Podcasts about Glycosylation

In this heartfelt and powerful conversation, Matty J speaks with Broome local Lee, mother of 9-year-old Kelsey Stewart, who lives with a rare genetic condition called CDG1A (Congenital Disorder of Glycosylation). With only around 1,200 cases worldwide, CDG is a complex and often misunderstood disorder that affects physical and intellectual development. Lee shares her family's story—raising Kelsey in Broome, navigating therapy and travel challenges, and advocating for greater inclusion and support through the Sybree Rose Button Foundation. She also highlights Kelsey's incredible milestones: learning to crawl, speak with the help of a $7,500 communication device, and her love for music, school, and community. This interview builds toward the CDG Awareness Family Fundraiser, held at St Mary's Primary School on Saturday, May 17 from 2 PM. Featuring: A Wonka movie screening

Dr Matthew Wilson, Postdoctoral Fellow at the Centre for Human Genetics, KU Leuven, joins hosts Silvia Radenkovic and Rodrigo Starosta to discuss a scintillating selection of CDG papers in our first ever research round-up. The papers discussed include: A pseudoautosomal glycosylation disorder prompts the revision of dolichol biosynthesis. Wilson et al Clinical and biochemical footprints of congenital disorders of glycosylation: Proposed nosology. Ng et al Rft1 catalyzes lipid-linked oligosaccharide translocation across the ER membrane. Chen et al Molecular characterization of Rft1, an ER membrane protein associated with congenital disorder of glycosylation RFT1-CDG. Hirata et al Genome and RNA sequencing were essential to reveal cryptic intronic variants associated to defective ATP6AP1 mRNA processing. Morales-Romero et al N-glycoproteomic and proteomic alterations in SRD5A3-deficient fibroblasts. Garapati et al In vitro treatment with liposome-encapsulated Mannose-1-phosphate restores N-glycosylation in PMM2-CDG patient-derived fibroblasts. Shirakura et al Liposome-encapsulated mannose-1-phosphate therapy improves global N-glycosylation in different congenital disorders of glycosylation. Budhraja et al D-mannose as a new therapy for fucokinase deficiency-related congenital disorder of glycosylation (FCSK-CDG). Starosta et al Glycoproteomics in Cerebrospinal Fluid Reveals Brain-Specific Glycosylation Changes. Baerenfaenger et al Neural and metabolic dysregulation in PMM2-deficient human in vitro neural models. Radenkovic et al

Send us a textShort Summary: The science of glycosylation, its impact on health, and potential treatments for congenital disorders of glycosylation (CDGs).About the guest: Dr. Eva Morava is a pediatric geneticist originally from Hungary, who has specialized in inborn errors of metabolism, particularly CDGs. She has a background in pediatrics and genetics from training in Hungary and the U.S., and currently leads the genetics and genomics department at Mount Sinai.Note: Podcast episodes are fully available to paid subscribers on the M&M Substack and to everyone on YouTube. Partial versions are available elsewhere.Episode Summary: Dr. Eva Morava discusses the critical role of glycosylation in biology, where sugars are not just used for energy but in modifying proteins to perform their functions. She explains how defects in this process lead to CDGs, a group of rare genetic disorders. The conversation covers the mechanics of glycosylation, clinical presentations of CDGs, current research on treatments including dietary interventions and gene therapy, and the broader implications of glycosylation in health, such as in liver disease and cancer.Key Takeaways:Glycosylation Basics: Sugars are attached to proteins (glycosylation) to modify their structure and function, influencing everything from clotting to immune response.CDGs: These disorders are caused by genetic defects in the glycosylation process, leading to a wide array of symptoms because many proteins require glycosylation to function correctly.Clinical Variability: CDGs can range from severe, multi-systemic presentations to relatively mild cases, affecting life expectancy and quality of life variably.Therapeutic Approaches: Current research includes drug repurposing for enzyme activity enhancement, dietary interventions with special sugars like mannose, and gene therapy, with some trials underway.Liver Connection: A significant portion of glycosylation occurs in the liver; thus, liver diseases like non-alcoholic fatty liver disease can impact glycosylation.*Not medical advice.Support the showAll episodes (audio & video), show notes, transcripts, and more at the M&M Substack Affiliates: MASA Chips—delicious tortilla chips made from organic corn and grass-fed beef tallow. No seed oils or artificial ingredients. Use code MIND for 20% off. Lumen device to optimize your metabolism for weight loss or athletic performance. Use code MIND for 10% off. Athletic Greens: Comprehensive & convenient daily nutrition. Free 1-year supply of vitamin D with purchase. KetoCitra—Ketone body BHB + potassium, calcium & magnesium, formulated with kidney health in mind. Use code MIND20 for 20% off any subscription. Learn all the ways you can support my efforts

Send us a Text Message.Dr. Robert Kiltz is a physician who promotes eating fatty meat for overall health and wellness. He believes that eating this way can help improve fertility and he encourages people to share their success stories on social media. When asked about the dangers of hormones in dairy, he states that it is not a big concern compared to the hormones found in plants. He also mentions that inflammation caused by plant sugars and chemicals can damage cells and affect fertility. Richard and Stephen emphasize how ancient humans were omnivorous with their protein sources predominantly from animals and hunting, not plants. Modern doctors and dieticians give misguided advice as they don't know man's true evolution. People lack macro-nutrients and focus on micro-nutrients too much. Dr. Kiltz stresses that fat is essential for one's body and should be prioritized in a diet, as nature provides protein and fat in perfectly packaged proportions. He also challenges the idea that obesity is a disease, stating that it is a natural state that is only harmful if paired with a low-fat diet that inflames the body's cells.https://www.instagram.com/doctorkiltz/https://www.doctorkiltz.com/Thank you so much for listening to my podcast. I hope you enjoyed it. Your support means the absolute world to me. And if you're enjoying the show, I've got a small favor to ask you. I'd be incredibly grateful if you would consider becoming a supporter and make a small monthly donation. Your contribution will really help to improve the show. It's a small monthly contribution. You can cancel at any time, and the link is in the show notes. Tune in to the Patient Advocacy Voices Podcast todayJoin Sanofi's Eric Racine to meet the unsung heroes leading patient advocacy organizationsListen on: Apple Podcasts SpotifySupport the Show.All my links in 1 easy list, including booking and personal training workout plans at LINKTREE You can now download the carnivore experience appApple direct link for apple devices Google play store direct link to app for Android Coach Stephen's Instagram Book me for coaching My growing UK carnivore YouTube channel I have set up a community that is all about eating low-carb and specifically carnivore. CLICK HERE Support my podcast from just £3 per monthBECOME A SUPPORTER Success stories Optimal Health 5 Star reviews All my facebook and other reviews are here Thanks to www.audionautix.com for any music included. Ple...

In the latest shortcast, Inez Bosnyak describes a case of ALG1-CDG presenting without an abnormal isoelectric focusing pattern. Normal transferrin glycosylation does not rule out severe ALG1 deficiency Inez Bosnyak, et al https://doi.org/10.1002/jmd2.12415

Dr. Ryan Flynn is a Principal Investigator at Boston Children's Hospital and an Assistant Professor in the Stem Cell and Regenerative Biology Department at Harvard University. His lab explores the interface between RNA and glycans. He talks about the discovery of glycoRNA and the chemistry of the RNA-glycan connection. He also discusses the effects of glycans on stem cell function and deciding between a career in medicine and basic science.

TIME STAMPS: 00:08 Dr. Kiltz's credentials, accomplishments, and backstory. 02:40 The “BBEBIS” diet! 04:00 Dr. Kiltz's famous ICE CREAM recipe! It's a TREAT, not a CHEAT. 07:02 HIGHLY ADDICTIVE CHEMICALS found in most plants. 09:38 GLYCATION: the opposite of GLYCOSYLATION, like “rust” to our body, especially proteins. 12:02 Plants + lean protein = ALCOHOL in your GUT! 14:04 ADIPOSE TISSUE: Why it's the most nutritious part of the animal. 18:08 Occam's Razor: problem-solving principle that recommends searching for explanations constructed with the smallest possible set of elements. 23:03 The value of a COMMUNITY in your EXERCISE and in your FAITH! 30:40 PLANTS are the ULTIMATE PREDATORS. 39:00 SPIRITUAL & PHYSICAL FITNESS: Strength to overcome just about anything in this universe. 43:02 Dr. Kiltz's BOOKS & WRITING HABITS. 51:18 The misunderstood SCIENCE OF METABOLISM. Books referenced: Essentials of Glycobiology, Third Edition, by Ajit Varki & Introduction to Glycobiology by Maureen E. Taylor and Kurt Drickamer 57:01 INTRAMUSCULAR FAT and INSULIN INJECTIONS: Is this why so many bodybuilders are DYING YOUNG?

As seen on TV Richard Smith, nutritionist to the celebrities and Coach Stephen, the most famous Carnivore in the UK, take your questions liveMAIN POINTSAvoid lectin-rich nuts, consider OMAD, and use tallow for dry skin.ROUGH TIMESTAMPS0:00 Introduction07:27 Ketone production is 400% more efficient at creating and utilizing energy than glucose.14:54 Glycosylation and glycation have different effects on the body.22:21 A low-carb keto carnivore lifestyle can have various positive effects on the body, including improved health, mental clarity, and skin conditions.29:48 Grounding is beneficial for the body as it allows electrons to enter, improving sleep and overall well-being.37:15 You don't need to be in ketosis all the time and testing ketones at certain times can give false readings.44:42 The body requires 5 to 10 grams of glucose per hour and does not need exogenous carbohydrate52:09 The importance of incrementally transitioning to a carnivore diet and the benefits of electrolytes59:35 Avoid consuming lectins and phytic acid in nuts to prevent intestinal permeability and related issues.Mighty Networks https://keto-low-carb-carnivore-co.mn.co/users/onboarding/choose_plan?plan_id=527958&bundle_token=eae3214aca9e93fca943d927fd73fc77&utm_source=manualTry to join in via a computer rather than a mobile deviceThank you so much for listening to my podcast. I hope you enjoyed it. Your support means the absolute world to me. And if you're enjoying the show, I've got a small favor to ask you. I'd be incredibly grateful if you would consider becoming a supporter and make a small monthly donation. Your contribution will really help to improve the show. It's a small monthly contribution. You can cancel at any time, and the link is in the show notes. Support the showAll my links in 1 easy list, including booking and personal training workout plans at LINKTREE You can now download the carnivore experience appApple direct link for apple devices Google play store direct link to app for Android Coach Stephen's Instagram Book me for coaching My growing UK carnivore YouTube channel I have set up a community that is all about eating low-carb and specifically carnivore. CLICK HERE Support my podcast from just £3 per monthBECOME A SUPPORTER Success stories Optimal Health 5 Star reviews All my facebook and other reviews are here Thanks to www.audionautix.com for any music included. Ple...

