O Papel do ensino do português como língua estrangeira na defesa do multicultarismo

As política actuais existentes a nível oficial para a implementação e defesa do ensino da Língua Portuguesa como Língua Estrangeira (L. E.) na Europa e no resto do mundo levam-nos a pensar que são, sobretudo, os casos isolados de leitores portugueses pioneiros, inspirados e marginais que na sua missão individual e afastada lutam pela implementação e defesa desta língua nos seus países de acolhimento. Segundo Volfgram, “cabe ensinar a alguns que o multiculturalismo não está apenas na teoria e sim ao nosso redor, nos elevando realmente à condição de seres humanos” (2005), e o mesmo é dizer que o multiculturalismo começa nas suas bases pela aprendizagem desinteressada e não interesseira das crianças na sua mais tenra idade. Não é impunemente que em países multiculturais como a Bélgica, a Língua Portuguesa ensinada como segunda língua ou como língua estrangeira desempenha um papel preponderante na defesa e na preservação do Português e, em simultâneo, pugna pela defesa incontestável da necessidade incontornável que o multiculturalismo é hoje. É indubitável que a luta contra a xenofobia, a luta pela tolerância e o respeito mútuo, bem como o diálogo profícuo biunívoco não podem sobreviver actualmente sem uma consciencialização da importância das línguas minoritárias, da crioulização, da relação com as línguas maioritárias e da conquista da defesa do multiculturalismo hic et nunc. Abordando algumas opiniões avisadas, esperamos trazer à discussão temas importantes, tais como, a necessidade de articulação de políticas de difusão da língua portuguesa na Europa e no Mundo concertadamente com o Brasil e outros Países Lusófonos, a necessidade de implementação de medidas concretas no terreno para defesa da Língua de Camões fora de Portugal, a sobrevivência do Português que embora sendo minoritária na Europa é uma das línguas mais faladas no mundo, a necessidade da consciencialização para a crescente importância geo-estratégica do Português paralelamente com o recrudescimento do multiculturalismo à escala global

Analysis of LexA binding sites and transcriptomics in response to genotoxic stress in Leptospira interrogans

We determined the effects of DNA damage caused by ultraviolet radiation on gene expression in Leptospira interrogans using DNA microarrays. These data were integrated with DNA binding in vivo of LexA1, a regulator of the DNA damage response, assessed by chromatin immunoprecipitation and massively parallel DNA sequencing (ChIP-seq). In response to DNA damage, Leptospira induced expression of genes involved in DNA metabolism, in mobile genetic elements and defective prophages. The DNA repair genes involved in removal of photo-damage (e.g. nucleotide excision repair uvrABC, recombinases recBCD and resolvases ruvABC) were not induced. Genes involved in various metabolic pathways were down regulated, including genes involved in cell growth, RNA metabolism and the tricarboxylic acid cycle. From ChIP-seq data, we observed 24 LexA1 binding sites located throughout chromosome 1 and one binding site in chromosome 2. Expression of many, but not all, genes near those sites was increased following DNA damage. Binding sites were found as far as 550 bp upstream from the start codon, or 1 kb into the coding sequence. Our findings indicate that there is a shift in gene expression following DNA damage that represses genes involved in cell growth and virulence, and induces genes involved in mutagenesis and recombination.Conselho Nacional de Pesquisas (Brazil)Fundacao de Amparo a Pesquisa do Estado de Sao PauloFundacao ButantanNational Institutes of Health (U.S.) (National Institute of General Medical Sciences (U.S.) Award GM41934

Phenotypic analyses of <i>L. interrogans</i> serovar Copenhageni after UV-C irradiation.

<p>(A) Relationship between survival after treatment with increasing doses of UV-C (left axis) and the mean size of the persistent cells (right axis). Cultures of bacteria were treated with increasing doses of UV-C, diluted in fresh medium and incubated during 24h in the dark. Surviving bacteria were counted and the frequency was calculated as the ratio of irradiated to non-irradiated cells. (B) Fluorescent microscopy of leptospires in cultures treated or not with UV-C, after staining with DAPI (1000x magnification). The bar represents 20 µm. (C) Frequencies of the size categories from treated and non-treated samples. Categories were determined by the size values (expressed in µm) that divided the non-irradiated culture in its four quartiles. All the data represent the average of three independent experiments.</p

Genomic and transcriptional organization of the <i>lexA1</i> region.

