that same individual would find themselves inebriated on their ass.THC is a chemical compound that fits into a receptor already found in your body from birth called, CB1. This CB1 receptor is located in the cerebral cortex (brain).  Fill too many receptors (intake a crapton of THC) and you will find yourself on your couch all day.There are currently 85 active cannabinoids identified in cannabis. One of which is called, Cannabidiol (CBD). This accounts for 40% of a plants total cannabinoid content.Whether you have seen the South Park episode where everyone becomes weed farmers or not; in these hemp fields, cannabis plants grow in a manner that the male plants are able to fertilize the female plants. When the male plants are quarantined from the female plants, they are no longer able to fertilize them. When females cannot be pollinated, they produce copious amounts of THC. When a female is happily fertilized, she only produces less than 1% THC. I am sure there is some joke that can be associated with this but I will let it go for now and leave it up to you the reader to come up with something good.Production of CBD (international standards require less than 1% THC), is dependent on good relations between the male and female cannabis plants. Chemistry, Biochemistry, Organic Chemistry and Inoragnic Chemistry???I have been asked why would you want to take CBD without the fun of THC? other than being concerned about progress of your day and appearing as a coherent professional, there are a crapton of reasons why.But first, let us get into the text book stuff so you can understand the physiology of your body and what CBD’e effects truly are.As previously discussed, THC attaches primarily to CB1 receptors. CBD however, has a low affinity for both CB1 and CB2 receptors and acts as an indirect antagonist of the agonist. This means that what activates CB1 or CB2 are turned off by CBD.An example of this is CBD can increasing CB1 receptor density so that there’s too many CB1 receptors for THC to bind to, thus taking the edge off the potential psychoactivity of weed, while still retaining all the opioid-like painkilling effects.This means you have to increase the consumption of THC if you are using CBD separately. It also means that CBD will prolong the duration of effects of THC in your system as it inhibits cytochrome P-450. This enzyme causes the rapid metabolism of THC. As it is inhibited, the THC lasts longer and therefore you feel the effects longer.In using CBD, plasma concentrations of THC increase, causing increased amounts of THC to be available to CB1 receptors. With the increase, CBD acts as an antagonist at a cannabinoid receptor named GPR55, located in the caudate nucleus of the putamen (brain). This reduces the effects of THC. What this boils down to is by using CBD on it’s own, THC lasts longer but reduces any psychoactive effects. An actual receptor CBD effects is called 5-HT1A. This is why CBD has anti-depressant and anti-anxiety effects and is neuroprotective. It is also an “allosteric modulator” of opioid receptors, which is why it reduces pain and chronic inflammation.CBD was found to eliminate memory loss problems from marijuana (agonist/ antagonist relationship). CBD has really strong anti-oxidant and anti-inflammatory properties, due primarily to its effects on your adenosine receptors and cytochrome P-450 and 2C enzymes.Research has shown as with weed that includes THC, it is not possible to overdose on CBD. CBD has an unusually low level of toxicity. In over 6,000 years, not one individual has overdosed on pure CBD.Did you know:-NSAIDS (ibuprofen, Advil, Tylenol, and aspirin) kill over 1,000 people per year?-Alcohol kills over 110,000 people each year?-marijuana kills 0 people each year.  Nada. Not one.CBD and AddictionThe reason CBD is not addictive is because CBD does not act on any receptors in your brain that produce an addiction. We discussed this previously...remember?Former national administrator of the US governments marijuana research programs, Dr. Tod Mikriya stated, “no other single drug or substance has as many therapeutic benefits as cannabis”. Dr. Mikriya never found or discussed any evidence of addiction to cannabis. I hear all the time though individuals state that cannabis is addictive. I hear all the time cannabis is a gateway drug and the federal government even locks people away for decades because of canabis. The Boggs Act of 1951 established mandatory sentences for drug users and stated that cannabis was addictive and has held it’s effect despite what other government officials and researchers such as Dr. Mikriya have discovered. Even Dr. Harris Isbell the Director of Research at the Public Health Service Hospital located in Lexington, Kentucky have villified the idea that cannabidols are not physically addictive. Oriental medicine has been used for over 6,000 years and used cannabis as a popular remedy. There have been no reported cases of addiction. The research study, Ganja in Jamaica: A Medical Anthropological Study of Chronic Marijuana Use, which was published in the Journal of the American Medical Association in 1975, disclosed zero concerns with addiction, even after patients who had used cannabis for decades had stopped. The 1980 study Cannabis in Costa Rica: A Study in Chronic Marijuana Use backed this up. Most interestingly, studies like this are not finding any addictive potential with CBD even in the presence of THC.Cannabis Safety​For over 40 years, Dr. Lester Grinspoon, Professor Emeritus at Harvard Medical School, spent the majority of his professional life studying cannabis. The result was “Marijuana: The Forbidden Medicine“. As you can see, Dr. Grinspoon didn’t find one single case of death, stating, “There are no deaths from cannabis use. Anywhere. You can’t find one.”Francis Young, an administrative DEA judge, took medical testimony for over two weeks, and at the end of it, he said, “Marijuana, in its natural form, is one of the safest therapeutically active substances known to man.”In proper doses, CBD may produce the same pain reduction compared to opioid prescription drugs, such as morphine, hydrocodone, and oxycodone, and when combined with these drugs, allows you to use far less of the actual prescription, thus reducing the toxic load on your liver and kidneys.CBD and Your HormonesYour choice at this point is to stop reading, read a bunch of journal articles and do your own peer reviews, read a bunch of opinions online and at your local grocery store, or keep reading…...good...glad you are still here.Your endocrine system consists of glands throughout your body which regulates literally every function in your body from metabolism to sex and so forth. One function of your hormones is that in response to the fight or flight response. When your sympathetic system is activated, you produce excess cortisol levels. However, when cortisol levels continue to rise, you feel exhausted at all times and your performance decreases. CBD decreases plasma cortisol levels and as a result, can increase your performance and quality of life. But CBD has other effects on your endocrine system, particularly your appetite. You may simply think that marijuana produces the munchies and therefore makes you fat, and although this makes