Join us as we take questions from listeners 'live and unedited'Of course Richard Smith (the keto pro) and Coach Stephen will also be answering but do make the most of having the hour with our esteemed guest Dr Robert Kiltzhttps://www.cnyfertility.com/our-team-members/dr-robert-kiltz/https://www.instagram.com/doctorkiltz/APPROX TIMESTAMPS00:04 INTRO: Dr Kiltz is enjoying a beautiful day in Upstate New York06:01 Keto and carnivore lifestyles, along with fasting and faith, can help live a more vibrant life and possibly increase lifespan.18:33 Ketosis is a fasting state that can be achieved by eating one meal a day or less, primarily consisting of fat.24:36 Exercise can increase blood glucose levels even without consuming carbohydrates.35:54 Glycosylation is crucial for the human body41:28 Reducing dietary fat can help in leaning up for competition.52:20 Exercise is crucial for maintaining mobility and overall health.57:36 Fasting helps with autophagy, which is the process of cells cleaning up dead or damaged cells.1:08:22 Weight loss and fat loss are different things.1:13:34 Listen to your body, eat healthy, and everything else will fall into place.1:23:56 Extended fasting can trigger autophagy after 30 days, but it is not recommended for humans.1:28:58 Optimal health doesn't necessarily rely on specific ranges or measurements.1:39:55 Standard diets are causing cancer and harming our health1:45:40 Keto / Carnivore and fasting are important for health and wellness.Thank you so much for listening to my podcast. I hope you enjoyed it. Your support means the absolute world to me. And if you're enjoying the show, I've got a small favor to ask you. I'd be incredibly grateful if you would consider becoming a supporter and make a small monthly donation. Your contribution will really help to improve the show. It's a small monthly contribution. You can cancel at any time, and the link is in the show notes. Support the showAll my links in 1 easy list, including booking and personal training workout plans at LINKTREE You can now download the carnivore experience appApple direct link for apple devices Google play store direct link to app for Android Coach Stephen's Instagram Book me for coaching My growing UK carnivore YouTube channel I have set up a community that is all about eating low-carb and specifically carnivore. CLICK HERE Support my podcast from just £3 per monthBECOME A SUPPORTER Success stories Optimal Health 5 Star reviews All my facebook and other reviews are here Thanks to www.audionautix.com for any music included. Ple...

This is one hot tech discussion!  Protein therapeutics have great potential, but significant limitations to their utility. Dr. Dan Mandrell of Gro Biosciences explains how they are using a non-standard library of amino acids to increase protein presentation. Along with glycosylation and other modifications, the proteins are more therapeutically active. 

Episode Description: In the latest Grow Everything podcast episode, hosts Karl and Erum explore the exciting world of biotechnology, sustainability, and healthcare with Josh Robinson, CEO of Cocoon Bioscience. The trio delve into the pivotal role of small-scale production in diverse sectors, such as semiconductors and biology, hinting at the revolutionary potential of MRNA vaccines that can be mass-produced using minimal enzyme, despite its significant cost. Robinson emphasizes the need for natural, innovative solutions like fermentation, known for yielding high-value molecules. He piques listeners' curiosity about Cocoon's future and the broader biotech landscape in the forthcoming 3 to 10 years. The conversation concludes with a book recommendation - Andy Weir's "Hail", a space-set fictional adventure sure to intrigue the audience of science enthusiasts, researchers, and professionals alike. Grow Everything brings to life the bioeconomy when hosts Karl Schmieder and Erum Azeez Khan share stories from the field and interview leaders and influencers in the space.  Life is a powerful force and it can be engineered. What are we creating? Learn more at www.messaginglab.com/groweverything Topics Covered: 00:00:00 GMOs: Unforeseen Impacts 00:01:51 Microbes: Turning Fruit Waste to Leather 00:05:18 Philadelphia Flooding: Lack of Preparedness 00:09:55 Cocoon Biosciences: Insects in Biotech Ecosystem 00:16:01 Enzyme Efficiency: Role of Folding and Glycosylation 00:17:38 Lab to Table: Cultivated Meat and Cocoon's Role 00:22:02 Bio Airport Facility: Upscaling Insect Rearing 00:27:14 Balancing Sustainability and Healthcare Access 00:34:01 Future of Cocoon and Biotech Industry 00:38:24 Book Recommendation: 'Hail' by Andy Weir 00:43:03 Navigating Spanish and Importance of Food Innovation 00:44:22 Next: Biotech Progress Discussion with Josh 00:51:25 Gratitude: Acknowledging Feedback and Engagement Episode Links: LinkedIn Cocoon Bioscience Algenex (Cocoon's Tech Partner) Pivot Bio  Hail Mary by Andy Weir Successful Science Writing by Janice Matthews Call or Text the Grow Everything Hotline:  +1 804-505-5553 Have a question or comment? Message us here: Instagram / TikTok / Twitter / LinkedIn / Youtube / GrowEverything website Email: groweverything@messaginglab.com Support here: Patreon Music by: Nihilore Production by: Amplafy Media --- Send in a voice message: https://podcasters.spotify.com/pod/show/messaginglab/message

Glycosylation, a biological process vital to life, is likely something you've never heard of before. It involves the addition of sugar molecules, known as glycans, to proteins and lipids which enable a number of crucial physiological functions, from immune regulation to cell-to-cell communication. This episode explores what happens when genetic mutations disrupt the glycosylation pathway as well as the detective work and collaboration required to diagnose and treat the resulting conditions. Hudson Freeze is the Director of the Sanford Children's Health Research Centre and the Human Genetics Programme at Sanford Burnham Prebys Medical Discovery Institute, his research focuses on Congenital Disorders of Glycosylation, or CDG for short. Listen in as he reflects on the adventurous road to discovery of these rare conditions, the unlikely therapeutic options, the knowledge gap within the medical and scientific communities, and how a single discovery at Yellowstone National Park changed the course of history.We Discuss:Hudson's Background [02:11]Glycosylation vs. Glycation [03:37]Congenital Disorders of Glycosylation (CDG) [06:10]The Accelerating Pace of CDG Research [15:50]Hudson's Groundbreaking Discovery [21:02]Utilising Animal Models for Glycosylation Research [25:36]Including Families and Patients in CDG Research [34:51]The Future of Diagnostic Options [40:55] The Role of Dietary Supplementation in CDG Treatment [45:37]Overcoming Funding Challenges [57:11]View the Show Notes Page for This Episode Purchase a GlycanAge Test Engage with us on social media: Instagram LinkedInFacebookTwitter Please be advised that this show is for information only and should not be considered as a replacement or equal to medical advice. 

Link to bioRxiv paper: http://biorxiv.org/cgi/content/short/2023.03.29.534715v1?rss=1 Authors: van Deventer, S. J., Hoogvliet, I. A., van de Voort, M., Arnold, F., van Spriel, A. Abstract: Tetraspanin proteins play an important role in many cellular processes as they are key organizers of different plasma membrane receptors. Most tetraspanins are highly glycosylated, but the function of this post-translational modification remains largely unstudied. In this study we investigated the glycosylation of CD37 and CD53, two tetraspanins important for cellular and humoral immunity. Broad and cell-specific repertoires of N-glycosylated CD37 and CD53 were observed in human B cells. We generated different glycosylation mutants and analyzed their localization, nanoscale organization and protein interactions. Abrogation of glycosylation in CD37 revealed the importance of this modification for CD37 surface expression, whereas neither surface expression nor nanoscale organization of CD53 was affected by its glycosylation. CD37 interaction with its known partner proteins, CD20 and IL-6R, was not affected by glycosylation, other than via its changed subcellular localization. Surprisingly, glycosylation was found to inhibit the interaction between CD53 and its partner proteins CD45 and CD20. Together, our data show that tetraspanin glycosylation affects their function in immune cells, which adds another layer of regulation to tetraspanin-mediated membrane organization. Copy rights belong to original authors. Visit the link for more info Podcast created by Paper Player, LLC

How much control do we have over our health? The study of epigenetics made us aware of the remarkable malleability of our phenotype, prompting closer investigation on how lifestyle and environment impact our health and longevity. In this context, glycans - a class of historically understudied biological polymers may hold the key to a better understanding of the interplay between genetic and epigenetic factors contributing to disease development. Olga Zaytseva is a post-doctoral researcher at Genos Glycoscience Research Laboratory in Croatia, her research focuses on the relationship between human diseases and the regulation of antigen-specific IgG glycans. Listen in as she unravels the latest research on genetic and epigenetic regulation of IgG glycosylation and introduces us to the role of environmental factors in common diseases, from obesity to depression. We Discuss: Genetics versus Epigenetics: Understanding the Differences [02:22]Epigenetic Changes in Disease: Exploring the Example of Depression [07:37]Good vs. Bad Glycans: Debunking the Myths [12:31]Measuring Glycans on IgG: Benefits and Applications [15:40]GWAS and Glycosylation: Unravelling the Genetic Regulation [21:21]The Role of Environment in Epigenetic Changes: Examining the Learning Process Example [24:46]Mutations make sense in the Context: A Closer Look at Obesity [30:08]Environmental Impact on IgG Glycome: Insights from Smoking and Diet [32:44]Behaviour and IgG Glycome: Is There a Connection? [34:46]Pleiotropy and Poor Hearing in White Cats: What's the Link? [36:18]Glycan Changes and Disease: Causative or Consequential? [40:59]IgG Glycans and Country Development: Investigating the Association [47:51]Glycans as Biomarkers: Challenges and Opportunities [51:51]Exciting Questions Yet to Be Answered: Future Directions in Glycan Research [57:46] Follow Olga on social mediaFacebookLinkedInView the Show Notes Page for This Episode Purchase a GlycanAge Test Engage with us on social media: Instagram LinkedInFacebookTwitter Please be advised that this show is for information only and should not be considered as a replacement or equal to medical advice. 

Link to bioRxiv paper: http://biorxiv.org/cgi/content/short/2023.02.25.530047v1?rss=1 Authors: Millhauser, G. L. Abstract: The C-terminal domain of cellular prion protein (PrP-C) contains two N-linked glycosylation sites, the occupancy of which impacts disease pathology. In this study, we demonstrate that glycans at these sites are required to maintain an intramolecular interaction with the N-terminal domain, mediated through a previously identified copper-histidine tether, which suppresses the neurotoxic activity of PrP-C. 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/2022.11.15.516651v1?rss=1 Authors: Zhang, C., Shafaq-Zadah, M., Pawling, J., Hesketh, G. G., Ng, D., Dransart, E., Pacholczyk, K., Longo, J., Gingras, A.-C., Penn, L. Z., Johannes, L., Dennis, J. W. Abstract: SLC3A2 (4F2hc, CD98) is an adaptor to the SLC7A exchangers and has undergone extensive repositioning of N-glycosylation sites with vertebrate evolution, presumably in synchrony with the species-specific demands of metabolism. The SLC3A2*SLC7A5 heterodimer imports essential amino acids (AA) and thereby stimulates mTOR signaling, while SLC3A2*SLC7A11 imports cystine required for glutathione synthesis and mitigation of oxidative stress. Analysis of SLC3A2 N-glycans revealed stable site-specific profiles of Golgi remodeling, apart from the conserved N365 site where branching and poly-N-acetylglucosamine content were sensitive to the insertion of lost ancestral sites and to metabolism. N-glycans at N381 and N365 stabilized SLC3A2 in the galectin lattice and opposed endocytosis, while N365 which is nearest the membrane, also promoted down-regulation by galectin-driven clathrin-independent endocytosis (glycolipid-lectin GL-Lect). This is the first report of both positive and negative regulation by galectin binding to N-glycans that are strategically positioned in the same membrane glycoproteins. Proteomics analysis in SLC3A2 mutant HeLa cells with induced re-expression of SLC3A2 as bait revealed the canonical non-N-glycosylated interactors, SLC7A5 and SLC7A11 exchangers, but also AA transporters that were dependent on SLC3A2 N-glycosylation, and are themselves, N-glycosylated AA/Na+ symporters (SLC7A1, SLC38A1, SLC38A2, SLC1A4, SLC1A5). The results suggest that the N-glycans on SLC3A2 regulate clustering of SLC7A exchangers with AA/Na+ symporters, thereby promoting Gln/Glu export-driven import of essential AA and cystine, with the potential to adversely impact redox balance. The evolution of modern birds (Neoaves) led to improved control of bioenergetics with the loss of genes including SLC3A2, SLC7A-5, -7, -8, -10, BCAT2, KEAP1, as well as duplications of SLC7A9, SLC7A11 and the Golgi branching enzymes MGAT4B and MGAT4C known to enhance affinities for galectins. Analyzing the fate of these and other genes in the down-sized genomes of birds, spanning ~10,000 species and greater than 100 Myr of evolution, may reveal the mystery of their longevity with prolonged vitality. 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/2022.10.15.512359v1?rss=1 Authors: Karas, B. F., Terez, K. R., Battula, N., Gural, B. M., Flannery, K. P., Aboussleman, G., Mubin, N., Manzini, C. Abstract: Biallelic mutations in Protein O-mannosyltransferase 1 (POMT1) are among the most common causes of a severe group of congenital muscular dystrophies (CMDs) known as dystroglycanopathies. POMT1 is a glycosyltransferase responsible for the attachment of a functional glycan mediating interactions between the transmembrane glycoprotein dystroglycan and its binding partners in the extracellular matrix (ECM). Disruptions in these cell-ECM interactions lead to multiple developmental defects causing brain and eye malformations in addition to CMD. Removing Pomt1 in the mouse leads to early embryonic death due to the essential role of dystroglycan in embryo implantation in rodents. Here, we characterized and validated a model of pomt1 loss of function in the zebrafish showing that developmental defects found in individuals affected by dystroglycanopathies can be recapitulated in the fish. We also discovered that pomt1 mRNA provided by the mother in the oocyte supports dystroglycan glycosylation during the first few weeks of development. Muscle disease, retinal synapse formation deficits, and axon guidance defects can only be uncovered during the first week post-fertilization by generating knock-out embryos from knock-out mothers. Conversely, maternal pomt1 from heterozygous mothers was sufficient to sustain muscle, eye, and brain development only leading to detectable muscle disease and loss of photoreceptor synapses at 30 days post fertilization. Our findings show that it is important to define the contribution of maternal mRNA while developing zebrafish models of dystroglycanopathies and that offspring generated from heterozygous and knock-out mothers can be used to differentiate the role of dystroglycan glycosylation in tissue formation and maintenance. Copy rights belong to original authors. Visit the link for more info Podcast created by Paper Player, LLC

