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https://www.readbyqxmd.com/read/28671821/the-t-cell-receptor-can-bind-to-the-peptide-bound-major-histocompatibility-complex-and-uncomplexed-beta-2-microglobulin-through-distinct-binding-sites
#1
Patrick Sascha Merkle, Melita Irving, Song Hongjian, Mathias Ferber, Thomas J D Jørgensen, Kirsten Scholten, Immanuel Florian Luescher, George Coukos, Vincent Zoete, Michel A Cuendet, Olivier Michielin, Kasper D Rand
T-cell receptor (TCR)-mediated recognition of peptide-bound major histocompatibility complex (pMHC) initiates an adaptive immune response against antigen-presenting target cells. The recognition events take place at the TCR-pMHC interface and their implications on TCR conformation and dynamics are controversial. Here, we have measured the time-resolved hydrogen/deuterium exchange (HDX) of a soluble TCR in the presence and absence of its cognate pMHC by mass spectrometry to delineate the impact of pMHC binding on solution-phase structural dynamics in the TCR...
July 3, 2017: Biochemistry
https://www.readbyqxmd.com/read/28614804/in-vivo-kinetics-and-nonradioactive-imaging-of-rapidly-proliferating-cells-in-graft-versus-host-disease
#2
Nataliya P Buxbaum, Donald E Farthing, Natella Maglakelidze, Martin Lizak, Hellmut Merkle, Andrea C Carpenter, Brittany U Oliver, Veena Kapoor, Ehydel Castro, Gregory A Swan, Liliane M Dos Santos, Nicolas J Bouladoux, Catherine V Bare, Francis A Flomerfelt, Michael A Eckhaus, William G Telford, Yasmine Belkaid, Remy J Bosselut, Ronald E Gress
Hematopoietic stem cell transplantation (HSCT) offers a cure for cancers that are refractory to chemotherapy and radiation. Most HSCT recipients develop chronic graft-versus-host disease (cGVHD), a systemic alloimmune attack on host organs. Diagnosis is based on clinical signs and symptoms, as biopsies are risky. T cells are central to the biology of cGVHD. We found that a low Treg/CD4+ T effector memory (Tem) ratio in circulation, lymphoid, and target organs identified early and established mouse cGVHD. Using deuterated water labeling to measure multicompartment in vivo kinetics of these subsets, we show robust Tem and Treg proliferation in lymphoid and target organs, while Tregs undergo apoptosis in target organs...
June 15, 2017: JCI Insight
https://www.readbyqxmd.com/read/28459440/thiolutin-is-a-zinc-chelator-that-inhibits-the-rpn11-and-other-jamm-metalloproteases
#3
Linda Lauinger, Jing Li, Anton Shostak, Ibrahim Avi Cemel, Nati Ha, Yaru Zhang, Philipp E Merkl, Simon Obermeyer, Nicolas Stankovic-Valentin, Tobias Schafmeier, Walter J Wever, Albert A Bowers, Kyle P Carter, Amy E Palmer, Herbert Tschochner, Frauke Melchior, Raymond J Deshaies, Michael Brunner, Axel Diernfellner
Thiolutin is a disulfide-containing antibiotic and anti-angiogenic compound produced by Streptomyces. Its biological targets are not known. We show that reduced thiolutin is a zinc chelator that inhibits the JAB1/MPN/Mov34 (JAMM) domain-containing metalloprotease Rpn11, a deubiquitinating enzyme of the 19S proteasome. Thiolutin also inhibits the JAMM metalloproteases Csn5, the deneddylase of the COP9 signalosome; AMSH, which regulates ubiquitin-dependent sorting of cell-surface receptors; and BRCC36, a K63-specific deubiquitinase of the BRCC36-containing isopeptidase complex and the BRCA1-BRCA2-containing complex...