logical sense, science has shown that it’s not the case that marijuana makes you fat, especially when CBD is present.The idea behind smoking weed is that you get the munchies afterwards and as a result- gain weight. The opposite is true though. CBD stabilizes insulin, regulates your appetite and decreases cortisol. These are all aspects of weight gain/ loss. But it is actually the THC that increases your appetite. On the flip side, CBD can suppress your appetite. Hmmm….Physiologically speaking, your pancreas secretes the hormones glucagon and insulin to regulate blood sugar by signaling your liver to break down fat into sugar (glucagon) or to store sugar as fat (insulin). These hormones work as a pair to maintain homeostasis, and they stimulate the release of each other through a complex feedback mechanism. While THC primarily increases glucagon and blood sugar, CBD lowers insulin levels, and it is this CBD action that helps to explain why marijuana users tend to eat more calories but do not gain any extra weight, have less obesity and have lower rates of type II diabetes than non-users, and is also why some diabetics find that marijuana makes it easier to manage their blood sugar.Type II diabetics (whose pancreas still functions) tend to have very high levels of insulin, but the liver is unable to use that insulin, so blood sugar stays high, and the pancreas eventually damages itself by trying to continually produce more and more insulin, eventually leading to organ failure if the diabetes is unmanaged. By lowering pancreatic insulin release, CBD may alleviate or prevent the progression of type II diabetes and blood sugar disorders. Cannabinoid antagonists such as CBD have been shown to reduce obesity, and not only do rodents given these antagonists eat less, but they also lose more weight than their reduced feeding can account for.So the summary of the biggest effects of CBD on the endocrine system? Lower cortisol and better blood sugar control. Yep...Your Anxiety and Stress and CBDWe discussed the effects of CBD and cortisol. When it comes to the full plant, it is THC that causes stress and anxiety unless there is enough CBD present in order to negate the effects. (recall the agonist/ antagonist relationship). This is due to CBD’s direct action on your 5HT1A and TRPV1 receptors (responsible for anxiolytic response to stress).Effective dosing appears  to fall within the ranges from 10 mg to 100 mg, depending on the individual and the situation and even time of day. This is far healthier than the alternatives of addictive drugs such as Valium or Xanax. CBD and Your SleepIt is believed that in the United States, approximately 70 million people suffer from insomnia, insufficient sleep or another sleep disorder. It’s ironic that Cannabidiol actually activates the adenosine receptors as does caffeine, a stimulant. It has been found though that CBD has a calming effect that research shows allows people to sleep.You can use larger amounts of CBD to help drift blissfully to sleep (ex. 100 mg) or use smaller amounts and combine it with melatonin, lemon balm, valerian root, magnesium, chamomile or your favorite bedtime relaxation herb.CBD and InflammationInflammation is all the rage now-a-days, isn’t it? The answer to pre-mature aging and chronic disease is inflammation. It’s not really a rage or the trend but the truth. Low level inflammation is the enemy and easily caused by our faced paced, high stress lives with great amounts of pollution in our environment and pro-inflammatory foods that are consumed.Let’s nerd out for a moment. Signaling proteins called, cytokines are synthesized and secreted by your immune cells (macrophages, T-lymphocytes, B-lymphocytes, mast cells, endothelial cells, fibroblasts and stromal cells) when stimulated by a pathogen. CBD inhibits cytokine and IL-6 (pro-inflammatory) production.In one interesting study, researchers decided to test the effect of CBD on four cell signaling or mediating molecules associated with intestinal inflammation and oxidative damage to the gut. Their findings were as follows:Inducible nitric oxide synthase (iNOS) – CBD reduced the overexpression of iNOS in response to colitis. iNOS overexpression is well correlated with disease activity with colitis, and inhibitors of iNOS lead to improvement in experimental models of IBD. iNOS results in high-output production of NO, which results in oxidative damage to the intestine via reactive oxygen species (ROS).Interleukin-1β – levels significantly increased with experimental colitis. CBD was shown to decrease levels. IL-1β is shown to have potent pro-inflammatory activity and thus heightens the inflammatory response that leads to intestinal injury. IL-1β amplifies the production of inflammatory leukocytes (immune system cells), resulting in an increase of inflammation.Interleukin-10 – levels significantly decreased with experimental colitis. CBD was shown to restore levels. IL-10 has anti-inflammatory activity by inhibiting the release of pro-inflammatory cytokines. Restoration of IL-10 activity is critical to intestinal health.The reduction of iNOS and reactive oxygen species by CBD, along with the reduction of lipid peroxidation, shows the important therapeutic action of CBD in reduction of colonic inflammation by indirect reduction of oxidative damage. In addition, the dysregulation of the interleukins IL-1B and IL-10 is a well-known disruption caused by irritable bowel disease (IBD). The restoration of these interleukins to normal behavior by CBD, although the specific pathway is unknown, is another important therapeutic action that CBD has on reduction of colonic inflammation.It is the CBD in marijuana that is being researched to treat mTBI and other head traumas. Athletes in the US have been using marijuana for decades, often to relax them but without (or with) factual knowledge been using it to recover their bodies from inflammation and the pain they undergo in order to perform at their peak right away.DosageRegarding dosage, most clinical trials show CBD dosing ranging from 10-800 mg of CBD per day (although to treat schizophrenia, doses may be as high as 1,300mg). But everyone is different and you’ll likely need to experiment with a dosage range that works for you. A lot of this also depends on absorption. Not all CBD supplements are created equal. AbsorptionWhat is the purpose of consuming something beneficial if your body cannot absorb it? Most hemp CBD carries a particle size of 150 nm to 5,000 nm. This makes it nearly impossible to fully absorb and even to cross the blood/ brain barrier. CBD Oils such as Naked Jane use a nano-emulsion system that bring the particle size down to 25 nm-60 nm making this in of itself better bioavailable.  This means that a hybrid-nanoengineered CBD is over 10x more bioavailable in the body than any other oil based CBD and that just 10mg of a nanoparticle CBD is comparable to 100 mg of standard CBD.Combining a nano-emulsion system in cannabinoids and terpenoids in CBD with the isolated curcuminoids of a high-curcumin containing turmeric plant, the bioavailability of the CBD increases even greater!!! As curcumin in of itself is poorly absorbed, combining it with piperine will create the mack daddy of all absorption. Read the Full Show Notes  

