A smart polymer for sequence-selective binding, pulldown, and release of DNA targets

Selective isolation of DNA is crucial for applications in biology, bionanotechnology, clinical diagnostics and forensics. We herein report a smart methanol-responsive polymer (MeRPy) that can be programmed to bind and separate single- as well as double-stranded DNA targets. Captured targets are quickly isolated and released back into solution by denaturation (sequence-agnostic) or toehold-mediated strand displacement (sequence-selective). The latter mode allows 99.8% efficient removal of unwanted sequences and 79% recovery of highly pure target sequences. We applied MeRPy for the depletion of insulin, glucagon, and transthyretin cDNA from clinical next-generation sequencing (NGS) libraries. This step improved the data quality for low-abundance transcripts in expression profiles of pancreatic tissues. Its low cost, scalability, high stability and ease of use make MeRPy suitable for diverse applications in research and clinical laboratories, including enhancement of NGS libraries, extraction of DNA from biological samples, preparative-scale DNA isolations, and sorting of DNA-labeled non-nucleic acid targets. © 2020, The Author(s)

Toward Functional Synthetic Cells: In-Depth Study of Nanoparticle and Enzyme Diffusion through a Cross-Linked Polymersome Membrane

Understanding the diffusion of nanoparticles through permeable membranes in cell mimics paves the way for the construction of more sophisticated synthetic protocells with control over the exchange of nanoparticles or biomacromolecules between different compartments. Nanoparticles postloading by swollen pH switchable polymersomes is investigated and nanoparticles locations at or within polymersome membrane and polymersome lumen are precisely determined. Validation of transmembrane diffusion properties is performed based on nanoparticles of different origin—gold, glycopolymer protein mimics, and the enzymes myoglobin and esterase—with dimensions between 5 and 15 nm. This process is compared with the in situ loading of nanoparticles during polymersome formation and analyzed by advanced multiple-detector asymmetrical flow field-flow fractionation (AF4). These experiments are supported by complementary i) release studies of protein mimics from polymersomes, ii) stability and cyclic pH switches test for in polymersome encapsulated myoglobin, and iii) cryogenic transmission electron microscopy studies on nanoparticles loaded polymersomes. Different locations (e.g., membrane and/or lumen) are identified for the uptake of each protein. The protein locations are extracted from the increasing scaling parameters and the decreasing apparent density of enzyme-containing polymersomes as determined by AF4. Postloading demonstrates to be a valuable tool for the implementation of cell-like functions in polymersomes

Clinical and molecular implications of NAB2-STAT6 fusion variants in solitary fibrous tumour

Solitary fibrous tumour (SFT) is a mesenchymal neoplasm characterised by pathognomonic NAB2-STAT6 gene fusions. The clinical implications and prognostic value of different fusion variants has not been clarified. In the current study, we explore the clinicopathological, prognostic and molecular differences between tumours with different fusions. Thirty-nine patients with localised, extrameningeal SFT were included, of whom 20 developed distant recurrence and 19 were without recurrence after long term follow-up. Capture-based RNA sequencing identified 12 breakpoint variants, which were categorised into two groups based on the STAT6 domain composition in the predicted chimeric proteins. Twenty-one of 34 (62%) sequenced tumours had fusions with most of the STAT6 domains intact and were classified as STAT6-Full. Thirteen tumours (38%) contained only the transactivation domain of STAT6 and were classified as STAT6-TAD. Tumours with STAT6-TAD fusions had a higher mitotic count (p=0.016) and were associated with inferior recurrence-free interval (p=0.004) and overall survival (p=0.012). Estimated 10-year recurrence-free survival was 25% for patients with STAT6-TAD tumours compared to 78% for the STAT6-Full group. Distinct transcriptional signatures between the fusion groups were identified, including higher expression of FGF2 in the STAT6-TAD group and IGF2, EGR2, PDGFRB, STAT6 and several extracellular matrix genes in STAT6-Full tumours. In summary, we demonstrate that NAB2-STAT6 fusion variants are associated with distinct clinicopathological and molecular characteristics and have prognostic significance in extrameningeal SFT.publishedVersio

Dry-jet wet spinning of thermally stable lignin-textile grade polyacrylonitrile fibers regenerated from chloride-based ionic liquids compounds

In this paper, we report on the use of amorphous lignin, a waste by-product of the paper industry, for the production of high performance carbon fibers (CF) as precursor with improved thermal stability and thermo-mechanical properties. The precursor was prepared by blending of lignin with polyacrylonitrile (PAN), which was previously dissolved in an ionic liquid. The fibers thus produced offered very high thermal stability as compared with the fiber consisting of pure PAN. The molecular compatibility, miscibility, and thermal stability of the system were studied by means of shear rheological measurements. The achieved mechanical properties were found to be related to the temperature-dependent relaxation time (consistence parameter) of the spinning dope and the diffusion kinetics of the ionic liquids from the fibers into the coagulation bath. Furthermore, thermogravimetric analysis (TGA), differential scanning calorimetry (DSC), and dynamic mechanical tests (DMA) were utilized to understand in-depth the thermal and the stabilization kinetics of the developed fibers and the impact of lignin on the stabilization process of the fibers. Low molecular weight lignin increased the thermally induced physical shrinkage, suggesting disturbing effects on the semi-crystalline domains of the PAN matrix, and suppressed the chemically induced shrinkage of the fibers. The knowledge gained throughout the present paper allows summarizing a novel avenue to develop lignin-based CF designed with adjusted thermal stability

Avidin Localizations in pH-Responsive Polymersomes for Probing the Docking of Biotinylated (Macro)molecules in the Membrane and Lumen