In this week's episode we'll discuss the efficacy of tranexamic acid prophylaxis in patients with hematological malignancies, learn more about N-glycosylation as a therapeutic vulnerability in CALR-mutant MPN, and discuss how early initiation of disease-modifying therapy may be able to reduce myocardial fibrosis in sickle cell anemia.

In honour of CDG Awareness Day, Dr Hudson Freeze and Dr Jaak Jaeken revisit a discussion from the Scientific CDG Symposium 2021, considering what makes a Congenital Disorder of Glycosylation. CDG or not CDG Hudson H. Freeze, Jaak Jaeken and Gert Matthijs https://doi.org/10.1002/jimd.12498

Episode 16 of the Modern Chemistry podcast dives back into glycobiology and its practical application with Benjamin Schumann. Ben is a chemical biologist who studies the biology of carbohydrates (glycans). After completing his undergraduate biochemistry studies in Tübingen, Germany, he was trained in synthetic carbohydrate chemistry in the lab of Peter H. Seeberger at the Max Planck Institute of Colloids and Interfaces Potsdam and the FU Berlin. Developing vaccines against pathogenic bacteria based on synthetic glycans, Ben learned to apply his compounds in biological settings in vivo and in vitro. For his achievements, he received the Award for Excellence in Glycosciences and, in 2017, the prestigious Otto Hahn Medal by the Max Planck Society. During his postdoctoral work in the lab of Carolyn R. Bertozzi at Stanford University as an Alexander von Humboldt foundation Feodor Lynen fellow, Ben developed an interest in "precision tools" to study glycosylation of human cells in great detail. He started as a Group Leader at the Crick and Imperial College London in 2018.Towards the end of the show, Ben mentions a prize. Ben and colleagues from Imperial College and Stanford University were awarded the Royal Society of Chemistry's new Chemistry Biology Interface Division Horizon Prize, the Rita and John Cornforth Award. For more details, please check out this link on the Francis Crick Institute's website - https://www.crick.ac.uk/news/2021-06-08_research-prize-for-chemical-toolbox-to-study-the-role-of-cell-surface-sugars. You will hear the following terms used during the interview. I've included some descriptions here. Glycobiology - the study of complex sugar molecules in biological systems and cells.Glycobiome/glycoproteome – the total glycan make-up of a cell or organismGlycosylation – the addition of sugar molecules to other substances, often proteins.Glycans – complex sugar molecules composed of single sugar sub-units linked by chemical bondsChemical biology – the application of chemical techniques, and often small molecules produced by chemistry to study biological processes.Biochemistry – the study of biomolecules and pathways in living systemsLectins – highly specific carbohydrate-binding proteinsAntibody – a 'Y'-shaped protein produced by the immune system which identifies and binds to a specific substanceMonosaccharide – a single sugar subunitBioorthogonal chemistry – a chemical reaction that occurs in a living system without interfering with natural biochemistryGlycosyl transferase – enzymes which add a sugar (simple or complex) to an acceptor moleculeChemoenzymatic synthesis – producing chemical using biochemical pathways within cellsCRISPR - CRISPR stands for Clustered Regularly Interspaced Short Palindromic Repeats, a family of specific DNA sequences in prokaryotes. Enzymes, such as 'Cas9' can modify DNA at these CRISPR sequences, editing in or out new DNA sequences. Although this technique is known correctly as CRISPR-Cas9, the discovery and development of a range of CRISPR-associated enzymes has led to the term 'CRISPR' being used as a generic term for this suite of gene editing technologies.Isoenzymes – enzymes that catalyse the same reaction, but have different amino acid sequencesProteolytic digest – breaking up a protein, or peptide, chain into smaller fragmentsFluorophore – a fluorescent molecule (in this case, used as a reporter molecule)Biotin tag – a chemical molecule used as a reporter for other biomoleculesBio-organic chemistry – scientific studies combining organic chemistry and biochemistry, which studies biological processes using chemical methods.

Researchers at the Council for Scientific and Industrial Research (CSIR) briefed the media on Friday on some of the organisation's research, development and innovation (RDI) efforts aimed at addressing the HIV/AIDS and Tuberculosis (TB) epidemics in South Africa. CSIR researcher Dr Advaita Singh shed light on an innovative plant-based, highly potent anti-HIV antibodies research initiative. “The CSIR's plant-based anti-HIV antibodies research is aimed at developing a cost-effective pre prophylaxis vaccine which will passively immunise against HIV/AIDS, pre and post exposure. This initiative was inspired by the desire to empower young girls and women to protect themselves against the virus as studies have indicated that women are theoretically, at higher risk of HIV acquisition,” said Singh. Working with the National Institute of Communicable Diseases (NICD), the CSIR is developing broadly neutralising antibodies against HIV-1, the most widespread human immunodeficiency virus, for the prevention and treatment of HIV/AIDS. CSIR scientists and their research partners were able to demonstrate the ability to engineer tobacco plants to produce unique glycosylation and rare sulfation post-translational modifications, a result that is not usually seen in plants. Glycosylation is a biochemical process during which a glycan – a large carbohydrate molecule – attaches to a protein, a lipid, or another organic molecule. The glycoengineered Nicotiana benthamiana (a relative of the tobacco plant) expressed two broadly neutralising antibodies named CAP256-VRC26 08 and CAP256-VRC26 09. These two antibodies were originally isolated by collaboration Aids research centre CAPRISA and the NICD from a South African HIV positive patient during a trial conducted by CAPRISA. The outcome showed that the antibodies from the CAP256 lineage were the most active, with high potency and broad specificity. “We still have a number of steps to get through, as the research process commonly requires, before this technology is available in the market, however, the CSIR and its partners intends on making it accessible to patients through an industry partner once it is ready,” added Singh. Another technology presented by the CSIR at the media briefing aims to provide rural areas with a diagnostic assay that will test for HIV and TB simultaneously using one device in the comfort of the patient's home. CSIR lab technician Kanyane Malatji provided insight into a multiplex HIV and TB point-of-care diagnostic test being developed for use in remote and rural areas. “The effective management of patients infected with HIV and TB is restricted by separate diagnoses. The situation is exacerbated in remote areas where patients must either travel long distances to reach a healthcare facility or wait a long time before obtaining their TB diagnostic results. Therefore, through our multiplex HIV and TB point-of-care diagnostic assay, we hope to mitigate these challenges with the objective of improving treatment outcomes for patients in rural areas,” said Malatji. The locally developed diagnostic test uses a solid surface that couples the HIV and TB antigens and makes use of a locally produced fluorescence detector. “The benefits of this technology are that it offers a cost-effective solution with a low turnaround time of approximately 2 hours. Additionally, no laboratory infrastructure is required, the device is easy to use and can be performed at point-of-care without highly trained personnel,” added Malatji. The CSIR's multiplex HIV and TB point-of-care diagnostic assay is funded by government and the plant-based anti-HIV antibodies research work is funded by the Department of Science and Innovation and the South African Medical Research Council.

Elisa Fadda obtained her PhD in 2004 from the Department of Chemistry at the Université de Montréal under Professor Dennis R. Salahub. From May 2004 to May 2008, she worked as a post-doctoral fellow in Dr Régis Pomès group in Molecular Structure and Function at the Hospital for Sick Children (Sickkids) Research Institute in Toronto. From June 2008 until May 2013, Elisa worked as a research associate and honorary research lecturer in Prof Robert J. Woods group in the School of Chemistry at NUI Galway. In 2013 she was awarded a Post-Graduate Certificate in Teaching and Learning in Higher Education from the Centre for Learning and Teaching (CELT) at NUI Galway. In August 2013, Elisa became an Assistant Lecturer in the Department of Chemistry at Maynooth University, taking on a Lecturer position since 2014.You will hear the following terms used during the interview. I've included some descriptions here. Quantum chemistry -The branch of chemistry that apply quantum mechanics to chemical systems, including electronic structure, molecular dynamics and Schrödinger equations.Biophysics – And approach to science that applies methods typically used in physics to study biology and biological systems.Glycoproteins – Proteins which contain oligosaccharide chains (glycans), attached to amino acid side-chains via a covalent bond.Carbohydrates – Molecules (typically biological) composed of Carbon, Hydrogen and Oxygen, typically with a 2:1 Hydrogen:Oxygen atom ratio.Glycan (or polysaccharide) – Compounds made of many monosaccharide subunits, linked via a glycoside bond.N-Glycans – Glycans attached to a protein at an Asparagine residue via an N-glycosidic bond.Sequon – A sequence of amino acids in a protein that serve as a carbohydrate binding site.The carbohydrate is often an N-linked-Glycan.Asparagine, proline, serine, threonine. – Amino acids found naturally in biological proteins. Asparagine, serine and threonine are required in specific combinations to form a sequon, proline must be absent from a sequon.Glycosaminoglycans or mucopolysaccharides- Long, linear glucans consisting of repeating disaccharide units – most commonly uronic acid and an amino sugar.Glycosylation – A reaction in which a carbohydrate molecule is attached to a functional group of another molecule (such as a hydroxyl).  In biology the term typically refers to the carbohydrate being attached to a protein molecule.Folded protein – Proteins have several levels of structure, secondary, tertiary (and arguably quaternary) levels of structure describe how the polypeptide chain forms into specific structures that typically confer functional properties.Cryo-EM – Cryogenic Electron Microscopy studies samples cooled to cryogenic temperatures (-153 oC or lower), while embedded in vitreous water.X-Ray crystallography – A technique which uses X-rays to determine crystal structures, but studying the X-ray diffraction patterns.NMR – Nuclear Magnetic Resonance subjects samples to a strong magnetic fields and measures the resonance pattern of the nuclei. It is widely used to study the structure and dynamics of organic molecules.Spike proteins – More properly ‘Peplomers', spike proteins are glycoproteins that project from the surface of a virus particle lipid bilayer and play an important part in viral infectivity.Coronavirus – One of a group of related RNA viruses that cause respiratory tract infections in birds and mammals. These infections lead to diseases that can have mild effects, or be lethal. The Covid-19 pandemic was caused by a coronavirus, the SARS-CoV-2 virus. The 2002/4 SARS outbreak was caused by the SARS-CoV-1 virus.HIV – The Human Immunoseficiency Virus is two species of lentivirus that if left untreated cause Acquired Immunodeficiency Syndrome (AIDS) in humans.Receptor – A protein embedded in a cell membrane which binds to a specific molecule, or class of molecules.  Once the target molecule is bound, there is typically and effect within the cell to trigger some form of biological process.(viral) Pathogensis – The process by which a disease progresses. Viral pathogensis is specific to a disease caused by a virus.Computer node – Each computer in a connected cluster that are working together.GPUs – Graphics Processing Units are specific electronic circuits that rapidly address memory in order to output images to a display device. Their highly parallel structure makes them efficient at processing algorithms that process large data blocks in parallel.Glycoanalytics – Scientific study of glycosylated molecules, often biological in nature.Neuraminidase, or Sialidase – Are enzymes that cut the glycosidic bonds of neuraminic acids. This action helps viruses move through the respiratory tract mucus and infect host cells. The publication we refer to early on in the discussion is available at https://www.sciencedirect.com/science/article/pii/B9780128194751000560?via%3Dihub.  A full list of Elisa's publications is available at her group website. Elisa is contactable on social media, and you can find her on LinkedIn https://www.linkedin.com/in/elisa-fadda-a012b194/ (although, Elisa admits, she's rarely on LinkedIn)On Twitter, search @ElisaTelisaThe group website is https://efadda73.wixsite.com/elisafadda Our theme music is "Wholesome" by Kevin MacLeod (https://incompetech.com)Music from https://filmmusic.ioLicense: CC BY (http://creativecommons.org/licenses/by/4.0/) Connect with me (Paul) at https://www.linkedin.com/in/paulorange/H.E.L. group can be found at www.helgroup.com online,on LinkedIn at https://www.linkedin.com/company/hel-group/ on Twitter, we're @hel_group, https://twitter.com/hel_groupor search for us on Facebook