Paul Wang:         Welcome to the monthly podcast, On the Beat for Circulation: Arrhythmia and Electrophysiology. I'm Dr Paul Wang, editor-in-chief, with some of the key highlights from this month's issue. In our first paper from the Finland Group Registry, Nordenswan and her colleagues have further defined the risk of sudden cardiac death in ventricular arrhythmias in patients with cardiac sarcoidosis. While it has been observed that there's a high risk of lethal arrhythmias in cardiac sarcoidosis patients with high grade AV block, the incidents in these patients with lone AV block, in the absence of known VT or left ventricular dysfunction, has not been described. From the patients with myocardial inflammatory diseases in their registry, the authors identified 143 cardiac sarcoidosis patients with high grade AV block. Of these patients, 90 had lone AV block. For these patients, the five-year composite incidence of sudden cardiac death or VT was 24%, in comparison with 54% in AV block with ventricular tachycardia or severe left ventricular dysfunction, and 24% with AV block in non-severe left ventricular dysfunction.                                 These findings support implementation of an implantable cardioverter defibrillator for cardiac sarcoidosis patients with lone AV block. A recent study by Gold and Colleagues suggests atrial ventricular optimization improves reverse remodeling in patients with significant intraventricular electrical delay. In this sub-study of the Smart AV Trial, 275 subjects were randomized to either electrogram-based AV optimization, smart delay, or nominal AV delay, 120 milliseconds. In this mostly male cohort, with a mean age of 65 years, and a left ventricular ejection fraction of 28% at follow up, at six months the benefit of AV optimization increased as intraventricular electrical delay prolonged, defined as the time between the peaks of the right and left ventricular electrograms. The longest cohort trial of intraventricular conduction delay had 4.26 times greater odds of remodeling response with AV optimization compared with nominal AV delay. This suggests that the left ventricular lead position in AV optimization might have a synergistic effect in reverse remodeling in cardiac resynchronization patients.                                 Concerns have long been raised that anorexia nervosa causes QT interval prolongation predisposing these individuals to life-threatening ventricular arrhythmias and sudden cardiac death. Fredrickson and Associates made progress in challenging this conclusion, demonstrating that despite a slightly increased incidence in borderline mean QTC, greater than 440 milliseconds, in 430 female anorexia nervosa patients, compared to 123 healthy controls from the Danish Civil Register, there was no difference in the risk of prolonged corrected QT interval greater than 460 milliseconds between the two groups.                                 However, during a 10-year follow up, anorexia nervosa patients had a slightly increased risk of ventricular tachycardiac, aborted cardiac arrest, and cardiac arrest compared to healthy controls. Incidence of 0.5% in the anorexia nervosa patients versus 0.07% in healthy controls. In addition, anorexia nervosa patients had a significantly increased risk of all-cause mortality with a hazard ratio of 11.2 over 10 years compared to population-based cohort, suggesting that anorexia nervosa increases the risk of medical complications, including cardiac complications, but these are not directly related to anorexia nervosa's effects on corrected QT interval.                                 In our next paper, Ahmed Karim Talib and Associates reported on the outcomes of endocardial ablation of drug-resistant ventricular fibrillation in Brugada syndrome. Endocardial mapping ablation was performed in 21 patients with one or more episodes of ventricular fibrillation. Endocardial mapping revealed low voltage fractionated endocardial electrograms in 19% of patients. All patients underwent ablation of ventricular fibrillation triggers, followed by endocardial substrate modification. Noninducibility of ventricular fibrillation and normalization of Brugada ECG was documented in 14, in three patients, respectively. During a mean follow up of 54 months, seven patients, or 33%, experienced ventricular fibrillation recurrences, presence of cruris notching in D1 was associated with the presence of low-voltage fractionated endocardial electrograms in ablation failure.                                 In our next paper, Jae Yeong Cho and Associates tried to elucidate the mechanism of sudden cardiac death in patients with heart failure with preserved ejection fraction using an ambulatory recording in a rat model. They showed that rats who had a clinical phenotype of heart failure with preserved ejection fraction had prolonged QT and QTC intervals and reduced heart rate variability compared to the controls. Importantly, the rats with heart failure and preserved ejection fraction had significantly higher rates of spontaneous ventricular arrhythmias, and about a third died suddenly with ventricular arrhythmia being the terminal rhythm.                                 In our next paper, Elad Anter and Colleagues used a novel mapping algorithm which integrated atrial activation mapping, vector analysis, and the physiologic constraints of atrial excitation to determine an atrial arrhythmias mechanism in circuitry. Ander's group initially applied this technique to historical controls and found that atrial tachycardias were diagnosed and their termination site predicted with 92.5% accuracy. When this algorithm was then applied prospectively to mapping of unknown atrial arrhythmias in 20 patients, this technique was able to identify macroreentrant, localized reentrant in focal tachycardias, even when standard mapping techniques had failed. This enhanced mapping capability allowed for significantly shorter ablation time when compared to historical controls, 3.2 +/- 1.7 minutes versus 17.3 +/- 6.6 minutes.                                 In our next paper Richard Walton and Colleagues explored the anatomical and electrical characteristics of the moderator band which provides a substrate for macroreentrant ventricular tachycardia. Ventricular wedge preparations with intact moderator bands were studied from humans in two cases and sheep in 15 cases. The moderator band structure was remarkably organized as two excitable yet uncoupled compartments of myocardium in [inaudible 00:07:45]. In humans, actual potential duration was significantly shorter in the moderator band than the right ventricular myocardium. S1, S2 moderator band pacing induced unidirectional propagation via the moderator band myocardium, permitting sustained macroreentrant ventricular tachycardia.                                 In sheep, the instance of ventricular tachycardia for right ventricular moderator band, or S1 right ventricular, and S2 moderator band pacing was 1.3%, 5.1%, and 10.3%, respectively. Severing the moderator band led to ventricular tachycardiac termination, confirming a primary arrhythmic role. Inducible preparations had shorter actual potential durations in the moderator band than right ventricle, whereas noninducible preparations showed no difference.                                 In our final paper, Witt and Associates studied the outcomes of 1421 patients who underwent ICD generator placement at Mayo Clinic, Minnesota and Beth Israel Deaconess Medical Center. During a mean follow up of 2.7 years appropriate ICD therapy occurred in 30.6% of the patients, predicted by low left ventricular ejection fraction and history of appropriate ICD therapy prior to generator replacement. On the other hand, 23.7% died prior to appropriate ICD therapy, predicted by old age, low left ventricular ejection fraction, diabetes mellitus, chronic lung disease, peripheral vascular disease, low hemoglobin level, and low glomerular filtration rate. The progressive increase in mortality after ICD generator change was observed in those who had aggregation of non-cardiac co-morbidities.                                 That's it for this month. We hope that you'll find the Journal to be the go-to place for everyone interested in the field. See you next time.  

