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2011 wurden Gräben der Oberrheinebene auf ihren Makrophytenbestand untersucht. Die Mehrzahl der Gräben wies keine bemerkenswerten Pflanzenvorkommen auf. Callitriche obtusangula gehört allerdings zu den im Gebiet offenbar relativ weit verbreiteten Arten. Bemerkenswert ist der Bereich innerhalb einer Altrheinschlinge zwischen Geinsheim, Leeheim und Wallerstädten, wo in kleinen Gräben, deren Umgebung intensiv ackerbaulich genutzt wird, bemerkenswerte Arten wie Nitella capillaris, Ranunculus lingua, Samolus valerandi und Utricularia vulgaris vorkommen. Die Neuanlage von Gräben in diesem Bereich wird empfohlen.
Im Rahmen einer hessenweiten Untersuchung wurden etwa 70 künstliche Stillgewässer – Kiesgruben, Tagebaurestseen, Fischteiche – auf das Vorkommen von Wasserpflanzen untersucht.
Die Untersuchung erbrachte den Nachweis von 78 Taxa, darunter 59 Arten Höherer Pflanzen und 19 Arten Characeen. Rund 25 % der nachgewiesnen Arten sind in der Roten Liste des Landes Hessen aufgeführt. Einige der nachgewiesenen Arten galten als verschollen. Von herausragender Bedeutung sind Kiesgruben in der Untermain- und Oberrhein-ebene, wo bis zu 10 Characeen-Arten in einem Gewässer nachgewiesen werden konnten. Von bundesweiter Bedeutung sind Funde von Nitella confervacea, N. tenuissima, Tolypella glomerata, T. intricata und T. prolifera. Als weit häufiger als erwartet erwies sich Potamogeton trichoides, die in allen Untersuchungsbereichen festgestellt wurde. Elodea nuttallii ist in den untersuchten Gewässern deutlich häufiger als E. canadensis. Sehr stark als Vogelrastplatz genutzte Teiche in der Wetterau zeigen, offenbar bedingt durch den Nährstoffeintrag durch Wasservögel, eine deutliche Eutrophierung.
Mahn, Detlef ; Hodvina, Sylvain ; Ottich, Indra ; König, Andreas ; Gregor, Thomas ; Baumann, Kurt ; Uebeler, Michael ; Angersbach, Rolf
Zur Schließung von Kenntnislücken zur Flora der Gewässer Hessens wurden 2009 von der BVNH neun Altarme von Rhein und Main untersucht. Die Ergeb-nisse zeigen, dass viele als sehr selten oder verschollen geltende Gewässermakrophyten noch oder wieder in den Altarmen des hessischen Oberrheinabschnitts zu finden sind. Insgesamt wurden 51 Arten nachgewiesen, darunter fünf Armleuchteralgen.
Bacterial and fungal toll-like receptor activation elicits type I IFN responses in mast cells
(2021)
Kornstädt, Lisa ; Pierre, Sandra ; Weigert, Andreas ; Ebersberger, Stefanie ; Schäufele, Tim J. ; Kolbinger, Anja ; Schmid, Tobias ; Cohnen, Jennifer ; Thomas, Dominique Jeanette ; Ferreirós Bouzas, Nerea ; Brüne, Bernhard ; Ebersberger, Ingo ; Scholich, Klaus
Next to their role in IgE-mediated allergic diseases and in promoting inflammation, mast cells also have antiinflammatory functions. They release pro- as well as antiinflammatory mediators, depending on the biological setting. Here we aimed to better understand the role of mast cells during the resolution phase of a local inflammation induced with the Toll-like receptor (TLR)-2 agonist zymosan. Multiple sequential immunohistology combined with a statistical neighborhood analysis showed that mast cells are located in a predominantly antiinflammatory microenvironment during resolution of inflammation and that mast cell-deficiency causes decreased efferocytosis in the resolution phase. Accordingly, FACS analysis showed decreased phagocytosis of zymosan and neutrophils by macrophages in mast cell-deficient mice. mRNA sequencing using zymosan-induced bone marrow-derived mast cells (BMMC) revealed a strong type I interferon (IFN) response, which is known to enhance phagocytosis by macrophages. Both, zymosan and lipopolysaccharides (LPS) induced IFN-β synthesis in BMMCs in similar amounts as in bone marrow derived macrophages. IFN-β was expressed by mast cells in paws from naïve mice and during zymosan-induced inflammation. As described for macrophages the release of type I IFNs from mast cells depended on TLR internalization and endosome acidification. In conclusion, mast cells are able to produce several mediators including IFN-β, which are alone or in combination with each other able to regulate the phagocytotic activity of macrophages during resolution of inflammation.