The Congenital Disorders of Glycosylation are a rapidly growing group of IMDs but can present a number of diagnostic challenges. In this podcast, Dr Julien Park, Dr Robert Mealer, and Professor Thorsten Marquardt discuss an additional technique for assessing glycosylation and its role in the diagnosis and management of SLCC39A-CDG. N‐glycome analysis detects dysglycosylation missed by conventional methods in SLC39A8 deficiency Julien H. Park et al. https://doi.org/10.1002/jimd.12306

Link to bioRxiv paper: http://biorxiv.org/cgi/content/short/2020.11.16.384594v1?rss=1 Authors: Brun, J., Vasiljevic, S., Gangadharan, B., Hensen, M., Chandran, A. V., Hill, M. L., Kiappes, J. L., Dwek, R. A., Alonzi, D. S., Struwe, W. B., Zitzmann, N. Abstract: Severe acute respiratory syndrome coronavirus 2 is the causative pathogen of the COVID-19 pandemic which as of Nov 15, 2020 has claimed 1,319,946 lives worldwide. Vaccine development focuses on the viral trimeric spike glycoprotein as the main target of the humoral immune response. Viral spikes carry glycans that facilitate immune evasion by shielding specific protein epitopes from antibody neutralisation. Immunogen integrity is therefore important for glycoprotein-based vaccine candidates. Here we show how site-specific glycosylation differs between virus-derived spikes and spike proteins derived from a viral vectored SARS-CoV-2 vaccine candidate. We show that their cellular secretion pathways are unique, resulting in different protein glycosylation and secretion, which may have implications for the resulting immune response and future vaccine design. Copy rights belong to original authors. Visit the link for more info

Link to bioRxiv paper: http://biorxiv.org/cgi/content/short/2020.10.01.322537v1?rss=1 Authors: Williams, S. W., Noel, M., Lehoux, S. D., Cetinbas, M., Xavier, R. J., Sadreyev, R., Scolnick, E. M., Smoller, J. W., Cummings, R. D., Mealer, R. G. Abstract: Glycosylation is essential to brain development and function, though prior studies have often been limited to a single analytical technique. Using several methodologies, we analyzed Asn-linked (N-glycans) and Ser/Thr/Tyr-linked (O-glycans) protein glycosylation between brain regions and sexes in mice. Brain N-glycans were surprisingly less complex in sequence and variety compared to other tissues, consisting predominantly of high-mannose precursors and fucosylated/bisected structures. Most brain O-glycans were unbranched, sialylated O-GalNAc and O-mannose structures. A consistent pattern was observed between regions, and sex differences were minimal compared to those observed in plasma. Brain glycans correlate with RNA expression of their synthetic enzymes, and analysis of all glycosylation genes in humans showed a global downregulation in the brain compared to other tissues. We hypothesize that the restricted repertoire of protein glycans arises from their tight regulation in the brain. These results provide a roadmap for future studies of glycosylation in neurodevelopment and disease. Copy rights belong to original authors. Visit the link for more info

Link to bioRxiv paper: http://biorxiv.org/cgi/content/short/2020.09.15.291732v1?rss=1 Authors: Sakson, R., Beedgen, L., Bernhard, P., Alp, K. M., Luebbehusen, N., Roeth, R., Niesler, B., Mayer, M. P., Thiel, C., Ruppert, T. Abstract: Protein glycosylation is essential in all domains of life and its impairment can result in severe human diseases named Congenital Disorders of Glycosylation (CDGs). Studies on molecular level are however challenging, because many glycosyltransferases in the endoplasmic reticulum (ER) are low abundance membrane proteins. We established a comprehensive multiple reaction monitoring (MRM) assay to quantify most human glycosyltransferases involved in the processes of N-glycosylation, O- and C-mannosylation in the ER. To increase reproducibility, a membrane protein fraction of isotopically labeled HEK 293T cells was used as an internal standard. This HEK 293T cells-derived standard could be used to reliably quantify 22 glycosyltransferases in HeLa cells and skin fibroblast cell lines. In addition, we showed that the MRM assay is easily transferable between laboratories. We then analyzed fibroblasts derived from CDG type I patients with defects in the ALG1, ALG2 or ALG11 gene. Mutations in ALG1 or ALG2 gene strongly reduced the levels of the ALG1 and ALG2 protein, respectively. In contrast, the levels of ALG proteins not directly affected by a genetic defect remained unchanged, which was unexpected given evidence that the ALG1, ALG2 and ALG11 proteins form a stable complex. This study describes an efficient workflow for the development of MRM assays for low abundance proteins, establishes a ready-to-use tool for the comprehensive quantification of ER-localized glycosyltransferases and provides new insight into the organization of disease-relevant glycosylation processes. Copy rights belong to original authors. Visit the link for more info

Link to bioRxiv paper: http://biorxiv.org/cgi/content/short/2020.09.12.294504v1?rss=1 Authors: Rahnama, S., Azimzadeh Irani, M., Amininasab, M., Ejtehadi, R. Abstract: SARS-COV-2 is a strain of Coronavirus family which caused the extensive pandemic of COVID-19, which is still going on. Several studies showed that the glycosylation of virus spike (S) protein and the Angiotensin-Converting Enzyme 2 (ACE2) receptor on the host cell is critical for the virus infectivity. Molecular Dynamics (MD) simulations were used to explore the role of a novel mutated O-glycosylation site (D494S) on the Receptor Binding Domain (RBD) of S protein. This site was suggested as a key mediator of virus-host interaction. We showed that the decoration of S494 with core and elongated O-glycans results in stabilized interactions on the direct RBD-ACE2 interface with more favorable binding free energies for longer oligosaccharides. Hence, the further drug design attempts should take this crucial factor into account, while suggesting any novel therapeutic candidate. Copy rights belong to original authors. Visit the link for more info

Link to bioRxiv paper: http://biorxiv.org/cgi/content/short/2020.09.09.289835v1?rss=1 Authors: Zolotarov, Y., Ma, C., Gonzalez-Recio, I., Hardy, S., Franken, G., Uetani, N., Latta, F., Kostantin, E., Boulais, J., Thibault, M.-P., Cote, J.-F., Diaz Moreno, I., Diaz Quintana, A., Hoenderop, J. G. J., Martinez-Cruz, L. A., Tremblay, M. L., de Baaij, J. H. F. Abstract: Cyclin M (CNNM1-4) proteins maintain cellular and body magnesium (Mg2+) homeostasis. Using various biochemical approaches, we have identified members of the CNNM family as direct interacting partners of ADP-ribosylation factor-like protein 15 (ARL15), a small GTP-binding protein. ARL15 interacts with CNNMs at their carboxyl-terminal conserved cystathionine-{beta}- synthase (CBS) domains. In silico modeling of the interaction using the reported structures of both CNNM2 and ARL15 supports that the small GTPase specifically binds the CBS1 domain. Immunocytochemical experiments demonstrate that CNNM2 and ARL15 co-localize in the kidney, with both proteins showing subcellular localization in the Golgi-apparatus. Most importantly, we found that ARL15 is required for forming complex N-glycosylation of CNNMs. Overexpression of ARL15 promotes complex N-glycosylation of CNNM3. Mg2+ uptake experiments with a stable isotope demonstrate that there is a significant increase of 25Mg2+ uptake upon knockdown of ARL15 in multiple kidney cancer cell lines. Altogether, our results establish ARL15 as a novel negative regulator of Mg2+ transport by promoting the complex N-glycosylation of CNNMs. Copy rights belong to original authors. Visit the link for more info

Professor Eva Morava of the Mayo Clinic takes us through the recently published consensus statement on the diagnosis and management of PGM1-CDG. Professor Morava provides a concise background to Congenital Disorders of Glycosylation and PGM1 disease specifically, and she explains how to recognise and diagnose this rare but treatable condition. International consensus guidelines for phosphoglucomutase 1 deficiency (PGM1‐CDG): diagnosis, follow‐up and management Altassan et al https://doi.org/10.1002/jimd.12286 Interested listeners may also want to read the consensus statement for PMM2-CDG published at the start of this year (https://doi.org/10.1002/jimd.12024).