The TWiP Titans solve the case of the Man from Queens with a Blister Burster, and explain the role of inflammatory monocytes during Leishmania infection of the skin. Hosts: Vincent Racaniello, Dickson Despommier, and Daniel Griffin Become a patron of TWiP. Links for this episode: Inflammatory monocytes and skin Leishmania (PLoS Path) Dracunculiasis (TWiP 37) Photo: Daniel using a LifeStraw Letters read on TWiP 136 This episode is brought to you by Blue Apron. Blue Apron is the #1 fresh ingredient and recipe delivery service in the country. See what’s on the menu this week and get your first 3 meals free with your first purchase – WITH FREE SHIPPING – by going to blueapron.com/twip. Case Study for TWiP 136 Patient seen by Daniel in India, 18 yo Islamic college student, left home, living in dorms in south, Hindu couple prepare meals, called dorm parents. He is being seen because developed lump in left side of neck, 1-2 cm mass. Previously completely healthy, no med/surg, no allergies. Prays multiple times a day, observes dietary restrictions. Afebrile, normal, but has 2 cm firm nontender lump inside interior portion of sternocleido mastoid muscle. Not tender. End of November, rainy season. No screens on dorm windows. No animal contact. Ultrasound done, and was helpful. Noticed in his neck over several weeks. Send your case diagnosis, questions and comments to twip@microbe.tv Music by Ronald Jenkees