Winter, Andreas ; Nepper, Pascal ; Hermann, Marcus ; Bayer, Franziska ; Rieß, Stephanie ; Marinos, Spiros Lukas ; Linden, Arnaud van ; Holubec, Tomas ; Steinbicker, Andrea U. ; Zacharowski, Kai ; Walther, Thomas ; Emrich, Fabian Christoph
Background Vasoplegic syndrome is frequently observed during cardiac surgery and resembles a complication of high mortality and morbidity. There is a clinical need for therapy and prevention of vasoplegic syndrome during complex cardiac surgical procedures. Therefore, we investigated different strategies in a porcine model of vasoplegia.
Methods We evaluated new medical therapies and prophylaxis to avoid vasoplegic syndrome in a porcine model. After induction of anesthesia, cardiopulmonary bypass was established through median sternotomy and central cannulation. Prolonged aortic cross-clamping (120 min) simulated a complex surgical procedure. The influence of sevoflurane-guided anesthesia (sevoflurane group) and the administration of glibenclamide (glibenclamide group) were compared to a control group, which received standard anesthesia using propofol. Online hemodynamic assessment was performed using PiCCO® measurements. In addition, blood and tissue samples were taken to evaluate hemodynamic effects and the degree of inflammatory response.
Results Glibenclamide was able to break through early vasoplegic syndrome by raising the blood pressure and systemic vascular resistance as well as less need of norepinephrine doses. Sevoflurane reduced the occurrence of the vasoplegic syndrome in the mean of stable blood pressure and less need of norepinephrine doses.
Conclusion Glibenclamide could serve as a potent drug to reduce effects of vasoplegic syndrome. Sevoflurane anesthesia during cardiopulmonary bypass shows less occurrence of vasoplegic syndrome and therefore could be used to prevent it in high-risk patients.
Clinical Perspective; what is new?
* to our knowledge, this is the first randomized in vivo study evaluating the hemodynamic effects of glibenclamide after the onset of vasoplegic syndrome
* furthermore according to literature research, there is no study showing the effect of sevoflurane-guided anesthesia on the occurrence of a vasoplegic syndrome
Clinical Perspective; clinical implications?
to achieve better outcomes after complex cardiac surgery there is a need for optimized drug therapy and prevention of the vasoplegic syndrome
Lange, Christian ; Miki, Daiki ; Ochi, Hidenori ; Nischalke, Hans-Dieter ; Bojunga, Jörg ; Bibert, Stéphanie ; Morikawa, Kenichi ; Gouttenoire, Jérôme ; Cerny, Andreas ; Dufour, Jean-François ; Gorgievski-Hrisoho, Meri ; Heim, Markus H. ; Malinverni, Raffaele ; Müllhaupt, Beat ; Negro, Francesco ; Semela, David ; Kutalik, Zoltán ; Müller, Tobias ; Spengler, Ulrich ; Berg, Thomas ; Chayama, Kazuaki ; Bochud, Pierre-Yves ; Moradpour, Darius
Background: Vitamin D insufficiency has been associated with the occurrence of various types of cancer, but causal relationships remain elusive. We therefore aimed to determine the relationship between genetic determinants of vitamin D serum levels and the risk of developing hepatitis C virus (HCV)-related hepatocellular carcinoma (HCC).