July 2017: Nature Chemical Biology
https://www.readbyqxmd.com/read/28445466/human-pluripotent-stem-cells-recurrently-acquire-and-expand-dominant-negative-p53-mutations
#4
Florian T Merkle, Sulagna Ghosh, Nolan Kamitaki, Jana Mitchell, Yishai Avior, Curtis Mello, Seva Kashin, Shila Mekhoubad, Dusko Ilic, Maura Charlton, Genevieve Saphier, Robert E Handsaker, Giulio Genovese, Shiran Bar, Nissim Benvenisty, Steven A McCarroll, Kevin Eggan
Human pluripotent stem cells (hPS cells) can self-renew indefinitely, making them an attractive source for regenerative therapies. This expansion potential has been linked with the acquisition of large copy number variants that provide mutated cells with a growth advantage in culture. The nature, extent and functional effects of other acquired genome sequence mutations in cultured hPS cells are not known. Here we sequence the protein-coding genes (exomes) of 140 independent human embryonic stem cell (hES cell) lines, including 26 lines prepared for potential clinical use...
May 11, 2017: Nature
https://www.readbyqxmd.com/read/28442771/double-stranded-dna-induces-a-prothrombotic-phenotype-in-the-vascular-endothelium
#5
Erik Gaitzsch, Thomas Czermak, Andrea Ribeiro, Yvonn Heun, Monica Bohmer, Monika Merkle, Hanna Mannell, Christian Schulz, Markus Wörnle, Joachim Pircher
Double-stranded DNA (dsDNA) constitutes a potent activator of innate immunity, given its ability to bind intracellular pattern recognition receptors during viral infections or sterile tissue damage. While effects of dsDNA in immune cells have been extensively studied, dsDNA signalling and its pathophysiological implications in non-immune cells, such as the vascular endothelium, remain poorly understood. The aim of this study was to characterize prothrombotic effects of dsDNA in vascular endothelial cells. Transfection of cultured human endothelial cells with the synthetic dsDNA poly(dA:dT) induced upregulation of the prothrombotic molecules tissue factor and PAI-1, resulting in accelerated blood clotting in vitro, which was partly dependent on RIG-I signalling...
April 25, 2017: Scientific Reports
https://www.readbyqxmd.com/read/28088176/ttca-an-r-package-for-the-identification-of-differentially-expressed-genes-in-time-course-microarray-data
#6
Marco Albrecht, Damian Stichel, Benedikt Müller, Ruth Merkle, Carsten Sticht, Norbert Gretz, Ursula Klingmüller, Kai Breuhahn, Franziska Matthäus
BACKGROUND: The analysis of microarray time series promises a deeper insight into the dynamics of the cellular response following stimulation. A common observation in this type of data is that some genes respond with quick, transient dynamics, while other genes change their expression slowly over time. The existing methods for detecting significant expression dynamics often fail when the expression dynamics show a large heterogeneity. Moreover, these methods often cannot cope with irregular and sparse measurements...
January 14, 2017: BMC Bioinformatics
https://www.readbyqxmd.com/read/28004241/the-arabidopsis-tho-trex-component-tex1-functionally-interacts-with-mos11-and-modulates-mrna-export-and-alternative-splicing-events
#7
Brian B Sørensen, Hans F Ehrnsberger, Silvia Esposito, Alexander Pfab, Astrid Bruckmann, Judith Hauptmann, Gunter Meister, Rainer Merkl, Thomas Schubert, Gernot Längst, Michael Melzer, Marion Grasser, Klaus D Grasser
We identify proteins that associate with the THO core complex, and show that the TEX1 and MOS11 components functionally interact, affecting mRNA export and splicing as well as plant development. TREX (TRanscription-EXport) is a multiprotein complex that plays a central role in the coordination of synthesis, processing and nuclear export of mRNAs. Using targeted proteomics, we identified proteins that associate with the THO core complex of Arabidopsis TREX. In addition to the RNA helicase UAP56 and the mRNA export factors ALY2-4 and MOS11 we detected interactions with the mRNA export complex TREX-2 and multiple spliceosomal components...
