Rubbing induced reversible fluorescence switching in thiophene-based organic semiconductor films by mechanical amorphisation

Here, we applied rubbing on thiophene-basedorganic semiconductor thin films to induce a reversible mechanical amorphisation. Amorphisation is associated with fluorescence switching, which is regulated by the polymorphic nature of the film. Thermal annealing of rubbed films produces an opposite effect with respect to rubbing, inducing film crystallization. Notably, thermal crystallisation starts at a low temperature but generates the polymorph stable at a high temperature in the bulk. The mechanism of mechanical transformation is explained considering the mechanical properties of the material and demonstrated through combined X-ray diffraction, atomic force microscopy and photoluminescence at confocal microscopy. This journal i

Platelet Activation in Ovarian Cancer Ascites: Assessment of GPIIb/IIIa and PF4 in Small Extracellular Vesicles by Nano-Flow Cytometry Analysis

In ovarian cancer, ascites represent the microenvironment in which the platelets extravasate to play their role in the disease progression. We aimed to develop an assay to measure ascites’ platelet activation. We enriched small extracellular vesicles (EVs) (40–200 nm) from ascites of high-grade epithelial ovarian cancer patients (n = 12) using precipitation with polyethylene glycol, and we conducted single-particle phenotyping analysis by nano-flow cytometry after labelling and ultra-centrifugation. Atomic force microscopy single-particle nanomechanical analysis showed heterogeneous distributions in the size of the precipitated particles and their mechanical stiffness. Samples were fluorescently labelled with antibodies specific to the platelet markers GPIIb/IIIa and PF4, showing 2.6 to 18.16% of all particles stained positive for the biomarkers and, simultaneously, the EV membrane labelling. Single-particle phenotyping analysis allowed us to quantify the total number of non-EV particles, the number of small-EVs and the number of platelet-derived small-EVs, providing a platelet activation assessment independent of the ascites volume. The percentage of platelet-derived small-EVs was positively correlated with platelet distribution width to platelet count in sera (PDW/PLT). Overall, we presented a high-throughput method that can be helpful in future studies to determine the correlation between the extent of platelet activation in ascites and disease status

Image Retrieval through Abstract Shape Indication

Abstract A new system for image retrieval is presented. The query is indicated by means of three pictures, which should span the abstract shape concept that the user has in mind. The search is accomplished by using a set of size functions, and giving them different weights, computed on the base of the three input images. The system has been tested on a database of 2976 synthetic images

Effect of Spermidine on Misfolding and Interactions of Alpha-Synuclein

Alpha-synuclein (α-Syn) is a 140 aa presynaptic protein which belongs to a group of natively unfolded proteins that are unstructured in aqueous solutions. The aggregation rate of α-Syn is accelerated in the presence of physiological levels of cellular polyamines. Here we applied single molecule AFM force spectroscopy to characterize the effect of spermidine on the very first stages of α-Syn aggregation – misfolding and assembly into dimers. Two α-Syn variants, the wild-type (WT) protein and A30P, were studied. The two protein molecules were covalently immobilized at the C-terminus, one at the AFM tip and the other on the substrate, and intermolecular interactions between the two molecules were measured by multiple approach-retraction cycles. At conditions close to physiological ones at which α-Syn misfolding is a rare event, the addition of spermidine leads to a dramatic increase in the propensity of the WT and mutant proteins to misfold. Importantly, misfolding is characterized by a set of conformations, and A30P changes the misfolding pattern as well as the strength of the intermolecular interactions. Together with the fact that spermidine facilitates late stages of α-Syn aggregation, our data demonstrate that spermidine promotes the very early stages of protein aggregation including α-Syn misfolding and dimerization. This finding suggests that increased levels of spermidine and potentially other polyamines can initiate the disease-related process of α-Syn

Common Features at the Start of the Neurodegeneration Cascade

A single-molecule study reveals that neurotoxic proteins share common structural features that may trigger neurodegeneration, thus identifying new targets for therapy and diagnosis

