1,721,006 research outputs found
Custom-built optical tweezers for locally probing the viscoelasticproperties of cancer cells
No full textWe report a home built optical tweezers setup to investigate the mechanism of the mem- brane tether formation from single cells in vitro. Using an optically trapped microbead as probe, we have determined the force-elongation curve during tether formation and extracted several parameters characterizing the viscoelastic behavior of the cell membrane: tether stiffness, force, and viscosity. Breast cancer MDA-MB-231 cells have been studied in two different conditions, at room and physiological temperatures, showing a strong tem- perature dependence of the visoelastic properties of the cell membrane. To get detailed inside information about the tether formation mechanism we have extended the analysis of the force-elongation curves fitting them with a Kelvin model. These preliminary results are part of a larger project of whose goal is to compare the viscoelastic properties of several types of cancer cell lines, characterized by different aggressiveness and metastatic potential
Cdc42 and RhoA reveal different spatio-temporal dynamics upon local stimulation with Semaphorin-3A
Get PDFSmall RhoGTPases, such as Cdc42 and RhoA, are key players in integrating external cues and intracellular signaling pathways that regulate growth cone (GC) motility. Indeed, Cdc42 is involved in actin polymerization and filopodia formation, whereas RhoA induces GC collapse and neurite retraction through actomyosin contraction. In this study we employed Förster Resonance Energy Transfer (FRET) microscopy to study the spatio-temporal dynamics of Cdc42 and RhoA in GCs in response to local Semaphorin-3A stimulation obtained with lipid vesicles filled with Semaphorin-3A and positioned near the selected GC using optical tweezers. We found that Cdc42 and RhoA were activated at the leading edge of NG108-15 neuroblastoma cells during spontaneous cycles of protrusion and retraction, respectively. The release of Semaphorin-3A brought to a progressive activation of RhoA within 30 seconds from the stimulus in the central region of the GC that collapsed and retracted. In contrast, the same stimulation evoked waves of Cdc42 activation propagating away from the stimulated region. A more localized stimulation obtained with Sema3A coated beads placed on the GC, led to Cdc42 active waves that propagated in a retrograde manner with a mean period of 70 seconds, and followed by GC retraction. Therefore, Semaphorin-3A activates both Cdc42 and RhoA with a complex and different spatial-temporal dynamics
Going Beyond Counting First Authors in Author Co-citation Analysis
Get PDFThe present study examines one of the fundamental aspects of author co-citation analysis (ACA) - the way co-citation
counts are defined. Co-citation counting provides the data on which all subsequent statistical analyses and mappings
are based, and we compare ACA results based on two different types of co-citation counting - the traditional type that
only counts the first one among a cited work's authors on the one hand and a non-traditional type that takes into
account the first 5 authors of a cited work on the other hand. Results indicate that the picture produced through this non-traditional author co-citation counting contains more coherent author groups and is therefore considerably clearer. However, this picture represents fewer specialties in the research field being studied than that produced through the traditional first-author co-citation counting when the same number of top-ranked authors is selected and analyzed. Reasons for these effects are discussed
Actin instability alters red blood cell mechanics and Piezo1 channel activity
No full textThe organization and dynamics of the spectrin-actin membrane cytoskeleton play a crucial role in determining the mechanical properties of red blood cells (RBC). RBC are subjected to various forces that induce deformation during blood microcirculation. Such forces also regulate membrane tension, leading to Piezo1 channel activation, which is functionally linked to RBC dehydration through calcium influx and subsequent activation of Gardos channels, ultimately resulting in variations in RBC volume. In this study, we investigated how actin instability affects Piezo1 channel gating, in relation to RBC deformation and mechanical properties, using micropipette aspiration and optical tweezers. Actin instability, induced by 0.5 μM Cytochalasin-D (Cyt-D), led to a 22% reduction in the activation pressure. Additionally, we observed a decreasing trend in Young's modulus, membrane tension, and viscosity. By measuring the time required for cell shape recovery after deformation in an optical trap, we found that Cyt-D-treated RBC took approximately 14% longer to recover compared to untreated cells. The bimodal imaging feature of our experimental approach allowed us to simultaneously measure and correlate activation pressure with mechanical properties at the single-cell level. A significant correlation was found between these parameters in both treated and untreated RBC. Our findings demonstrate the influence of actin instability on both Piezo1 activation and RBC mechanics. These results offer new insights into the interplay between F-actin and Piezo1 in RBC mechanobiology