Methodology/Principal Findings: Associations between CYP2R1, GC, and DHCR7 genotypes that are determinants of reduced 25-hydroxyvitamin D (25[OH]D3) serum levels and the risk of HCV-related HCC development were investigated for 1279 chronic hepatitis C patients with HCC and 4325 without HCC, respectively. The well-known associations between CYP2R1 (rs1993116, rs10741657), GC (rs2282679), and DHCR7 (rs7944926, rs12785878) genotypes and 25(OH)D3 serum levels were also apparent in patients with chronic hepatitis C. The same genotypes of these single nucleotide polymorphisms (SNPs) that are associated with reduced 25(OH)D3 serum levels were found to be associated with HCV-related HCC (P = 0.07 [OR = 1.13, 95% CI = 0.99–1.28] for CYP2R1, P = 0.007 [OR = 1.56, 95% CI = 1.12–2.15] for GC, P = 0.003 [OR = 1.42, 95% CI = 1.13–1.78] for DHCR7; ORs for risk genotypes). In contrast, no association between these genetic variations and liver fibrosis progression rate (P>0.2 for each SNP) or outcome of standard therapy with pegylated interferon-α and ribavirin (P>0.2 for each SNP) was observed, suggesting a specific influence of the genetic determinants of 25(OH)D3 serum levels on hepatocarcinogenesis.
Conclusions/Significance: Our data suggest a relatively weak but functionally relevant role for vitamin D in the prevention of HCV-related hepatocarcinogenesis.
Altınçekiç, Nadide ; Korn, Sophie Marianne ; Qureshi, Nusrat Shahin ; Dujardin, Marie ; Ninot-Pedrosa, Martí ; Abele, Rupert ; Abi Saad, Marie Jose ; Alfano, Caterina ; Almeida, Fabio C. L. ; Alshamleh, Islam ; Amorim, Gisele Cardoso de ; Anderson, Thomas K. ; Anobom, Cristiane D. ; Anorma, Chelsea ; Bains, Jasleen Kaur ; Bax, Adriaan ; Blackledge, Martin ; Blechar, Julius ; Böckmann, Anja ; Brigandat, Louis ; Bula, Anna ; Bütikofer, Matthias ; Camacho-Zarco, Aldo R. ; Carlomagno, Teresa ; Caruso, Icaro Putinhon ; Ceylan, Betül ; Chaikuad, Apirat ; Chu, Feixia ; Cole, Laura ; Crosby, Marquise G. ; De Jesus, Vanessa ; Dhamotharan, Karthikeyan ; Felli, Isabella C. ; Ferner, Jan ; Fleischmann, Yanick ; Fogeron, Marie-Laure ; Fourkiotis, Nikolaos K. ; Fuks, Christin ; Fürtig, Boris ; Gallo, Angelo ; Gande, Santosh L. ; Gerez, Juan Atilio ; Ghosh, Dhiman ; Gomes-Neto, Francisco ; Gorbatyuk, Oksana ; Guseva, Serafima ; Hacker, Carolin ; Häfner, Sabine ; Hao, Bing ; Hargittay, Bruno ; Henzler-Wildman, Katherine ; Hoch, Jeffrey C. ; Hohmann, Katharina Felicitas ; Hutchison, Marie T. ; Jaudzems, Kristaps ; Jović, Katarina ; Kaderli, Janina ; Kalnins, Gints ; Kanepe, Iveta ; Kirchdoerfer, Robert N. ; Kirkpatrick, John ; Knapp, Stefan ; Krishnathas, Robin ; Kutz, Felicitas ; Zur Lage, Susanne ; Lambertz, Roderick ; Lang, Andras ; Laurents, Douglas ; Lecoq, Lauriane ; Linhard, Verena ; Löhr, Frank ; Malki, Anas ; Bessa, Luiza Mamigonian ; Martin, Rachel W. ; Matzel, Tobias ; Maurin, Damien ; McNutt, Seth W. ; Mebus-Antunes, Nathane Cunha ; Meier, Beat H. ; Meiser, Nathalie ; Mompeán, Miguel ; Monaca, Elisa ; Montserret, Roland ; Perez, Laura Mariño ; Moser, Celine ; Muhle-Goll, Claudia ; Neves-Martins, Thais Cristtina ; Ni, Xiamonin ; Norton-Baker, Brenna ; Pierattelli, Roberta ; Pontoriero, Letizia ; Pustovalova, Yulia ; Ohlenschläger, Oliver ; Orts, Julien ; Poian, Andrea T. da ; Pyper, Dennis Joshua ; Richter, Christian ; Riek, Roland ; Rienstra, Chad M. ; Robertson, Angus ; Pinheiro, Anderson S. ; Sabbatella, Raffaele ; Salvi, Nicola ; Saxena, Krishna ; Schulte, Linda ; Schiavina, Marco ; Schwalbe, Harald ; Silber, Mara ; Almeida, Marcius da Silva ; Sprague-Piercy, Marc A. ; Spyroulias, Georgios A. ; Sreeramulu, Sridhar ; Tants, Jan-Niklas ; Tars, Kaspars ; Torres, Felix ; Töws, Sabrina ; Trevino, Miguel A. ; Trucks, Sven ; Tsika, Aikaterini C. ; Varga, Krisztina ; Wang, Ying ; Weber, Marco E. ; Weigand, Julia E. ; Wiedemann, Christoph ; Wirmer-Bartoschek, Julia ; Wirtz Martin, Maria Alexandra ; Zehnder, Johannes ; Hengesbach, Martin ; Schlundt, Andreas
The highly infectious disease COVID-19 caused by the Betacoronavirus SARS-CoV-2 poses a severe threat to humanity and demands the redirection of scientific efforts and criteria to organized research projects. The international COVID19-NMR consortium seeks to provide such new approaches by gathering scientific expertise worldwide. In particular, making available viral proteins and RNAs will pave the way to understanding the SARS-CoV-2 molecular components in detail. The research in COVID19-NMR and the resources provided through the consortium are fully disclosed to accelerate access and exploitation. NMR investigations of the viral molecular components are designated to provide the essential basis for further work, including macromolecular interaction studies and high-throughput drug screening. Here, we present the extensive catalog of a holistic SARS-CoV-2 protein preparation approach based on the consortium’s collective efforts. We provide protocols for the large-scale production of more than 80% of all SARS-CoV-2 proteins or essential parts of them. Several of the proteins were produced in more than one laboratory, demonstrating the high interoperability between NMR groups worldwide. For the majority of proteins, we can produce isotope-labeled samples of HSQC-grade. Together with several NMR chemical shift assignments made publicly available on covid19-nmr.com, we here provide highly valuable resources for the production of SARS-CoV-2 proteins in isotope-labeled form.