Minimal information for studies of extracellular vesicles (MISEV2023): From basic to advanced approaches

© 2024 The Authors. Journal of Extracellular Vesicles, published by Wiley Periodicals, LLC on behalf of the International Society for Extracellular Vesicles. This is an open access article distributed under the terms of the Creative Commons Attribution License (CC BY), https://creativecommons.org/licenses/by/4.0/Extracellular vesicles (EVs), through their complex cargo, can reflect the state of their cell of origin and change the functions and phenotypes of other cells. These features indicate strong biomarker and therapeutic potential and have generated broad interest, as evidenced by the steady year-on-year increase in the numbers of scientific publications about EVs. Important advances have been made in EV metrology and in understanding and applying EV biology. However, hurdles remain to realising the potential of EVs in domains ranging from basic biology to clinical applications due to challenges in EV nomenclature, separation from non-vesicular extracellular particles, characterisation and functional studies. To address the challenges and opportunities in this rapidly evolving field, the International Society for Extracellular Vesicles (ISEV) updates its 'Minimal Information for Studies of Extracellular Vesicles', which was first published in 2014 and then in 2018 as MISEV2014 and MISEV2018, respectively. The goal of the current document, MISEV2023, is to provide researchers with an updated snapshot of available approaches and their advantages and limitations for production, separation and characterisation of EVs from multiple sources, including cell culture, body fluids and solid tissues. In addition to presenting the latest state of the art in basic principles of EV research, this document also covers advanced techniques and approaches that are currently expanding the boundaries of the field. MISEV2023 also includes new sections on EV release and uptake and a brief discussion of in vivo approaches to study EVs. Compiling feedback from ISEV expert task forces and more than 1000 researchers, this document conveys the current state of EV research to facilitate robust scientific discoveries and move the field forward even more rapidly.Peer reviewe

Minimal information for studies of extracellular vesicles (MISEV2023): From basic to advanced approaches.

Extracellular vesicles (EVs), through their complex cargo, can reflect the state of their cell of origin and change the functions and phenotypes of other cells. These features indicate strong biomarker and therapeutic potential and have generated broad interest, as evidenced by the steady year-on-year increase in the numbers of scientific publications about EVs. Important advances have been made in EV metrology and in understanding and applying EV biology. However, hurdles remain to realising the potential of EVs in domains ranging from basic biology to clinical applications due to challenges in EV nomenclature, separation from non-vesicular extracellular particles, characterisation and functional studies. To address the challenges and opportunities in this rapidly evolving field, the International Society for Extracellular Vesicles (ISEV) updates its 'Minimal Information for Studies of Extracellular Vesicles', which was first published in 2014 and then in 2018 as MISEV2014 and MISEV2018, respectively. The goal of the current document, MISEV2023, is to provide researchers with an updated snapshot of available approaches and their advantages and limitations for production, separation and characterisation of EVs from multiple sources, including cell culture, body fluids and solid tissues. In addition to presenting the latest state of the art in basic principles of EV research, this document also covers advanced techniques and approaches that are currently expanding the boundaries of the field. MISEV2023 also includes new sections on EV release and uptake and a brief discussion of in vivo approaches to study EVs. Compiling feedback from ISEV expert task forces and more than 1000 researchers, this document conveys the current state of EV research to facilitate robust scientific discoveries and move the field forward even more rapidly

Minimal information for studies of extracellular vesicles (MISEV2023): From basic to advanced approaches

