Apéndice I: vocabulario del Ingeniero nacional don Juan Pelleschi

Estos dos importantes Vocabularios son extractados de las carteras del Sr. Pelleschi, y él los apuntó de lo que oía hablar á los indios que lo rodeaban. Su valor para la fonología Chaquense es grande, pues nos hace leer lo que oía un italiano que apuntaba con signos y fonología castellana. Las curiosidades que contiene son muchas.Facultad de Ciencias Naturales y Muse

Apéndice I: vocabulario del Ingeniero nacional don Juan Pelleschi

Estos dos importantes Vocabularios son extractados de las carteras del Sr. Pelleschi, y él los apuntó de lo que oía hablar á los indios que lo rodeaban. Su valor para la fonología Chaquense es grande, pues nos hace leer lo que oía un italiano que apuntaba con signos y fonología castellana. Las curiosidades que contiene son muchas.Facultad de Ciencias Naturales y Muse

Apéndices a la gramática Mocoví

Estos dos importantes Vocabularios son extractados de las carteras del Sr. Pelleschi, y él los apuntó de lo que oía hablar á los indios que lo rodeaban. Su valor para la fonología Chaquense es grande, pues nos hace leer lo que oía un italiano que apuntaba con signos y fonología castellana.Las curiosidades que contiene son muchas.Chico         — mezca lúh — es literalmente «no grande» Mocovitas  — Mo-co-uit.   (Aquí se vé que esos indios oyende tales).Tobas        — Ntocu-it.El vocabulario de la india Fulgencia es aún mas interesante porque contiene frases.¿Me conoces?    —    Mediadini?Aquí está la M interrogante, el Di caso régimen de 1a, y adini 2a persona del verbo.No te conozco    —    Misi   — S-adini.incluye  el Misi negación   S sujeto de 1a y tema  verbal de 2a persona.Nosotros   —    Nojeum.r     rda el valor fonético de la c doble en oceom.¿Me quieres?   —   Mujutir-ibá?Este ejemplo á la interrogante M agrega la terminación tran sitiva de 1aiba = «me».No te quiero    — Mesirujutir-abá.La Me ó Mes es la negación, el sujeto yo está en el Sir y la terminación abá es el caso régimen te. Estos ejemplos confir man lo escrito en el texto del Arte.Si te quiero    —    Eeh-há tirabau

Evaluation of the cell growth, sensitivity of the <i>Pbccp</i>-aRNA mutant to oxidative stress and survival in macrophages.

<p><b>(A)</b> Curve of cell growth in BHI medium with the strains <i>Pb</i>18-WT (wild type <i>Pb</i>18 strain), <i>Pb</i>339-WT (wild type <i>Pb</i>339 strain), EV: yeast cells (<i>Pb</i>339 strain containing the empty vector); <i>Pbccp</i>-aRNA1 and <i>Pbccp</i>-aRNA2: independent colonies of yeast cells (<i>Pb</i>339 strain) containing the cassette with the CCP-AS fragment. <b>(B)</b>The <i>Pbccp-</i>aRNA sensitivity to oxidative stress was examined in the presence of 40 μM and 80 μM of menadione. <i>P</i>. <i>brasiliensis Pb</i>18-WT, <i>Pb</i>339-WT and EV were used as controls. <b>(C)</b> Interaction assay of <i>P</i>. <i>brasiliensis</i> and macrophage cells. The experiments were performed in biological triplicates. <i>Pb</i>18-WT: wild type (<i>Pb</i>18 strain); <i>Pb</i>339-WT: wild type (<i>Pb</i>339) EV: yeast cells (<i>Pb</i>339 strain) containing the empty vector without CCP-AS; <i>Pbccp</i>-aRNA1 and <i>Pbccp</i>-aRNA2: independent colonies of yeast cells (<i>Pb</i>339 strain) containing the cassette with the CCP-AS fragment. The asterisk denotes <i>p</i> < 0.01(Student’s t-test).</p

Interaction of <i>P</i>. <i>brasiliensis</i> yeast cells with macrophages and evaluation of phagolysosome maturation.

<p>The interaction assay was performed using two <i>P</i>. <i>brasiliensis</i> isolates (<i>Pb</i>18 and <i>Pb</i>339) and are shown in the lanes named macrophages + <i>Pb</i>18 yeast cells and macrophages + <i>Pb</i>339 yeast cells, respectively. The pictures were taken in bright field (shown in the bright field column), at 395/420 nm for Calcofluor probe (shown in the Calcofluor white column) and at 579/599 nm for Lysotracker probe (shown in the Lysotracker column). Fungal control cells are shown in the lanes named <i>Pb</i>18 yeast cells and <i>Pb</i>339 yeast cells. Control macrophage cells were also performed and are shown in the lane named macrophage (control). All representative pictures were taken using an Axioscope microscope (Carl Zeiss) and magnified 1000X.</p

Molecular mechanism used by <i>P</i>. <i>brasiliensis</i> to survive inside macrophages.

