Small angle X-ray scattering (SAXS), an increasingly popular method for structural analysis of biological macromolecules in solution, is often hampered by inherent sample polydispersity. We developed ...an all-in-one system combining in-line sample component separation with parallel biophysical and SAXS characterization of the separated components. The system coupled to an automated data analysis pipeline provides a novel tool to study difficult samples at the P12 synchrotron beamline (PETRA-3, EMBL/DESY, Hamburg).
The Czech Republic has been developing its motorway network since the 1970s, while efforts to upgrade its railway system from the 1990s have been limited to improvements of existing major lines. Only ...recently has the government decided to construct new “speed connection” rail lines. This article investigates the possible territorial benefits from the future development of planned motorways and of various speed connection railway options. The modelling is based on Huff’s gravity model that calculates the benefits from improved accessibility, to job and service centres for residents of each municipality. The modelling outcomes are used to compare planned motorway development and rail development options with respect to their efficiency, related to the investment and potential numbers of users.
Lipid nanoparticles (LNPs) have gained great attention as carriers for mRNA-based therapeutics, finding applications in various indications, extending beyond their recent use in vaccines for ...infectious diseases. However, many aspects of LNP structure and their effects on efficacy are not well characterized. To further exploit the potential of mRNA therapeutics, better control of the relationship between LNP formulation composition with internal structure and transfection efficiency in vitro is necessary. We compared two well-established ionizable lipids, namely DODMA and MC3, in combination with two helper lipids, DOPE and DOPC, and two polymer-grafted lipids, either with polysarcosine (pSar) or polyethylene glycol (PEG). In addition to standard physicochemical characterization (size, zeta potential, RNA accessibility), small-angle X-ray scattering (SAXS) was used to analyze the structure of the LNPs. To assess biological activity, we performed transfection and cell-binding assays in human peripheral blood mononuclear cells (hPBMCs) using Thy1.1 reporter mRNA and Cy5-labeled mRNA, respectively. With the SAXS measurements, we were able to clearly reveal the effects of substituting the ionizable and helper lipid on the internal structure of the LNPs. In contrast, pSar as stealth moieties affected the LNPs in a different manner, by changing the surface morphology towards higher roughness. pSar LNPs were generally more active, where the highest transfection efficiency was achieved with the LNP formulation composition of MC3/DOPE/pSar. Our study highlights the utility of pSar for improved mRNA LNP products and the importance of pSar as a novel stealth moiety enhancing efficiency in future LNP formulation development. SAXS can provide valuable information for the rational development of such novel formulations by elucidating structural features in different LNP compositions.
The wars of the Staufen emperors with the Lombard League have attracted a lot of attention in recent years, and modern scholars have spent much effort correcting the romantic mistakes of ...nineteenth-century national scholarship. Yet a good deal of work remains to be done. A detailed comparison of the two decisive battles in these conflicts allows us to examine serious issues in medieval warfare, including medieval generals’ decision-making processes, the role of chivalry in those processes, the external conditions that shaped military operations, and the utility of force, perceived and actual, in achieving political objectives. Both Frederick I and his grandson Frederick II emerge as capable commanders who pursued rational military policies and made the best decisions possible under the circumstances, despite the different outcomes of these two campaigns.
Small‐angle X‐ray scattering (SAXS) is an established method for studying nanostructured systems and in particular biological macromolecules in solution. To obtain element‐specific information about ...the sample, anomalous SAXS (ASAXS) exploits changes of the scattering properties of selected atoms when the energy of the incident X‐rays is close to the binding energy of their electrons. While ASAXS is widely applied to condensed matter and inorganic systems, its use for biological macromolecules is challenging because of the weak anomalous effect. Biological objects are often only available in small quantities and are prone to radiation damage, which makes biological ASAXS measurements very challenging. The BioSAXS beamline P12 operated by the European Molecular Biology Laboratory (EMBL) at the PETRA III storage ring (DESY, Hamburg) is dedicated to studies of weakly scattering objects. Here, recent developments at P12 allowing for ASAXS measurements are presented. The beamline control, data acquisition and data reduction pipeline of the beamline were adapted to conduct ASAXS experiments. Modelling tools were developed to compute ASAXS patterns from atomic models, which can be used to analyze the data and to help designing appropriate data collection strategies. These developments are illustrated with ASAXS experiments on different model systems performed at the P12 beamline.
Hardware and software for anomalous small‐angle X‐ray scattering on biological macromolecules in solution implemented at the P12 beamline of the EMBL (PETRA III storage ring, DESY, Hamburg) are described.
Despite the efforts of developed countries to protect undeveloped land, development continues to expand beyond urban boundaries. High-quality land needed for food production is often consumed. This ...study aims to verify possible causes of undeveloped land and high-quality land consumption within regions (NUTS3) using a new approach to building growth monitoring. It investigates residential (RBs) and commercial buildings (retail and industrial buildings, RIBs). The development between 2006 and 2016 in the Czech Republic, a country in Central Europe, is used as a case study. Population growth and gross domestic product per capita (GDP) within regions are considered two potential causes of land consumption; this hypothesis is verified using a linear regression model. Only GDP showed statistically significant results. It correlated negatively with RBs and RBs + RIBs built on undeveloped land and positively with RBs + RIBs and either RBs or RIBs built on high-quality land. Based on the results, we recommend that land protection policies be differentiated according to regional specifics to be more effective. Regions with lower GDPs should obtain more support in protecting undeveloped land against residential development. The protection of high-quality land should be emphasized by supporting residential and commercial development on brownfield sites in regions with higher GDPs.
Logging experiments with the laboratory‐information management system ISPyB (Information System for Protein crystallography Beamlines) enhances the automation of small‐angle X‐ray scattering of ...biological macromolecules in solution (BioSAXS) experiments. The ISPyB interface provides immediate user‐oriented online feedback and enables data cross‐checking and downstream analysis. To optimize data quality and completeness, ISPyBB (ISPyB for BioSAXS) makes it simple for users to compare the results from new measurements with previous acquisitions from the same day or earlier experiments in order to maximize the ability to collect all data required in a single synchrotron visit. The graphical user interface (GUI) of ISPyBB has been designed to guide users in the preparation of an experiment. The input of sample information and the ability to outline the experimental aims in advance provides feedback on the number of measurements required, calculation of expected sample volumes and time needed to collect the data: all of this information aids the users to better prepare for their trip to the synchrotron. A prototype version of the ISPyBB database is now available at the European Synchrotron Radiation Facility (ESRF) beamline BM29 and is already greatly appreciated by academic users and industrial clients. It will soon be available at the PETRA III beamline P12 and the Diamond Light Source beamlines I22 and B21.
Oil-in-water (o/w) and water-in-oil (w/o) Pickering emulsions were prepared using ferritin-polymer conjugates. UV cross-linking stabilised the particle assembly around the fluid droplets. The ...resulting soft protein-polymer capsules were transferred to a medium of equal polarity as the inside of the capsule, creating water-in-water (w/w) and oil-in-oil (o/o) capsules.
This festschrift in Richard Kaeuper's honor brings together scholars from across disciplines to engage with three salient concerns of medieval society - knightly prowess and violence, lay and ...religious piety, and public order and government - from a variety of perspectives.
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