Isolation of extracellular vesicles with combined enrichment methods

Extracellular vesicles (EVs) are currently of tremendous interest in many research disciplines and EVs have potential for development of EV diagnostics or therapeutics. Most well-known single EV isolation methods have their particular advantages and disadvantages in terms of EV purity and EV yield. Combining EV isolation methods provides additional potential to improve the efficacy of both purity and yield. This review assesses the contribution and efficacy of using combined EV isolation methods by performing a two-step systematic literature analysis from all papers applying EV isolation in the year 2019. This resulted in an overview of the various methods being applied for EV isolations. A second database was generated for all studies within the first database that fairly compared multiple EV isolation methods by determining both EV purity and EV yield after isolation. From these databases it is shown that the most used EV isolation methods are not per definition the best methods based on EV purity or EV yield, indicating that more factors play a role in the choice which EV isolation method to choose than only the efficacy of the method. From the included studies it is shown that ~60% of all the included EV isolations were performed with combined EV isolation methods. The majority of EV isolations were performed with differential ultracentrifugation alone or in combination with differential ultrafiltration. When efficacy of EV isolation methods was determined in terms of EV purity and EV yield, combined EV isolation methods clearly outperformed single EV isolation methods, regardless of the type of starting material used. A recommended starting point would be the use of size-exclusion chromatography since this method, especially when combined with low-speed centrifugation, resulted in the highest EV purity, while still providing a reasonable EV yield

Wie viel Globalisierung verträgt die Welt?

Der Begriff der Globalisierung beherrscht seit Jahren die öffentliche Debatte. Für die einen geht die Öffnung der Märkte noch nicht weit genug, für die anderen führt die Globalisierung zur weiteren ungleichen Entwicklung der Nationalökonomien und vergrößert den Abstand zwischen den Industrie- und Entwicklungsländern. Prof. Dr. Dr. h.c. Hans-Werner Sinn zeigt in seinem Beitrag die ökonomischen Kräfte auf, die durch den Prozess der Globalisierung freigesetzt werden. Nach Meinung von Prof. Dr. Michael Rauscher, Universität Rostock, wird die Globalisierung ohne Frage weiter voranschreiten. Es bedarf aber nicht nur einer internationalen Koordination. Im Gegenteil: »Auch in einer sich globalisierenden Welt gibt es gute Argumente für das Subsidiaritätsprinzip: Probleme sollten auf der nied-rigstmöglichen Ebene geregelt und gelöst werden. Viele der Probleme, die im Zusammenhang mit der Globalisierung auftreten, können auf nationaler Ebene angegangen werden.« Auch für Prof. Dr. Karl Wohlmuth, Universität Bremen, ist die Gestaltung der Globalisierung die entscheidende Frage. Wichtig ist für ihn vor allen Dingen, »inwieweit eine Bereitschaft zur Anpassung der nationalen und internationalen Rahmenbedingungen an das Tempo der Globalisierung besteht«. Für Prof. Dr. Rainer Bartel, Universität Linz, muss die Globalisierung vor allem »effizient« und »nachhaltig« sein, »aber Wissenschaft und Politik dürften dazu noch nicht bereit sein«.Globalisierung, Entwicklung, Volkswirtschaft, Internationale wirtschaftspolitische Koordination, Subsidiaritätsprinzip

A tabletop Optically Pumped Magnetometer setup for the monitoring of magnetic nanoparticle clustering and immobilization using Thermal Noise Magnetometry

Many characterization techniques for magnetic nanoparticles depend on the usage of external fields. This is not the case in Thermal Noise Magnetometry (TNM), where thermal fluctuations in the magnetic signal of magnetic nanoparticle ensembles are measured without any external excitation. This can provide valuable information about the fundamental dynamical properties of the particles, due to the purely observative experiments of this relatively new technique. Until now, TNM signals have been detected only by a superconducting quantum interference device (SQUID) sensor. We present a tabletop setup using Optically Pumped Magnetometers (OPMs) in a small magnetic shield, offering a flexible and accessible alternative and show the agreement between both measurement systems for two different commercially available nanoparticle samples. We argue that the OPM setup with high accessibility complements the SQUID setup with high sensitivity and bandwidth. Furthermore, because of its excellent sensitivity in the lower frequencies, the OPM tabletop setup is well suited to monitor aggregation processes where the magnetization dynamics of the particles tend to slow down, e.g. in biological processes. As a proof of concept, we show for three different immobilization and clustering processes the changes in the noise spectrum measured in the tabletop setup: 1) the aggregation of particles due to the addition of ethanol, 2) the formation of polymer structures in the sample due to UV exposure, and 3) the cellular uptake of the particles by THP-1 cells. From our results we conclude that the tabletop setup offers a flexible and widely adoptable sensor measurement unit to monitor the immobilization and clustering of magnetic nanoparticles over time for different applications

Proteome analysis of vaccinia virus IHD-W-infected HEK 293 cells with 2-dimensional gel electrophoresis and MALDI-PSD-TOF MS of on solid phase support N-terminally sulfonated peptides