February 2017: Plant Molecular Biology
https://www.readbyqxmd.com/read/27871100/biomechanical-response-of-military-booted-and-unbooted-foot-ankle-tibia-from-vertical-loading
#8
Frank A Pintar, Michael B Schlick, Narayan Yoganandan, Liming Voo, Andrew C Merkle, Michael Kleinberger
A new anthropomorphic test device (ATD) is being developed by the US Army to be responsive to vertical loading during a vehicle underbody blast event. To obtain design parameters for the new ATD, a series of non-injurious tests were conducted to derive biofidelity response corridors for the foot-ankle complex under vertical loading. Isolated post mortem human surrogate (PMHS) lower leg specimens were tested with and without military boot and in different initial foot-ankle positions. Instrumentation included a six-axis load cell at the proximal end, three-axis accelerometers at proximal and distal tibia, and calcaneus, and strain gages...
November 2016: Stapp Car Crash Journal
https://www.readbyqxmd.com/read/27863662/surface-assembled-poly-i-c-on-pegylated-plga-microspheres-as-vaccine-adjuvant-apc-activation-and-bystander-cell-stimulation
#9
Annina M Hafner, Blaise Corthésy, Marcus Textor, Hans P Merkle
Biodegradable poly(lactic-co-glycolic acid) (PLGA) microspheres are potential vehicles to deliver antigens for vaccination. Because they lack the full capacity to activate professional antigen presenting cells (APCs), combination with an immunostimulatory adjuvant may be considered. A candidate is the synthetic TLR3 ligand polyriboinosinic acid-polyribocytidylic acid, poly(I:C), which drives cell-mediated immunity. However, poly(I:C) has also been linked to the pathogenesis of autoimmunity, as affected by widespread stimulation of non-hematopoietic bystander cells...
November 30, 2016: International Journal of Pharmaceutics
https://www.readbyqxmd.com/read/27806710/gene-silencing-pathways-found-in-the-green-alga-volvox-carteri-reveal-insights-into-evolution-and-origins-of-small-rna-systems-in-plants
#10
Anne Dueck, Maurits Evers, Stefan R Henz, Katharina Unger, Norbert Eichner, Rainer Merkl, Eugene Berezikov, Julia C Engelmann, Detlef Weigel, Stephan Wenzl, Gunter Meister
BACKGROUND: Volvox carteri (V. carteri) is a multicellular green alga used as model system for the evolution of multicellularity. So far, the contribution of small RNA pathways to these phenomena is not understood. Thus, we have sequenced V. carteri Argonaute 3 (VcAGO3)-associated small RNAs from different developmental stages. RESULTS: Using this functional approach, we define the Volvox microRNA (miRNA) repertoire and show that miRNAs are not conserved in the closely related unicellular alga Chlamydomonas reinhardtii...
November 2, 2016: BMC Genomics
https://www.readbyqxmd.com/read/27692847/molecular-complexity-of-taxane-induced-cytotoxicity-in-prostate-cancer-cells
#11
Josef Mang, Konstanze Merkle, Martina Heller, Julia Schüler, Yanis Tolstov, Jielin Li, Markus Hohenfellner, Stefan Duensing
BACKGROUND: Taxanes are routinely used to treat men with advanced prostate cancer, yet their molecular mode of action is poorly characterized. Taxanes stabilize microtubules and may hence interfere with a plethora of cellular processes, most notably mitosis. However, prostate cancer is typically a slowly growing tumor suggesting that additional processes play a role in the response to taxanes. METHODS: Here, we analyzed the potential effect of taxanes on microtubuli-dependent intracellular transport and signaling processes, specifically, nuclear translocation of the androgen receptor and modulation of the RAS-RAF-MEK-ERK signaling cascade...
January 2017: Urologic Oncology
https://www.readbyqxmd.com/read/27576713/analysis-of-s-cerevisiae-rna-polymerase-i-transcription-in-vitro
#12
Michael Pilsl, Philipp E Merkl, Philipp Milkereit, Joachim Griesenbeck, Herbert Tschochner
RNA polymerase I (Pol I) activity is crucial to provide cells with sufficient amounts of ribosomal RNA (rRNA). Synthesis of rRNA takes place in the nucleolus, is tightly regulated and is coordinated with synthesis and assembly of ribosomal proteins, finally resulting in the formation of mature ribosomes. Many studies on Pol I mechanisms and regulation in the model organism S. cerevisiae were performed using either complex in vitro systems reconstituted from more or less purified fractions or genetic analyses...
