Podcasts about Glycoprotein

Go online to PeerView.com/CTH860 to view the activity, download slides and practice aids, and complete the post-test to earn credit. Myelin oligodendrocyte glycoprotein antibody-associated disorder (MOGAD) was identified as a distinct demyelinating disease entity only recently. Prior to this characterization of MOGAD, many patients were considered to have a variant of MS, NMOSD, or other neurologic condition. Now an international panel of experts has proposed diagnostic criteria that, when validated, promise to improve MOGAD diagnostic accuracy and confidence. This is particularly important, given that several clinical trials are investigating potential treatments for MOGAD. This unique PeerView activity will use problem-based educational interventions to expose learners to the science and clinical experience behind accurate diagnosis and best practices in management of MOGAD. You will be able to employ optimal clinical decision-making for your patients with MOGAD that is based on the latest evidence, best practice recommendations, and effective interdisciplinary collaboration. Upon completion of this activity, participants should be better able to: Explain the diagnostic criteria for MOGAD as proposed by the International MOGAD Panel; Employ recommended diagnostic tools (eg, the MOG antibody test, relevant MRI findings) to identify MOGAD and distinguish it from other demyelinating diseases; and Implement therapeutic strategies to provide acute treatment and relapse prevention in patients with MOGAD.

Go online to PeerView.com/CTH860 to view the activity, download slides and practice aids, and complete the post-test to earn credit. Myelin oligodendrocyte glycoprotein antibody-associated disorder (MOGAD) was identified as a distinct demyelinating disease entity only recently. Prior to this characterization of MOGAD, many patients were considered to have a variant of MS, NMOSD, or other neurologic condition. Now an international panel of experts has proposed diagnostic criteria that, when validated, promise to improve MOGAD diagnostic accuracy and confidence. This is particularly important, given that several clinical trials are investigating potential treatments for MOGAD. This unique PeerView activity will use problem-based educational interventions to expose learners to the science and clinical experience behind accurate diagnosis and best practices in management of MOGAD. You will be able to employ optimal clinical decision-making for your patients with MOGAD that is based on the latest evidence, best practice recommendations, and effective interdisciplinary collaboration. Upon completion of this activity, participants should be better able to: Explain the diagnostic criteria for MOGAD as proposed by the International MOGAD Panel; Employ recommended diagnostic tools (eg, the MOG antibody test, relevant MRI findings) to identify MOGAD and distinguish it from other demyelinating diseases; and Implement therapeutic strategies to provide acute treatment and relapse prevention in patients with MOGAD.

Go online to PeerView.com/CTH860 to view the activity, download slides and practice aids, and complete the post-test to earn credit. Myelin oligodendrocyte glycoprotein antibody-associated disorder (MOGAD) was identified as a distinct demyelinating disease entity only recently. Prior to this characterization of MOGAD, many patients were considered to have a variant of MS, NMOSD, or other neurologic condition. Now an international panel of experts has proposed diagnostic criteria that, when validated, promise to improve MOGAD diagnostic accuracy and confidence. This is particularly important, given that several clinical trials are investigating potential treatments for MOGAD. This unique PeerView activity will use problem-based educational interventions to expose learners to the science and clinical experience behind accurate diagnosis and best practices in management of MOGAD. You will be able to employ optimal clinical decision-making for your patients with MOGAD that is based on the latest evidence, best practice recommendations, and effective interdisciplinary collaboration. Upon completion of this activity, participants should be better able to: Explain the diagnostic criteria for MOGAD as proposed by the International MOGAD Panel; Employ recommended diagnostic tools (eg, the MOG antibody test, relevant MRI findings) to identify MOGAD and distinguish it from other demyelinating diseases; and Implement therapeutic strategies to provide acute treatment and relapse prevention in patients with MOGAD.

Go online to PeerView.com/CTH860 to view the activity, download slides and practice aids, and complete the post-test to earn credit. Myelin oligodendrocyte glycoprotein antibody-associated disorder (MOGAD) was identified as a distinct demyelinating disease entity only recently. Prior to this characterization of MOGAD, many patients were considered to have a variant of MS, NMOSD, or other neurologic condition. Now an international panel of experts has proposed diagnostic criteria that, when validated, promise to improve MOGAD diagnostic accuracy and confidence. This is particularly important, given that several clinical trials are investigating potential treatments for MOGAD. This unique PeerView activity will use problem-based educational interventions to expose learners to the science and clinical experience behind accurate diagnosis and best practices in management of MOGAD. You will be able to employ optimal clinical decision-making for your patients with MOGAD that is based on the latest evidence, best practice recommendations, and effective interdisciplinary collaboration. Upon completion of this activity, participants should be better able to: Explain the diagnostic criteria for MOGAD as proposed by the International MOGAD Panel; Employ recommended diagnostic tools (eg, the MOG antibody test, relevant MRI findings) to identify MOGAD and distinguish it from other demyelinating diseases; and Implement therapeutic strategies to provide acute treatment and relapse prevention in patients with MOGAD.

Go online to PeerView.com/CTH860 to view the activity, download slides and practice aids, and complete the post-test to earn credit. Myelin oligodendrocyte glycoprotein antibody-associated disorder (MOGAD) was identified as a distinct demyelinating disease entity only recently. Prior to this characterization of MOGAD, many patients were considered to have a variant of MS, NMOSD, or other neurologic condition. Now an international panel of experts has proposed diagnostic criteria that, when validated, promise to improve MOGAD diagnostic accuracy and confidence. This is particularly important, given that several clinical trials are investigating potential treatments for MOGAD. This unique PeerView activity will use problem-based educational interventions to expose learners to the science and clinical experience behind accurate diagnosis and best practices in management of MOGAD. You will be able to employ optimal clinical decision-making for your patients with MOGAD that is based on the latest evidence, best practice recommendations, and effective interdisciplinary collaboration. Upon completion of this activity, participants should be better able to: Explain the diagnostic criteria for MOGAD as proposed by the International MOGAD Panel; Employ recommended diagnostic tools (eg, the MOG antibody test, relevant MRI findings) to identify MOGAD and distinguish it from other demyelinating diseases; and Implement therapeutic strategies to provide acute treatment and relapse prevention in patients with MOGAD.

Go online to PeerView.com/CTH860 to view the activity, download slides and practice aids, and complete the post-test to earn credit. Myelin oligodendrocyte glycoprotein antibody-associated disorder (MOGAD) was identified as a distinct demyelinating disease entity only recently. Prior to this characterization of MOGAD, many patients were considered to have a variant of MS, NMOSD, or other neurologic condition. Now an international panel of experts has proposed diagnostic criteria that, when validated, promise to improve MOGAD diagnostic accuracy and confidence. This is particularly important, given that several clinical trials are investigating potential treatments for MOGAD. This unique PeerView activity will use problem-based educational interventions to expose learners to the science and clinical experience behind accurate diagnosis and best practices in management of MOGAD. You will be able to employ optimal clinical decision-making for your patients with MOGAD that is based on the latest evidence, best practice recommendations, and effective interdisciplinary collaboration. Upon completion of this activity, participants should be better able to: Explain the diagnostic criteria for MOGAD as proposed by the International MOGAD Panel; Employ recommended diagnostic tools (eg, the MOG antibody test, relevant MRI findings) to identify MOGAD and distinguish it from other demyelinating diseases; and Implement therapeutic strategies to provide acute treatment and relapse prevention in patients with MOGAD.

In today's episode I give a gigantic high five to our scientists/discoveres! Figuring out yet another way to stop clot formation by working out the puzzle of how platelets work to form a clot. With that worked out they then make drugs that put a stop

Looking 4 Healing Radio with Dr. Henry Ealy – SGP is a cellular wrecking ball. It devastates Mitochondria leading to diminished energy production. It disrupts Nitric Oxide production exacerbating cardiac symptomatology. SGP severely disrupts cell communication and cell signaling cascades. SGP severely dysregulates coordinated immune responses. It binds to...

Link to bioRxiv paper: http://biorxiv.org/cgi/content/short/2023.06.26.546143v1?rss=1 Authors: Bucher, M. L., Dunn, A. R., Bradner, J. M., Egerton, K. S., Burkett, J. P., Johnson, M. A., Miller, G. W. Abstract: Dopaminergic neurons of the substantia nigra exist in a persistent state of vulnerability resulting from high baseline oxidative stress, high energy demand, and broad unmyelinated axonal arborizations. Impairments in the storage of dopamine compound this stress due to cytosolic reactions that transform the vital neurotransmitter into an endogenous neurotoxicant, and this toxicity is thought to contribute to the dopamine neuron degeneration that occurs Parkinson's disease. We have previously identified synaptic vesicle glycoprotein 2C (SV2C) as a modifier of vesicular dopamine function, demonstrating that genetic ablation of SV2C in mice results in decreased dopamine content and evoked dopamine release in the striatum. Here, we adapted a previously published in vitro assay utilizing false fluorescent neurotransmitter 206 (FFN206) to visualize how SV2C regulates vesicular dopamine dynamics and determined that SV2C promotes the uptake and retention of FFN206 within vesicles. In addition, we present data indicating that SV2C enhances the retention of dopamine in the vesicular compartment with radiolabeled dopamine in vesicles isolated from immortalized cells and from mouse brain. Further, we demonstrate that SV2C enhances the ability of vesicles to store the neurotoxicant 1-methyl-4-phenylpyridinium (MPP+) and that genetic ablation of SV2C results in enhanced 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine (MPTP)-induced vulnerability in mice. Together, these findings suggest that SV2C functions to enhance vesicular storage of dopamine and neurotoxicants, and helps maintain the integrity of dopaminergic neurons. Copy rights belong to original authors. Visit the link for more info Podcast created by Paper Player, LLC

Dr. Sara Mariotto discusses her paper, "Significance of Myelin Oligodendrocyte Glycoprotein Antibodies in CSF: A Retrospective Multicenter Study". Show references: https://n.neurology.org/content/early/2022/12/16/WNL.0000000000201662

Dr. Melissa Wright discusses her abstract, "MOGAD in the Mountain West: Epidemiology and Outcomes in Pediatric and Adult Patients at Two Large Academic Referral Centers". You can find Dr. Wright's abstract, along with all of our summer conference abstracts here: https://www.aan.com/MSA/Public/Events/Index/44 This podcast is sponsored by argenx. Visit www.vyvgarthcp.com for more information.

In part 3 of a 3-part series, Erin Longbrake, MD, Ph.D., FAAN, discusses her article, "Myelin Oligodendrocyte Glycoprotein–Associated Disorders" from the Continuum August Multiple Sclerosis and Related Disorders issue. This article and the accompanying Continuum Audio interview are available to subscribers at continpub.com/MOGAD. This podcast is sponsored by argenx. Visit www.vyvgarthcp.com for more information.

In part 2 of a 3-part series, Erin Longbrake, MD, PhD, FAAN, discusses her article, "Myelin Oligodendrocyte Glycoprotein–Associated Disorders" from the Continuum August Multiple Sclerosis and Related Disorders issue. This article and the accompanying Continuum Audio interview are available to subscribers at continpub.com/MOGAD. This podcast is sponsored by argenx. Visit www.vyvgarthcp.com for more information.