<p>Abstract</p> <p>Background</p> <p>Despite the successful eradication of smallpox by the WHO-led vaccination programme, pox virus infections remain a considerable health threat. The possible use of smallpox as a bioterrorism agent as well as the continuous occurrence of zoonotic pox virus infections document the relevance to deepen the understanding for virus host interactions. Since the permissiveness of pox infections is independent of hosts surface receptors, but correlates with the ability of the virus to infiltrate the antiviral host response, it directly depends on the hosts proteome set. In this report the proteome of HEK293 cells infected with Vaccinia Virus strain IHD-W was analyzed by 2-dimensional gel electrophoresis and MALDI-PSD-TOF MS in a bottom-up approach.</p> <p>Results</p> <p>The cellular and viral proteomes of VACV IHD-W infected HEK293 cells, UV-inactivated VACV IHD-W-treated as well as non-infected cells were compared. Derivatization of peptides with 4-sulfophenyl isothiocyanate (SPITC) carried out on ZipTipμ-C18 columns enabled protein identification via the peptides' primary sequence, providing improved s/n ratios as well as signal intensities of the PSD spectra. The expression of more than 24 human proteins was modulated by the viral infection. Effects of UV-inactivated and infectious viruses on the hosts' proteome concerning energy metabolism and proteins associated with gene expression and protein-biosynthesis were quite similar. These effects might therefore be attributed to virus entry and virion proteins. However, the modulation of proteins involved in apoptosis was clearly correlated to infectious viruses.</p> <p>Conclusions</p> <p>The proteome analysis of infected cells provides insight into apoptosis modulation, regulation of cellular gene expression and the regulation of energy metabolism. The confidence of protein identifications was clearly improved by the peptides' derivatization with SPITC on a solid phase support. Some of the identified proteins have not been described in the context of poxvirus infections before and need to be further characterised to identify their meaning for apoptosis modulation and pathogenesis.</p

VLBI Observations of NGC6240: resolving the double nuclei and radio supernovae

The European VLBI Network (EVN) has been used at two epochs in 2003 and 2009 to obtain multi-frequency high-resolution images of the merger galaxy NGC6240 in order to study the radio properties of all compact high-brightness components in the galaxy. Our observations at milli-arcsecond resolution detected the northern and southern nuclei and two radio components, which we interpret as long-lived luminous supernovae associated with the circum-nuclear starburst activity at the southern nucleus. The new VLBI data support the presence of an active galactic nucleus (AGN) together with starburst activity at the southern nucleus and provides some evidence for an AGN at the northern nucleus. The two nuclei both display an inverted spectrum at lower GHz frequencies. The spectrum of the southern nucleus indicates thermal free-free absorption on parsec scales, consistent with the presence of an AGN.Comment: 19 pages, 6 figures, accepted for publication in the Astronomical Journa

Opportunities for farming in alpine countries – pathways to truly grassland-based beef and milk production in Austria and Switzerland

Farming in the alpine countries of Austria and Switzerland fulfils important economic, socio-cultural and ecological functions for society. At the same time, it is responsible for important environmental impacts, whereas nitrogen balance surpluses and related impacts play a central role. It is crucial to reduce nitrogen inputs and site-adapted production and closing material cycles are core elements of ecologically sustainable land use. The study analysed the effects of adapted beef and dairy systems on the environmental impact and the food production with the help of the SOL mass-flow model. This includes higher reliance on grassland-based feed by abandoning the use of concentrate feed and forage maize, locally adapted reduction of livestock numbers, increased use of nitrogen-fixing legumes, reduction in mineral nitrogen fertilization, site-specific plant production and increase in nitrogen efficiency in both animal husbandry and crop production. The implementation of such a grassland-based beef and milk production results in lower ammonia emissions, reduction of nitrogen balance surpluses and lower total greenhouse gas emissions from agriculture. These environmental improvements exceed the effects of the agricultural policy since the 1990s, even though the latter has increasingly focussed on environmental impacts. Moreover, the reduction in concentrate feed and forage maize releases arable land for alternative use. This allows for increased plant-based food production and therefore minimizes the competition between food and feed production. Other options for the use of the released land are less intensive farm operations, ecological compensation areas and/or nature conservation. Finally, the reduction in animal-based food production could be offset by changed dietary patterns and the increase of plant-based food production. The suggested transformation from a production focussed to an ecologically-oriented land use and food system requires a political framework and market conditions which cannot be implemented quickly but need awareness raising and fundamental societal change

Assessing agro-ecological practices using a combination of three sustainability assessment tools

This paper is based on results from the research project UNISECO, which is funded by European Union’s Horizon 2020 research and innovation programme under grant agreement No. 773901.We would like to thank all the farmers, farm organisations and stakeholders of the multi-actor platforms of the UNISECO project who contributed to the data collection. We also like to thank Andreas Basler whose corrections improved the comprehensibility of the paper.Peer reviewedPublisher PD

Differential Glucose-Regulation of MicroRNAs in Pancreatic Islets of Non-Obese Type 2 Diabetes Model Goto-Kakizaki Rat

The Goto-Kakizaki (GK) rat is a well-studied non-obese spontaneous type 2 diabetes (T2D) animal model characterized by impaired glucose-stimulated insulin secretion (GSIS) in the pancreatic beta cells. MicroRNAs (miRNAs) are short regulatory RNAs involved in many fundamental biological processes. We aim to identify miRNAs that are differentially-expressed in the pancreatic islets of the GK rats and investigate both their short- and long term glucose-dependence during glucose-stimulatory conditions

Potential Relevance of α1-Adrenergic Receptor Autoantibodies in Refractory Hypertension

-AAB might have a mechanistic role and could represent a therapeutic target. in cardiomyocytes and induce mesentery artery segment contraction.-AAB in hypertensive patients, and the notion of immunity as a possible cause of hypertension