Link to bioRxiv paper: http://biorxiv.org/cgi/content/short/2020.07.29.227785v1?rss=1 Authors: Zhang, Y., Zhao, W., Mao, Y., Chen, Y., Hu, L., Zhu, J., Gong, M., Cheng, J., Yang, H. Abstract: The densely glycosylated spike (S) proteins that are highly exposed on the surface of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) facilitate viral attachment, entry, and membrane fusion. We have previously reported all the 22 N-glycosites and site-specific N-glycans in the S protein protomer. Herein, we report the comprehensive and precise site-specific O-glycosylation landscapes of SARS-CoV-2 S proteins, which were characterized using high-resolution mass spectrometry. Following digestion using trypsin and trypsin/Glu-C, and de-N-glycosylation using PNGase F, we determined the mucin-type (GalNAc-type) O-glycosylation pattern of S proteins, including unambiguous O-glycosites and the 6 most common O-glycans occupying them, via Byonic identification and manual validation. Finally, 43 O-glycosites were identified in the insect cell-expressed S protein. Most glycosites were modified by non-sialylated O-glycans such as HexNAc(1) and HexNAc(1)Hex(1). In contrast, 30 O-glycosites were identified in the human cell-expressed S protein S1 subunit. Most glycosites were modified by sialylated O-glycans such as HexNAc(1)Hex(1)NeuAc(1) and HexNAc(1)Hex(1)NeuAc(2). Our results are the first to reveal that the SARS-CoV-2 S protein is a mucin-type glycoprotein; clustered O-glycans often occur in the N- and the C-termini of the S protein, and the O-glycosite and O-glycan compositions vary with the host cell type. These site-specific O-glycosylation landscapes of the SARS-CoV-2 S protein are expected to provide novel insights into the viral binding mechanism and present a strategy for the development of vaccines and targeted drugs. Copy rights belong to original authors. Visit the link for more info

Link to bioRxiv paper: http://biorxiv.org/cgi/content/short/2020.07.29.225110v1?rss=1 Authors: Sun, G., Putkaradze, N., Bohnacker, S., Joncyk, R., Fida, T., Hoffmann, T., Bernhardt, R., Härtl, K., Schwab, W. Abstract: C13-apocarotenoids (norisoprenoids) are carotenoid-derived oxidation products, which perform important physiological functions in plants. Although their biosynthetic pathways have been extensively studied, their metabolism including glycosylation remains elusive. Candidate uridine-diphosphate glycosyltransferase genes (UGTs) were selected for their high transcript abundance in comparison with other UGTs in vegetative tissues of Nicotiana benthamiana and Mentha x piperita, as these tissues are rich sources of apocarotenoid glucosides. Hydroxylated C13-apocarotenol substrates were produced by P450-catalyzed biotransformation and microbial/plant enzyme systems were established for the synthesis of glycosides. Natural substrates were identified by physiological aglycone libraries prepared from isolated plant glycosides. In total, we identified six UGTs that catalyze the unprecedented glucosylation of C13-apocarotenols, where glucose is bound either to the cyclohexene ring or butane side chain. MpUGT86C10 is a superior novel enzyme that catalyzes the glucosylation of allelopathic 3-hydroxy--damascone, 3-oxo--ionol, 3-oxo-7,8-dihydro--ionol (Blumenol C) and 3-hydroxy-7,8-dihydro-{beta}-ionol, while a germination test demonstrated the higher phytotoxic potential of a norisoprenoid glucoside in comparison to its aglycone. Glycosylation of C13-apocarotenoids has several functions in plants, including increased allelopathic activity of the aglycone, facilitating exudation by roots and allowing symbiosis with arbuscular mycorrhizal fungi. The results enable in-depth analyses of the roles of glycosylated norisoprenoid allelochemicals, the physiological functions of apocarotenoids during arbuscular mycorrhizal colonization and the associated maintenance of carotenoid homeostasis. Copy rights belong to original authors. Visit the link for more info

A group of more than 130 disorders share a common problem: a glycosylation issue in development where the necessary sugar chains are fully or partly missing from their needed location on protein surfaces. Patients with one of the congenital disorders of glycosylation (CDG) called SLC35A2 CDG are missing galactose, the sugar in milk, on their proteins. Without this sugar chain building block, SLC35A CDG patients often suffer from severe epilepsy, liver dysfunction and decreased coagulation among other symptoms. Tune in to this month’s GenePod to hear Dr. Eva Morava, a professor of medical genetics at the Mayo Clinic, discuss how moonlighting enzymes and metabolic adaptation from a galactose supplement benefited patients in a small pilot study published in Genetics in Medicine. See acast.com/privacy for privacy and opt-out information.

In den aktuellen Zeiten ist es so wichtig wie noch nie, ein intaktes Immunsystem zu haben. Du möchtest wissen, wie du dein Immunsystem stärken kannst, um dich vor dem Coronavirus und anderen Krankheitserregern zu schützen? Oder fehlen dir Antworten auf deine Fragen rund um COVID-19? Es lässt sich nicht leugnen: Der Wissensdurst rund um das Thema Immunsystem und Corona ist unersättlich. Wir haben uns Immunologe Dr. med. Kurt Mosetter vors Mikro geholt. Dr. Mosetter ist Humanmediziner, Autor und Betreuer zahlreicher Spitzensportvereine. Im ersten Teil geht es darum, wie gefährlich der Virus COVID-19 wirklich ist und warum es zahlreiche Infizierte ohne Symptome gibt. Anschließend geht es für dich in den Praxisteil: wie stärkst du dein Immunsystem? Welche Mittel machen Sinn, welche nicht? Warum ist trotz Quarantäne Sport extrem wichtig für dich? Mit dieser Episode wollen wir keinesfalls Panik verbreiten. Wir möchten mit dir durch diese fordernde Corona-Zeit gehen und dich bestmöglichst unterstützen. Also sei gespannt und hör rein! Du willst NOCH MEHR für dein Schutzschild tun? Hol dir das IMMUNE BUNDLE von BRAINEFFECT: https://www.brain-effect.com/immune-bundle Übersichtsarbeit zu Vitamin D Teymoori-Rad, M. / Shokri, F. / Salimi, V. / Marashi, S.M. (2019). The interplay between vitamin D and viral infections. Rev Med Virol 29(2). e2032. > https://www.ncbi.nlm.nih.gov/pubmed/30614127 Übersichtsarbeit zu den Myokinen Fiuza-Luces, C. / Garatachea, N. / Berger, N.A. / Lucia, A. (2013). Exercise is the real polypill. Physiology (Bethesda) 28(5). 330-58. > https://www.ncbi.nlm.nih.gov/pubmed/23997192 Übersichtarbeit Glycobiologie / Glycomedicine Reily, C. / Stewart, T.J. / Renfrow, M.B. / Novak, J. (2019). Glycosylation in health and disease. Nat Rev Nephrol 15(6). 346-366. > https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6590709/pdf/nihms-1035910.pdf in der Website / Praxis von Dr. med. Kurt Mosetter http://www.myoreflex.de/aktuelles/aktuelles-interview-von-kurt-mosetter http://www.myoreflex.de/ueber-uns/kurt-und-reiner-mosetter

A brief preview of the upcoming full episode, featuring upcoming topics (macrophages in joints, sugared RNA, lab models, and more on scoring misspellings)—plus word dissections, a book club recommendation (The Miniature Guide to Critical Thinking Concepts and Tools), and more! 00:19 | Topics 01:24 | Sponsored by HAPI Online Graduate Program 01:53 | Word Dissection 18:36 | Sponsored by HAPS 18:57 | Book Club 21:00 | Sponsored by AAA 21:16 | Staying Connected If you cannot see or activate the audio player click here. Questions & Feedback: 1-833-LION-DEN (1-833-546-6336) Follow The A&P Professor on Twitter, Facebook, Blogger, Nuzzel, Tumblr, or Instagram! Upcoming Topics 1 minute Update on new type of RNA called glycoRNA Update on barrier macrophages in joints Safely labeling anatomy models for lab practicals Continuing the conversation about grading misspellings Sponsored by HAPI Online Graduate Program 0.5 minute The Master of Science in Human Anatomy & Physiology Instruction—the MS-HAPI—is a graduate program for A&P teachers. A combination of science courses (enough to qualify you to teach at the college level) and courses in contemporary instructional practice, this program helps you power up  your teaching. Kevin Patton is a faculty member in this program. Check it out! nycc.edu/hapi Word Dissections 16.5 minutes formative, summative (lab) practical rheumatoid arthritis (RA) Y RNA glycan oligosaccharide, monosaccharide N-glycan, O-glycan glycosylation   Sponsored by HAPS 0.5 minute The Human Anatomy & Physiology Society (HAPS) is a sponsor of this podcast.  You can help appreciate their support by clicking the link below and checking out the many resources and benefits found there. There are a bunch of 1-day regional workshops scattered all over the continent. There's probably one near you coming up this year (or next)! Anatomy & Physiology Society theAPprofessor.org/haps Book Club 2 minutes The Miniature Guide to Critical Thinking by Richard Paul, Linda Elder amzn.to/2QjIGYN Special opportunity Contribute YOUR book recommendation for A&P teachers! First five submitted and used will be in a drawing for a Kindle Fire HD 10 tablet amzn.to/2WwLZvb Any contribution used will receive a $25 gift certificate The best contribution is one that you have recorded in your own voice (or in a voicemail at 1-833-LION-DEN) Check out The A&P Professor Book Club Sponsored by AAA 0.5 minutes   A searchable transcript for this episode, as well as the captioned audiogram of this episode, are sponsored by the American Association for Anatomy (AAA) at anatomy.org. Searchable transcript Captioned audiogram    If the hyperlinks here are not active, go to TAPPradio.org to find the episode page. More details at the episode page. Transcript available at the script page. Listen to any episode on your Alexa device. Need help accessing resources locked behind a paywall? Check out this advice from Episode 32 to get what you need! https://youtu.be/JU_l76JGwVw?t=440   Tools & Resources  Amazon TextExpander Rev.com Snagit & Camtasia The A&P Professor Logo Items   Sponsors   Transcript and captions for this episode are supported by the  American Association for Anatomy. anatomy.org   The Human Anatomy & Physiology Society  also provides marketing support for this podcast.  theAPprofessor.org/haps   Distribution of this episode is supported by  NYCC's online graduate program in  Human Anatomy & Physiology Instruction (HAPI)  nycc.edu/hapi   Clicking on sponsor links  helps let them know you appreciate their support of this podcast!   Follow The A&P Professor on  Twitter, Facebook, Blogger, Nuzzel, Tumblr, or Instagram!   The A&P Professor® and Lion Den® are registered trademarks of Lion Den Inc. (Kevin Patton)  

Obesity is on the rise worldwide and it’s a risk factor for systemic hypertension, hyperlipidemia, diabetes mellitus, and left ventricular hypertrophy, all of which are conditions associated with an increased prevalence of heart failure.  However, the applications of BNP and NT-proBNP as biomarkers in obese patients are limited, as the relationship between their levels and myocardial stiffness is complex.  An Editorial appearing in the September 2019 issue of Clinical Chemistry examines the interrelationship between obesity and BNP and NT-proBNP measurements.  

What happens when biotechnology and cannabis collide? Here to answer this is Ronan Levy of Trait Bio, a biotech company that just cracked the code on how to create flavorless, water-soluble cannabinoids - and without using nanoemulsion.  Learn more at https://www.traitbio.com  Key Takeaways: Ronan’s background in cannabis and how he came to start Trait Bio An inside look at Trait Bio and its mission to make hemp and cannabis into purer, safer products  The value of water-soluble cannabinoid products and where Trait Bio is in the process of achieving this The risks versus benefits of using nanoemulsion in the manufacturing of cannabis products  Glycosylation and how Trait Bio is using this instead of nanoemulsion to create water-soluble cannabinoid products How trait amplification works to increase the yield of cannabinoids in hemp and cannabis plants Trait Bio’s work with minor cannabinoids and why the industry is beginning to take more interest in them How yield differs between yeast and trait application Ronan’s long-term goals for Trait Bio and where the company currently is in the capital-raising process Where Ronan sees biotech and cannabis heading in the next five years