In part 1 of a 3-part series, Erin Longbrake, MD, PhD, FAAN, discusses her article, "Myelin Oligodendrocyte Glycoprotein–Associated Disorders" from the Continuum August Multiple Sclerosis and Related Disorders issue. This article and the accompanying Continuum Audio interview are available to subscribers at continpub.com/MOGAD. This podcast is sponsored by argenx. Visit www.vyvgarthcp.com for more information.

In the 71st episode of Lexman, we hear the fascinating story of how Scott Aaronson discovered melodrama in chelone urine.

Dr. Rae Bacharach discusses the Neurology Today article, "Maintenance Intravenous Immunoglobulin Prevents Relapse in Adult Myelin Oligodendrocyte Glycoprotein Antibody–Associated Disease". Show references: https://journals.lww.com/neurotodayonline/Fulltext/2022/05190/Maintenance_Intravenous_Immunoglobulin_Prevents.1.aspx

Linoleic acid is an essential free fatty acid in the human body and its metabolic pathway is central to immune regulation and inflammation – which are also key symptoms in COVID-19. Using cryo-electron microscopy, Christine Toelzer’s research identified linoleic acid bound to a hydrophobic pocket of the SARS-CoV-2 glycoprotein. Christine shares her thoughts on how these findings will contribute to the fight against COVID-19 and how her lab work has been altered by the pandemic. Christine also discusses the future of other young scientists coming up in the protein science space. Christine Toelzer is currently a Research Associate at the University of Bristol. After a M.Sc. in biology and an additional M.Sc. in physics she continued with PhD work in biochemistry at the University of Cologne. Her research has always focused on structure function relationships, starting with structure determination of biotechnologically important proteins by x-ray crystallography, magnetic structure determination of inorganic compounds by neutron diffraction and recently using electron cryo-microscopy to obtain the structure of large protein complexes involved in transcription and diseases. In the last year (2020) she started coronavirus related work to contribute to the global effort aimed at better understanding the virus and uncover its potential weaknesses.About the Young Scientist Keynote Award:This recognition honors a young scientist from the international protein science community who has contributed to scientific advancement and innovation in this field. Nominations were solicited from across academic and industry research groups in the fall of 2020, and the finalists were determined through the votes and input of our 15-person advisory panel.

My AP Biology Thoughts Unit 2 Cell Structure and Function Welcome to My AP Biology Thoughts podcast, my name is Nikki Evich and I am your host for episode #52 called Unit 2 Cell Structure and Function: Macromolecules that make up the Cell Membrane. Today we will be discussing the structure of the cell membrane.  Segment 1: Introduction to the fluid mosaic model So what is a fluid mosaic model?  A fluid mosaic model describes the structure of the plasma membrane as a mosaic of components that gives the membrane a fluid character  fluid combination of phospholipids, cholesterol, and proteins.  All in all- made up of a bunch of different molecules that are distributed across the membrane. If you were to zoom in on the cell membrane, you would see a pattern of different types of molecules put together, also known as a mosaic. These molecules are constantly moving in two dimensions, in a fluid fashion, similar to icebergs floating in the ocean.  Segment 2: More About each macromolecule Lipids  Phospholipid bilayer- made of of hydrophobic tails and hydrophilic heads  Saturated fatty acids are chains of carbon atoms that have only single bonds between them. As a result, the chains are straight and easy to pack tightly. Unsaturated fats are chains of carbon atoms that have double bonds between some of the carbons. The double bonds create kinks in the chains, making it harder for the chains to pack tightly  Double or triple bonds- not organized  Saturated better organized  Cholesterol-help with structure and fluidity of the because they prevent the phospholipid bilayer from separating too far  The cholesterol molecules are randomly distributed across the phospholipid bilayer, helping the bilayer stay fluid in different environmental conditions.  Without cholesterol, the phospholipids in your cells will start to get closer together when exposed to cold, making it more difficult for small molecules, like gases to squeeze in between the phospholipids like they normally do. Without cholesterol, the phospholipids start to separate from each other, leaving large gaps.  Carbons  Glycolipid- lipid with a carb attached outside of cell membranes  Help stabilize membrane structure  Glycoprotein-protein with a carb attached  Help stabilize membrane structure  Proteins  Many different proteins  Channel proteins  acts like a pore in the membrane that lets water molecules or small ions through quickly ○ Peripheral proteins (hang on side)  transport or communication Integral protein (all the way through)  transporting larger molecules, like glucose, across the cell membrane. They have regions, called “polar” and “nonpolar” regions, that correspond with the polarity of the phospholipid bilayer  What affects cell fluidity besides  Temperature-high lipids spread, low they get too close  Segment 3: Connection to the Course  Why is the cell membrane so important?  Fluid form makes it not totally closed off but permeable-permeability is very important so the cell can import and export needed materials  Plays a role in homeostasis  Allows cell to survive in diverse environments  We learned about all the different macromolecules and how they make up cells- this is an example of how macromolecules make up an integral part of the cell Thank you for listening to this episode of My AP Biology Thoughts. For more student-ran podcasts and digital content, make sure that you visit http://www.hvspn.com/ (www.hvspn.com). See you next time! Music Credits:“Ice Flow” Kevin MacLeod (incompetech.com) Licensed under Creative Commons: By Attribution 4.0 License http://creativecommons.org/licenses/by/4.0/ Subscribe to our Podcasthttps://podcasts.apple.com/us/podcast/my-ap-biology-thoughts/id1549942575 (Apple Podcasts)...

Link to bioRxiv paper: http://biorxiv.org/cgi/content/short/2020.11.15.383794v1?rss=1 Authors: Kapoor, k., Pant, S., Tajkhorshid, E. Abstract: P-glycoprotein (Pgp) is a major efflux pump in humans, overexpressed in a variety of cancers and associated with the development of multi-drug resistance. Allosteric modulation induced by binding of various ligands (e.g., transport substrates, inhibitors, and ATP) has been bio-chemically shown to directly influence the function of Pgp. However, the molecular details of such effects are not well established. In particular, the role and involvement of the sur-rounding lipid environment on ligand-induced modulation of the conformational dynamics of the transporter have not been investigated at any level. Here, we employ all-atom molecular dynamics (MD) simulations to study the conformational landscape of Pgp in the presence of a high-affinity, third-generation inhibitor, tariquidar, in comparison to the nucleotide-free (APO) and the ATP-bound states, in order to shed light on and to characterize how the inhibitor blocks the function of the transporter. Simulations in a multi-component lipid bi-layer show a dynamic equilibrium between open and closed inward-facing (IF) conformations in the APO-state, with binding of ATP shifting the equilibrium towards conformations feasible for ATP hydrolysis and subsequent completion of the transport cycle. In the presence of the inhibitor bound to the drug-binding pocket in the transmembrane domain (TMD), the transporter samples more open IF conformations, and the nucleotide binding domains (NBDs) are observed to become highly dynamic. Interestingly, and reproduced in multiple independent simulations, the inhibitor is observed to recruit lipid molecules into the Pgp lumen through the two proposed drug-entry portals, where the lipid head groups from the lower leaflet translocate inside the TMD, while the lipids tails remain extended into the bulk lipid environment. These 'wedge-lipid' molecules likely enhance the inhibitor-induced conformational changes in the TMD leading to the differential modulation of coupling pathways observed with the NBDs downstream. We suggest a novel inhibitory mechanism for tariquidar, and for related third-generation Pgp inhibitors, where lipids are seen to enhance the inhibitory role in the catalytic cycle of membrane transporters. Copy rights belong to original authors. Visit the link for more info

Link to bioRxiv paper: http://biorxiv.org/cgi/content/short/2020.10.22.350777v1?rss=1 Authors: McCormick, J. W., Ammerman, L., Chen, G., Vogel, P. D., Wise, J. G. Abstract: P-glycoprotein (P-gp) is a critical membrane transporter in the blood brain barrier (BBB) and is implicated in Alzheimer's disease (AD). However, previous studies on the ability of P-gp to directly transport the Alzheimer's associated amyloid-{beta} (A{beta}) protein have produced contradictory results. Here we use molecular dynamics (MD) simulations, transport substrate accumulation studies in cell culture, and biochemical activity assays to show that P-gp actively transports A{beta}. We observed transport of A{beta}40 and A{beta}42 monomers by P-gp in explicit MD simulations of a putative catalytic cycle. In in vitro assays with P-gp overexpressing cells, we observed enhanced accumulation of fluorescently labeled A{beta}42 in the presence of Tariquidar, a potent P-gp inhibitor. We also showed that A{beta}42 stimulated the ATP hydrolysis activity of isolated P-gp in nanodiscs. Our findings expand the substrate profile of P-gp, and suggest that P-gp may contribute to the onset and progression of AD. Copy rights belong to original authors. Visit the link for more info

Link to bioRxiv paper: http://biorxiv.org/cgi/content/short/2020.10.12.336016v1?rss=1 Authors: Ni, D., Lau, K., Lehmann, F., Fraenkli, A., Hacker, D., Pojer, F., Stahlberg, H. Abstract: The human membrane protein Angiotensin-converting enzyme 2 (hACE2) acts as the main receptor for host cells invasion of the new coronavirus SARS-CoV-2. The viral surface glycoprotein Spike binds to hACE2, which triggers virus entry into cells. As of today, the role of hACE2 for virus fusion is not well understood. Blocking the transition of Spike from its prefusion to post-fusion state might be a strategy to prevent or treat COVID-19. Here we report a single particle cryo-electron microscopy analysis of SARS-CoV-2 trimeric Spike in presence of the human ACE2 ectodomain. The binding of purified hACE2 ectodomain to Spike induces the disassembly of the trimeric form of Spike and a structural rearrangement of its S1 domain to form a stable, monomeric complex with hACE2. This observed hACE2 dependent dissociation of the Spike trimer suggests a mechanism for the therapeutic role of recombinant soluble hACE2 for treatment of COVID-19. Copy rights belong to original authors. Visit the link for more info

Link to bioRxiv paper: http://biorxiv.org/cgi/content/short/2020.10.08.331751v1?rss=1 Authors: Tiwari, V., Tandon, R., Sankaranarayanan, N. V., Beer, J. C., Kohlmeir, E. K., Swanson-Mungerson, M., Desai, U. R. Abstract: The COVID-19 pandemic caused by SARS-CoV-2 is in immediate need of an effective antidote. Although the Spike glycoprotein (SgP) of SARS-CoV-2 has been shown to bind to heparins, the structural features of this interaction, the role of a plausible heparan sulfate proteoglycan (HSPG) receptor, and the antagonism of this pathway through small molecules remain unaddressed. Using an in vitro cellular assay, we demonstrate HSPGs modified by the 3-O-sulfotransferase isoform-3, but not isoform-5, preferentially increased SgP-mediated cell-to-cell fusion in comparison to control, unmodified, wild-type HSPGs. Computational studies support preferential recognition of the receptor-binding domain of SgP by 3-O-sulfated HS sequences. Competition with either fondaparinux, a 3-O-sulfated HS-binding oligopeptide, or a synthetic, non-sugar small molecule, blocked SgP-mediated cell-to-cell fusion. Finally, the synthetic, sulfated molecule inhibited fusion of GFP-tagged pseudo SARS-CoV-2 with human 293T cells with sub-micromolar potency. Overall, overexpression of 3-O-sulfated HSPGs contribute to fusion of SARS-CoV-2, which could be effectively antagonized by a synthetic, small molecule. Copy rights belong to original authors. Visit the link for more info

