IL-1R and inflammasomes mediate early pulmonary protective mechanisms in respiratory brucella abortus infection

Brucella spp. infection is frequently acquired through contaminated aerosols. The role of interleukin-1 beta (IL-1β) in the early pulmonary response to respiratory Brucella infection is unknown. As shown here, IL-1β levels in lung homogenates and bronchoalveolar lavage fluid (BALF) of mice intratracheally inoculated with B. abortus were increased at 3 and 7 days p.i. At 7 days p.i., pulmonary CFU numbers were higher in IL-1 receptor (IL-1R) knockout (KO) mice than in wild type (WT) mice. At different times p.i. CFU in lungs and BALF were higher in mice lacking some inflammasome components (caspase-1, AIM2, NLRP3) than in WT mice. At 2 days p.i. pulmonary levels of IL-1b and CXCL1 (neutrophils chemoattractant) were lower in caspase-1/11 KO mice. At day 3 p.i., neutrophils counts in BALF were lower in caspase-1/11 KO mice than in WT mice. During in vitro infections, IL-1β secretion was lower in alveolar macrophages from caspase-1/11, NLRP3 or AIM2 KO mice than in WT controls. Similarly, IL-1β production by B. abortus-infected alveolar epithelial cells was reduced by pretreatment with a specific caspase-1 inhibitor. This study shows that IL-1R, probably through IL-1β action, and the NLRP3 and AIM2 inflammasomes are involved in pulmonary innate immune protective mechanisms against respiratory B. abortus infection.Fil: Hielpos, María Soledad. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Houssay. Instituto de Estudios de la Inmunidad Humoral Prof. Ricardo A. Margni. Universidad de Buenos Aires. Facultad de Farmacia y Bioquímica. Instituto de Estudios de la Inmunidad Humoral Prof. Ricardo A. Margni; Argentina. Universidad de Buenos Aires. Facultad de Farmacia y Bioquímica; ArgentinaFil: Fernandez, Andrea Giselle. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Houssay. Instituto de Estudios de la Inmunidad Humoral Prof. Ricardo A. Margni. Universidad de Buenos Aires. Facultad de Farmacia y Bioquímica. Instituto de Estudios de la Inmunidad Humoral Prof. Ricardo A. Margni; ArgentinaFil: Falivene, Juliana. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Houssay. Instituto de Estudios de la Inmunidad Humoral Prof. Ricardo A. Margni. Universidad de Buenos Aires. Facultad de Farmacia y Bioquímica. Instituto de Estudios de la Inmunidad Humoral Prof. Ricardo A. Margni; ArgentinaFil: Alonso Paiva, Iván Mathias. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Houssay. Instituto de Estudios de la Inmunidad Humoral Prof. Ricardo A. Margni. Universidad de Buenos Aires. Facultad de Farmacia y Bioquímica. Instituto de Estudios de la Inmunidad Humoral Prof. Ricardo A. Margni; ArgentinaFil: Muñoz González, Florencia. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Houssay. Instituto de Estudios de la Inmunidad Humoral Prof. Ricardo A. Margni. Universidad de Buenos Aires. Facultad de Farmacia y Bioquímica. Instituto de Estudios de la Inmunidad Humoral Prof. Ricardo A. Margni; ArgentinaFil: Ferrero, Mariana Cristina. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Houssay. Instituto de Estudios de la Inmunidad Humoral Prof. Ricardo A. Margni. Universidad de Buenos Aires. Facultad de Farmacia y Bioquímica. Instituto de Estudios de la Inmunidad Humoral Prof. Ricardo A. Margni; ArgentinaFil: Campos, Priscila C.. Universidade Federal de Minas Gerais; BrasilFil: Vieira, Angelica T.. Universidade Federal de Minas Gerais; BrasilFil: Oliveira, Sergio Costa. Universidade Federal de Minas Gerais; BrasilFil: Baldi, Pablo Cesar. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Houssay. Instituto de Estudios de la Inmunidad Humoral Prof. Ricardo A. Margni. Universidad de Buenos Aires. Facultad de Farmacia y Bioquímica. Instituto de Estudios de la Inmunidad Humoral Prof. Ricardo A. Margni; Argentin