Extracellular vesicles (EVs), through their complex cargo, can reflect the state of their cell of origin and change the functions and phenotypes of other cells. These features indicate strong biomarker and therapeutic potential and have generated broad interest, as evidenced by the steady year-on-year increase in the numbers of scientific publications about EVs. Important advances have been made in EV metrology and in understanding and applying EV biology. However, hurdles remain to realising the potential of EVs in domains ranging from basic biology to clinical applications due to challenges in EV nomenclature, separation from non-vesicular extracellular particles, characterisation and functional studies. To address the challenges and opportunities in this rapidly evolving field, the International Society for Extracellular Vesicles (ISEV) updates its ‘Minimal Information for Studies of Extracellular Vesicles’, which was first published in 2014 and then in 2018 as MISEV2014 and MISEV2018, respectively. The goal of the current document, MISEV2023, is to provide researchers with an updated snapshot of available approaches and their advantages and limitations for production, separation and characterisation of EVs from multiple sources, including cell culture, body fluids and solid tissues. In addition to presenting the latest state of the art in basic principles of EV research, this document also covers advanced techniques and approaches that are currently expanding the boundaries of the field. MISEV2023 also includes new sections on EV release and uptake and a brief discussion of in vivo approaches to study EVs. Compiling feedback from ISEV expert task forces and more than 1000 researchers, this document conveys the current state of EV research to facilitate robust scientific discoveries and move the field forward even more rapidly

Nanoscale Molecular Scale Assembly by Design: Building Flexible or Rigid, Static or Dynamic Nanostructures Thanks to the Controlled Self-assembly of DNA Molecules.

DNA is the molecule that encodes the hereditary information in living organisms. In the last years, the specific recognition abilities and the possibility to encode information that are intrinsic in the DNA molecule have been used to assemble nanoscale structures by design. As suggested by Ned Seeman, the recognized pioneer of this field,[1] the Holliday junction is the fundamental structural element around which a great variety of structures can be designed and implemented: this is a branching point where 4 chains of double-stranded DNA meet. By organizing a number of junctions in a proper fashion, it is possible to create rigid structures that overtake the intrinsic flexibility and stochasticity of polymers to create DNA nanoscale objects with the desired size and shape. Using this type of approach a number of monomeric or polymeric nano-objects can be assembled, with the added possibility of introducing tunable elements, that can change their geometry on an external signal, opening the way towards the construction of nanostructures with controllable dynamics. Using synthetic oligodeoxynucleotides (ODN), in our laboratory we have assembled parallelogram shaped nanostructures made of 4 blocked Holliday junctions that have “sticky ends” on their side. Programmed assembly of these ends brings to the construction of polymers that can be either flexible or rigid (100 nm of persistence length or more). Proper mixing and assembly of different monomer structures can yield different topologies: we can obtain linear, branched or circular nanostructures up to several hundred nanometers in size. The biochemical and structural characterization of these has been performed using gel eletrophoresis and atomic force microscopy. By proper functionalization of the ODNs used for the assembly, it is possible to include non-DNA objects on the structures: this seems a clever strategy to assemble (a desired number of) objects at a controlled distance on a nanostructure, with the possibility of also modulating their dynamics. As an example of this paradigm, we have assembled interacting fluorophores on a rigid parallelogram made of DNA, and we have measured a significant FRET. This does not take place if the fluorophores are, instead, free in solution, or even if they are assembled on incomplete (more flexible) parallelogram structures. Furthermore, by assemblying oligonucleotides in a pH-controlled triple helix, we have recently introduced a novel structural motif to the toolbox for DNA-based molecules constructions.[2] This tool is expected to expand further the possibilities of assemblying and controlling nanostructures made of DNA. [1] N. C. Seeman, Nature 2003, 421, 427. [2] M. Brucale, G. Zuccheri, B. Samorì, Org Biomol Chem 2005, 3, 575

Monitoring the conformational equilibria of monomeric intrinsically disordered proteins by single molecule force spectroscopy

This chapter contains sections titled: Abstract Introduction Basic Elements of the SMFS Methodology for Protein Folding Studies the SMFS Methodology in the Studies of the Mechanical Properties and Interactions of IDPs Probing the Conformational Equilibria of the IDPs Involved in Neurodegenerative Diseases: the Case of \u3b1-Synuclein Future Perspectives Reference