Link to bioRxiv paper: http://biorxiv.org/cgi/content/short/2020.09.28.317206v1?rss=1 Authors: Ghorbani, M., Brooks, B. R., Klauda, J. B. Abstract: The ongoing pandemic caused by coronavirus SARS-COV-2 continues to rage with devastating consequences on human health and global economy. The spike glycoprotein on the surface of coronavirus mediates its entry into host cells and is the target of all current antibody design efforts to neutralize the virus. The glycan shield of the spike helps the virus to evade the human immune response by providing a thick sugar-coated barrier against any antibody. To study the dynamic motion of glycans in the spike protein, we performed microsecond-long MD simulation in two different states that correspond to the receptor binding domain in open or closed conformations. Analysis of this microsecond-long simulation revealed a scissoring motion on the N-terminal domain of neighboring monomers in the spike trimer. Role of multiple glycans in shielding of spike protein in different regions were uncovered by a network analysis, where the high betweenness centrality of glycans at the apex revealed their importance and function in the glycan shield. Microdomains of glycans were identified featuring a high degree of intra-communication in these microdomains. An antibody overlap analysis revealed the glycan microdomains as well as individual glycans that inhibit access to the antibody epitopes on the spike protein. Overall, the results of this study provide detailed understanding of the spike glycan shield, which may be utilized for therapeutic efforts against this crisis. Copy rights belong to original authors. Visit the link for more info

Link to bioRxiv paper: http://biorxiv.org/cgi/content/short/2020.09.24.312355v1?rss=1 Authors: Marchan, J. G. Abstract: Coronavirus Disease 2019 (COVID-19) represents a new global threat demanding a multidisciplinary effort to fight its etiological agent, severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2). In this regard, immunoinformatics may aid to predict prominent immunogenic regions from critical SARS-CoV-2 structural proteins, such as the spike (S) glycoprotein, for their use in prophylactic or therapeutic interventions against this rapidly emerging coronavirus. Accordingly, in this study, an integrated immunoinformatics approach was applied to identify cytotoxic T cell (CTC), T helper cell (THC), and Linear B cell (BC) epitopes from the S glycoprotein in an attempt to design a high-quality multi-epitope vaccine. The best CTC, THC, and BC epitopes showed high viral antigenicity, lack of allergenic or toxic residues, and suitable HLA-viral peptide interactions. Remarkably, SARS-CoV-2 receptor-binding domain (RBD) and its receptor-binding motif (RBM) harbour several potential epitopes. The structure prediction, refinement, and validation data indicate that the multi-epitope vaccine has an appropriate conformation and stability. Three conformational epitopes and an efficient binding between Toll-like receptor 4 (TLR4) and the vaccine model were observed. Importantly, the population coverage analysis showed that the multi-epitope vaccine could be used globally. Notably, computer-based simulations suggest that the vaccine model has a robust potential to evoke and maximize both immune effector responses and immunological memory to SARS-CoV-2. Further research is needed to accomplish with the mandatory international guidelines for human vaccine formulations. Copy rights belong to original authors. Visit the link for more info

Link to bioRxiv paper: http://biorxiv.org/cgi/content/short/2020.09.21.307439v1?rss=1 Authors: Wang, X., Zhang, S., Qiao, S., Yu, J., Zeng, J., Shan, S., Lan, J., Tian, L., Zhang, L. Abstract: In recognizing the host cellular receptor and mediating fusion of virus and cell membranes, the spike (S) glycoprotein of coronaviruses is the most critical viral protein for cross-species transmission and infection. Here we determined the cryo-EM structures of the spikes from bat (RaTG13) and pangolin (PCoV_GX) coronaviruses, which are closely related to SARS-CoV-2. All three receptor-binding domains (RBDs) of these two spike trimers are in the "down" conformation, indicating they are more prone to adopt this receptor-binding inactive state. However, we found that the PCoV_GX, but not the RaTG13, spike is comparable to the SARS-CoV-2 spike in binding the human ACE2 receptor and supporting pseudovirus cell entry. Through structure and sequence comparisons, we identified critical residues in the RBD that underlie the different activities of the RaTG13 and PCoV_GX/SARS-CoV-2 spikes and propose that N-linked glycans serve as conformational control elements of the RBD. These results collectively indicate that strong RBD-ACE2 binding and efficient RBD conformational sampling are required for the evolution of SARS-CoV-2 to gain highly efficient infection. Copy rights belong to original authors. Visit the link for more info

Link to bioRxiv paper: http://biorxiv.org/cgi/content/short/2020.09.09.287771v1?rss=1 Authors: Mohammed, A. A., Elkhalifa, M. E., Elamin, K. E., Mohammed, R. A., Ibrahim, M. E., Dirar, A. I., Migdar, S. H., Hamid, M. A., Elawad, E. H., Abdelsalam, S. O., Hassan, M. A. Abstract: Background: Lujo virus LUJV is a highly fatal human pathogen belonging to the Arenaviridae family. Lujo virus causes viral hemorrhagic fever VHF. An In silico molecular docking was performed on the GPC domain of Lujo virus in complex with the first CUB domain of neuropilin-2. The aim of this study is to predict effective epitope-based vaccine against glycoprotein GPC precursor of Lujo virus using immunoinformatics approaches. Methods and Materials: glycoprotein GPC precursor of Lujo virus Sequence was retrieved from NCBI. Different prediction tools were used to analyze the nominees epitopes in BepiPred-2.0: Sequential B-Cell Epitope Predictor for B-cell, T-cell MHC class II & I. Then the proposed peptides were docked using Autodock 4.0 software program. Results and Conclusions: The proposed and promising peptides FWYLNHTKL and YMFSVTLCI show a very strong binding affinity to MHC class I & II alleles with high population coverage for the world, South Africa and Sudan. This indicates a strong potential to formulate a new vaccine especially with the peptide YMFSVTLCI which is likely to be the first proposed epitope-based vaccine against glycoprotein GPC of the Lujo virus. This study recommends an in-vivo assessment for the most promising peptides especially FWYLNHTKL, YMFSVTLCI and LPCPKPHRLR. Copy rights belong to original authors. Visit the link for more info

Link to bioRxiv paper: http://biorxiv.org/cgi/content/short/2020.08.27.269456v1?rss=1 Authors: Agarwal, V., Tiwari, A., Varadwaj, P. K. Abstract: The emergence of COVID-19 as a pandemic with a high morbidity rate is posing serious global concern. There is an urgent need to design a suitable therapy or vaccine that could fight against SARS-CoV-2 infection. As spike glycoprotein of SARS-CoV-2 plays a crucial role in receptor binding and membrane fusion inside the host, it could be a suitable target for designing of an epitope-based vaccine. SARS-CoV-2 is an RNA virus and thus has a property to mutate. So, a conserved peptide region of spike glycoprotein was used for predicting suitable B cell and T cell epitopes. 4 T cell epitopes were selected based on stability, antigenicity, allergenicity and toxicity. Further, MHC-I were found from the immune database that could best interact with the selected epitopes. Population coverage analysis was also done to check the presence of identified MHC-I, in the human population of the affected countries. The T cell epitope that binds with the respective MHC-I with highest affinity was chosen. Molecular dynamic simulation results show that the epitope is well selected. This is an in-silico based study that predicts a novel T cell epitope from the conserved spike glycoprotein that could act as a target for designing of the epitope-based vaccine. Further, B cell epitopes have also been found but the main work focuses on T cell epitope as the immunity generated by it is long lasting as compared to B cell epitope. Copy rights belong to original authors. Visit the link for more info

Link to bioRxiv paper: http://biorxiv.org/cgi/content/short/2020.08.19.256693v1?rss=1 Authors: Li, Y., Mullin, M., ZHANG, Y., Drews, F., Welch, L., Showalter, A. Abstract: Hydroxyproline-rich glycoproteins (HRGPs) are a superfamily of plant cell wall structural proteins that function in various aspects of plant growth and development, including pollen tube growth. We have previously characterized HRGP superfamily into three family members: the hyperglycosylated arabinogalactan-proteins, the moderately glycosylated extensins, and the lightly glycosylated proline-rich proteins. However, the mechanism of pollen-specific HRGP expression remains untouched. To this end, we developed an integrative analysis pipeline combining RNA-seq gene expression and promoter sequences that identified 15 transcriptional cis-regulatory motifs responsible for pollen-specific expression of HRGP in Arabidopsis Thaliana. Specifically, we mined the public RNA-seq datasets and identified 13 pollen-specific HRGP genes. Ensemble motif discovery with various filters identified 15 conserved promoter elements between Thaliana and Lyrata. Known motif analysis revealed pollen related transcription factors of GATA12 and brassinosteroid (BR) signaling pathway regulator BZR1. Lastly, we performed a machine learning regression analysis and demonstrated that the identified 15 motifs well captured the HRGP gene expression in pollen (R=0.61). In conclusion, we performed the integrative analysis as the first-of-its-kind study to identify cis-regulatory motifs in pollen-specific HRGP genes and shed light on its transcriptional regulation in pollen. Copy rights belong to original authors. Visit the link for more info

Link to bioRxiv paper: http://biorxiv.org/cgi/content/short/2020.07.31.230888v1?rss=1 Authors: Malaspina, D. C., Faraudo, J. Abstract: A prominent feature of coronaviruses is the presence of a large glycoprotein spike protruding from a lipidic membrane. This glycoprotein spike determines the interaction of coronaviruses with the environment and the host. In this paper, we perform all atomic Molecular Dynamics simulations of the interaction between the SARS-CoV-2 trimeric glycoprotein spike and surfaces of materials. We considered a material with high hydrogen bonding capacity (cellulose) and a material capable of strong hydrophobic interactions (graphite). Initially, the spike adsorbs to both surfaces through essentially the same residues belonging to the receptor binding subunit of its three monomers. Adsorption onto cellulose stabilizes in this configuration, with the help of a large number of hydrogen bonds developed between cellulose and the three receptor binding domains (RBD) of the glycoprotein spike. In the case of adsorption onto graphite, the initial adsorption configuration is not stable and the surface induces a substantial deformation of the glycoprotein spike with a large number of adsorbed residues not pertaining to the binding subunits of the spike monomers. Copy rights belong to original authors. Visit the link for more info

Link to bioRxiv paper: http://biorxiv.org/cgi/content/short/2020.07.27.223107v1?rss=1 Authors: Vallet, S. D., Davis, M. N., Barque, A., Ricard-Blum, S., Naba, A. Abstract: The extracellular matrix (ECM) protein SNED1 has been shown to promote breast cancer metastasis and control neural crest cell-specific craniofacial development, but the cellular and molecular mechanisms by which it does so remain unknown. ECM proteins exert their functions by binding to cell surface receptors, sequestering growth factors, and interacting with other ECM proteins, actions that can be predicted using knowledge of protein sequence, structure and post-translational modifications. Here, we combined in-silico and in-vitro approaches to characterize the physico-chemical properties of SNED1 and infer its putative functions. To do so, we established a mammalian cell system to produce and purify SNED1 and its N-terminal fragment, which contains a NIDO domain. We have determined experimentally the potential of SNED1 to be glycosylated, phosphorylated, and incorporated into insoluble ECM produced by cells. In addition, we used biophysical and computational methods to determine the secondary and tertiary structures of SNED1 and its N-terminal fragment. The tentative ab-initio model we built of SNED1 suggests that it is an elongated protein presumably able to bind multiple partners. Using computational predictions, we identified 114 proteins as putative SNED1 interactors. Pathway analysis of the newly-predicted SNED1 interactome further revealed that binding partners of SNED1 contribute to signaling through cell surface receptors, such as integrins, and participate in the regulation of ECM organization and developmental processes. Altogether, we provide a wealth of information on an understudied yet important ECM protein with the potential to decipher its functions in physiology and diseases. Copy rights belong to original authors. Visit the link for more info