2016: Methods in Molecular Biology
https://www.readbyqxmd.com/read/27532820/comprehensive-protocols-for-crispr-cas9-based-gene-editing-in-human-pluripotent-stem-cells
#13
David P Santos, Evangelos Kiskinis, Kevin Eggan, Florian T Merkle
Genome editing of human pluripotent stem cells (hPSCs) with the CRISPR/Cas9 system has the potential to revolutionize hPSC-based disease modeling, drug screening, and transplantation therapy. Here, we aim to provide a single resource to enable groups, even those with limited experience with hPSC culture or the CRISPR/Cas9 system, to successfully perform genome editing. The methods are presented in detail and are supported by a theoretical framework to allow for the incorporation of inevitable improvements in the rapidly evolving gene-editing field...
August 17, 2016: Current Protocols in Stem Cell Biology
https://www.readbyqxmd.com/read/27494133/identification-of-cell-type-specific-differences-in-erythropoietin-receptor-signaling-in-primary-erythroid-and-lung-cancer-cells
#14
Ruth Merkle, Bernhard Steiert, Florian Salopiata, Sofia Depner, Andreas Raue, Nao Iwamoto, Max Schelker, Helge Hass, Marvin Wäsch, Martin E Böhm, Oliver Mücke, Daniel B Lipka, Christoph Plass, Wolf D Lehmann, Clemens Kreutz, Jens Timmer, Marcel Schilling, Ursula Klingmüller
Lung cancer, with its most prevalent form non-small-cell lung carcinoma (NSCLC), is one of the leading causes of cancer-related deaths worldwide, and is commonly treated with chemotherapeutic drugs such as cisplatin. Lung cancer patients frequently suffer from chemotherapy-induced anemia, which can be treated with erythropoietin (EPO). However, studies have indicated that EPO not only promotes erythropoiesis in hematopoietic cells, but may also enhance survival of NSCLC cells. Here, we verified that the NSCLC cell line H838 expresses functional erythropoietin receptors (EPOR) and that treatment with EPO reduces cisplatin-induced apoptosis...
August 2016: PLoS Computational Biology
https://www.readbyqxmd.com/read/27197161/downregulation-of-the-tgf%C3%AE-pseudoreceptor-bambi-in-non-small-cell-lung-cancer-enhances-tgf%C3%AE-signaling-and-invasion
#15
Sebastian Marwitz, Sofia Depner, Dmytro Dvornikov, Ruth Merkle, Magdalena Szczygieł, Karin Müller-Decker, Philippe Lucarelli, Marvin Wäsch, Heimo Mairbäurl, Klaus F Rabe, Christian Kugler, Ekkehard Vollmer, Martin Reck, Swetlana Scheufele, Maren Kröger, Ole Ammerpohl, Reiner Siebert, Torsten Goldmann, Ursula Klingmüller
Non-small cell lung cancer (NSCLC) is characterized by early metastasis and has the highest mortality rate among all solid tumors, with the majority of patients diagnosed at an advanced stage where curative therapeutic options are lacking. In this study, we identify a targetable mechanism involving TGFβ elevation that orchestrates tumor progression in this disease. Substantial activation of this pathway was detected in human lung cancer tissues with concomitant downregulation of BAMBI, a negative regulator of the TGFβ signaling pathway...
July 1, 2016: Cancer Research
https://www.readbyqxmd.com/read/27150039/irak-m-expression-in-tumor-cells-supports-colorectal-cancer-progression-through-reduction-of-antimicrobial-defense-and-stabilization-of-stat3
#16
Rebecca Kesselring, Joachim Glaesner, Andreas Hiergeist, Elisabeth Naschberger, Helmut Neumann, Stefan M Brunner, Anja K Wege, Caroline Seebauer, Gudrun Köhl, Susanne Merkl, Roland S Croner, Christina Hackl, Michael Stürzl, Markus F Neurath, André Gessner, Hans-Juergen Schlitt, Edward K Geissler, Stefan Fichtner-Feigl
Colorectal cancer (CRC) is associated with loss of epithelial barrier integrity, which facilitates the interaction of the immunological microenvironment with the luminal microbiome, eliciting tumor-supportive inflammation. An important regulator of intestinal inflammatory responses is IRAK-M, a negative regulator of TLR signaling. Here we investigate the compartment-specific impact of IRAK-M on colorectal carcinogenesis using a mouse model. We demonstrate that IRAK-M is expressed in tumor cells due to combined TLR and Wnt activation...