The effects of riboflavin and ultraviolet light on keratocytes cultured in vitro

ABSTRACT Purpose: To culture quiescent human keratocytes and evaluate the effects of ultraviolet light and riboflavin on human corneal keratocytes in vitro. Methods: Keratocytes were obtained from remaining corneoscleral ring donor corneas previously used in corneal transplant surgeries and cultured in DMEM/F12 with 2% FBS until confluence. Characterization of cultured cells was performed by immunofluorescence analysis for anti-cytokeratin-3, anti-Thy-1, anti-α-smooth muscle actin, and anti-lumican. Immunofluorescence was performed before and after treatment of cultured cells with either ultraviolet light or riboflavin. Corneal stromal cells were covered with collagen (200 µL or 500 µL) and 0.1% riboflavin, and then exposed to ultraviolet light at 370 nm for 30 minutes. After 24 hours, cytotoxicity was determined using MTT colorimetric assays, whereas cell viability was assessed using Hoechst 33342 and propidium iodide. Results: Cell cultures achieved confluence in approximately 20 days. Expression of the lumican was high, whereas no expression of CK3, Thy-1, and α-SMA was observed. After crosslinking, MTT colorimetric assays demonstrated a low toxicity rate, whereas Hoechst 33342/propidium iodide staining demonstrated a low rate of apoptosis and necrosis, respectively, in all collagen-treatment groups. Conclusion: Keratocytes can be successfully cultured in vitro and characterized by immunofluorescence using lumican. MTT colorimetric assays, and Hoechst 33342, and propidium iodide staining demonstrated a higher rate of cell death in cells cultured without collagen, indicating collagen protects keratocytes from the cytotoxic effects of ultraviolet light

O LÚDICO NO PROCESSO PEDAGÓGICO DA EDUCAÇÃO INFANTIL: IMPORTANTE, PORÉM AUSENTE

This study observed and analyzed the presence of playful teaching practice on the educational action of teachers in the kindergarten school both from the public and private sectors in the city of São José do Rio Preto. By means of a bibliographical and empirical research, schools from four different parts of the city were evaluated and systematically observed. Later, a questionnaire was applied to the teachers. The main objective was to verify whether or not there were situations in which playful activities would be part of the teacher’s daily agenda, either as a planned activity or even used randomly, as well as the understanding of the teachers concerning this matter in the infantile education.Este estudio analisó la presencia del juguete en el hacer pedagógico de las maestras en guarderías de la red pública de la ciudad de São José do Rio Preto. Por medio de encuestas bibliográficas y empíricas, fueran abordadas escuelas distribuidas en los cuatro sectores de la ciudad en los cuales se realizó una observación sistematizada, y después la aplicación de una encuesta con los maestros. El objectivo primero fue verificar si en el cotidiano de las escuelas tenía o no situaciones de utilización de actividades lúdicas, planeamento de las actividades o empleo obligatorio, como también la comprensión de las mismas acerca de los juguetes infantiles.Este estudo observou e analisou a presença do lúdico no fazer educacional das professoras das escolas infantis das redes pública e particular da cidade de São José do Rio Preto. Por meio de pesquisas bibliográfica e empírica, foram abordadas escolas distribuídas nos quatro setores da cidade, nas quais se realizou, primeiramente, uma observação sistematizada e posteriormente a aplicação de um questionário junto às professoras. O principal objetivo foi verificar se no cotidiano escolar das unidades de ensino infantil envolvidas na pesquisa havia ou não situações de utilização de atividades lúdicas, planejamento das atividades ou seu uso aleatório, assim como a compreensão das professoras acerca do brincar na educação infantil. Os dados coletados foram tratados de modo comparativo entre os dois modelos de escolas compreendidas nos setores público e privado, o que possibilitou traçar um percurso do lúdico na educação infantil e no fazer pedagógico das professoras que, na época, atuavam nos distintos segmentos com a respectiva faixa etária

IL-1R and Inflammasomes Mediate Early Pulmonary Protective Mechanisms in Respiratory Brucella Abortus Infection

Brucella spp. infection is frequently acquired through contaminated aerosols. The role of interleukin-1 beta (IL-1β) in the early pulmonary response to respiratory Brucella infection is unknown. As shown here, IL-1β levels in lung homogenates and bronchoalveolar lavage fluid (BALF) of mice intratracheally inoculated with B. abortus were increased at 3 and 7 days p.i. At 7 days p.i., pulmonary CFU numbers were higher in IL-1 receptor (IL-1R) knockout (KO) mice than in wild type (WT) mice. At different times p.i. CFU in lungs and BALF were higher in mice lacking some inflammasome components (caspase-1, AIM2, NLRP3) than in WT mice. At 2 days p.i. pulmonary levels of IL-1β and CXCL1 (neutrophils chemoattractant) were lower in caspase-1/11 KO mice. At day 3 p.i., neutrophils counts in BALF were lower in caspase-1/11 KO mice than in WT mice. During in vitro infections, IL-1β secretion was lower in alveolar macrophages from caspase-1/11, NLRP3 or AIM2 KO mice than in WT controls. Similarly, IL-1β production by B. abortus-infected alveolar epithelial cells was reduced by pretreatment with a specific caspase-1 inhibitor. This study shows that IL-1R, probably through IL-1β action, and the NLRP3 and AIM2 inflammasomes are involved in pulmonary innate immune protective mechanisms against respiratory B. abortus infection