More from Dr. Corsi's article about the probability of a COVID-20: https://corsination.com/russia-gets-hydroxychloroquine/ Discovering the Engineering of COVID-19 as a Bioweapon, Part 1 In January 2020, I realized that the scientific sequencing of COVID-19 strongly suggested a key element of the HIV-1 pathogen causing the AIDS epidemic worldwide, Glycoprotein 120 (GP12) had be spliced into the genome of a SARS virus “chassis” to create COVID-19. This was reported first in a study sequencing COVID-19 by a group of medical scientists in India who published their findings article published by a group of medical scientists in India in the medical science journal BioRXiv in January 2020 was entitled “Uncanny similarity of unique inserts in the 2019-nCov spike protein to HIV-1 and Gag.” In a highly controversial finding, the medical scientists in India reported: “We found 4 insertions in the spike glycoprotein which are unique to the 2019-nCoV and are not present in other coronaviruses. Importantly, amino acid residues in all 4 inserts have identity or similarity to those in the HIV-1 gp120 or HIV-Gag, all of which have identity/similarity to amino acid residues in key structural proteins of HIV-1 is unlikely to be fortuitous in nature.” The FAKE NEWS continues to attack HCQ as more doctors around the world tout their success with treating patients with a 70 year old proven and relatively safe medication. Acting DNI Rick Grenell is carpet bombing the Deep Sate on a daily basis and the truth pours out instead of lies leaking out. Refreshing change. President Trump must refer the General Flynn case directly to the United States Supreme Court to stop the madness and #FreeFlynn Tune in Monday through Friday at 11:00am EDT for another show. Visit our sister website, http://www.theprayerfulpatriot.com dedicated to faith and prayer. https://www.patreon.com/jeromecorsi/ https://www.subscribestar.com/jerome_corsi/

Get Signed Copies of Dr. Corsi’s Two Most Recent Books for Donations of $100 or more Donations to Corsi Nation are essential to allow us to continue the Fight Against the Deep State, to continue our efforts to re-elect President Donald J. Trump, and to support our efforts to report on important political and economic events that affect the United States of America. While you're at corsination.com, read the three articles by Dr. Corsi about virusGate: The first article references the list of patents Fauci has been awarded as well as applied for, arguing that important components of the HIV-1 virus attacking the immune system show up in patents naming Fauci as one of the inventors. The article also notes that a glycoprotein found in the HIV-1 virus, identified as Glycoprotein 120, or simply as GP120, has also been found to be a key component of the spikes in the current COVID-19 to produce this new disease virus that appears to combine a HIV-1 attack on the human immune system, with SARS CoV-1, the pathogen from the original SARS (“Severe Acute Respiratory System”) that created an international pandemic in 2002-2003. The second article documents that Fauci’s name also appears on patents involving the C4 receptor. HIV-1 binds to healthy cells through the CD4 receptor while COVID-19 enters healthy cells through the ACE2 receptor, producing SARS-CoV-2 pathogen. It is then hypothesized that by re-engineering COVID-19 to allow the SARS-CoV-2 pathogen through the CD4 receptors, COVID-20 would have a new SARS-CoV-3 pathogen that would enter the body faster, producing a virulent attack on the immune system to accompany the SARS attack on the lungs that would follow. The third article, "Coup de Grâce" hypothesizes how COVID-20 will use Fauci's patents to deliver a devastating HIV-1 attack to the intestines preceding a SARS attack to the Lungs. The COVID-20 re-engineering will be complete when a third element from HIV-1 research leading to patents bearing Fauci’s name will be added: namely an attack involving the mechanism whereby integrin α4β7 is incorporated into the  envelope of HIV-1 virions that becomes functionally active as it binds with MAdCAM-1 allowing the HIV-1 pathogen enters the body through the high endothelial venules in the intestinal mucosa. Acting DNI Rick Grenell is continuing to expose the Deep State traitors and the elements of the Coup d'Etat against President Trump. It is time for AG Barr to act, and for indictments to be released. #LockThemAllUP Tune in Monday through Friday at 11:00am EDT for another show. Visit our sister website, http://www.theprayerfulpatriot.com dedicated to faith and prayer. https://www.patreon.com/jeromecorsi/ https://www.subscribestar.com/jerome_corsi/

In this podcast, George Karpouzas, MD, discusses his research that evaluated whether β-2 glycoprotein 1 IgA antibodies can predict coronary plaque burden and progression, and whether the antibodies moderate the effect of inflammation on atherosclerosis among patients with rheumatoid arthritis. More at: www.consultan360.com/rheumatology

Nels and Vincent examine SARS-CoV-2 from an evolutionary viewpoint, examining what the spike glycoprotein sequence informs us about the origin of the virus. Hosts: Nels Elde and Vincent Racaniello Subscribe (free): iTunes, Google Podcasts, RSS, email Become a patron of TWiEVO Distinct CoV name needed (Lancet) Proximal origin of SARS-COV-2 (virological) CoV spillovers not rare (Goats and Soda) SARS-CoV-2 phylodynamics analysis (virological) Bat CoV surveillance in China (PLoS Path) Image credit Time stamps by Jolene. Thanks! Science Picks Nels - Alejandro Sanchez TED Talk Vincent - WHO SARS-CoV-2 Sitrep Music on TWiEVO is performed by Trampled by Turtles Send your evolution questions and comments to twievo@microbe.tv

The TWiV team returns this week to SARS-CoV-2019 coverage to review the latest epi curves, the fatality rate, furin cleavage site and receptor binding domain in the spike glycoprotein, related CoV recovered from pangolins, evidence that the virus did not escape from a laboratory, and many more questions sent in by listeners. Hosts: Vincent Racaniello, Dickson Despommier, Rich Condit, and Kathy Spindler Subscribe (free): iTunes, Google Podcasts, RSS, email Become a patron of TWiV! Links for this episode WHO CoV sitreps Epidemiology of COVID-19 (China CDC) COVID-19 incubation period (J Med Virol) China clinical trials for COVID-19 (Nature) Furin cleavage site in SARS-CoV-2 spike (virology blog) Proximal origin of SARS-CoV-2 (Virological) Pangolin photos (Nat Geo) Letters read on TWiV 588 Timestamps by Jolene. Thanks! Weekly Science Picks Kathy - The deep sea Rich - World's oldest message in a bottle Dickson - Coronavirus: A visual guide Vincent - Recent NSABB meeting Videocasts Day 1 Day 2 Listener Picks Stephen - This podcast will kill you Neva - China's surveillance Sophia - WHO online courses Intro music is by Ronald Jenkees. Send your virology questions and comments to twiv@microbe.tv

The TWiV team returns this week to SARS-CoV-2019 coverage to review the latest epi curves, the fatality rate, furin cleavage site and receptor binding domain in the spike glycoprotein, related CoV recovered from pangolins, evidence that the virus did not escape from a laboratory, and many more questions sent in by listeners. Hosts: Vincent Racaniello, Dickson Despommier, Rich Condit, and Kathy Spindler Subscribe (free): iTunes, Google Podcasts, RSS, email Become a patron of TWiV! Links for this episode WHO CoV sitreps Epidemiology of COVID-19 (China CDC) COVID-19 incubation period (J Med Virol) China clinical trials for COVID-19 (Nature) Furin cleavage site in SARS-CoV-2 spike (virology blog) Proximal origin of SARS-CoV-2 (Virological) Pangolin photos (Nat Geo) Letters read on TWiV 588 Timestamps by Jolene. Thanks! Weekly Science Picks Kathy - The deep sea Rich - World's oldest message in a bottle Dickson - Coronavirus: A visual guide Vincent - Recent NSABB meeting Videocasts Day 1 Day 2 Listener Picks Stephen - This podcast will kill you Neva - China's surveillance Sophia - WHO online courses Intro music is by Ronald Jenkees. Send your virology questions and comments to twiv@microbe.tv

Vincent speaks with Félix Rey about his career and his work on solving structures of a variety of viruses and the insights learned about viral membrane fusion and antibody-mediated neutralization. Subscribe (free): iTunes, Google Podcasts, RSS, email Become a patron of TWiV! Links for this episode Unité de Virologie Structurale Tick-borne encephalitis glycoprotein structure (Nature) HCV RNA polymerase structure (PNAS) Flavivirus structural heterogeneity (Curr Op Viral) Antibody responses to flaviviruses (EMBO Rep) Covalently linked dengue envelope dimers (Nat Comm) Timestamps by Jolene. Thanks! Intro music is by Ronald Jenkees. Send your virology questions and comments to twiv@microbe.tv

Vincent speaks with Félix Rey about his career and his work on solving structures of a variety of viruses and the insights learned about viral membrane fusion and antibody-mediated neutralization. Subscribe (free): iTunes, Google Podcasts, RSS, email Become a patron of TWiV! Links for this episode Unité de Virologie Structurale Tick-borne encephalitis glycoprotein structure (Nature) HCV RNA polymerase structure (PNAS) Flavivirus structural heterogeneity (Curr Op Viral) Antibody responses to flaviviruses (EMBO Rep) Covalently linked dengue envelope dimers (Nat Comm) Timestamps by Jolene. Thanks! Intro music is by Ronald Jenkees. Send your virology questions and comments to twiv@microbe.tv

Team TWiV reveals DNA polymerases that do not require a primer, and packaging of hepatitis delta virus by the envelope glycoproteins of diverse viruses. Hosts: Vincent Racaniello, Dickson Despommier, Alan Dove, Rich Condit, and Brianne Barker Guest: Kiki Warren Subscribe (free): iTunes, Google Podcasts, RSS, email Become a patron of TWiV! Links for this episode Bacteriophage DNA polymerase is primer-independent (PNAS) Primer-independent DNA polymerase from mobile elements (Cell Rep) Non-HBV helper viruses for HDV (Nat Comm) Waning of measles vaccine immunity (one, two, three) Letters read on TWiV 552 Timestamps by Jolene. Thanks! Weekly Science Picks Alan - Seek Rich - Want to See My Genes? Get a Warrant and NIH director will no longer speak on all-male science panels Brianne- Unique scientists Dickson- Vaccines - Calling the Shots Vincent- NY eliminates religious vaccine exemptions and Jessica Biel reveals that she does not understand vaccines Kiki- Converting type A to type O blood Listener Picks Justin - An Apple a Day Intro music is by Ronald Jenkees. Send your virology questions and comments to twiv@microbe.tv

Team TWiV reveals DNA polymerases that do not require a primer, and packaging of hepatitis delta virus by the envelope glycoproteins of diverse viruses. Hosts: Vincent Racaniello, Dickson Despommier, Alan Dove, Rich Condit, and Brianne Barker Guest: Kiki Warren Subscribe (free): iTunes, Google Podcasts, RSS, email Become a patron of TWiV! Links for this episode Bacteriophage DNA polymerase is primer-independent (PNAS) Primer-independent DNA polymerase from mobile elements (Cell Rep) Non-HBV helper viruses for HDV (Nat Comm) Waning of measles vaccine immunity (one, two, three) Letters read on TWiV 552 Timestamps by Jolene. Thanks! Weekly Science Picks Alan - Seek Rich - Want to See My Genes? Get a Warrant and NIH director will no longer speak on all-male science panels Brianne- Unique scientists Dickson- Vaccines - Calling the Shots Vincent- NY eliminates religious vaccine exemptions and Jessica Biel reveals that she does not understand vaccines Kiki- Converting type A to type O blood Listener Picks Justin - An Apple a Day Intro music is by Ronald Jenkees. Send your virology questions and comments to twiv@microbe.tv