Dr Carolyn Lam:                Welcome to Circulation on the Run, your weekly podcast summary and backstage pass to the journal and its editors. We're your co-hosts, I'm Dr Carolyn Lam, Associate Editor from the National Heart Center and Duke National University of Singapore. Dr Greg Hundley:             And I'm Greg Hundley, Associate Editor at the Pauley Heart Center at VCU Health in Richmond, Virginia.                                                 Well Carolyn, did you ever wonder whether cardiovascular drug effects could be investigated through natural variation in the genes for the protein targets? In our feature discussion today, investigators from the British Isles, Germany, and the United States use this approach to explore the potential side effects and repurposing potential of antihypertensive drugs. Sound interesting? Well listeners, we look forward to the results later in our program, but Carolyn, how about we chat about some of the other papers in this issue? Dr Carolyn Lam:                You bet Greg. So, have you ever asked yourself "What is the role of protein glycosylation in regulating LDL metabolism?" Dr Greg Hundley:             That was going through my mind when we were playing basketball just the other night. Dr Carolyn Lam:                Well this is truly a great study from Dr Holleboom at Academic Medical Center Amsterdam and Dr Lefeber from Radboud University Medical Center, both in the Netherlands. And their colleagues will study 29 patients of the two most prevalent types of Type 1 Congenital Disorder of Glycosylation, and these are the ALG6 and PMM2 types. They also study 23 first and second-degree relatives with a heterozygote mutation and measured their plasma cholesterol levels. LDL metabolism was studied in three cell models. They found that patients with type 1 congenital disorder of glycosylation have hypobetalipoproteinemia through increased LDL receptor expression. Carriers of the mutation in glycosylation enzymes affected in this syndrome had decreased LDL cholesterol levels compared to controls, and defects in glycosylation enzymes could play, therefore, an important role in LDL cholesterol metabolism. Dr Greg Hundley:             Boy, this is pretty insightful I think, Carolyn. So, what are the clinical implications? Dr Carolyn Lam:                Well, given that LDL cholesterol was also reduced in a group of clinically unaffected heterozygotes, the authors propose that increasing LDL receptor mediated cholesterol clearance, by targeting N-glycosylation in the LDL pathway, may therefore represent a novel therapeutic strategy to reduce LDL cholesterol, and of course prevent cardiovascular disease. Dr Greg Hundley:             Very interesting work. You know, we just keep learning more and more about LDL. I'm going to switch and jump back with Empagliflozin. And this is a study in diabetic mice that really has an interesting in-vivo imaging component. As an imager, I was really excited about this. The article is from Dr Kengo Kidokoro from Kawasaki Medical School. And we don't often talk about it, but listeners, if you have a chance, there's a very interesting video-enhanced file associated with this article, and if you can download it, it's really just so cool with multiple image clips demonstrating an operative mechanism of SGLT2 inhibition on renal function. And it really gives us an opportunity to revisit renal function.                                                 Quick quiz Carolyn. In diabetic kidney disease, is glomerular hyperfiltration good or bad? Dr Carolyn Lam:                Bad. Dr Greg Hundley:             Yeah, absolutely. So, hyperfiltration is characteristically observed at earlier stages of diabetic kidney disease and involves activation of the renin-angiotensin-aldosterone system at the efferent arteriole and tubuloglomerular feedback mechanisms, especially at the afferent arteriole. So, as they go through this, just picture in your mind that glomerulus and afferent is arriving, and efferent is leaving.                                                 So, SGLT2 upregulation in diabetes is thought to play an important role in TGF signaling by increasing sodium reabsorption at the proximal tubule, thereby decreasing distal delivery to the sodium sensing macula densa at the juxtaglomerular apparatus. This decline in distal sodium delivery is interpreted as a decline in effective circulating volume, leading to inappropriate afferent vasodilation in an effort to preserve intra-glomerular pressure and GFR.                                                 In diabetes, these TGF effects lead to intra-glomerular hypertension and hyperfiltration. You got that quiz right, Carolyn. Which promotes diabetic kidney disease progression and impaired kidney function, ultimately increasing overall cardiovascular risk and mortality. Conversely, blocking SGTL2 pharmacologically reduces renal hyperperfusion and hyperfiltration in animals and humans, which may preserve renal function, thereby reducing risk associated with diabetic kidney disease progression. Dr Carolyn Lam:                You know what, Greg? I kind of had an unfair advantage in this quiz. I work with a lot with the SGLT2 inhibitors, but I just love that you asked us to picture it and look at that video. Anyways, so this article really allows us to review SGLT2 inhibition at the glomerular level, which is truly hot. So, tell us what did they find? Dr Greg Hundley:             So, this is the first report of changes in renal hemodynamic function by SGLT2 inhibition using direct in-vivo visualization techniques in a diabetic animal model. The videos, they're spectacular, and they're excellent so that you can download them for educational purposes. Afferent arteriolar vasoconstriction, and reduced hyperfiltration occurred within a few hours after a single dose of a SGLT2 inhibitor. And Adenosine signaling, through tubuloglomerular feedback, is a key pathway to prevent diabetic hyperfiltration via SGLT2 inhibition.                                                 Clinically, Carolyn, now I know you would ask me about that, so I got ready, this study highlights another potential mechanism for the benefits of SGTL2 inhibition. The SGLT2 inhibitor-related mechanism's responsible for reducing cardiovascular risk in clinical trials may be due to protection against diabetic kidney disease progression, thereby attenuating risk factors for heart failure, such as volume overload and hypertension. Dr Carolyn Lam:                Ah. That is just so cool, and really just so consistent with the clinical data that's emerging too. Thank you, Greg. So, have you ever asked yourself this other question, what role do platelets play in ischemia reperfusion injury? So, I'm not going to quiz you. I'm actually kind and loving and a good person. And so, I will tell you about ischemia reperfusion injury, which is a common complication of cardiovascular disease.                                                 Now, resolution of the detrimental effects of ischemia reperfusion injury generated prothrombotic and proinflammatory responses, is essential to restore homeostasis. Now, although platelets are known to play a crucial role in the integration of thrombosis and inflammation, their role as participants in the resolution of thrombo-inflammation is really under-appreciated. And hence, this other paper that I chose today, and it's from Dr Gavins from Louisiana State University Health Sciences Center Shreveport, and her colleagues, who used pharmacological and genetic approaches, coupled with murine and clinical samples to uncover key concepts underlying this role for platelets. Dr Greg Hundley:             So Carolyn, what did they find? Dr Carolyn Lam:                Well, they found that exacerbation of thrombo-inflammatory responses occurred in ischemia reperfusion injury mouse models of middle cerebral arterial occlusion, as well as lower plasma levels of the anti-inflammatory pro-resolving protein Annexin A1. And this was a lower plasma level of this Annexin A1 among patients with acute ischemic stroke.                                                 Administration of Annexin A1 promoted cerebral protection against thrombo-inflammation and the development of subsequent thrombotic events post-stroke. Annexin A1 was also able to reduce platelet activation and thrombosis, via the suppression of integrins. So, overall, these data reveal a novel multi-faceted role for Annexin A1 to act both as therapeutic and prophylactic drug via its ability to promote endogenous pro-resolving anti-thrombo, anti-inflammatory circuits in the cerebral ischemia reperfusion injury. And collectively, these results further enhance our understanding in the field of platelet and ischemia reperfusion injury biology. Dr Greg Hundley:             Oh wow. So, another important insight from this author group on platelet activation and thrombosis in key clinically relevant syndromes. Well, my last paper is going to be talking about a risk prediction score for life-threatening ventricular tachyarrhythmias. And they're going to study this in laminopathies, and the lead investigator is Dr Karim Wahabi from Cochin Hospital in France.                                                 To estimate the risk of life-threatening ventricular tachyarrhythmia in patients with LMNA mutations, and thus select candidates for implantable cardiac defibrillators, the investigators evaluated 444 patients of about 40 years in age in a derivation sample. And then, 145 patients that are about the same age, 38 years, in a validation sample, for the occurrence of a) sudden cardiac death or b) ICD-treated or hemodynamically unstable ventricular tachyarrhythmias. Dr Carolyn Lam:                Oh. Very important. These laminopathies are really not that uncommon. So what did they find, Greg? Dr Greg Hundley:             Carolyn, predictors of events included male sex, non-missense LMNA mutations, first-degree and higher AV block, non-sustained ventricular tachycardia, and LVEF. The authors developed a new score to estimate the 5-year risk of life-threatening ventricular tachyarrhythmias in patients with LMNA mutations. And compared to the current standard of care, the proposed risk prediction model offered more accurate prediction of life-threatening ventricular tachyarrhythmias, and correctly re-classified almost 30% of the patients in the study.                                                 Nicely, the authors have made this available, and the score can be derived from readily collected clinical and genetic parameters and estimated using an online calculator that's provided in the journal. But, it's https://lmna-risk-vta.fr.                                                 Future prospective studies should focus on the estimation of the clinical benefit conferred by the use of this score in terms of sudden cardiac death prevention. Dr Carolyn Lam:                That is super cool, Greg. But, I am so excited now to move to our feature discussion. Shall we? Dr Greg Hundley:             You bet. Dr Carolyn Lam:                Can we use natural variations in our genes for the protein targets as a way to look at cardiovascular drug effects? Man, this is going to be such an important and exciting discussion, because this is what our feature paper talks about. I am so pleased to have with us our corresponding author, Dr Dipender Gill from Imperial College London, as well as our Associate Editor, Dr Wendy Post from Johns Hopkins.                                                 So, first of all, Dipender, please, could you give us a background on what you did? This is really very novel in approach. Dr Dipender Gill:               It was also a lot of fun to conduct. I think, currently, we're living in an era where there's been a recent explosion in the availability of genetic data, and this really inspired us to think about how we could use that to learn more about commonly prescribed drugs. The implementation of genetics, or genetic variance, to study drug effects isn't entirely novel. It's actually been undertaken for some years now.                                                 Most of the work has been related to lipid lowering drugs, for example, statins, where people can take genetic variance, or versions of genes, corresponding to the drug effect, and study these to investigate what effects these drugs might have, both on the intended target, but also potential side effects. To my knowledge, this hadn't previously been done for anti-hypertensive drugs. But yet, the data for this was available. And therefore, we thought that actually we could very well go ahead and do this, and perhaps find some interesting things. Dr Carolyn Lam:                Oh, that's so interesting thing, Dipender. You know, there was this term in your abstract, and mentioned multiple times, Mendelian randomization. Now, for those of us that don't think about this every day, could you tell us a little bit what that means? Dr Dipender Gill:               Yeah. So, I'll actually give a little bit of background. One of the main limitations of traditional epidemiological research is that any association, it's sometimes difficult to infer causation. They can be confounded by environmental factors, lifestyle factors. In the Mendelian randomization technique, what we do is we use randomly allocated genetic variants to study the effect to an exposure.                                                 So, we select these genetic variants because they are related to the exposure of interest. And because these genes are randomly allocated at conception, they're not subject to confounding from environmental or lifestyle factors. Whether you have a gene or not, is not necessarily related to your lifestyle or your environment. And therefore, the association of these genetic variants with certain outcomes isn't subject to confounding. Dr Carolyn Lam:                That makes so much sense, and I suppose that, not to allow cause and effect to be determined. So please, tell us, in this particular case of the anti-hypertensive drugs, what did you do and what did you find? Dr Dipender Gill:               First, we decided specifically which drugs we wanted to look at, and we thought, actually, let's start off with the most commonly prescribed anti-hypertensive drugs. So, we short-listed these based on recent consensus guidelines, and we looked at ACE inhibitors, beta-blockers, calcium channel blockers, thiazide type diuretics. And then, we went back to various online databases to identify which genes correspond to the target protein of these drugs.                                                 We took these genes, and we then identified genetic variants at their specific genetic loci, their specific region of the genome, and we identified the variants in these regions that were also related to systolic blood pressure. And in this way, we inferred that genetic variants, at the protein coding targets of these genes, that were also related to systolic blood pressure, likely represented the effect of variations in these proteins that also implicated blood pressure, and therefore, could serve as proxies, or instruments, to study the effect of these drug targets.                                                 We then went ahead to validate the selection of these genetic variants by forming Mendelian randomization, and specifically, we checked whether people that have genetic variants that correspond to, say, ACE inhibitor activity, or beta-blocker activity, or calcium channel blocker activity, if they also have correspondingly lower risk of coronary heart disease and stroke, to the same degree that we would observe in randomized control trials against placebo.                                                 And indeed, we found that actually, the results were fairly similar, and this gave us confidence. And studying these genetic variants that mimic the effect of these drugs could be used as a proxy or as a surrogate to study their clinical effect of taking these drugs. So, that was the first phase. Dr Wendy Post:                 Dipender, congratulations to you and our team. This is a really exciting paper, and the editors were especially interested in the novelty, and the potentially impactful findings, especially of the second part of the study, which I think you'll describe briefly next. And that was using an approach that many who are listening may not have heard about too much before called PheWAS, or a phenome wide association study. And maybe you could tell us briefly what you found in that part of the analysis. Dr Dipender Gill:               The first part, it was very cool, because it allowed us to identify versions of genes that corresponded to the effect of these drugs. But in itself, it didn't tell us anything novel. It didn't tell us anything new. So, the real question was, how could we use this new information to make progress towards helping patients? So, we went back and we thought, "So okay." So, we knew that these drugs are used for certain conditions already to prevent heart disease, to prevent stroke.                                                 But, what about their side effects? What about their repurposing potential? How could we use our new approach to study that a little bit more carefully? As you alluded to, when we used this new technique, relatively new technique called phenome wide association study, and we essentially investigated the association of our genetic variants for each respective anti-hypertensive target with all clinically relevant outcomes throughout the phenome, using the UK bio-back cohort, which was the main population used for this PheWAS, this phenome wide association study.                                                 We were actually able to rapidly investigate over 900 disease outcomes, and their association with our genetic risk score for these drugs. And this was very exciting for us, because it allowed us to very rapidly, efficiently, and cost-effectively explore the potential repurposing opportunity and side effects of these very commonly prescribed drugs, which to our mind, offered significant advantage over previous approaches.                                                 We all know that sometimes randomized control trials can be very expensive and time-consuming, and of course, traditional observational research can be limited by reverse causation, assessment-vise confounding. And so, what we were able to do here had several important advantages, and not to mention the efficiency by which it allowed study of these outcomes. Dr Wendy Post:                 Dipender, tell us what you found in your PheWAS study. Dr Dipender Gill:               We identified genetic variants for 3 commonly prescribed anti-hypertensive targets. The first were ACE inhibitors, second, beta blockers, and the third were calcium channel blockers. When performing PheWAS for all of these drug targets, we identified associations with common cardiovascular disease that are related, or implicated in hypertension, specifically hypertension itself, but also circulatory diseases, things like atrial fibrillation, coronary heart disease. They all came up.                                                 And this actually gave us a lot of confidence because that's exactly what we'd expect. We know that these medications prevent or reduce risk of these diseases, and therefore, this served as kind of a positive control that our approach was doing it what it was supposed to do. The novel finding came when we investigated the genetic risk score, or the genetic variants for calcium channel blockers, in this PheWAS approach.                                                 And we actually identified an association which we weren't expecting. We showed that blood pressure reduction through the genetic risk score for calcium channel blockers was an association with an increased risk of diverticulosis, a condition not conventionally thought to be associated with blood pressure. We were very excited and interested by this, and we went on to investigate it further using some other techniques as well. Dr Wendy Post:                 The really impactful part of this, many things, but especially this association with diverticulosis. So, maybe you can briefly summarize what you think the potential clinical implications are, and what the next step should be. Dr Dipender Gill:               The first question we had was whether this was related to blood pressure alone, the effect of calcium channel blockers, or perhaps some other effect of these drugs. We investigated the genetic risk score for systolic blood pressure generally and found that this itself wasn't associated with risk of diverticulosis, which suggested that the effect isn't really mediated by blood pressure alone, but it's some other property of calcium channel blockers.                                                 We know that sometimes calcium channel blockers can be associated with constipation, and it may be through this mechanism that they're having consequent effects on risk of diverticulosis. Other possible mechanisms might be through effects on blood flow, through the vasa recta in the bowel. But, what was very interesting was that we went forward with this finding, and investigated, observed, drug use in the UK bio-bank.                                                 Specifically, we looked at people taking non-dihydropyridine, and dihydropyridine calcium channel blockers at baseline, and found that those taking non-dihydropyridine calcium blockers only were known to have a higher risk of diverticulosis as compared to those taking other anti-hypertensive classes, which further added support for our findings. The interesting point here is that looking at the genetics doesn't allow us to discriminate between these drug classes.                                                 That was only possible with the observed data, and that was because the genes for these drug classes were the same. Dr Carolyn Lam:                Well, congratulations. Wow. I'm just so intrigued listening to all of this. Wendy, I would love if you could help put all of this in context for us. The US, the novel information, and the approach that could potentially go way beyond just anti-hypertensive. Dr Wendy Post:                 So, this is a very exciting new approach to doing genetic studies that can help us to understand potential targets for therapy in the future, and understanding more about causality, which as Dipender explained, can sometimes be confusing, as it may be confounded by environmental factors. So, using these genetic approaches through Mendelian randomization, and what we heard about today, which is PheWAS, or phenome wide association study, we can learn much more about how the potential observational analyses can be related to new discoveries through mechanisms, or potential side effects, as we heard about here of calcium channel blockers.                                                 So, wanted to congratulate Dipender again with his impactful paper here. Dr Carolyn Lam:                Thanks, Wendy. And then if I could, I'm just going to steal minutes here, because this is so interesting. Where do you think the field's going to go next? And Dipender, with these findings of diverticulitis and diverticulosis, what next? How do we apply this? Dr Dipender Gill:               There's 2 main points to cover here. The first is what we do specifically with the findings we got for calcium channel blockers and diverticulosis. I should emphasize that on their own, I don't think that this should currently change practice. But, I think it should inspire and capitalize further research into this association. If we're able to replicate and validate it further, then perhaps there might be some implications for the drugs that we prescribe with patients at risk of diverticulosis.                                                 The second point I wanted to make is more generally, what does this mean for research, and particularly, genetic research. I think we're living in very exciting times, and there's a lot of really great work that's going to come out using these types of approaches. I think 2 areas that we could expand further is what else we can do with our genetic instruments, or our genetic variants that proxied these drugs. How do we look at other targeted refocusing potential? Can we try and explore other side effects? Can we investigate efficacy for other disease outcomes? Specifically, for these anti-hypertensives.                                                 And the other thing is, which other drugs can we identify genetic variants to proxy? We've been thinking about looking at diabetes medicines. There's a variety of other drugs that correspond to specific gene targets, and proteins. And in theory, these could also be studied using genetics. So, there's a lot more work to come out from this. Dr Carolyn Lam:                Thanks so much, both of you, for joining us today. This was just such an exciting discussion.                                                 Thank you for listening to Circulation on the Run. Don't forget to tune in again next week. This program is copyright American Heart Association, 2019.  

