1,720,960 research outputs found

    Capacitance immunosensors for the early detection of circulating cancer biomarkers

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    I have been successful in improving and developing a homemade device based on a three-electrodes electrochemical redout setup thanks to it, we are able to perform measurements of DNA-hybridization, in real-time, from probe ssDNA-SAMs coupled to gold coated sensor surfaces. The measurements were carried out, in pure saline buffer solution, on a large range of concentrations of complementary-DNA strands (from 1 pM to 100 nM), monitoring the differential capacitance at the Working Electrode versus the incubation time. The studies on kinetics, modeled using the Langmuir adsorption model, not only give us important information on the kinetics itself but they allow us to detect eventual mismatches along the DNA-sequence target proving to be sensitive to the position of the mismatch with respect to the surface of the device or to define, in human extract and plasma, the unknown concentration of a specific miRNA-target connected to the heart failure taking into account the hindrances carried by the Argonaute proteins in which the miRNA are inglobed. This goal was achieved by performing a calibration curve on experiments of DNA/DNA hybridization performed in a simple saline buffer. The results were then confirmed using a real time qPCR by our partners in MD D. Cesselli's and MD A.P. Beltrami's group at University of Udine. Another strand of my PhD project concerns the detection of more complex components such as proteins or single-domain antibodies (e.g. VHH fragments)–DNA conjugates, with the final purpose of the detection of circulating tumor cells (CTCs) not only in pure saline buffer but also in human serum. In particular, we have focused on the detection of the protein HER2 whose overexpression is connected to certain aggressive types of breast cancer. In addition, the systematic characterization of the device caught our attention, and it was developed by performing measurements of Self Assembled Monolayer (SAM) detection, carried out in different physiological buffers (KCl, NaCl, MgCl2, PBS, etc.), in different probe-density conditions and applying different potential in order to have a more comprehensive understanding of the phenomena occurring at the electrode/electrolyte interface. Studies that have led to the implementation of a theoretical model, able to provide an acceptable physical explanation of the biorecognition events of interest

    Miniaturized Aptamer-Based Assays for Protein Detection

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    The availability of devices for cancer biomarker detection at early stages of the disease is one of the most critical issues in biomedicine. Towards this goal, to increase the assay sensitivity, device miniaturization strategies empowered by the employment of high affinity protein binders constitute a valuable approach. In this work we propose two different surface-based miniaturized platforms for biomarker detection in body fluids: the first platform is an atomic force microscopy (AFM)-based nanoarray, where AFM is used to generate functional nanoscale areas and to detect biorecognition through careful topographic measurements; the second platform consists of a miniaturized electrochemical cell to detect biomarkers through electrochemical impedance spectroscopy (EIS) analysis. Both devices rely on robust and highly-specific protein binders as aptamers, and were tested for thrombin detection. An active layer of DNA-aptamer conjugates was immobilized via DNA directed immobilization on complementary single-stranded DNA self-assembled monolayers confined on a nano/micro area of a gold surface. Results obtained with these devices were compared with the output of surface plasmon resonance (SPR) assays used as reference. We succeeded in capturing antigens in concentrations as low as a few nM.We put forward ideas to push the sensitivity further to the pM range, assuring low biosample volume (L range) assay conditions

    Grating-Coupled Surface Plasmon Resonance (GC-SPR) Optimization for Phase-Interrogation Biosensing in a Microfluidic Chamber

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    Surface Plasmon Resonance (SPR)-based sensors have the advantage of being label-free, enzyme-free and real-time. However, their spreading in multidisciplinary research is still mostly limited to prism-coupled devices. Plasmonic gratings, combined with a simple and cost-effective instrumentation, have been poorly developed compared to prism-coupled system mainly due to their lower sensitivity. Here we describe the optimization and signal enhancement of a sensing platform based on phase-interrogation method, which entails the exploitation of a nanostructured sensor. This technique is particularly suitable for integration of the plasmonic sensor in a lab-on-a-chip platform and can be used in a microfluidic chamber to ease the sensing procedures and limit the injected volume. The careful optimization of most suitable experimental parameters by numerical simulations leads to a 30–50% enhancement of SPR response, opening new possibilities for applications in the biomedical research field while maintaining the ease and versatility of the configuration

    A compact diffractive sorter for high-resolution demultiplexing of orbital angular momentum beams

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    The design and fabrication of a compact diffractive optical element is presented for the sorting of beams carrying orbital angular momentum (OAM) of light. The sorter combines a conformal mapping transformation with an optical fan-out, performing demultiplexing with unprecedented levels of miniaturization and OAM resolution. Moreover, an innovative configuration is proposed which simplifies alignment procedures and further improves the compactness of the optical device. Samples have been fabricated in the form of phase-only diffractive optics with high-resolution electron-beam lithography (EBL) over a glass substrate. A soft-lithography process has been optimized for fast and cheap replica production of the EBL masters. Optical tests with OAM beams confirm the designed performance, showing excellent efficiency and low cross-talk, with high fidelity even with multiplexed input beams. This work paves the way for practical OAM multiplexing and demultiplexing devices for use in classical and quantum communication

    Going Beyond Counting First Authors in Author Co-citation Analysis

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    The 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

    Variations on the Author

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    “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

    Appropriate Similarity Measures for Author Cocitation Analysis

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    We 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

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    We 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

    Author Index

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