The TWiPsters solve the case of the Brazilian Immigrant With Heart Problems, and describe how genome organization controls trypanosome antigenic variation. Hosts: Vincent Racaniello, Dickson Despommier, and Daniel Griffin Subscribe (free): iTunes, Google Podcasts, RSS, email Become a patron of TWiP. Links for this episode: Trypanosome genome organization and antigenic variation (Nature) Hero: Arthur Looss Letters read on TWiP 161 Case Study for TWiP 161 Daniel was asked to see 30 yo female from Bolivia, had to travel back during 3rd trimester. Was there for most of 3rd trimester. Child born in US, pericardial effusion, ascites, moderate PDA. Heart function is ok. Woman was healthy, no issues during pregnancy. Baby’s  white count elevated, diagnostic evaluation. It is a parasite. Send your case diagnosis, questions and comments to twip@microbe.tv Music by Ronald Jenkees

At Tufts University Dental School in Boston, Vincent speaks with Katya Heldwein and Sean Whelan about their careers and their work on herpesvirus structure and replication of vesicular stomatitis virus. Host: Vincent Racaniello Guests: Katya Heldwein and Sean Whelan Become a patron of TWiV! Links for this episode Crystal structure of HSV gB (Science) Crystal structure of HSV fusion regulator gH-gL (Nat Struct Mol Biol) Nuclear Exodus: Herpesviruses Lead the Way (Ann Rev Virol) VSV Pseudotypes Bearing gB, gD, gH, and gL (J Virol) Recovery of infectious VSV from cDNA clones (PNAS) Structure of the L Protein of VSV (Cell) Unique strategy for mRNA cap methylation by VSV (PNAS) Molecular architecture of VSV RNA polymerase (PNAS) This episode is brought to you by the Defense Threat Reduction Agency. Part of the U.S. Department of Defense, the Agency’s Chemical and Biological Technologies Department hosts the 2017 Chemical and Biological Defense Science & Technology Conference to exchange information on the latest and most dynamic developments for countering chemical and biological weapons of mass destruction. Find out more at http://www.cbdstconference.com Intro music is by Ronald Jenkees. Send your virology questions and comments to twiv@microbe.tv

Hosts: Vincent Racaniello, Dickson Despommier, Alan Dove, Rich Condit, and Kathy Spindler The TWiVirions reveal bacteriophage genes that control eukaryotic reproduction, and the biochemical basis for increased Ebolavirus glycoprotein activity during the recent outbreak.   Become a patron of TWiV! Links for this episode Nido2017 Meeting ASM Microbe 2017 Mark Buller obituary RS virus pipeline (pdf) Prophage WO genes and cytoplasmic incompatibility (Nature) Vanderbilt Virology (TWiV 332) Basis for increased Ebolavirus glycoprotein activity (Cell Host Micr) Letters read on TWiV 431 This episode is brought to you by Blue Apron. Blue Apron is the #1 fresh ingredient and recipe delivery service in the country. See what’s on the menu this week and get your first 3 meals free with your first purchase – WITH FREE SHIPPING – by going to blueapron.com/twiv Weekly Science Picks Rich - Germ Theory by Robert P. GaynesAlan - Student Scientist Partnerships Kathy - Symbiartic Dickson - PaleoBioDB Navigator Vincent - Kusama Infinity Room Listener Pick Chaim - How to fight back against the backfire effect Intro music is by Ronald Jenkees. Send your virology questions and comments to twiv@microbe.tv

Hosts: Vincent Racaniello, Alan Dove, Rich Condit, and Kathy Spindler Guests: Jeremy Luban, Aaron Lin, and Ted Diehl Jeremy, Aaron, and Ted join the TWiV team to discuss their work on identifying a single amino acid change in the Ebola virus glycoprotein from the West African outbreak that increases infectivity in human cells.   Become a patron of TWiV! Links for this episode Ebola virus glycoprotein with increased infectivity (Cell) Effect in cells of Ebola virus mutations from West African outbreak (Cell) Ebola virus in semen for over 500 days (Clin Inf Dis) Puzzling origin of 2014 Ebola virus outbreak (J Virol) Mutant Ebola virus may have caused explosive outbreak (Goats and Soda) Virus Genomics and Evolution website This episode is brought to you by CuriosityStream, a subscription streaming service that offers over 1,400 documentaries and non­fiction series from the world's best filmmakers. Get unlimited access starting at just $2.99 a month, and for our audience, the first two months are completely free if you sign up at curiositystream.com/microbe and use the promo code MICROBE. 0:25, 1:05:40 Check out the graduate and postdoctoral programs at the Department of Microbiology at the Icahn School of Medicine at Mount Sinai. Deadline for applying to the graduate program is 1 December 2016. For more information about the Department, please visit http://bit.ly/micromssm 1:35 Register for the 2017 ASM Grant Writing Online course. Weekly Science Picks Jeremy – Real time tracking of Ebola virus evolutionTed - The Brain Scoop Aaron - Kate Rubins sequencing DNA on ISS (video) Alan – A Sand County Almanac by Aldo Leopold Rich –  Vendee GlobeKathy – Axios Vincent – Vendors from above by Loes Heerinck Send your virology questions and comments to twiv@microbe.tv

Hosts: Vincent Racaniello, Rich Condit, and Kathy Spindler Guests: Ralph Baric and Vineet Menachery Vincent, Rich, and Kathy speak with Ralph and Vineet about their research on the potential of SARS-like bat coronaviruses  to infect human cells and cause disease in mice.   Links for this episode Zoonotic potential of SARS-like bat viruses (Nature) SARS-CoV susceptibility loci via collaborative cross (PLoS Genetics) 47:00, 57:25 Ebright and Wain-Hobson quotes (Nature) 1:17:20 Misleading press release (UNC) 1:24:45 Super powerful SARS 2.0 (not!) (Motherboard) SARS-like bat virus can jump without mutating (not!) (Daily Mail) 1:26:30 Photo credit Letters read on TWiV 364 1:47:40 This episode is sponsored by Mt. Sinai Department of Microbiology (2:10), ASM Education (33:25), and the 32nd Clinical Virology Symposium (1:51:30) Timestamps by Jennifer. Thanks! Weekly Science Picks 1:53:00 Kathy - California Dept of Public HealthRich - Leafy greens on space stationVincent - Mechanical keyboards and TWiV on Perfect your podcast Listener Picks Jesse - Networking for Nerds by Alaina G. LevineRussell - Worldmapper Send your virology questions and comments to twiv@twiv.tv

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.

Human cytomegalovirus (HCMV) is the major viral cause of congenital infection and birth defects. Primary maternal infection often results in virus transmission, and symptomatic babies can have permanent neurological deficiencies and deafness. Congenital infection can also lead to intrauterine growth restriction, a defect in placental transport. HCMV replicates in primary cytotrophoblasts (CTBs), the specialized cells of the placenta, and inhibits differentiation/invasion. Human trophoblast progenitor cells (TBPCs) give rise to the mature cell types of the chorionic villi, CTBs and multi-nucleated syncytiotrophoblasts (STBs). Here we report that TBPCs are fully permissive for pathogenic and attenuated HCMV strains. Studies with a mutant virus lacking a functional pentamer complex (gH/gL/pUL128-131A) showed that virion entry into TBPCs is independent of the pentamer. In addition, infection is blocked by a potent human neutralizing monoclonal antibody (mAb), TRL345, reactive with glycoprotein B (gB), but not mAbs to the pentamer proteins pUL130/pUL131A. Functional studies revealed that neutralization of infection preserved the capacity of TBPCs to differentiate and assemble into trophospheres composed of CTBs and STBs in vitro. Our results indicate that mAbs to gB protect trophoblast progenitors of the placenta and could be included in antibody treatments developed to suppress congenital infection and prevent disease.

Introduction: This study aimed to characterize the glycophenotype of osteoarthritic cartilage and human chondrocytes. Methods: Articular knee cartilage was obtained from nine osteoarthritis (OA) patients. mRNA levels for 27 glycosyltransferases were analyzed in OA chondrocytes using RT-qPCR. Additionally, N- and O-glycans were quantified using mass-spectrometry. Histologically, two cartilage areas with Mankin scores (MS) either

Complete knowledge of autoantigen spectra is crucial for understanding pathomechanisms of autoimmune diseases like equine recurrent uveitis (ERU), a spontaneous model for human autoimmune uveitis. While several ERU autoantigens were identified previously, no membrane protein was found so far. As there is a great overlap between glycoproteins and membrane proteins, the aim of this study was to test whether pre-enrichment of retinal glycoproteins by ConA affinity is an effective tool to detect autoantigen candidates among membrane proteins. In 1D Western blots, the glycoprotein preparation allowed detection of IgG reactions to low abundant proteins in sera of ERU patients. Synaptotagmin-1, a Ca2+-sensing protein in synaptic vesicles, was identified as autoantigen candidate from the pre-enriched glycoprotein fraction by mass spectrometry and was validated as a highly prevalent autoantigen by enzyme-linked immunosorbent assay. Analysis of Syt1 expression in retinas of ERU cases showed a downregulation in the majority of ERU affected retinas to 24%. Results pointed to a dysregulation of retinal neurotransmitter release in ERU. Identification of synaptotagmin-1, the first cell membrane associated autoantigen in this spontaneous autoimmune disease, demonstrated that examination of tissue fractions can lead to the discovery of previously undetected novel autoantigens. Further experiments will address its role in ERU pathology.

Bovine herpesvirus type 1 (BoHV-1), an alphaherpesvirus, is a major pathogen of cattle causing different syndromes such as infectious bovine rhinotracheitis (IBR), infectious pustular vulvovaginitis (IPV) and infectious balanoposthitis (IBP). BoHV-1 control programs have been initiated in several European countries including Germany. One of the major components is the vaccination with inactivated or attenuated glycoprotein E (gE)-deleted live marker vaccines. The aim of this study was the development of a triplex real-time polymerase chain reaction (qPCR) assay for the sensitive, specific and reliable BoHV-1 detection. A BoHV-1-specific glycoprotein D (gD) assay was combined with a gE-specific qPCR system for differentiation between wild-type strains and gE-negative vaccine virus strains. Finally, an internal control based on amplification of the bovine beta-actin gene was introduced to verify efficient DNA extraction and PCR amplification. The analytical sensitivity of the triplex BoHV-1 qPCR enables the detection of 10 genome copies per reaction. Furthermore, the sensitivity of the newly developed qPCR assay was compared to an OIE-validated qPCR and the “gold standard” method of virus isolation in cell culture using 10-fold dilution series of BoHV-1 in extended semen as well as in cell culture medium. For all preparations, the tested qPCR assays showed comparable results and the sensitivity of the triplex qPCR was equal or even greater than that of virus isolation. A broad spectrum of reference strains and field isolates was detected reliably. The specificity of the test was confirmed using nasal swabs, semen and different organ materials of BoHV-1-negative cattle. Bovine herpesviruses type 2, 4 and 5 and further ruminant herpesviruses, namely bubaline herpesvirus (BuHV-1), caprine herpesvirus type 1 and cervine herpesvirus type 1 and 2 (CvHV-1, -2) were tested as well. The close genetic and serological relationship between these viruses combined with their ability to infect bovines may interfere with diagnostics resulting in false-positive results. All non-BoHV-1 herpesviruses were negative in the gD-specific assay, while BuHV-1, CvHV-1 and -2 were tested positive by the gE-specific qPCR. Consequently, the triplex qPCR offers for the first time the possibility to detect some related herpesviruses and distinguish them from BoHV-1 in addition to the simultaneous differentiation of BoHV-1 wild-type and gE-deleted vaccine strains.

