76 research outputs found

    Hydroxylated fluorescent dyes for live-cell labeling: Synthesis, spectra and super-resolution STED.

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    Hydroxylated rhodamines, carbopyronines, silico- and germanorhodamines with absorption maxima in the range of 530-640 nm were prepared and applied in specific labeling of living cells. The direct and high-yielding entry to germa- and silaxanthones tolerates the presence of protected heteroatoms and may be considered for the syntheses of various sila- and germafluoresceins, as well as -rhodols. Application in stimulated emission depletion (STED) fluorescence microscopy revealed a resolution of 50-75 nm in one- and two-color imaging of vimentin-HaloTag fused protein and native tubulin. The established structure-property relationships allow prediction of the spectral properties and the positions of spirolactone/zwitterion equilibria for the new analogs of rhodamines, carbo-, silico- and germanorhodamines using simple additive schemes

    Modeling Bacterial DNA: Simulation of Self-avoiding Supercoiled Worm-Like Chains Including Structural Transitions of the Helix

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    Under supercoiling constraints, naked DNA, such as a large part of bacterial DNA, folds into braided structures called plectonemes. The double-helix can also undergo local structural transitions, leading to the formation of denaturation bubbles and other alternative structures. Various polymer models have been developed to capture these properties, with Monte-Carlo (MC) approaches dedicated to the inference of thermodynamic properties. In this chapter, we explain how to perform such Monte-Carlo simulations, following two objectives. On one hand, we present the self-avoiding supercoiled Worm-Like Chain (ssWLC) model, which is known to capture the folding properties of supercoiled DNA, and provide a detailed explanation of a standard MC simulation method. On the other hand, we explain how to extend this ssWLC model to include structural transitions of the helix.Comment: Book chapter to appear in The Bacterial Nucleoid, Methods and Protocols, Springer serie

    Giant worm-shaped ESCRT scaffolds surround actin-independent integrin clusters

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    Endosomal Sorting Complex Required for Transport (ESCRT) proteins can be transiently recruited to the plasma membrane for membrane repair and formation of extracellular vesicles. Here, we discovered micrometer-sized worm-shaped ESCRT structures that stably persist for multiple hours at the plasma membrane of macrophages, dendritic cells, and fibroblasts. These structures surround clusters of integrins and known cargoes of extracellular vesicles. The ESCRT structures are tightly connected to the cellular support and are left behind by the cells together with surrounding patches of membrane. The phospholipid composition is altered at the position of the ESCRT structures, and the actin cytoskeleton is locally degraded, which are hallmarks of membrane damage and extracellular vesicle formation. Disruption of actin polymerization increased the formation of the ESCRT structures and cell adhesion. The ESCRT structures were also present at plasma membrane contact sites with membrane-disrupting silica crystals. We propose that the ESCRT proteins are recruited to adhesion-induced membrane tears to induce extracellular shedding of the damaged membrane

    Quasi-local conserved charges and spin transport in spin-1 integrable chains

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    We consider the integrable one-dimensional spin-1 chain defined by the Zamolodchikov-Fateev (ZF) Hamiltonian. The latter is parametrized, analogously to the XXZ spin-1/2 model, by a continuous anisotropy parameter and at the isotropic point coincides with the well-known spin-1 Babujian-Takhtajan Hamiltonian. Following a procedure recently developed for the XXZ model, we explicitly construct a continuous family of quasi-local conserved operators for the periodic spin-1 ZF chain. Our construction is valid for a dense set of commensurate values of the anisotropy parameter in the gapless regime where the isotropic point is excluded. Using the Mazur inequality, we show that, as for the XXZ model, these quasi-local charges are enough to prove that the high-temperature spin Drude weight is non-vanishing in the thermodynamic limit, thus establishing ballistic spin transport at high temperature

    Update on potential medical treatments for encapsulating peritoneal sclerosis; human and experimental data

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    Encapsulating peritoneal sclerosis (EPS) is an infrequent but serious complication of peritoneal dialysis (PD). The pathogenesis is unknown but speculation is ongoing. The current management of EPS focuses on prevention and treatment of the inflammatory and fibrotic changes at the level of the peritoneal membrane with immunosuppressive and antifibrotic agents, respectively. This article reviews the currently available human and animal data on potential agents to prevent and/or treat EPS. We propose a strategy for early diagnose EPS in an attempt to avoid the development of the full-blown and potentially life-threatening clinical syndrome of EPS. Future research should focus on studying potential prophylactic and therapeutic agents in humans in large, multicenter, randomized trials but also on early detection of EPS in the inflammatory phase by means of biomarkers and the establishment of a composite EPS score

    Assembling a single-molecule view on nucleosome dynamics

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    The main focus of this thesis is a better understanding of the basic compaction mechanism of our DNA using multiple single-molecule techniques. The stretched-out length of our DNA is enormous compared with the dimensions of a cell. To make DNA fit within a cell it is systematically wrapped around proteins (histone octamers), forming nucleosomes. Besides compacting, nucleosomes also decrease the necessary accessibility of DNA for important processes like transcription and replication. The dynamical interplay between DNA and proteins ensures the right balance between compact and accessible DNA, but is not yet fully understood. Therefore, we study the process of nucleosome assembly as well as the dynamics of nucleosomes at the single-molecule level. Furthermore, we investigate proteins that probe the conformation of the DNA helical structure.BionanoscienceApplied Science
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