Phytosociological survey on native pasture of Paspalum notatum

Todos os textos, informa??es e resultados apresentados s?o de inteira responsabilidade dos autores.Coordenador/Orientador do Projeto: M?rcia Vit?ria Santos.Parte da inicia??o cientifica do primeiro autor, financiado pelo Conselho Nacional de Desenvolvimento Cient?fico e Tecnol?gico (CNPq), Funda??o de Amparo ? Pesquisa do Estado de Minas Gerais (FAPEMIG) e Coordena??o de Aperfei?oamento de Pessoal de N?vel Superior (CAPES).Objetivou-se com esse trabalho, realizar o levantamento fitossociol?gico em pastagem nativa de Paspalum notatum no munic?pio de Couto de Magalh?es de Minas-MG. O levantamento fitossociol?gico foi realizado em pastagem nativa de Paspalum notatum no munic?pio de Couto de Magalh?es de Minas MG. Para a identifica??o das plantas daninhas foi utilizado o m?todo do quadrado invent?rio, lan?ando ao acaso na ?rea 40 vezes um quadro de um metro de lado, totalizando uma ?rea amostral de 40m2. Avaliou-se a frequ?ncia (Fre), densidade (Den) e abund?ncia (Abu) das esp?cies encontradas na pastagem. Foram identificadas esp?cies distribu?das em seis fam?lias, destacando-se a fam?lia das Poaceae com cinco esp?cies. As esp?cies que apresentaram maior frequ?ncia (Fre) foram Sidastrum micranthum (malva-preta), Rhynchelystrum repens (capim-favorito) e Crotalaria incana (xique-xique), respectivamente. As esp?cies Eupatorium maximilianii (mata-pasto), Paspalum notatum (grama batatais) e Brachiaria decumbens (braquiarinha), apresentaram os maiores valores de densidade (Den), respectivamente. Paspalum notatum (grama batatais), Eupatorium maximilianii (mata-pasto) e Brachiaria decumbens (braquiarinha) foram ?s esp?cies que apresentaram maior abund?ncia (Abu). A pastagem de Paspalum notatum encontra-se infestada com esp?cies daninhas necessitando controle e corre??o do manejo do pasto para maior desempenho animal.The aim of this study was to do the phytosociological evaluation on a native pasture of Paspalum notatum in Couto de Magalh?es de Minas-MG. The phytosociological evaluation was made on native pasture of Paspalum notatum in Couto de Magalh?es de Minas-MG. To identification of the weed, the method used was the square inventory, hurled randomly in the area 40 times a square of one meter of side, totaling 40m2 of sample area. It was evaluate the frequency (Fre), density (Den), and abundancy (Abu) of the species found in the area. It was identified species of six families, standing out the family of the Poaceae including five species. The species that presented more frequency (Fre) were Sidastrum micranthum (dainty sandmallow), Rhynchelystrum repens (natal grass), and Crotalaria incana (woolly rattlepod), respectively. The species Eupatorium maximilianii ("mata-pasto"), Paspalum notatum (pensacola), and Brachiaria decumbens (signal grass), presented the higher values of density (Den), respectively. Paspalum notatum (pensacola), Eupatorium maximilianii ("mata-pasto"), and Brachiaria decumbens (signal grass), were the species that presented higher abundancy (Abu). The pasture of Paspalum notatum has found infested by weed, requiring control and correction of the management of the area to increase the animal performance