Background: Murine gammaherpesvirus 68 (MHV-68) is used as a model to study the function of gammaherpesvirus glycoproteins. gp150 of MHV-68, encoded by open reading frame M7, is a positional homolog of gp350/220 of EBV and of gp35/37 of KSHV. Since it had been proposed that gp350/220 of EBV might be a suitable vaccine antigen to protect from EBV-associated diseases, gp150 has been applied as a model vaccine in the MHV-68 system. When analyzing the function of gp150, previous studies yielded conflicting results on the role of gp150 in latency amplification, and disparities between the mutant viruses which had been analyzed were blamed for the observed differences. Results: To further develop MHV-68 as model to study the function of gammaherpesvirus glycoproteins in vivo, it is important to know whether gp150 contributes to latency amplification or not. Thus, we re-evaluated this question by testing a number of gp150 mutants side by side. Our results suggest that gp150 is dispensable for latency amplification. Furthermore, we investigated the effect of vaccination with gp150 using gp150-containing exosomes. Vaccination with gp150 induced a strong humoral and cellular immune response, yet it did not affect a subsequent MHV-68 challenge infection. Conclusions: In this study, we found no evidence for a role of gp150 in latency amplification. The previously observed contradictory results on the role of gp150 in latency amplification were not related to differences between the mutant viruses which had been used.

Background: Ehrlichia species are the etiological agents of emerging and life-threatening tick-borne human zoonoses that inflict serious and fatal infections in companion animals and livestock. The aim of this paper was to phylogeneticaly characterise a new species of Ehrlichia isolated from Rhipicephalus (Boophilus) microplus from Minas Gerais, Brazil. Methods: The agent was isolated from the hemolymph of Rhipicephalus (B.) microplus engorged females that had been collected from naturally infested cattle in a farm in the state of Minas Gerais, Brazil. This agent was then established and cultured in IDE8 tick cells. The molecular and phylogenetic analysis was based on 16S rRNA, groEL, dsb, gltA and gp36 genes. We used the maximum likelihood method to construct the phylogenetic trees. Results: The phylogenetic trees based on 16S rRNA, groEL, dsb and gltA showed that the Ehrlichia spp isolated in this study falls in a clade separated from any previously reported Ehrlichia spp. The molecular analysis of the ortholog of gp36, the major immunoreactive glycoproteins in E. canis and ortholog of the E. chaffeensis gp47, showed a unique tandem repeat of 9 amino acids (VPAASGDAQ) when compared with those reported for E. canis, E. chaffeensis and the related mucin-like protein in E. ruminantium. Conclusions: Based on the molecular and phylogenetic analysis of the 16S rRNA, groEL, dsb and gltA genes we concluded that this tick-derived microorganism isolated in Brazil is a new species, named E. mineirensis (UFMG-EV), with predicted novel antigenic properties in the gp36 ortholog glycoprotein. Further studies on this new Ehrlichia spp should address questions about its transmissibility by ticks and its pathogenicity for mammalian hosts.

In Part 2 of this Episode on Acute Coronary Syndromes Risk Stratification & Management, the evidence for various medications for ACS, from supplemental oxygen to thrombolytics are debated, and decision making around reperfusion therapy for STEMI as well as NSTEMI are discussed. Finally, there is a discussion on risk stratification of low risk chest pain patients and all it's attendant challenges as well as disposition and follow-up decisions. Dr. Eric Letovsky, the Head of the CCFP(EM) Program at the University of Toronto, Dr. Mark Mensour & Dr. Neil Fam, an interventional cardiologist answer questions like: What is the danger of high flow oxygen in the setting of ACS? When, if ever, should we be using IV B-blockers in AMI patients? How can you predict, in the ED, who might go on to have an urgent CABG, in which case Clopidogrel is contra-indicated? Which anticoagulant is best for unstable angina, NSTEMI and STEMI - unfractionated heparin (UFH), low molecular weight heparin (LMWH), or fonduparinux? Is there currenly any role for Glycoprotein 2b3a Inhibitors in ACS in the ED? When is thrombolysis better than PCI for STEMI? When should we consider facilitated angioplasty and rescue angioplasty? Which low risk chest pain patients require an early stress test? CT coronary angiography? Stress Echo? Admission to a Coronary Decision Unit (CDU)? and many more.......

In Part 2 of this Episode on Acute Coronary Syndromes Risk Stratification & Management, the evidence for various medications for ACS, from supplemental oxygen to thrombolytics are debated, and decision making around reperfusion therapy for STEMI as well as NSTEMI are discussed. Finally, there is a discussion on risk stratification of low risk chest pain patients and all it's attendant challenges as well as disposition and follow-up decisions. Dr. Eric Letovsky, the Head of the CCFP(EM) Program at the University of Toronto, Dr. Mark Mensour & Dr. Neil Fam, an interventional cardiologist answer questions like: What is the danger of high flow oxygen in the setting of ACS? When, if ever, should we be using IV B-blockers in AMI patients? How can you predict, in the ED, who might go on to have an urgent CABG, in which case Clopidogrel is contra-indicated? Which anticoagulant is best for unstable angina, NSTEMI and STEMI - unfractionated heparin (UFH), low molecular weight heparin (LMWH), or fonduparinux? Is there currenly any role for Glycoprotein 2b3a Inhibitors in ACS in the ED? When is thrombolysis better than PCI for STEMI? When should we consider facilitated angioplasty and rescue angioplasty? Which low risk chest pain patients require an early stress test? CT coronary angiography? Stress Echo? Admission to a Coronary Decision Unit (CDU)? and many more....... The post Episode 15 Part 2: Acute Coronary Syndromes Management appeared first on Emergency Medicine Cases.

In this NewsFlash, we find out how lasers can cut complications in cataract surgery, why some people are allergic to wine and discover how scientists have solved a long-standing mystery relating the structure of the placenta. Plus, we get a rundown of the highlights from the world's biggest neuroscience meeting, Neuroscience 2010 in San Diego.

Hintergrund Die Schwangerschaft ist ein natürliches, erfolgreiches Modell immunologischer Toleranz [1]. Das Kind, dessen genetisches Material zu 50% allogen ist, wird während der Zeit seiner intrauterinen Entwicklung vom mütterlichen Immunsystem akzeptiert. Ein Zustand, der fundamentalen Regeln der Transplantationsimmunologie (Selbst-Fremd Erkennung) widerspricht. Beim Aufbau der fetomaternalen Grenzfläche wachsen fetale Zellen (sog. Trophoblasten) in die mütterliche Uterusschleimhaut ein, arrodieren mütterliche Blutgefäße und bilden in der reifen Plazenta die Auskleidung eines mütterlichen Blutsees [2]. Dieses trophoblastäre Synzytium ist also gleichermaßen fetales Epithel wie plazentares Endothel und interagiert mit mütterlichen Leukozyten [3]. Die Frage immunologischer Toleranz ist jedoch auch in der Kanzerogenese und in der Etablierung des Tumormikromilieus von entscheidender Bedeutung [4]. Die Entstehung und immunologische Etablierung eines malignen Tumors ist die gemeinsame Endstrecke eines letztendlich ungerichteten Prozesses. Die Charakteristika einer malignen Erkrankung sind daher in hohem Maße individuell. Ausdruck dessen ist die zunehmende Hinwendung zu individualisierten Krebstherapien (sog. targeted therapies) wie sie z.B. auch immuntherapeutische Ansätze darstellen [5]. Der spezifische Aufbau immunologischer Toleranz an der Tumor-Stroma Grenzfläche ist auf Grund der großen interindividuellen Unterschiede im humanen System nur schwer nachzuvollziehen. Demgegenüber verläuft der Aufbau des spezifischen immunologischen Mikromilieus an der fetomaternalen Grenzfläche entlang geordneter Bahnen, deren Erforschung allgemeine Prinzipien der Toleranzentwicklung im humanen System zu Tage fördern könnte. Das vorliegende Habilitationsprojekt widmet sich Mechanismen immunologischer Toleranz und ihrer Durchbrechung am Plazenta- und Tumor-Modell. Bisher bearbeitete Fragestellungen Dendritische Zellen (DC) besetzen eine zentrale Schaltstelle des Immunsystems und können einerseits antigenspezifische cytotoxische T-Zell Immunantworten induzieren, andererseits im steady state für immunologische Toleranz sorgen [6, 7]. Ihre Eigenschaft der spezifischen Immuninduktion prädestinieren DC für eine individualisierten Krebs-Immuntherapie, deren immunogene Eigenschaften wir in Zellkultur-Modellen beurteilen konnten [8]. Apoptose als der physiologische Zelluntergang induziert peripher (d.h. außerhalb lymphatischer Organe) vermittelt über DC immunologische Toleranz. Apoptotisch zu Grunde gegangene Zellen werden dabei von DC aufgenommen und so aufbereitet, dass ihre charakteristische Proteinstruktur von cytotoxischen T-Zellen erkannt wird. Zusätzliche Signale bestimmen nun, ob diesen T-Zellen angezeigt wird, die betreffende Proteinstruktur zu tolerieren oder dagegen eine Immunantwort zu induzieren [9, 10]. Eine solche Immunantwort ist hochspezifisch und bietet sich daher als targeted therapy in der Krebstherapie an [11]. Wir konnten in diesem Zusammenhang den Weg apoptotischen Tumormaterials in Zellkultur-DC genauer verfolgen und als Einflussfaktor der folgenden Immunantwort näher charakterisieren [12]. Neben der Charakteristik des aufgenommen Zellmaterials ist die Eigenart jener zusätzlichen Signale (den von P. Matzinger erstmals so genannten „Gefahrensignalen“) von entscheidender Bedeutung für die Immunantwort. Gefahrensignale sind immunologische Muster, die eine Infektion oder Zellschädigung kennzeichnen und eine pathogen- und gewebsspezifische Immunreaktion nach sich ziehen. So konnten wir mit Adenosin-Triphosphat ein obligat intrazelluläres Molekül als ein solches Gefahrensignal charakterisieren [13]. An die Stelle der klassischen Unterscheidung zwischen Selbst- und Fremd tritt damit die Unterscheidung zwischen Gefahr und Nicht-Gefahr. Der Zustand der Nicht-Gefahr der sog. steady state wird in diesem Modell mit der Induktion einer gewebsspezifischen Toleranz andererseits jede Schädigung durch ein Pathogen durch eine auf Pathogen und Gewebe maßgeschneiderte Immunreaktion beantwortet. Das lokale Gewebe ist in diesem Modell Auslöser und Ziel der Immunantwort während im klassischen Selbst Fremd Modell das Immunsystem der Auslöser und das Gewebe lediglich das Zielorgan darstellt [14]. Bonney und Matzinger konnten im Maus-Modell zeigen, dass diese Unterscheidung zwischen intakter systemischer Immunantwort und lokaler Immuntoleranz auch auf das klassische Paradoxon der Fortpflanzung zutrifft [15]. Hieran anknüpfend konnten wir im humanen in vitro System Glycodelin, ein progesteronabhängiges Glycoprotein der fetomaternalen Grenzfläche, als einen solchen lokalen Faktor im Hinblick auf eine Toleranzinduktion in DC nachweisen [16]. Im Rahmen hypertensiver Schwangerschaftserkrankungen gelang es zudem erstmals, eine Rolle des Aktivierungszustandes dendritischer Zellen am Patientenmaterial zu zeigen [17]. In der Frühschwangerschaft konnten wir außerdem nachweisen, dass eine verminderte Expression von Glycodelin mit einem Abortgeschehen assoziiert ist [18]. Das ansonsten schwangerschaftsspezifische lokal immunsuppressive Glycodelin wird jedoch auch von gynäkologischen Tumoren im Rahmen der Karzinogenese zur lokalen Immunsuppression benutzt. Im Ovarialkarzinom konnten wir Glycodelin-abhängige Immunsupression auf Zellkultur-DC ebenso nachweisen wie eine Korrelation mit dem Östrogen- und Progesteronrezeptorstatus als prädiktivem Faktor in histologischen Schnitten des Mammakarzinoms [19; 21] Eine der zentralen Aufgaben der fetomaternalen Grenzfläche ist die Trennung des mütterlichen und kindlichen Blutkreislaufes. Bei einem Leck dieser Trennung kann es zum Ausbluten des Feten in den Kreislauf der Mutter kommen. Mechanische Belastung wurde lange Zeit als ein Hauptriskofaktor für dieses seltene, jedoch in seinem Verlauf oftmals sehr dramatische Krankheitsbild gesehen. In einer Beobachtungsstudie konnten wir mit einem sehr sensitiven durchflußzytometrischen Testverfahren jedoch eine plazentare Entzündungsreaktion als bislang nicht beschriebenen Risikofaktor etablieren [22]. Ein lange Zeit mit besonderer Aufmerksamkeit verfolgter Risikofaktor einer fetomaternalen Transfusion Besonderes war die mechanische Belastung im Rahmen der sog. äußeren Wendung, bei der ein Kind am Ende der Schwangerschaft aus Beckenendlage durch Manipulation von außen in eine Schädellage gedreht wird, um eine vaginale Geburt aus Schädellage zu ermöglichen. Die Sicherheit des Kindes steht dabei naturgemäß an oberster Stelle. In einer klinischen Beobachtungs-Studien konnten wir mit einem sehr sensitiven durchflußzytometrischen Testverfahren dazu beitragen die Volumina der fetomaternalen Transfusion im Rahmen einer äußeren Wendung mit o.g. Testverfahren genauer zu quantifizieren und den Einfluss der mechanischen Belastung auf die fetomaternale Transfusion damit zu relativieren [23].