Théry, Clotilde ; Witwer, Kenneth W. ; Aikawa, Elena ; Alcaraz, Maria Jose ; Anderson, Johnathon D. ; Andriantsitohaina, Ramaroson ; Antoniou, Anna ; Arab, Tanina ; Archer, Fabienne ; Atkin-Smith, Georgia K. ; Ayre, D. Craig ; Bach, Jean-Marie ; Bachurski, Daniel ; Baharvand, Hossein ; Balaj, Leonora ; Baldacchino, Shawn ; Bauer, Natalie N. ; Baxter, Amy A. ; Bebawy, Mary ; Beckham, Carla ; Zavec, Apolonija Bedina ; Benmoussa, Abderrahim ; Berardi, Anna C. ; Bergese, Paolo ; Bielska, Ewa ; Blenkiron, Cherie ; Bobis-Wozowicz, Sylwia ; Boilard, Eric ; Boireau, Wilfrid ; Bongiovanni, Antonella ; Borràs, Francesc E. ; Bosch, Steffi ; Boulanger, Chantal M. ; Breakefield, Xandra ; Breglio, Andrew M. ; Brennan, Meadhbh Aín ; Brigstock, David R. ; Brisson, Alain ; Broekman, Marike L. D. ; Bromberg, Jacqueline F. ; Bryl-Gorecka, Paulina ; Buch, Shilpa ; Buck, Amy H. ; Burger, Dylan ; Busatto, Sara ; Buschmann, Dominik ; Bussolati, Benedetta ; Buzás, Edit I. ; Byrd, James Bryan ; Camussi, Giovanni ; Carter, David R. F. ; Caruso, Sarah ; Chamley, Lawrence W. ; Chang, Yu-Ting ; Chen, Chihchen ; Chen, Shuai ; Cheng, Lesley ; Chin, Andrew R. ; Clayton, Aled ; Clerici, Stefano P. ; Cocks, Alex ; Cocucci, Emanuele ; Coffey, Robert J. ; Cordeiro-da-Silva, Anabela ; Couch, Yvonne ; Coumans, Frank A. W. ; Coyle, Beth ; Crescitelli, Rossella ; Criado, Miria Ferreira ; D’Souza-Schorey, Crislyn ; Das, Saumya ; Chaudhuri, Amrita Datta ; de-Candia, Paola ; De Santana, Eliezer F. ; De Wever, Olivier ; del Portillo, Hernando A. ; Demaret, Tanguy ; Deville, Sarah ; Devitt, Andrew ; Dhondt, Bert ; Di Vizio, Dolores ; Dieterich, Lothar C. ; Dolo, Vincenza ; Dominguez Rubio, Ana Paula ; Dominici, Massimo ; Dourado, Mauricio R. ; Driedonks, Tom A. P. ; Duarte, Filipe V. ; Duncan, Heather M. ; Eichenberger, Ramon Marc ; Ekström, Karin ; El Andaloussi, Samir ; Elie-Caille, Celine ; Erdbrügger, Uta ; Falcón-Pérez, Juan M. ; Fatima, Farah ; Fish, Jason E. ; Flores-Bellver, Miguel ; Försönits, András ; Frelet-Barrand, Annie ; Fricke, Fabia ; Fuhrmann, Gregor ; Gabrielsson, Susanne ; Gámez-Valero, Ana ; Gardiner, Chris ; Gärtner, Kathrin ; Gaudin, Raphael ; Gho, Yong Song ; Giebel, Bernd ; Gilbert, Caroline ; Gimona, Mario ; Giusti, Ilaria ; Goberdhan, Deborah C. I. ; Görgens, André ; Gorski, Sharon M. ; Greening, David W. ; Groß, Julia Christina ; Gualerzi, Alice ; Gupta, Gopal N. ; Gustafson, Dakota ; Handberg, Aase ; Haraszti, Réka Ágnes ; Harrison, Paul ; Hegyesi, Hargita ; Hendrix, An ; Hill, Andrew F. ; Hochberg, Fred H. ; Hoffmann, Karl F. ; Holder, Beth ; Holthofer, Harry ; Hosseinkhani, Baharak ; Hu, Guoku ; Huang, Yiyao ; Huber, Veronica ; Hunt, Stuart ; Ibrahim, Ahmed Gamal-Eldin ; Ikezu, Tsuneya ; Inal, Jameel M. ; Isin, Mustafa ; Ivanova, Alena ; Jackson, Hannah K. ; Jacobsen, Soren ; Jay, Steven M. ; Jayachandran, Muthuvel ; Jenster, Guido ; Jiang, Lanzhou ; Johnson, Suzanne M. ; Jones, Jennifer C. ; Jong, Ambrose ; Jovanovic-Talisman, Tijana ; Jung, Stephanie ; Kalluri, Raghu ; Kano, Shin-ichi ; Kaur, Sukhbir ; Kawamura, Yumi ; Keller, Evan T. ; Khamari, Delaram ; Khomyakova, Elena ; Khvorova, Anastasia ; Kierulf, Peter ; Kim, Kwang Pyo ; Kislinger, Thomas ; Klingeborn, Mikael ; Klinke, David J. ; Kornek, Miroslaw ; Kosanović, Maja M. ; Kovács, Árpád Ferenc ; Krämer-Albers, Eva-Maria ; Krasemann, Susanne ; Krause, Mirja ; Kurochkin, Igor V. ; Kusuma, Gina D. ; Kuypers, Sören ; Laitinen, Saara ; Langevin, Scott M. ; Languino, Lucia R. ; Lannigan, Joanne ; Lässer, Cecilia ; Laurent, Louise C. ; Lavieu, Gregory ; Lázaro-Ibáñez, Elisa ; Le Lay, Soazig ; Lee, Myung-Shin ; Lee, Yi Xin Fiona ; Lemos, Debora S. ; Lenassi, Metka ; Leszczynska, Aleksandra ; Li, Isaac T. S. ; Liao, Ke ; Libregts, Sten F. ; Ligeti, Erzsebet ; Lim, Rebecca ; Lim, Sai Kiang ; Linē, Aija ; Linnemannstöns, Karen ; Llorente, Alicia ; Lombard, Catherine Ann ; Lorenowicz, Magdalena J. ; Lörincz, Ákos M. ; Lötvall, Jan ; Lovett, Jason ; Lowry, Michelle C. ; Loyer, Xavier ; Lu, Quan ; Lukomska, Barbara ; Lunavat, Taral Rameshchand ; Maas, Sybren L. N. ; Malhi, Harmeet ; Marcilla, Antonio ; Mariani, Jacopo ; Mariscal, Javier ; Martens-Uzunova, Elena S. ; Martin-Jaular, Lorena ; Martinez, M. Carmen ; Martins, Vilma Regina ; Mathieu, Mathilde ; Mathivanan, Suresh ; Maugeri, Marco ; McGinnis, Lynda K. ; McVey, Mark J. ; Meckes, David G. ; Meehan, Katie ; Mertens, Inge ; Minciacchi, Valentina R. ; Möller, Andreas ; Møller Jørgensen, Malene ; Morales-Kastresana, Aizea ; Morhayim, Jess ; Mullier, François ; Muraca, Maurizio ; Musante, Luca ; Mussack, Veronika ; Muth, Dillon C. ; Myburgh, Kathryn H. ; Najrana, Tanbir ; Nawaz, Muhammad ; Nazarenko, Irina ; Nejsum, Peter ; Neri, Christian ; Neri, Tommaso ; Nieuwland, Rienk ; Nimrichter, Leonardo ; Nolan, John P. ; Nolte-’t Hoen, Esther N. M. ; Hooten, Nicole Noren ; O’Driscoll, Lorraine ; O’Grady, Tina ; O’Loghlen, Ana ; Ochiya, Takahiro ; Olivier, Martin ; Ortiz, Alberto ; Ortiz, Luis A. ; Osteikoetxea, Xabier ; Østergaard, Ole ; Ostrowski, Matias ; Park, Jaesung ; Pegtel, Dirk Michiel ; Peinado, Hector ; Perut, Francesca ; Pfaffl, Michael ; Phinney, Donald G. ; Pieters, Bartijn C. H. ; Pink, Ryan Charles ; Pisetsky, David S. ; Pogge von Strandmann, Elke ; Polakovicova, Iva ; Poon, Ivan K. H. ; Powell, Bonita H. ; Prada, Ilaria ; Pulliam, Lynn ; Quesenberry, Peter ; Radeghieri, Annalisa ; Raffai, Robert L. ; Raimondo, Stefania ; Rak, Janusz ; Ramirez, Marcel I. ; Raposo, Graça ; Rayyan, Morsi S. ; Regev-Rudzki, Neta ; Ricklefs, Franz Lennard ; Robbins, Paul D. ; Roberts, David D. ; Rodrigues, Silvia C. ; Rohde, Eva ; Rome, Sophie ; Rouschop, Kasper M. ; Rughetti, Aurelia ; Russell, Ashley E. ; Saá, Paula ; Sahoo, Susmita ; Salas-Huenuleo, Edison ; Sánchez, Catherine ; Saugstad, Julie A. ; Saul, Meike J. ; Schiffelers, Raymond M. ; Schneider, Raphael ; Schøyen, Tine Hiorth ; Scott, Aaron ; Shahaj, Eriomina ; Sharma, Shivani ; Shatnyeva, Olga ; Shekari, Faezeh ; Shelke, Ganesh Vilas ; Shetty, Ashok K. ; Shiba, Kiyotaka ; Siljander, Pia R.