<p>The up regulated enzymes in <i>P</i>. <i>brasiliensis</i> during macrophage interaction are: PGM: phosphoglucomutase; PBPase: fructose 1,6-biphosphatase; GAPDH glyceraldehyde 3-phosphate dehydrogenase; PGAM: phosphoglycerate mutase; ENO: enolase; PDC: pyruvate decarboxylase; ADH: alcohol dehydrogenase; PDH: pyruvate dehydrogenase SDH: succinate dehydrogenase, ECH: enoyl-CoA hydratase; AGXT: alanine glyoxylate aminotransferase; GOT: aspartate aminotransferase; HDP: 4-hydroxyphenylpyruvate dioxygenase; ABAT: 4-aminobutyrate aminotransferase; GLUD: glutamate dehydrogenase; ASP: aspartyl protease; CXP: carboxypepetidase Y; APE: vacuolar aminopeptidase; ND: NADH ubiquinone oxidoreductase; and ATPase: ATP synthase. The numbers before enzyme names represent increasing rates in the protein expression during macrophage interaction. The asterisk represents the proteins detected in <i>P</i>. <i>brasiliensis</i> only during macrophage infection.</p

Evaluation of silencing efficiency of cytochrome c peroxidase knockdown mutants (<i>Pbccp</i>-aRNA).

<p>Relative quantification performed by real-time quantitative PCR to confirm CCP silencing. WT: wild type yeast cells (<i>Pb</i>339 strain); EV: yeast cells (<i>Pb</i>339 strain) containing the empty vector with no CCP-AS; <i>Pbccp</i>-aRNA1 and <i>Pbccp</i>-aRNA2: colonies from yeast cells (<i>Pb</i>339 strain) containing the cassette with the <i>ccp</i> antisense fragment. The Student's t-test was used for statistical comparisons. Error bars represent the standard deviation from three biological replicates, while * represents <i>p</i>≤0.05.</p

Up regulated proteins putatively related to cell rescue and defense in <i>P</i>. <i>brasiliensis</i> yeast cells during macrophage infection.

<p>¹ Accession number obtained in the <i>Paracoccidioides</i> database available at <a href="http://www.broadinstitute.org/annotation/genome/paracoccidioides_brasiliensis/MultiHome.html" target="_blank">http://www.broadinstitute.org/annotation/genome/paracoccidioides_brasiliensis/MultiHome.html</a>.</p><p>² PLGS score is the result of different mathematical models for peptide and fragment assign prediction.</p><p>³ Fold-change values were obtained by dividing the values of protein abundance (in fmol) from <i>P</i>. <i>brasiliensis</i> yeast cells during macrophage infection by the abundance in the control. Proteins with a minimum fold-change of 50% were considered to be regulated.</p><p>* Proteins detected in <i>P</i>. <i>brasiliensis</i> only during macrophage infection.</p><p>Up regulated proteins putatively related to cell rescue and defense in <i>P</i>. <i>brasiliensis</i> yeast cells during macrophage infection.</p

Virulence of the <i>Pb</i>ccp-aRNA mutant in the liver and spleen of infected BALB/c mice.

<p>Colony forming units recovered from the spleen (A) and liver (B) of mice infected with <i>P</i>. <i>brasiliensis</i> wild type (WT), <i>P</i>. <i>brasiliensis</i> containing the empty vector (EV) and the silenced mutant <i>Pb</i>ccp-aRNA1. The experiments were performed in biological triplicates. Error bars represent the standard deviation from biological replicates, while * represents <i>p</i>-values < 0.01.</p

Macrophage Interaction with <i>Paracoccidioides brasiliensis</i> Yeast Cells Modulates Fungal Metabolism and Generates a Response to Oxidative Stress

<div><p>Macrophages are key players during <i>Paracoccidioides brasiliensis</i> infection. However, the relative contribution of the fungal response to counteracting macrophage activity remains poorly understood. In this work, we evaluated the <i>P</i>. <i>brasiliensis</i> proteomic response to macrophage internalization. A total of 308 differentially expressed proteins were detected in <i>P</i>. <i>brasiliensis</i> during infection. The positively regulated proteins included those involved in alternative carbon metabolism, such as enzymes involved in gluconeogenesis, beta-oxidation of fatty acids and amino acids catabolism. The down-regulated proteins during <i>P</i>. <i>brasiliensis</i> internalization in macrophages included those related to glycolysis and protein synthesis. Proteins involved in the oxidative stress response in <i>P</i>. <i>brasiliensis</i> yeast cells were also up-regulated during macrophage infection, including superoxide dismutases (SOD), thioredoxins (THX) and cytochrome c peroxidase (CCP). Antisense knockdown mutants evaluated the importance of CCP during macrophage infection. The results suggested that CCP is involved in a complex system of protection against oxidative stress and that gene silencing of this component of the antioxidant system diminished the survival of <i>P</i>. <i>brasiliensis</i> in macrophages and in a murine model of infection.</p></div