Epilepsien zählen zu den häufigsten chronischen neurologischen Erkrankungen des Menschen, aber auch der Hunde und Katzen. Sie sind mit erheblichen Einschränkungen der Lebensqualität und Lebenserwartung verbunden und führen zu einer fortschreitenden Schädigung des gesamten Nervensystems. Gegenwärtig ist die wichtigste Therapieform der Epilepsie nach wie vor die Pharmakotherapie. Ungeachtet einer Vielzahl neuer Antiepileptika kann jedoch bei über 30% der Patienten mit Epilepsien durch eine medikamentelle Therapie keine ausreichende Kontrolle der Anfälle herbeigeführt werden, was neue Therapie- und Behandlungsstrategien unbedingt erforderlich macht. Eine, der im Zusammenhang mit einer Pharmakoresistenz am häufigsten erwähnten Thesen, ist die Multidrug-Transporter-Hypothese. Hier wird davon ausgegangen, dass es, durch an der Blut-Hirn-Schranke überexprimierte Efflux-Transporter, nicht zu einer ausreichend hohen Konzentration eines Antiepileptikums im neuronalen Gewebe kommt. Dem Multidrug-Transporter P-Glycoprotein (Pgp) wird dabei eine besondere Bedeutung beigemessen. Basierend auf der Multidrug-Transporter-Hypothese war es ein Ziel dieses Dissertationsvorhabenes die Regulation der anfallsinduzierten Pgp-Überexpression genauer zu charakterisieren, um daraus resultierend mögliche neue Angriffspunkte für eine Modulation der Pgp-Überexpression zu identifizieren. In einem ersten Teil der Dissertation wurde deshalb die Spezifität der molekularen Veränderungen des verwendeten Tiermodells untersucht. Hierfür wurde die Wirkung von spontanen Anfällen und eines SE auf die Pgp-Expression im Kaninen Gehirn untersucht. Die Untersuchungen zeigten, dass wie im Tiermodell auch nach einem spontan ablaufenden SE eine deutliche Induktion des Mutlidrug-Transporters Pgp an der Blut-Hirn-Schranke nachweisbar ist. Da bisherige Forschungsarbeiten unserer Arbeitsgruppe auf eine besondere Bedeutung des Glutamat/NMDA-Rezeptor/COX 2-Weges für die anfallsinduzierte Pgp-Überexpression in Endothelzellen der Blut-Hirn-Schranke hinweisen, wurden in diesem Dissertationsprojekt eingehende Untersuchungen durchgeführt, die darauf abzielten diesen Signalweg weiter aufzuklären. Durch vorausgegangene in vitro Untersuchungen wurde davon ausgegangen, dass die weitere Pgp-Induktion über den EP-1-Rezeptor von PGE2 abläuft. Diese Annahme konnte durch die Verwendung des EP-1-Rezeptor Hemmers SC- 51089 in vivo im Rahmen dieses Dissertationsprojektes bestätigt werden. Ein neuroprotektiver Effekt dieser Substanz konnte jedoch bei der verwendeten Konzentration von SC 51089 in diesem SE-Modell im Hilus nicht nachgewiesen werden. Weiterhin wurden Untersuchungen zu möglichen Downstream-Elementen des EP-1-Rezeptors angeschlossen, den Transkriptionsfaktoren YB-1 und ZONAB. Erstmalig konnte eine Expression dieser Transkriptionsfaktoren im adulten Gehirn unterschiedlicher Spezies nachgewiesen werden, die jedoch vermutlich nicht mit der anfallsinduzierten Pgp-Überexpression im Zusammenhang steht. Die bisherigen Ergebnisse demonstrieren jedoch, dass durch eine genaue Kenntnis, der an der Regulation der Pgp-Überexpression beteiligten Faktoren, eine Modulation bzw. Inhibtion der anfallsinduzierten Induktion von Pgp möglich ist und dadurch eine hoffnungsvolle Möglichkeit zur Behandlung und zur Prävention einer Multidrug-Transporter basierten Pharmakoresistenz geschaffen werden kann.

Audio Journal of Cardiovascular Medicine, November 5th, 2007 Reporting from: American Heart Association Scientific Sessions, 4-7 November, 2007, Orlando, Florida Eptifibatide After PCI: Brief Infusion Is Just As Good ANTHONY FUNG, University of British Columbia, Vancouver REFERENCE: Late Breaking Clinical Trials Session I, AHA 2007 Brief courses of antiplatelet therapy with the glycoprotein 2b 3a inhibitor are just as effective as longer infusions for patients receiving non-emergency percutaneous intervention. So said Anthony Fung at the first session of Late breaking Clinical Trials held at the American Heart Association's annual meeting in Orlando. He talked with Peter Goodwin about his findings.

Background: The pregnancy-specific glycoprotein (Psg) genes encode proteins of unknown function, and are members of the carcinoembryonic antigen (Cea) gene family, which is a member of the immunoglobulin gene (Ig) superfamily. In rodents and primates, but not in artiodactyls (even-toed ungulates/hoofed mammals), there have been independent expansions of the Psg gene family, with all members expressed exclusively in placental trophoblast cells. For the mouse Psg genes, we sought to determine the genomic organisation of the locus, the expression profiles of the various family members, and the evolution of exon structure, to attempt to reconstruct the evolutionary history of this locus, and to determine whether expansion of the gene family has been driven by selection for increased gene dosage, or diversification of function. Results: We collated the mouse Psg gene sequences currently in the public genome and expressed-sequence tag (EST) databases and used systematic BLAST searches to generate complete sequences for all known mouse Psg genes. We identified a novel family member, Psg31, which is similar to Psg30 but, uniquely amongst mouse Psg genes, has a duplicated N1 domain. We also identified a novel splice variant of Psg16 (bCEA). We show that Psg24 and Psg30/Psg31 have independently undergone expansion of N-domain number. By mapping BAC, YAC and cosmid clones we described two clusters of Psg genes, which we linked and oriented using fluorescent in situ hybridisation ( FISH). Comparison of our Psg locus map with the public mouse genome database indicates good agreement in overall structure and further elucidates gene order. Expression levels of Psg genes in placentas of different developmental stages revealed dramatic differences in the developmental expression profile of individual family members. Conclusion: We have combined existing information, and provide new information concerning the evolution of mouse Psg exon organization, the mouse Psg genomic locus structure, and the expression patterns of individual Psg genes. This information will facilitate functional studies of this complex gene family.