<p>(A) Schematic representation of the <i>lexA1</i> genomic region. The arrows indicate the direction of transcription. The fragments amplified by the primer pairs used for the RT-PCR analysis are indicated by numbered lines below the genes. (B) Composite image of agarose gels from resulting RT-PCR reactions, using either genomic DNA (DNA), RNA (RT-) or cDNA (RT+) as templates. The numbers refer to the respective fragments shown in (A).</p

Analysis of LexA2 binding.

<p>(A) LexA2 binding assays were carried out with 80 ng of purified recombinant protein, using 1.55 fmol DIG-labeled lexA2_up as probe. For the competition assays, 200 fold excess of unlabeled probes was added to the binding reaction. (B) Scheme representing the fragments used for LexA2 binding experiments. Genomic coordinates and size of each fragment (in nucleotides) are indicated. Box 1 indicates the palindrome ATTCN ₁₃GAAT, and box 2, TTGTAN₁₀TACAA. The putative <i>lexA2</i> promoter is indicated by an arrow. (C) Competition assays, where 200 fold excess of unlabeled probes was added to the binding reactions, corresponding to the two putative binding sites contained in lexA2up. The number of nucleotides flanking the palindromes are indicates in braces ({3} or{9}).</p

Expression of transcripts in (A) <i>lexA1</i> and (B) <i>lexA2</i> genomic regions post-treatment with UV-C.

<p>The graphic represents the fold change in gene expression in the irradiated sample <i>versus</i> the non-irradiated one 12h after irradiation with 4.5 J.m^-2 and analyzed by qPCR. Genes showing significant changes are indicated by *** (P value<0.001) or ** (P value>0.01). Below, the genomic organization is indicated.</p

Expression kinetics of <i>L. interrogans</i> serovar Copenhageni genes in response to UV-C.

<p>The <i>Leptospira</i> culture was irradiated with 4.5 J.m^-2, kept in the dark, and the RNA samples were obtained at the indicated time points. The fold change corresponds to gene expression in the irradiated sample <i>versus</i> the non-irradiated one at each time point, analyzed by qPCR. The 16S was used as normalizer. Error bars represent the standard deviation of the average of three independent experiments. Samples showing significant changes are indicated by *** (P value<0.001), ** (P value>0.01) or * (P value>0.05).</p

Genomic and transcriptional organization of the <i>lexA2</i> region.

<p>(A) Schematic representation of the <i>lexA2</i> genomic region from <i>L. interrogans</i> serovar Copenhageni (upper) compared to the equivalent region of serovar Lai (lower). Arrows represent predicted genes and transcription orientation. Light grey arrows represent genes orthologous between genomes, dark grey genes that are specific to Copenhageni and black arrows indicate genes encoding transposases. The white arrows represent genes with truncated versions in Lai genome (traced arrows) by insertion of IS elements. Remnants of a phage integrase are indicated by a traced line. The numbered bars below the genes indicate the amplified fragments corresponding to the primer pairs used in the RT-PCR analyses. (B) RT-PCR reactions, using either genomic DNA (gDNA), RNA (RT-) or cDNA (RT+) as templates, and primers flanking intergenic regions. The numbers refer to the respective fragments shown in (A).</p

Phylogenetic analysis of LexA.

<p>Phylogenetic analysis was performed using LexA amino acid sequences from several bacteria. The leptospiras clade is highlighted by a grey box, and the sequences of the LexA proteins present in <i>L. interrogans</i> serovar Copenhageni are indicated with arrows. Local bootstrap values are attached to the internal nodes. Species code description and sequences used are compiled in <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0076419#pone.0076419.s005" target="_blank">Table S1</a>.</p