-M. ; Silva, Andreia M. ; Skowronek, Agata ; Snyder, Orman L. ; Soares, Rodrigo Pedro ; Sódar, Barbara W. ; Soekmadji, Carolina ; Sotillo, Javier ; Stahl, Philip D. ; Stoorvogel, Willem ; Stott, Shannon L. ; Strasser, Erwin F. ; Swift, Simon ; Tahara, Hidetoshi ; Tewari, Muneesh ; Timms, Kate ; Tiwari, Swasti ; Tixeira, Rochelle ; Tkach, Mercedes ; Toh, Wei Seong ; Tomasini, Richard ; Torrecilhas, Ana Claudia ; Tosar, Juan Pablo ; Toxavidis, Vasilis ; Urbanelli, Lorena ; Vader, Pieter ; Balkom, Bastiaan Wilhelmus Maria van ; Grein, Susanne G. van der ; Van Deun, Jan ; Herwijnen, Martijn J. C. van ; Van Keuren-Jensen, Kendall R. ; Niel, Guillaume van ; Royen, Martin E. van ; Wijnen, Andre J. van ; Vasconcelos, M. Helena ; Vechetti, Ivan J. ; Veit, Tiago D. ; Vella, Laura J. ; Velot, Émilie ; Verweij, Frederik ; Vestad, Beate ; Viñas, Jose Luis ; Visnovitz, Tamás ; Vukman, Krisztina V. ; Wahlgren, Jessica ; Watson, Dionysios C. ; Wauben, Marca H. M. ; Weaver, Alissa ; Webber, Jason P. ; Weber, Viktoria ; Wehman, Ann M. ; Weiss, Daniel J. ; Welsh, Joshua A. ; Wendt, Sebastian ; Wheelock, Åsa M. ; Wiener, Zoltán ; Witte, Leonie ; Wolfram, Joy ; Xagorari, Angeliki ; Xander, Patricia ; Xu, Jing ; Yan, Xiaomei ; Yáñez-Mó, María ; Yin, Hang ; Yuana, Yuana ; Zappulli, Valentina ; Zarubova, Jana ; Žėkas, Vytautas ; Zhang, Jian-ye ; Zhao, Zezhou ; Zheng, Lei ; Zheutlin, Alexander R. ; Zickler, Antje Maria ; Zimmermann, Pascale ; Zivkovic, Angela M. ; Zocco, Davide ; Zuba-Surma, Ewa K.
The last decade has seen a sharp increase in the number of scientific publications describing physiological and pathological functions of extracellular vesicles (EVs), a collective term covering various subtypes of cell-released, membranous structures, called exosomes, microvesicles, microparticles, ectosomes, oncosomes, apoptotic bodies, and many other names. However, specific issues arise when working with these entities, whose size and amount often make them difficult to obtain as relatively pure preparations, and to characterize properly. The International Society for Extracellular Vesicles (ISEV) proposed Minimal Information for Studies of Extracellular Vesicles (“MISEV”) guidelines for the field in 2014. We now update these “MISEV2014” guidelines based on evolution of the collective knowledge in the last four years. An important point to consider is that ascribing a specific function to EVs in general, or to subtypes of EVs, requires reporting of specific information beyond mere description of function in a crude, potentially contaminated, and heterogeneous preparation. For example, claims that exosomes are endowed with exquisite and specific activities remain difficult to support experimentally, given our still limited knowledge of their specific molecular machineries of biogenesis and release, as compared with other biophysically similar EVs. The MISEV2018 guidelines include tables and outlines of suggested protocols and steps to follow to document specific EV-associated functional activities. Finally, a checklist is provided with summaries of key points.
