108 research outputs found

    Improved isolation of murine hepatocytes for in vitro malaria liver stage studies

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    <p>Abstract</p> <p>Background</p> <p>Primary hepatocyte cultures are a valuable tool for the understanding of cellular and molecular phenomena occurring during malaria liver stage. This paper describes an improved perfusion/dissociation procedure to isolate hepatocytes from mouse liver that is suitable for malaria studies and allows reproducible preparation of primary hepatocytes with consistent cell yields and controlled purity.</p> <p>Results</p> <p>This protocol is a detailed description of a technique to isolate and culture mouse hepatocytes and represents an improvement over previous descriptions of hepatocyte isolation for malaria studies, regarding three technical aspects: (1) dissociation reagents choice; (2) cell separation gradient and (3) cell purity control. Cell dissociation was optimized for a specific collagenase digestion media. The cell dissociation step was improved by using a three-layer discontinuous gradient. A cell purity check was introduced to monitor the expression of CD95 on hepatocytes using flow cytometry methods.</p> <p>Conclusion</p> <p>The procedure described allows reproducible recovery of one to three million hepatocytes per preparation with cell purity of about 90% as determined by FACS analysis. Completion of the protocol is usually achieved in about four hours per preparation and pooling is suggested for multiple preparations of larger number of cells.</p

    Intonation in European and Brazilian Portuguese

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    This chapter describes the intonation system of Portuguese, concentrating on the analysis of the three main functions of intonation: demarcation, highlighting, and distinction of utterance types. It is based on the description of a single variety: the Lisbon variety for European Portuguese (EP), and the Rio de Janeiro variety for Brazilian Portuguese (BP). The chapter describes the language-particular preferences in intonational grouping that characterize European and Brazilian Portuguese, the size of intonational phrases, the distribution of tonal events within the intonational phrase and the ways in which intonational boundaries are realized. The chapter describes the intonation of two major types of questions: wh-questions and yes-no questions, as types of pragmatically neutral information-seeking questions. It is concerned with the intonation of wh-questions with the question word in sentence initial position. Finally, the chapter presents a typological look at the intonation of Portuguese within the Romance space.info:eu-repo/semantics/publishedVersio

    Quantitative Evaluation of Artifact Removal in Real Magnetoencephalogram Signals with Blind Source Separation

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    The magnetoencephalogram (MEG) is contaminated with undesired signals, which are called artifacts. Some of the most important ones are the cardiac and the ocular artifacts (CA and OA, respectively), and the power line noise (PLN). Blind source separation (BSS) has been used to reduce the influence of the artifacts in the data. There is a plethora of BSS-based artifact removal approaches, but few comparative analyses. In this study, MEG background activity from 26 subjects was processed with five widespread BSS (AMUSE, SOBI, JADE, extended Infomax, and FastICA) and one constrained BSS (cBSS) techniques. Then, the ability of several combinations of BSS algorithm, epoch length, and artifact detection metric to automatically reduce the CA, OA, and PLN were quantified with objective criteria. The results pinpointed to cBSS as a very suitable approach to remove the CA. Additionally, a combination of AMUSE or SOBI and artifact detection metrics based on entropy or power criteria decreased the OA. Finally, the PLN was reduced by means of a spectral metric. These findings confirm the utility of BSS to help in the artifact removal for MEG background activity

    Correlated topographic analysis: estimating an ordering of correlated components

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    Abstract This paper describes a novel method, which we call correlated topographic analysis (CTA), to estimate non-Gaussian components and their ordering (topography). The method is inspired by a central motivation of recent variants of independent component analysis (ICA), namely, to make use of the residual statistical dependency which ICA cannot remove. We assume that components nearby on the topographic arrangement have both linear and energy correlations, while far-away components are statistically independent. We use these dependencies to fix the ordering of the components. We start by proposing the generative model for the components. Then, we derive an approximation of the likelihood based on the model. Furthermore, since gradient methods tend to get stuck in local optima, we propose a three-step optimization method which dramatically improves topographic estimation. Using simulated data, we show that CTA estimates an ordering of the components and generalizes a previous method in terms of topography estimation. Finally, to demonstrate that CTA is widely applicable, we learn topographic representations for three kinds of real data: natural images, outputs of simulated complex cells and text data
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