Variations on the Author
Get PDF“Variations on the Author” discusses two of Eduardo Coutinho’s recent films (Um Dia na Vida, from 2010, and Últimas Conversas, posthumously released in 2015) and their contribution to the general question of documentary authorship. The director’s filmography is characterized by a consistent yet self-effacing form of authorial self-inscription: Coutinho often features as an interviewer that rather than express opinions propels discourses; an interviewer that is good at listening. This mode of self-inscription characterizes him as an author who is not expressive but who is nonetheless markedly present on the screen. In Um Dia na Vida, however, Coutinho is completely absent form the image, while Últimas Conversas, on the contrary, includes a confessional prologue that moves the director from the margins to the center of his films. This article examines the ways in which these works stand out in the filmography of a director who offers new insights into the notion of cinematic authorship
A new approach to follow a single extracellular vesicle–cell interaction using optical tweezers
No full textExtracellular vesicles (EVs) are spherical membrane structures released by most cells. These highly conserved mediators of intercellular communication carry proteins, lipids, and nucleic acids, and transfer these cellular components between cells by different mechanisms, such as endocytosis, macropinocytosis, or fusion. However, the temporal and spatial dynamics of vesicle–cell interactions still remain largely unexplored. Here we used optical tweezers to drive single EVs produced by microglial cells onto the surface of astrocytes or microglia in primary culture. By visualizing single EV–cell contacts, we observed that microglial vesicles displayed different motilities on the surface of astrocytes compared with microglia. After contact, EVs positioned on astrocytes displayed some minor oscillatory motion around the point of adhesion, while vesicles dragged to microglia displayed quite regular directional movement on the plasma membrane. Both the adhesion and motion of vesicles on glial cells were strongly reduced by cloaking phosphatidylserine (PS) residues, which are externalized on the vesicle membrane and act as determinants for vesicle recognition by target cells. These data identify optical manipulation as a powerful tool to monitor in vitro vesicle–cell dynamics with high temporal and spatial resolution and to determine in a quantitative manner the contribution of surface receptors/extracellular protein ligands to the contact
Appropriate Similarity Measures for Author Cocitation Analysis
Get PDFWe provide a number of new insights into the methodological discussion about author cocitation analysis. We first argue that the use of the Pearson correlation for measuring the similarity between authors’ cocitation profiles is not very satisfactory. We then discuss what kind of similarity measures may be used as an alternative to the Pearson correlation. We consider three similarity measures in particular. One is the well-known cosine. The other two similarity measures have not been used before in the bibliometric literature. Finally, we show by means of an example that our findings have a high practical relevance.information science;Pearson correlation;cosine;similarity measure;author cocitation analysis
Dispelling the Myths Behind First-author Citation Counts
Get PDFWe conducted a full-scale evaluative citation analysis study of scholars in the XML research field to explore just how different from each other author rankings resulting from different citation counting methods actually are, and to demonstrate the capability of emerging data and tools on the Web in supporting more realistic citation counting methods. Our results contest some common arguments for the continued
use of first-author citation counts in the evaluation of scholars, such as high correlations between author rankings by first-author citation counts and other citation
counting methods, and high costs of using more realistic citation counting methods that are not well-supported by the ISI databases. It is argued that increasingly available digital full text research papers make it possible for citation analysis studies to go beyond what the ISI databases have directly supported and to employ more
sophisticated methods
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