To mimic organelles and cells and to construct next-generation therapeutics, asymmetric functionalization and location of proteins for artificial vesicles is thoroughly needed to emphasize the complex interplay of biological units and systems through spatially separated and spatiotemporal controlled actions, release, and communications. For the challenge of vesicle (= polymersome) construction, the membrane permeability and the location of the cargo are important key characteristics that determine their potential applications. Herein, an in situ and post loading process of avidin in pH-responsive and photo-cross-linked polymersomes is developed and characterized. First, loading efficiency, main location (inside, lumen, outside), and release of avidin under different conditions have been validated, including the pH-stable presence of avidin in polymersomes’ membrane outside and inside. This advantageous approach allows us to selectively functionalize the outer and inner membranes as well as the lumen with several bio(macro)molecules, generally suited for the construction of asymmetrically functionalized artificial organelles. In addition, a fluorescence resonance energy transfer (FRET) effect was used to study the permeability or uptake of the polymersome membrane against a broad range of biotinylated (macro)molecules (different typology, sizes, and shapes) under different conditions

Light-driven proton transfer for cyclic and temporal switching of enzymatic nanoreactors

CITATION: Moreno, S. et al. 2020. Light-driven proton transfer for cyclic and temporal switching of enzymatic nanoreactors. Small, 16:2002135, doi:10.1002/smll.202002135.The original publication is available at https://onlinelibrary.wiley.comTemporal activation of biological processes by visible light and subsequent return to an inactive state in the absence of light is an essential characteristic of photoreceptor cells. Inspired by these phenomena, light-responsive materials are very attractive due to the high spatiotemporal control of light irradiation, with light being able to precisely orchestrate processes repeatedly over many cycles. Herein, it is reported that light-driven proton transfer triggered by a merocyanine-based photoacid can be used to modulate the permeability of pH-responsive polymersomes through cyclic, temporally controlled protonation and deprotonation of the polymersome membrane. The membranes can undergo repeated light-driven swelling–contraction cycles without losing functional effectiveness. When applied to enzyme loaded-nanoreactors, this membrane responsiveness is used for the reversible control of enzymatic reactions. This combination of the merocyanine-based photoacid and pH-switchable nanoreactors results in rapidly responding and versatile supramolecular systems successfully used to switch enzymatic reactions ON and OFF on demand.Publisher's versio

Legislative Documents

Also, variously referred to as: House bills; House documents; House legislative documents; legislative documents; General Court documents

Moderne Fraktioniermethoden zur Charakterisierung von verzweigten Polymeren

Die Charakterisierung von verzweigten Polymeren stellt eine große Herausforderung dar. In dieser Arbeit wurden zum einen die Asymmetrische Fluss-Feldflussfraktionierung (AF4) als Alternative zur Größenausschlusschromatographie angewandt, um dendritische Polymere zu charakterisieren. Dabei wurden erstmals auch Komplexierungs-und Aggregationsstudien von dendritischen Glykopolymeren durchgeführt. Weiterhin ist es mittels Adaption der Phasenverteilungschromatographie gelungen, in Abhängigkeit des Verzweigungscharakters aufzutrennen. Hierbei konnten lineare und sternförmige Polystyrenproben separiert werden

Fra reproduktion til refleksion. En kvalitativ studie af læreres arbejde med udvikling af elevers kritisk reflekterende evner.

Internationale forskningsresultater viser, at lærere på tværs af uddannelsestrin møder udfordringer i deres arbejde med at få elever til at bevæge sig fra en faktapræget reproducerende tilnærmning til kundskabsudvikling til en mere kritisk reflekterende tilnærmning. Hensigten med mit projekt er at identifisere og forstå nogle af disse udfordringer i tilknytning til undervisning i samfundsfaglige temaer i norsk videregående skole. Målet er samtidig at synliggøre, hvordan læreres forståelse af kritisk refleksion, kundskab og kundskabsudvikling er en vigtig præmisleverandør for, hvad kritisk refleksion kan være i skolen. Indgangsvinkelen til projektet er følgende problemstilling: Hvad karakteriserer læreres erfaringer med arbejdet med udvikling af elevers kritiske reflekterende evner i samfundsfaglige temaer i norsk videregående skole? Projektets analytiske perspektiver omhandler forskellige forståelser af og tilnærmninger til kritisk refleksion som kundskab og kompetence. I diskusionkapitlet inddrager jeg dessuden perspektiver på professionsudøvelse og lærerens professionelle pædagogiske handlingsrum. Projektet er gennemført som en kvalitativ studie med interview af fem lærere i norsk videregående skole. Datamaterialet er analyseret gennem en induktiv metode med fokus på kategorisering af semantiske relationer. Resultaterne viser, at de elementer som indrammer deltagernes forståelse af sit eget pædagogiske arbejdsrum i forbindelse med undervisning med fokus på kritisk refleksion kan knyttes til henholdsvis skolen som institution, skolen om organisation og elevernes forudsætninger som overordnede momenter. Resultaterne viser også, hvordan den enkelte deltagers forståelse af og positionering i forhold til elementerne fungerer som betingelse for, hvordan kritisk refleksion kan fremdyrkes i skolen, samt hvordan dette kan ses i sammenhæng med, at der eksisterer forskellige forståelser af kritisk refleksion. Diskusionskapitlet synliggør særligt, hvordan deltagernes positionering i forhold til henholdsvis elevernes manglende faglige ballast og mødet mellem skolens og elevens virkelighed bliver en udfordrende rammebetingelse for, hvad kritisk refleksion kan være i skolen