Schrade, Katharina ; Tröger, Jessica ; Eldahshan, Adeeb ; Zühlke, Kerstin ; Abdul Azeez, Kamal R. ; Elkins, Jonathan M. ; Neuenschwander, Martin ; Oder, Andreas ; Elkewedi, Mohamed ; Jaksch, Sarah ; Andrae, Karsten ; Li, Jinliang ; Fernandes, Joao ; Müller, Paul Markus ; Grunwald, Stephan ; Marino, Stephen A. ; Vukićević, Tanja ; Eichhorst, Jenny ; Wiesner, Burkhard ; Weber, Marcus ; Kapiloff, Michael ; Rocks, Oliver ; Daumke, Oliver ; Wieland, Thomas ; Knapp, Stefan ; Kries, Jens Peter von ; Klussmann, Enno
Stimulation of renal collecting duct principal cells with antidiuretic hormone (arginine-vasopressin, AVP) results in inhibition of the small GTPase RhoA and the enrichment of the water channel aquaporin-2 (AQP2) in the plasma membrane. The membrane insertion facilitates water reabsorption from primary urine and fine-tuning of body water homeostasis. Rho guanine nucleotide exchange factors (GEFs) interact with RhoA, catalyze the exchange of GDP for GTP and thereby activate the GTPase. However, GEFs involved in the control of AQP2 in renal principal cells are unknown. The A-kinase anchoring protein, AKAP-Lbc, possesses GEF activity, specifically activates RhoA, and is expressed in primary renal inner medullary collecting duct principal (IMCD) cells. Through screening of 18,431 small molecules and synthesis of a focused library around one of the hits, we identified an inhibitor of the interaction of AKAP-Lbc and RhoA. This molecule, Scaff10-8, bound to RhoA, inhibited the AKAP-Lbc-mediated RhoA activation but did not interfere with RhoA activation through other GEFs or activities of other members of the Rho family of small GTPases, Rac1 and Cdc42. Scaff10-8 promoted the redistribution of AQP2 from intracellular vesicles to the periphery of IMCD cells. Thus, our data demonstrate an involvement of AKAP-Lbc-mediated RhoA activation in the control of AQP2 trafficking.
Tesch, Roberta ; Rak, Marcel ; Raab, Monika ; Berger, Lena Marie ; Kronenberger, Thales ; Jörger, Andreas C. ; Berger, Benedict-Tilman ; Abdi, Ismahan ; Hanke, Thomas ; Poso, Antti ; Strebhardt, Klaus ; Sanhaji, Mourad ; Knapp, Stefan
Salt-inducible kinases (SIKs) are key metabolic regulators. Imbalance of SIK function is associated with the development of diverse cancers, including breast, gastric and ovarian cancer. Chemical tools to clarify the roles of SIK in different diseases are, however, sparse and are generally characterized by poor kinome-wide selectivity. Here, we have adapted the pyrido[2,3-d]pyrimidin-7-one-based PAK inhibitor G-5555 for the targeting of SIK, by exploiting differences in the back-pocket region of these kinases. Optimization was supported by high-resolution crystal structures of G-5555 bound to the known off-targets MST3 and MST4, leading to a chemical probe, MRIA9, with dual SIK/PAK activity and excellent selectivity over other kinases. Furthermore, we show that MRIA9 sensitizes ovarian cancer cells to treatment with the mitotic agent paclitaxel, confirming earlier data from genetic knockdown studies and suggesting a combination therapy with SIK inhibitors and paclitaxel for the treatment of paclitaxel-resistant ovarian cancer.