Wed, 28 Jan 2015 12:00:00 +0100 https://edoc.ub.uni-muenchen.de/18655/ https://edoc.ub.uni-muenchen.de/18655/1/Herzog_Julia.pdf Herzog, Julia ddc:540, ddc:500, Fakultät fü

Macrophage-derived foam cells play a critical role in all stages of atherosclerosis, from the earliest discernable lesions to complex plaques. oxLDL is thought to be a main trigger for endothelial release of pro-inflammatory cytokines, subsequently causing transmigration of the monocytes into the vessel wall. Moreover, formation of macrophage-derived foam cells is mainly induced by oxLDL. Deposition of macrophage-derived foam cells in the lesions is induced by oxLDL uptake, as this uptake causes migratory arrest of the cells. Therefore, reversion of migratory arrest of macrophage-derived foam cells might enable these cells to leave the plaques resulting in reduction of plaque sizes. Our results show that iNOS participates in the mechanisms of oxLDL induced inhibition of macrophage-derived foam cell migration. Inhibition of iNOS expression completely reversed oxLDL mediated migratory arrest of macrophage-derived foam cells. Inhibition of iNOS was associated with enhanced phosphorylation of focal adhesion kinase (FAK) and subseqent actin polymerization. Furthermore, the p-FAK triggered increase in actin polymerization is dependent on iNOS mediated increased oxidative stress. Our results suggest that iNOS may be an interesting target gene to reverse the process of atherosclerosis.

