La renovación de la palabra en el bicentenario de la Argentina : los colores de la mirada lingüística

El libro reúne trabajos en los que se exponen resultados de investigaciones presentadas por investigadores de Argentina, Chile, Brasil, España, Italia y Alemania en el XII Congreso de la Sociedad Argentina de Lingüística (SAL), Bicentenario: la renovación de la palabra, realizado en Mendoza, Argentina, entre el 6 y el 9 de abril de 2010. Las temáticas abordadas en los 167 capítulos muestran las grandes líneas de investigación que se desarrollan fundamentalmente en nuestro país, pero también en los otros países mencionados arriba, y señalan además las áreas que recién se inician, con poca tradición en nuestro país y que deberían fomentarse. Los trabajos aquí publicados se enmarcan dentro de las siguientes disciplinas y/o campos de investigación: Fonología, Sintaxis, Semántica y Pragmática, Lingüística Cognitiva, Análisis del Discurso, Psicolingüística, Adquisición de la Lengua, Sociolingüística y Dialectología, Didáctica de la lengua, Lingüística Aplicada, Lingüística Computacional, Historia de la Lengua y la Lingüística, Lenguas Aborígenes, Filosofía del Lenguaje, Lexicología y Terminología

Expression of TcPARG throughout the <i>Trypanosoma cruzi</i> life-cycle.

<p>(A) Microarray expression data for TcPARG over the course of <i>T. cruzi</i> life-cycle. TcPARG mRNA relative abundance was evaluated by using the transcriptome analysis of different <i>T. cruzi</i> stages available at Gene Expression Omnibus database (<a href="http://www.ncbi.nlm.nih.gov/geo" target="_blank">www.ncbi.nlm.nih.gov/geo</a>, DataSets: GSE14641). Shown are mean microarray log₂ ratios (stage/reference) for TS significantly regulated in <i>Trypanosoma cruzi</i> amastigotes (AMA), trypomastigotes (TRYP), epimastigotes (EPI), and metacyclic trypomastigotes (META). (B) Western blot analysis of the three life-cycle stages of <i>T. cruzi</i>. Protein extracts (35 µg) of amastigote, epimastigote or trypomastigote stages of <i>T. cruzi</i> were solved in a 10% polyacrylamide gel, transfer to a nitrocellulose membrane and revealed with an anti-TcPARG (1:10000) specific antiserum. β-tubulin was used as loading control.</p

Role of TcPARG in <i>Trypanosoma cruzi</i> epimastigotes proliferation and cell cycle progression.

<p>A) Effect of the PARG inhibitors ADP-HPD and DEA on <i>T. cruzi</i> growth and survival was determined by incubating epimastigotes at an initial density of 10⁷ parasites/ml in the continuous presence of inhibitors at 1 µM. The number of epimastigotes was determined daily by counting formaldehyde-fixed parasites in a Neubauer chamber. All data points were determined in triplicates and shown as means with standard deviation. The significance of the results versus the control at day 4 was analyzed with t test and indicated in the figure (* p0.05). B) Effect of ADP-HPD at 1 µM concentration on cell cycle progression of epimastigotes was determined by adding the inhibitor at the indicated concentration to the culture media of hydroxyurea synchronized parasites after digitonin permeabilization. Samples were drawn every 2 hours for 14 hours and DNA content was determined by propidium iodide staining followed by flow cytometry analysis. The percentage of epimastigotes in each cell cycle phase was determined by setting gates according to the DNA content in the 0 hs of the control sample and maintained for all other samples. The data were analyzed using the Cyflogic software.</p

Effect of PARG inhibitors on <i>T. cruzi</i> infection on Vero or A549 host cells.

<p><i>T. cruzi</i> trypomastigotes were purified from the supernatant of previously infected cells and preincubated for 30 min in the respective culture medium in the absence (Control) or presence of 1 µM PARG inhibitor (DEA). Twenty-four hours Vero, A549 wild type or shPARG (hPARG silenced) cell monolayers were infected with 50 trypomastigotes/cell. The infection process was followed by microscopic direct visualization. At the indicated days (A and C) or at day 6 post-infection (B and D), percentage of infected cells and number of amastigotes intracellular were determined on May-Grünwald Giemsa stained samples. Amastigotes and cells were counted using the ImageJ software in at least 7 fields. The number of trypomastigotes/ml in the supernatant of infected cell cultures was determined by counting unfixed trypomastigotes in a Neubauer chamber at the indicated days (E) or at day 9 post-infection (F). All points were determined in triplicates and shown as means with standard deviation. Significance of the result versus the Control (***p0.001; two way ANOVA) or Wild Type Control (***p0.001; **, p0.01; two way ANOVA) is indicated.</p

Immunolocalization of PARG on <i>Trypanosoma cruzi</i>, CL Brener epimastigotes.

<p>The parasites were fixed for 25 min with 3.8% (W/V) formaldehyde in PBS at 4°C, permeabilized with fresh PBS - 0,1% Triton X-100 and blocked for 1 h at room temperature with 5% (W/V) BSA in PBS. (A) Differential interference contrast (DIC). (B) Cells were counterstained with DAPI to identify nuclear DNA and kinetoplastid DNA. (C) PARG was detected with 1:500 mouse polyclonal TcPARG antibody followed by 1:600 Alexa Fluor 488 goat anti-mouse IgG antibody. (D) Merge of PARG and DNA signals show the nuclear localization of this enzyme. Bar: 10 µm. (E–F) For electron microscopy, epimastigotes were fixed in PBS 2.5% glutaraldehyde, 4% formaldehyde, embedded in epoxy resin and PARG detected with 1:50 mouse polyclonal TcPARG antibody followed by 1:100 anti-mouse antibody conjugated with 10-nm gold particle. N: nucleus; K: kinetoplast. Bar: 0.2 µm.</p

Amino acid sequence alignment of the PARG signature from different organisms.

<p>The multiple alignment of the PARG signature amino acid sequences corresponding to <i>T. cruzi</i> PARG (accession number ABG73229); <i>T. brucei</i> PARG (GeneDB Systematic Name: Tb09.211.3760); <i>C. elegans</i>_1 PARG (accession number NP_501496) and <i>C. elegans</i>_2 PARG (accession number NP_501508); <i>T</i><i>. thermophila</i> (accession number EAR94344); <i>A. thaliana</i>_1 PARG (accession number NP_973578); <i>A. thaliana</i>_2 PARG (accession number AAK72256); <i>D. discoideum</i> PARG (accession number XP_642024); <i>D. melanogaster</i> PARG (accession number NP_477321); <i>C</i><i>. quinquefasciatus</i> PARG (accession number XP_001853435); <i>A. aegypti</i> PARG (accession number XP_001659301); <i>D. rerio</i> PARG (accession number XP_001338257); <i>X. laevis</i> PARG (accession number NP_001089602); <i>G. gallus</i> PARG (accession number XP_421502); <i>B. taurus</i> PARG (accession number NP_776563); <i>R. norvegicus</i> PARG (accession number NP_112629); <i>M. musculus</i> PARG (accession number NP_036090); <i>H. sapiens</i> PARG (accession number NP_003622); <i>P. abel</i>ii PARG (accession number NP_001125086); <i>P</i><i>. troglodytes</i> PARG (accession number XP_001139727) was generated with the ClustalW2 program and edited with the BOXSHADE (3.21) software. Colors used for amino acids background are as follow: white for different residues, black for identical residues, gray for similar and conserved residues. Asterisk: essential acidic residues D-E-E, underlined: key residues, G and two consecutive E, and black diamond: important Y residue.</p