Inborn errors of metabolism are rare diseases in which a single gene defect causes a clinically significant block in a metabolic pathway resulting either in an accumulation of substrate behind the block or a lack or deficiency of the product.  Profiling metabolites in the pathway could allow for accurate and timely identification of patients who have these diseases and help physicians to devise effective treatment.   Congenital disorders of glycosylation represent one of the largest groups of such metabolic disorders.  In May 2019, Clinical Chemistry published a study on the development and validation of a plasma protein N-glycan assay using a flow injection-electrospray ionization-quadrupole time-of-flight mass spectrometry.

Vincent and Elio discuss the reason for poor efficacy of one of the influenza virus vaccines, and using a hyperthermophilic anaerobe to produce hydrogen from fruit and vegetable wastes in seawater. Hosts:  Vincent Racaniello and Elio Schaechter. Subscribe to TWiM (free) on iPhone, Android, RSS, or by email. You can also listen on your mobile device with the ASM Podcast app. Become a patron of TWiM. Links for this episode Glycosylation site on influenza H3N2 viruses (PNAS) Biohydrogen production by Thermotoga (Waste Man) Image credit Letters read on TWiM 166 Send your microbiology questions and comments (email or recorded audio) to twim@microbe.tv

Intramembrane proteolysis - hydrolysis of membrane proteins within or close to their membrane-spanning regions - is a crucial cellular process that is conserved throughout all kingdoms of life. It is executed by distinct classes of polytopic membrane proteins, the intramembrane-cleaving proteases, that provide a hydrophilic, proteinaceous environment accommodating membrane protein substrates as well as water molecules within the hydrophobic membrane interior and catalyse peptide bond hydrolysis. In particular, intramembrane-cleaving aspartyl proteases have received attention as the presenilins, the catalytic subunits of the γ-secretase complex, were identified as key players in Alzheimer's disease pathophysiology. In addition to presenilins, mammalian genomes harbour presenilin homologues which include signal peptide peptidase (SPP) and SPP-like (SPPL) proteases. Among these, the Golgi-resident protease SPPL3 stands out as it is highly conserved among metazoa and SPPL3 orthologues are also found in plants. However, due to the lack of known substrates, SPPL3 has thus far hardly been characterised. Hence, the purpose of this study was to identify its substrates and elucidate its physiological function(s). In the first part of this study, the foamy virus envelope glycoprotein (FVenv) was identified as the first substrate of SPPL3. This allowed to study SPPL3's proteolytic activity in detail, with a focus on its substrate selectivity and sensitivity towards previously characterised inhibitors of intramembrane-cleaving aspartyl proteases. Importantly, this study revealed in addition that two other intramembrane-cleaving proteases, SPPL2a and SPPL2b, also endoproteolyse FVenv. SPPL2b in particular had been studied in detail before and therefore SPPL3- and SPPL2b- mediated endoproteolysis of FVenv were examined in parallel to directly compare these phylogenetically related intramembrane-cleaving proteases. This uncovered an unexpected idiosyncrasy of SPPL3 that clearly sets SPPL3 apart from other intramembrane-cleaving aspartyl proteases: SPPL3 endoproteolysed full-length FVenv and did not require the substrate's prior tailoring by another proteolytic activity - an otherwise common phenomenon among intramembrane-cleaving aspartyl proteases. In the second part, the physiological function of SPPL3 was investigated. Alterations in the cellular levels of proteolytically active SPPL3 turned out to impact the composition of N-glycans attached to endogenous cellular glycoproteins. SPPL3 over-expression was accompanied by a decrease in glycoprotein molecular weight, i.e. a hypoglycosylation phenotype, while loss of SPPL3 expression in cell culture models but also in vivo resulted in a hyperglycosylation phenotype. This led to the identification of Golgi glycan-modifying enzymes such as GnT-V and β3GnT1 as novel physiological substrates of SPPL3. Loss or reduction of SPPL3 expression, for instance, led to a marked intracellular accumulation of these enzymes, explaining the more extensive N-glycan elaboration and the hyperglycosylation phenotype observed under these conditions. At the same time secretion of these enzymes was reduced under these conditions. Together with additional observations such as the mapping of the SPPL3 cleavage site to the membrane-spanning region of GnT-V, this study demonstrates that SPPL3-mediated intramembrane proteolysis of such glycan-modifying enzymes liberates their active site-harbouring ectodomains. Acting in this manner, SPPL3 controls the intracellular pool of active glycan-modifying enzymes. Importantly, the finding that SPPL3 proteolytically cleaves full-length glycan-modifying enzymes and sheds their ectodomains is well in line with the observations made for FVenv and suggested that SPPL3 acts functionally equivalent to classical sheddases or rhomboid proteases but much unlike all other characterised mammalian intramembrane-cleaving aspartyl proteases. To examine whether these observations hold also true on a global cellular scale, a proteomic approach was undertaken in the third part of the study to define the SPPL3 degradome of HEK293 cells in conditions of SPPL3 over-expression. On the one hand, this led to the identification of numerous novel, mostly Golgi-resident candidate SPPL3 substrates and, considering the physiological implications, suggests that SPPL3 is very intricately linked to Golgi function. On the other hand, this approach supports the initial hypothesis that SPPL3 acts as a cellular type II membrane protein-selective sheddase. Taken together, this study provides the first in-depth characterisation of the intramembrane protease SPPL3 and reveals the cellular function of SPPL3. SPPL3 displays considerable and marked differences to other intramembrane-cleaving aspartyl proteases and emerges as a fundamental cellular sheddase that exhibits strong selectivity for type II-oriented, Golgi-resident membrane proteins. Products of SPPL3-mediated endoproteolysis of these Golgi factors are secreted and/or may be subject to intracellular degradation which compromises their catalytic activity. Thus, SPPL3 indirectly controls protein glycosylation in the Golgi apparatus.