Thu, 13 Dec 2012 12:00:00 +0100 https://edoc.ub.uni-muenchen.de/15208/ https://edoc.ub.uni-muenchen.de/15208/1/Hsieh_Chinyuan.pdf Hsieh, Chin-Yuan

Thu, 29 Nov 2012 12:00:00 +0100 https://edoc.ub.uni-muenchen.de/15100/ https://edoc.ub.uni-muenchen.de/15100/1/Hamm_Wolfgang.pdf Hamm, Wolfgang ddc:610, ddc:600, Med

Thu, 30 Jun 2011 12:00:00 +0100 https://edoc.ub.uni-muenchen.de/13221/ https://edoc.ub.uni-muenchen.de/13221/1/Schwerd_Tobias.pdf Schwerd, Tobias ddc:610, ddc:60

Wed, 2 Mar 2011 12:00:00 +0100 https://edoc.ub.uni-muenchen.de/13013/ https://edoc.ub.uni-muenchen.de/13013/1/Megerle_Judith.pdf Megerle, Judith ddc:530, ddc:500, Fakult

Because of the tremendous need for transgenic large animal models for human diseases, the process of SCNT is a crucial step in transgenic pig production. In our study, we evaluated the particular steps during the production for their impact on the efficiency of cloning transgenic pigs. For this purpose, statistical analysis was performed for all SCNT data from the years 2006 until June 2010. The RANKL transgenic osteoporosis model was chosen for an example for the production steps needed to finally achieve a disease model, to elucidate pitfalls and chances of SCNT procedure. In total 151 in vivo SCNT experiments using different transgenic cell lines were carried out, resulting in 243 piglets and fetuses. Statistical analysis revealed that donor cells treated exclusively in our laboratory had a significant better birth rate than donor cell originated of other laboratories. Furthermore, there was a significant relation between number of transferred NT embryos and later pregnancy checks, birth rate and abortion rate. The more NT embryos were transferred, the more pregnancies finished to terms. It was also elucidated that in our studies a different in vitro culture time of 24 or 48 hours had no significant impact on the outcome like pregnancy or birth rate. Seasonal changes during the years had no significant influence on pregnancy rate, birth or abortion. But there was a strong tendency that autumn showed best performance of all seasons, and most pregnancies were lost after embryo transfers during the summer. All these findings will be integrated in future in vivo SCNT experiments and embryo transfers. For the production of a transgenic osteoporosis model 17 in vivo experiments took place so far, with an outcome of 4 fetuses and 25 piglets. For gaining a controllable expression of RANKL, it was necessary to establish double transgenic pigs to sidestep harmful effects of RANKL overexpression during the fetal development. First attempts to integrate both genes, tetracycline controlled transactivator (Tet-On) and RANKL, in a single step of cell transfection and SCNT, had no satisfying result. We obtained 4 fetuses and stillborn recloned piglets carrying both genes, but they showed only expression of Tet-On and it was impossible to induce RANKL overexpression. Therefore the strategy was changed in favor to two rounds of transfection and nuclear transfer. First Tet-On transgenic piglets were established and screened for integration and expression. Piglet 9894 showed the best expression and severed as donor for next cell transfection step. These Tet-On + TARE RANKL cells were in vitro tested for their inducibility. Thereafter SCNT and embryo transfer of the best candidate were performed and they resulted in 4 pregnancies which all finished to term. One double transgenic piglet could be raised and will be kept until adulthood to establish a line of Tet-On +TARE RANKL transgenic pigs. Importantly, this founder animal showed inducible RANKL overexpression. Other constructs might be based on the existing Tet-On cell line in the future, offering an inducible system for a broad variety of different transgenes. Thus a functional Tet-On system in the pig is reported for the first time.