Pastures degradation assessment by infestation in plants weeds

Parte da inicia??o cientifica do primeiro autor, financiado pelo CNPq, FAPEMIG, CAPES.Todos os textos, informa??es e resultados apresentados s?o de inteira responsabilidade dos autores.Objetivou-se a partir desse trabalho avaliar a degrada??o de uma pastagem nativa de Paspalum notatum a partir da ocorr?ncia de plantas daninhas. O levantamento fitossociol?gico foi realizado em pastagem nativa de Paspalum notatum no munic?pio de Couto de Magalh?es de Minas - MG. Para a identifica??o das plantas daninhas foi utilizado o m?todo do quadrado invent?rio, lan?ando ao acaso na ?rea 40 vezes um quadro de um metro de lado, totalizando uma ?rea amostral de 40m2. Avaliou-se a frequ?ncia relativa (Frr), densidade relativa (Der), abund?ncia relativa (Abr) e o ?ndice de valor de import?ncia (IVI) das esp?cies encontradas na ?rea. Os maiores valores de frequ?ncia relativa (Frr) foram das esp?cies Sidastrum micrathum (malva-preta), Imperata brasiliensis (Sap?), Crotalaria incana (xique-xique) e Mimosa pudica (dormideira), respectivamente. As esp?cies Eupatorium maximilianii (mata-pasto), Paspalum notatum (grama batatais), Brachiaria decumbens (braquiarinha) e Cynodon dactylon (estrela roxa) apresentaram os maiores valores de densidade relativa, abund?ncia relativa e ?ndice de valor de import?ncia. A presen?a de Brachiaria decumbens, Cynodon dactylon, Eupatorium maximilianii e a baixa frequ?ncia relativa da esp?cie nativa s?o indicativos que a pastagem encontrase em est?dio de degrada??o avan?ado.ABSTRACT The objective of this study was to evaluate the degradation of a native pasture of Paspalum notatum starting from the weeds occurrence. The phytosociological evaluation was made in native pasture of Paspalum notatum in Couto de Magalh?es de Minas - MG. To identify the weed the method of the inventory square was utilized, hurled randomly in the area 40 times a square of one meter of side, totaling 40m2 of sample area. It was evaluate the relative frequency (Frr), relative density (Der), relative abundancy (Abr), and the value of importance index (IVI) of the species found in the area. The higher values of relative frequency (Frr) were of the species Sidastrum micrathum (dainty sandmallow), Imperata brasiliensis (cogon grass), Crotalaria incana (woolly rattlepod), and Mimosa pudica (sensitive plant), respectively. The species Eupatorium maximilianii ("mata-pasto"), Paspalum notatum (pensacola), Brachiaria decumbens (signal grass), and Cynodon dactylon (bernuda grass) presented the higher values of relative density (Drr), relative abundancy (Abr), and value of importance index (IVI). The presence of the Brachiaria decumbens, Cynodon dactylon, Eupatorium maximilianii and the lower frequency of the native specie, are indicatives that the pasture has found in advanced state of degradation

Analysis of expressed sequence tags from Trypanosoma cruzi amastigotes

A total of 880 expressed sequence tags (EST) originated from clones randomly selected from a Trypanosoma cruzi amastigote cDNA library have been analyzed. Of these, 40% (355 ESTs) have been identified by similarity to sequences in public databases and classified according to functional categorization of their putative products. About 11% of the mRNAs expressed in amastigotes are related to the translational machinery, and a large number of them (9% of the total number of clones in the library) encode ribosomal proteins. A comparative analysis with a previous study, where clones from the same library were selected using sera from patients with Chagas disease, revealed that ribosomal proteins also represent the largest class of antigen coding genes expressed in amastigotes (54% of all immunoselected clones). However, although more than thirty classes of ribosomal proteins were identified by EST analysis, the results of the immunoscreening indicated that only a particular subset of them contains major antigenic determinants recognized by antibodies from Chagas disease patients

Poly (A)+ Transcriptome Assessment of ERBB2-Induced Alterations in Breast Cell Lines

We report the first quantitative and qualitative analysis of the poly (A)+ transcriptome of two human mammary cell lines, differentially expressing (human epidermal growth factor receptor) an oncogene over-expressed in approximately 25% of human breast tumors. Full-length cDNA populations from the two cell lines were digested enzymatically, individually tagged according to a customized method for library construction, and simultaneously sequenced by the use of the Titanium 454-Roche-platform. Comprehensive bioinformatics analysis followed by experimental validation confirmed novel genes, splicing variants, single nucleotide polymorphisms, and gene fusions indicated by RNA-seq data from both samples. Moreover, comparative analysis showed enrichment in alternative events, especially in the exon usage category, in ERBB2 over-expressing cells, data indicating regulation of alternative splicing mediated by the oncogene. Alterations in expression levels of genes, such as LOX, ATP5L, GALNT3, and MME revealed by large-scale sequencing were confirmed between cell lines as well as in tumor specimens with different ERBB2 backgrounds. This approach was shown to be suitable for structural, quantitative, and qualitative assessment of complex transcriptomes and revealed new events mediated by ERBB2 overexpression, in addition to potential molecular targets for breast cancer that are driven by this oncogene