Catherine Wasser and Joachim Herz have found that normal synaptic function and fear learning require glycosylation of the apolipoprotein E receptor Apoer2.

Background: HIV-1 entry into host cells is mediated by interactions between the virus envelope glycoprotein (gp120/gp41) and host-cell receptors. N-glycans represent approximately 50% of the molecular mass of gp120 and serve as potential antigenic determinants and/or as a shield against immune recognition. We previously reported that N-glycosylation of recombinant gp120 varied, depending on the producer cells, and the glycosylation variability affected gp120 recognition by serum antibodies from persons infected with HIV-1 subtype B. However, the impact of gp120 differential glycosylation on recognition by broadly neutralizing monoclonal antibodies or by polyclonal antibodies of individuals infected with other HIV-1 subtypes is unknown. Methods: Recombinant multimerizing gp120 antigens were expressed in different cells, HEK 293T, T-cell, rhabdomyosarcoma, hepatocellular carcinoma, and Chinese hamster ovary cell lines. Binding of broadly neutralizing monoclonal antibodies and polyclonal antibodies from sera of subtype A/C HIV-1-infected subjects with individual gp120 glycoforms was assessed by ELISA. In addition, immunodetection was performed using Western and dot blot assays. Recombinant gp120 glycoforms were tested for inhibition of infection of reporter cells by SF162 and YU.2 Env-pseudotyped R5 viruses. Results: We demonstrated, using ELISA, that gp120 glycans sterically adjacent to the V3 loop only moderately contribute to differential recognition of a short apex motif GPGRA and GPGR by monoclonal antibodies F425 B4e8 and 447-52D, respectively. The binding of antibodies recognizing longer peptide motifs overlapping with GPGR epitope (268 D4, 257 D4, 19b) was significantly altered. Recognition of gp120 glycoforms by monoclonal antibodies specific for other than V3-loop epitopes was significantly affected by cell types used for gp120 expression. These epitopes included CD4-binding site (VRC03, VRC01, b12), discontinuous epitope involving V1/V2 loop with the associated glycans (PG9, PG16), and an epitope including V3-base-, N332 oligomannose-, and surrounding glycans-containing epitope (PGT 121). Moreover, the different gp120 glycoforms variably inhibited HIV-1 infection of reporter cells. Conclusion: Our data support the hypothesis that the glycosylation machinery of different cells shapes gp120 glycosylation and, consequently, impacts envelope recognition by specific antibodies as well as the interaction of HIV-1 gp120 with cellular receptors. These findings underscore the importance of selection of appropriately glycosylated HIV-1 envelope as a vaccine antigen.

Myelin Oligodendrocyte Glycoprotein (MOG) is one of the few proteins known to be localized on the outermost sheath of central nervous system (CNS) myelin. Due to this localization, MOG is accessible to antibodies. Anti-MOG antibodies are demyelinating and enhance clinical symptoms in a number of animal models of CNS inflammation. Autoantibodies recognizing conformationally intact MOG are found in different inflammatory diseases of the CNS, but their antigenic epitopes had not been mapped. In this work, 9 variants of MOG with an intracellular enhanced green fluorescent protein (EGFP) tag were expressed on the cell surface of human HeLa cells and used to analyze sera from 111 patients (104 children, 7 adults), who had antibodies recognizing cell-bound human MOG. These patients had different diseases, namely acute disseminated encephalomyelitis (ADEM), one episode of transverse myelitis or optic neuritis, multiple sclerosis (MS), anti-aquaporin-4 (AQP4)–negative neuromyelitis optica (NMO), and chronic relapsing inflammatory optic neuritis (CRION). The expression levels of the mutants were comparable and cells with a defined expression level (fluorescence intensity in the EGFP channel of 102-103) were gated. Each MOG-mutant was recognized by at least one MOG-specific mAb. This allowed the comparison of binding to the different mutants. In order to assess the reproducibility of the system, binding of the 111 sera to the mutants was analyzed up to three times in independent experiments, yielding a very good reproducibility of the binding percentage with an absolute SD of 7.8% in the case of low recognition of a mutant and a relative SD of 20% in the case of high recognition of a mutant. The applied variants of MOG gave insight into epitope recognition of 98 patients. All epitopes identified in this work were located at loops connecting the ß-strands of MOG. The immunodominant epitope of human anti-MOG antibodies was at the membrane-proximal CC’-loop containing aa42, which is not present in rodent MOG. This loop was recognized by about half of all patients. Overall, seven epitope patterns were distinguished, including the one mainly recognized by mouse mAbs at the FG-loop around aa104. Evidence from mouse models of CNS inflammation shows that anti-MOG antibodies recognizing different epitopes can be demyelinating and thus pathogenic. This suggests that not only those antibodies recognizing the same epitope of MOG as the pathogenic mAbs (i.e. the FG-loop), but also the ones recognizing the CC'-loop are pathogenic in humans, as both epitopes allow for the recognition of cell-bound MOG. In half of the patients, the anti-MOG response was directed to a single epitope. To analyze the effect of glycosylation on the recognition of MOG by human autoantibodies, a “non-glycosylation mutant” N31D was made. Digestion with PNGaseF and Western blot analysis confirmed that N31 was the only used N-glycosylation site of the MOG constructs in HeLa cells. Glycosylation of MOG was not needed for antibody binding, but 8% of the patients recognized deglycosylated MOG at least two-fold better. The epitope specificity was not linked to certain disease entities. The individual epitope recognition patterns stayed constant in 11 analyzed patients over an observation period of up to 5 years without evidence for intramolecular epitope spreading. Some patients with acute syndromes had anti-MOG IgG at disease onset, but rapidly lost their anti-MOG IgG reactivity. These patients were able to generate a long-lasting IgG response to measles and rubella virus vaccine indicating that the loss of anti-MOG reactivity was not reflective of a lack of capacity for longstanding IgG responses. Human anti-MOG antibodies are mainly of the IgG1 isotype, which can activate complement and antibody dependent cellular cytotoxicity. Upon binding to MOG in the CNS, human anti-MOG antibodies are hence expected to cause demyelination. Transfer experiments with purified human anti-MOG antibodies have not been performed yet. The fact that the majority of human anti-MOG antibodies did not recognize rodent MOG has implications for animal studies. Using the described assay will help to identify patient samples appropriate for these transfer experiments and finally lead to the formal proof of the pathogenicity of human anti-MOG antibodies. This work also gives important information for future detection of potential mimotopes and the development of anti-MOG antibody detection assays and might pave the way to antigen-specific depletion.

C. albicans is one of the most common fungal pathogen of humans, causing local and superficial mucosal infections in immunocompromised individuals. Given that the key structure mediating host-C. albicans interactions is the fungal cell wall, we aimed to identify features of the cell wall inducing epithelial responses and be associated with fungal pathogenesis. We demonstrate here the importance of cell wall protein glycosylation in epithelial immune activation with a predominant role for the highly branched N-glycosylation residues. Moreover, these glycan moieties induce growth arrest and apoptosis of epithelial cells. Using an in vitro model of oral candidosis we demonstrate, that apoptosis induction by C. albicans wild-type occurs in early stage of infection and strongly depends on intact cell wall protein glycosylation. These novel findings demonstrate that glycosylation of the C. albicans cell wall proteins appears essential for modulation of epithelial immunity and apoptosis induction, both of which may promote fungal pathogenesis in vivo.

Fri, 10 Dec 2010 12:00:00 +0100 https://edoc.ub.uni-muenchen.de/13146/ https://edoc.ub.uni-muenchen.de/13146/1/Yin_Ruohe.pdf Yin, Ruohe ddc:540, ddc:500, Fakultät für

The mannosyltransferase Och1 is the key enzyme for synthesis of elaborated protein N-glycans in yeast. In filamentous fungi genes implicated in outer chain formation are present, but their function is unclear. In this study we have analyzed the Och1 protein of Aspergillus fumigatus. We provide first evidence that poly-mannosylated N-glycans exist in A. fumigatus and that their synthesis requires AfOch1 activity. This implies that AfOch1 plays a similar role as S. cerevisiae ScOch1 in the initiation of an N-glycan outer chain. A Δafoch1 mutant showed normal growth under standard and various stress conditions including elevated temperature, cell wall and oxidative stress. However, sporulation of this mutant was dramatically reduced in the presence of high calcium concentrations, suggesting that certain proteins engaged in sporulation require N-glycan outer chains to be fully functional. A characteristic feature of AfOch1 and Och1 homologues from other filamentous fungi is a signal peptide that clearly distinguishes them from their yeast counterparts. However, this difference does not appear to have consequences for its localization in the Golgi. Replacing the signal peptide of AfOch1 by a membrane anchor had no impact on its ability to complement the sporulation defect of the Δafoch1 strain. The mutant triggered a normal cytokine response in infected murine macrophages, arguing against a role of outer chains as relevant Aspergillus pathogen associated molecular patterns. Infection experiments provided no evidence for attenuation in virulence; in fact, according to our data the Δafoch1 mutant may even be slightly more virulent than the control strains.

Lecture 10: In this continued discussion on protein translocation, Kaplan gives emphasis to glycosylation and the Scavenger pathway.

Biomarkers for detection of early stages of Alzheimer's disease, News from the Editorial Office, Differentially expressed serum proteins following skin irradiation in mice, Announcing: PROTEOMICS - Clinical Applications Neuroproteomics Special Issue and Practical Proteomics latest issue, Glycosylation profiling of IgG subclasses from human serum.

Biomarkers for detection of early stages of Alzheimer's disease, News from the Editorial Office, Differentially expressed serum proteins following skin irradiation in mice, Announcing: PROTEOMICS - Clinical Applications Neuroproteomics Special Issue and Practical Proteomics latest issue, Glycosylation profiling of IgG subclasses from human serum.

The squamous stratified epithelia contain a proliferative (harboring mitotic activity) and a differentiating compartment. Due to the potential of protein-carbohyd rate interactions to regulate cellular activities we introduced a mammalian lectin to cyto- and histochemical analysis. We answer the questions of whether and to what extent this new probe can pinpoint differentiation-dependent glycosylation changes in sections and in culture of keratinocytes. Material and Methods: Purification and labeling enabled monitoring of galectin-3 reactivity in frozen sections of human and pig epidermis and basal cell carcinomas as well as in culture of keratinocytes. The staining pattern of the lectin was correlated with the staining profile of other cell markers including desmosomal proteins, beta(1) integrin, and the proliferation marker Ki-67. The Dolichos biflorus agglutinin (DBA) sharing binding reactivity of galectin-3 to the A type histoblood group epitope was used for comparison. Results: Both lectins exhibit suprabasal binding. However, their profiles were not identical, substantiated by lack of coinhibition. Strong DBA reactivity was also observed in a limited number of basal layer cells, namely in cells without the expression of the proliferation marker Ki-67. Cultured mitotic epidermal cells have no reactivity for DBA. Presence of ligands for this plant lectin was connected with decreased positivity of nuclei for Ki-67 and the occurrence of ring-shaped nucleoli, micronucleoli or absence of nucleoli. Considering colocalization the pattern of galectin-3-binding sites coincided with the presence of desmosomal proteins such as desmoplakin-1 and desmoglein but not beta(1) integrin, a potential ligand. Interestingly, studied basal cell carcinomas expressed no binding sites for galectin-3, while a limited number of cells were DBA-reactive. Conclusion: The expression of galectin-3-binding sites and also DBA-reactive glycoligands correlates with an increased level of differentiation and/or cessation of proliferation in the examined squamous stratified epithelia. Further application of tissue lectins for characterizing ligand expression and its modulation is an important step to reveal functional relevance.