May 9, 2016: Cancer Cell
https://www.readbyqxmd.com/read/27086952/arterial-thrombosis-in-the-context-of-hcv-associated-vascular-disease-can-be-prevented-by-protein-c
#17
Philipp Blüm, Joachim Pircher, Monika Merkle, Thomas Czermak, Andrea Ribeiro, Hanna Mannell, Florian Krötz, Alexander Hennrich, Michael Spannagl, Simone Köppel, Erik Gaitzsch, Markus Wörnle
Hepatitis C virus (HCV) infection is a major problem worldwide. HCV is not limited to liver disease but is frequently complicated by immune-mediated extrahepatic manifestations such as glomerulonephritis or vasculitis. A fatal complication of HCV-associated vascular disease is thrombosis. Polyriboinosinic:polyribocytidylic acid (poly (I:C)), a synthetic analog of viral RNA, induces a Toll-like receptor 3 (TLR3)-dependent arteriolar thrombosis without significant thrombus formation in venules in vivo. These procoagulant effects are caused by increased endothelial synthesis of tissue factor and PAI-1 without platelet activation...
April 18, 2016: Cellular & Molecular Immunology
https://www.readbyqxmd.com/read/26711702/the-cholesterol-transporter-abca1-is-expressed-in-stallion-spermatozoa-and-reproductive-tract-tissues
#18
M Merkl, R Ertl, S Handschuh, C Aurich, S Schäfer-Somi
In the present study, we assessed the presence of the ATP-binding-cassette (ABC) transporter molecules ABCA1 in spermatozoa of adult stallions and in testicular and epididymal tissue of prepubertal and adult stallions. For this purpose, semen samples from six fertile Shetland pony stallions aged 4 to 19 years were collected. Semen was collected from each stallion on three consecutive days. Ejaculates were analyzed immediately after collection, and only ejaculates meeting minimal requirements for fertile stallions were further evaluated...
April 1, 2016: Theriogenology
https://www.readbyqxmd.com/read/26473009/collective-cell-migration-of-smooth-muscle-and-endothelial-cells-impact-of-injury-versus-non-injury-stimuli
#19
Kaitlyn R Ammann, Katrina J DeCook, Phat L Tran, Valerie M Merkle, Pak K Wong, Marvin J Slepian
BACKGROUND: Cell migration is a vital process for growth and repair. In vitro migration assays, utilized to study cell migration, often rely on physical scraping of a cell monolayer to induce cell migration. The physical act of scrape injury results in numerous factors stimulating cell migration - some injury-related, some solely due to gap creation and loss of contact inhibition. Eliminating the effects of cell injury would be useful to examine the relative contribution of injury versus other mechanisms to cell migration...
2015: Journal of Biological Engineering
https://www.readbyqxmd.com/read/26446017/bk-virus-infection-activates-the-tnf%C3%AE-tnf-receptor-system-in-polyomavirus-associated-nephropathy
#20
Andrea Ribeiro, Monika Merkle, Nasim Motamedi, Hans Nitschko, Simone Köppel, Markus Wörnle
Polyomavirus-associated nephropathy due to BK virus infection (BKVAN) is recognized as an important cause of significant kidney transplant dysfunction often leading to renal graft loss. The activation of innate immune defense mechanisms during BKVAN is still poorly understood and an altered regulation of inflammatory mediators by resident kidney cells upon viral infection can be expected to contribute to the onset and progression of disease. TNFα interacting with its receptors, TNF receptor 1 (TNFR1) and TNF receptor 2 (TNFR2), is largely accepted to be involved in viral responses, exhibiting both proinflammatory and immunosuppressive effects...
January 2016: Molecular and Cellular Biochemistry
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