Mon, 20 Dec 2010 12:00:00 +0100 https://edoc.ub.uni-muenchen.de/14028/ https://edoc.ub.uni-muenchen.de/14028/1/Saint_Paul_Veronica_2010.pdf Saint Paul, Veronica von ddc

Hepatocyte Growth Factor (HGF) is a pleiotropic factor acting on cells expressing the Met receptor tyrosine kinase. HGF/Met signaling has been described in detail and it is known to control cell migration, growth and differentiation in several embryonic organs and to be implicated in human cancer. Conversely, little is known about the transcriptional targets that lead to Met-mediated biological functions. Also, little is known about the physiological mechanisms that attenuate Met signaling. This work provides the results of a screen for genes transcriptionally regulated by Met in several cell lines and addresses the functions of the highly inducible gene Mig6 (Mitogen-inducible-gene6, also called Gene 33 and RALT). By the use of Met loss of function mutant mice Met is shown to be the major inducer of mig6 in hepatocytes and lungs of E13.5 embryos. Mig6 is shown in turn to negatively regulate HGF/Met-induced cell migration. The effect is observed by Mig6 overexpression and reversed by Mig6 siRNA knock down experiments indicating that endogenous Mig6 is part of a mechanism that inhibits Met signaling. Mig6 functions in cells of hepatic origin and in neurons suggesting a role for Mig6 in different cell lineages. Mechanistically, Mig6 requires an intact Cdc42/Rho interactive binding (CRIB) domain to exert its inhibitory action suggesting that Mig6 acts at least in part distally from Met possibly by sequestering Rho-like GTPases. Because Mig6 is also induced by HGF stimulation, this work provides evidence that Mig6 is part of a negative feedback loop that attenuates Met functions in different contexts and cell types.

Background: Hyperoxic exposures are often found in clinical settings of respiratory insufficient patients, although oxygen therapy (>50% O-2) can result in the development of acute hyperoxic lung injury within a few days. Upon hyperoxic exposure, the inducible nitric oxide synthase (iNOS) is activated by a variety of proinflammatory cytokines both in vitro and in vivo. In the present study, we used a murine hyperoxic model to evaluate the effects of iNOS deficiency on the inflammatory response. Methods: Wild-type and iNOS-deficient mice were exposed to normoxia, 60% O-2 or >95% O-2 for 72 h. Results: Exposure to >95% O-2 resulted in an increased iNOS mRNA and protein expression in the lungs from wild-type mice. No significant effects of iNOS deficiency on cell differential in bronchoalveolar lavage fluid were observed. However, hyperoxia induced a significant increase in total cell count, protein concentration, LDH activity, lipid peroxidation, and TNF-alpha concentration in the bronchoalveolar lavage fluid compared to iNOS knockout mice. Moreover, binding activity of NF-kappaB and AP-1 appeared to be higher in wild-type than in iNOS-deficient mice. Conclusion: Taken together, our results provide evidence to suggest that iNOS plays a proinflammatory role in acute hyperoxic lung injury.

The antibacterial effect of the soybean phytoalexin glyceollin was assayed using a liquid microculture technique. Log-phase cells of Bradyrhizobium japonicum and Sinorhizobium fredii were sensitive to glyceollin. As revealed by growth rates and survival tests, these species were able to tolerate glyceollin after adaptation. Incubation in low concentrations of the isoflavones genistein and daidzein induced resistance to potentially bactericidal concentrations of glyceollin. This inducible resistance is not due to degradation or detoxification of the phytoalexin. The inducible resistance could be detected in B. japonicum 110spc4 and 61A101, representing the two taxonomically divergent groups of this species, as well as in S. fredii HH103, suggesting that this trait is a feature of all soybean-nodulating rhizobia. Glyceollin resistance was also inducible in a nodDlD2YABC deletion mutant of B. japonicum 110spc4, suggesting that there exists another recognition site for flavonoids besides the nodD genes identified so far. Exudate preparations from roots infected with Phytophthora megasperma f. sp. glycinea exhibited a strong bactericidal effect toward glyceollin-sensitive cells of B. japonicum. This killing effect was not solely due to glyceollin since purified glyceollin at concentrations similar to those present in exudate preparations had a much lower toxicity. However, glyceollin-resistant cells were also more resistant to exudate preparations than glyceollin-sensitive cells. Isoflavonoid-inducible resistance must therefore be ascribed an important role for survival of rhizobia in the rhizosphere of soybean roots.