This study investigated the immunobiology of MOG-induced EAE in the DA rat, an animal model, which reproduces the immunopathology of the type II MS lesion (Lucchinetti et al., 2000). A newly established immunisation protocol results in a highly synchronised biphasic form of EAE, which mimics the disease course of secondary progressive MS, albeit in a strongly abbreviated time course (Figure 3.1.1). This study demonstrates that MOG-specific autoantibodies are responsible for initiating clinical relapse and driving disease progression. On the background of mild, sub-clinical inflammatory activity in the CNS, pathogenic antibodies enter the CNS and mediate demyelination, a process that in turn amplifies the local inflammatory response (Figure 3.1.14 A). It should however be noted that lethal clinical relapses may also occur in the absence of a pathogenic antibody response if an inflammatory lesion develops in a region of the CNS that is particularly sensitive to damage, or where it may perturb vital functions, such as the brain stem. Although antibodies have been shown to amplify the severity of ongoing clinical EAE (Schluesener et al., 1987; Linington et al., 1988; Lassmann et al., 1988), firm evidence for a role in driving relapse and disease progression was missing. This study has now established this principal, which in all probability is relevant to our understanding of the pathogenesis of severe, steroid non-responsive relapses in MS patients. However, this model of EAE is an artificial system, in which the role of antibody is only apparent because of the different kinetics of MOG-specific T and B cell responses. In MS we still have to answer two crucial questions, namely the identity of the autoantigens targeted by the demyelinating antibody response, and the factors that may trigger this response. MOG is the only myelin protein known to initiate a demyelinating antibody response in EAE, and MOG-induced EAE has provided a valuable tool to identify the role of pathogenic autoantibodies in immune mediated demyelination. However, there is a major discrepancy between the proportion of MS patients with pathogenic MOG-specific antibodies in their circulation (5%; Haase et al., 2000) and the frequency of patients with pathological changes suggestive of antibody-mediated pathomechanisms (>50%; Lucchinetti et al., 2000). This discrepancy may in part be accounted for by the absorption of the pathogenic antibodies into the CNS, which will lead to a dramatic reduction of the antibody titre in the periphery, as demonstrated in section 3.1.3.4 of this study. On the other hand, it is unlikely that MOG is the only target autoantigen, which is exposed on the myelin surface and can therefore initiate a demyelinating autoantibody response. The identification of potential targets is a prerequisite to develop diagnostic kits to identify those patients with pathogenic autoantibody responses and then provide an appropriate therapy such as plasma exchange, or immuno-absorption. As demonstrated in this study, DNA vaccination using a plasmid encoding a myelin antigen is one approach to generate high titre autoantibody responses directed against the native protein. The pathogenicity of this antibody response can then be assayed in the same animal by inducing EAE. This method circumvents problems such as purity, yield and denaturation, all of which complicate any study using antigens isolated from the CNS or generated using recombinant technologies. Coupling this approach to a proteomics based analysis of the myelin membrane and reverse genomics to identify candidate gene products provides the means to map out those protein antigens that can be targeted by a demyelinating autoantibody response. The feasibility of this concept is currently being tested in the rat using PLP and MAG as myelin components that may in certain circumstances provoke a pathogenic autoantibody response. Such an analysis will, however, not detect pathogenic antibody responses to glycolipid antigens, which are major target autoantigens in a number of diseases affecting the peripheral nervous system such as Guillain Barré syndrome (GBS). In GBS a pathogenic antibody response to gangliosides appears to be triggered by infections with particular serotypes of Campylobacter jejuni (Fredman, 1998; Willison and O´Hanlon, 1999). In the majority of patients these antibody responses are an acute phenomenon and disappear as the patients recover (Hahn, 1998). It is conceivable that a similar mechanism is responsible for the initiation of severe relapses in some MS patients, if an infection triggers a cross-reactive antibody response to a surface glycolipid epitope. This would induce an episode of acute CNS demyelination that would not be immediately responsive to immunosuppressive therapy, as tissue damage and amplification of the local inflammatory response would be driven by the pre- existing antibody response. Analysis of the autoantibody responses in MS should therefore be extended to examine lipid as well as protein autoantigens. Such studies should also not be restricted to myelin, but also address the question of responses to other structures such as the axon and oligodendrocyte progenitor cells. Such autoantibody responses are however only conditionally pathogenic, in other words their pathogenic potential is only expressed if they can enter the CNS across the blood brain barrier (BBB)(Litzenburger et al., 1998; Bourquin et al., 2000). In EAE the inflammatory insult to the CNS is responsible for the disruption of BBB function and the entry of antibody into the nervous system. MS is characterised by repeated episodes of CNS inflammation but what initiates and maintains this response is unclear. The observation, that DA rats develop a similar, although eventually self-limiting response in the CNS after immunisation with MOG-peptide in CFA provides a model to investigate the immuno-regulatory deficit(s) responsible for chronic CNS inflammation. The disease model is very reproducible with >90% of animals relapsing after peptide immunisation as opposed to

The sequence of the gene encoding glycoprotein H (gH) of murine cytomegalovirus (MCMV) strain Smith was determined and compared with the sequence of the gH of MCMV strain K181. Transcriptional analysis showed that gH is encoded by a large mRNA of 5.0 kb, which is synthesized late in infection. A recombinant vaccinia virus expressing the MCMV gH open reading frame was constructed (Vac-gH). Anti-MCMV serum precipitated a protein of 87K from Vac-gH-infected cells. Reactivity with a monoclonal antibody showed the identity of the MCMV gH with a 87K envelope glycoprotein described previously by Loh and Qualtiere. Immunization of mice with the Vac-gH recombinant gave rise to an anti-gH serum, which neutralized MCMV without complement in vitro.

As a basis towards a better understanding of the role of the pregnancy-specific glycoprotein (PSG) family in the maintenance of pregnancy, detailed investigations are described on the expression of a recently identified rat PSG gene (rnCGM1) at the mRNA and protein levels. Using specific oligonucleotide primers, rnCGM1 transcripts were identified after reverse transcription, polymerase chain reaction, and hybridization with a radiolabelled, internal oligonucleotide. Transcripts were only found in significant amounts in placenta. In situ hybridization visualized rnCGM1 transcripts at day 14 post coitum (p.c.), in secondary trophoblast giant cells and in the spongiotrophoblast. Only those secondary giant cells lining the maternal decidua were positive. In contrast, primary giant cells did not contain rnCGM1 mRNA. At day 18 p.c., rnCGM1. transcripts were almost exclusively detectable in the spongiotrophoblast. No rnCGM1 transcripts were found in rat embryos of these two developmental stages. Rabbit antisera were generated against the amino-terminal immunoglobulin variable-like domain and against a synthetic peptide containing the last 13 carboxy-terminal amino acids of rnCGM1. Bothe antisera recognized a 124 kDa protein in day 18 rat placental extracts as identified by Western blot analysis. The anti-peptide antiserum recognized a 116 kDa protein in the serum of a 14 day p.c. pregnant rat that is absent from the sera of non-pregnant females. Taken together, these results confirm exclusive expression of rnCGM1 in the rat trophoblast, but unlike human PSG, negligible or no expression is found in other organs, such as fetal liver or salivary glands, indicating a more specialized function of rnCGM1. Its spatiotemporal expression pattern is conducive with a potential role of PSG in protecting the fetus against the maternal immune system and/or in regulating the invasive growth of trophoblast cells.

Computer simulation of the conformations of short antigenic peptides (&lo residues) either free or bound to their receptor, the major histocompatibility complex (MHC)- encoded glycoprotein H-2 Ld, was employed to explain experimentally determined differences in the antigenic activities within a set of related peptides. Starting for each sequence from the most probable conformations disclosed by a pattern-recognition technique, several energyminimized structures were subjected to molecular dynamics simulations (MD) either in vacuo or solvated by water molecules. Notably, antigenic potencies were found to correlate to the peptides propensity to form and maintain an overall a-helical conformation through regular i,i + 4 hydrogen bonds. Accordingly, less active or inactive peptides showed a strong tendency to form i,i+3 hydrogen bonds at their Nterminal end. Experimental data documented that the C-terminal residue is critical for interaction of the peptide with H-2 Ld. This finding could be satisfactorily explained by a 3-D Q.S.A.R. analysis postulating interactions between ligand and receptor by hydrophobic forces. A 3-D model is proposed for the complex between a high-affinity nonapeptide and the H- 2 Ld receptor. First, the H-2 Ld molecule was built from X-ray coordinates of two homologous proteins: HLA-A2 and HLA-Aw68, energyminimized and studied by MD simulations. With HLA-A2 as template, the only realistic simulation was achieved for a solvated model with minor deviations of the MD mean structure from the X-ray conformation. Water simulation of the H-2 Ld protein in complex with the antigenic nonapeptide was then achieved with the template- derived optimal parameters. The bound peptide retains mainly its a-helical conformation and binds to hydrophobic residues of H-2 Ld that correspond to highly polymorphic positions of MHC proteins. The orientation of the nonapeptide in the binding cleft is in accordance with the experimentally determined distribution of its MHC receptor-binding residues (agretope residues). Thus, computer simulation was successfully employed to explain functional data and predicts a-helical conformation for the bound peptide

The gene encoding glycoprotein B (gB) of murine cytomegalovirus (MCMV) strain Smith was identified, sequenced, and expressed by recombinant vaccinia virus. The gB gene was found adjacent to the polymerase gene, as it is in the genome of human cytomegalovirus (HCMV). The open reading frame consists of 2,784 nucleotides capable of encoding a protein of 928 amino acids. Comparison with gB homologs of other herpesviruses revealed a high degree of homology. The similarity between the MCMV gB and the HCMV gB is most prominent, since 45% of the amino acids are identical. In addition, all cysteine residues are at homologous positions, indicating a similar tertiary structure of the two proteins. In contrast to HCMV, the MCMV gB mRNA is a true late transcript. A recombinant vaccinia virus expressing the MCMV gB gene has been constructed (Vac-gB). Antibodies raised against the Vac-gB recombinant precipitated proteins of 130, 105, and 52 kDa from MCMV-infected cells. The identity of the MCMV gB with the major envelope glycoprotein of MCMV described by Loh et al. was shown (L. C. Loh, N. Balachandran, and L. F. Qualtiere, Virology 166:206-216, 1988). Immunization of mice with the Vac-gB recombinant gave rise to neutralizing antibodies.

The nucleotide and complete amino acid sequence for the human ß2-glycoprotein I (ß,I) was derived by sequencing the cDNA clone pB2I- 1. In addition to the 326 amino acid residues of the mature protein this clone codes for a putative leader peptide and contains sequence representing 5' and 3' untranslated regions. When this amino acid sequence was compared with the previously published primary sequence, three major amino acid substitutions were found, two involving cysteine residues. These substitutions lead to a new alignment of the complement control protein (CCP) repeats present in fl2I and a prediction of the complete disulphide bond organization. Northern-blot analysis indicates that hepatocytes are a major site of biosynthesis for this protein. A transcription signal of about 1.5 kb was detected by using RNA from HepG2 cells

The amino acid sequence of the acrosin-trypsin inhibitor HUSI-II from human seminal plasma is presented which unequivocally identifies HUSI-II as being of Kazal-type. In addition, the HUSI-II sequence shows a striking similarity to the middle part of glycoprotein hormone β-subunits thus revealing a hitherto unknown structural and evolutionary relationship between Kazal-type inhibitors and glycoprotein hormones

The pregnancy-specific glycoprotein (PSG) genes encode a group of proteins which are found in large amounts in placenta and maternal serum. In situ hybridization analyses of metaphase chromosomes reveal that all the human pregnancy-specific glycoprotein (PSG) genes are located on the long arm of chromosome 19 (19q13.2–13.3), overlapping the region containing the closely-related carcinoembryonic antigen (CEA) gene subgroup. Higher resolution analyses indicate that the PSG genes are closely linked within an 800kb SacII restriction endonuclease fragment. This has been confirmed through restriction endonuclease mapping and DNA sequence analyses of isolated genomic clones, which show that at least some of these genes are located in very close proximity. Further, these studies have helped to identify a new member of the PSG gene sub-family (PSG7). DNA/RNA hybridization analyses, using gene-specific oligonucleotide probes based on published sequences, showed that five from six PSG genes tested are coordinately transcribed in the placenta. Due to the close proximity of these genes and their coordinated expression pattern, common transcriptional regulatory elements may exist.

The pregnancy-specific glycoprotein (PSG) genes constitute a subgroup of the carcinoembryonic antigen (CEA) gene family. Here we report the cloning of four cDNAs coding for different members of the PSG family from a human fetal liver cDNA library. They are derived from three closely related genes (PSG1, PSG4 and PSG6). Two of the cDNA clones represent splice variants of PSG1 (PSG1a, PSG1d) differing in their C-terminal domain and 3′-untranslated regions. All encoded proteins show the same domain arrangement (N-RA1-RA2-RB2-C). Transcripts of the genes PSG1 and PSG4 could be detected in placenta by hybridization with gene-specific oligonucleotides. Expression of cDNA in a mouse and monkey cell line shows that the glycosylated PSG1a protein has a Mr of 65–66 kD and is released from the transfected cells. Sequence comparisons in the C-terminal domain and the 3′-untranslated regions of CEA/PSG-like genes suggests a complex splicing pattern to exist for various gene family members and a common evolutionary origin of these regions