Educomunicação em Tempos de Pandemia:

Os textos que compõem esta obra são oriundos do VIII Colóquio Ibero-americano de Educomunicação (VIII CIEducom) e IX Colóquio Catarinense de Educomunicação (IX CCEducom), realizados em março de 2021. Em um ano no qual o vírus SARS-CoV-2 e variantes circularam por diversos territórios, Educomunicação em tempos de pandemia: práticas e desafios foi o tema discutido nos eventos. Este livro colocado à disposição do público é um modo de compartilhar caminhos e convidar pessoas curiosas a percorrerem, por meio das palavras e recursos gráficos, desafios identificados e estratégias para o enfrentamento deste inesperado período de pandemia

Evaluation of CD8+ T lymphocytes subpopulation in patients with paracoccidioidomycosis

Orientador: Maria Heloisa de Souza Lima BlottaDissertação (mestrado) - Universidade Estadual de Campinas, Faculdade de Ciências MédicasResumo: As células T CD8+ são caracterizadas pela sua capacidade citotóxica, mediada pela liberação de grânulos citotóxicos ou por meio da ligação da molécula FasL, que resulta na apoptose da célula alvo. Além da função citotóxica, essas células também secretam citocinas, principalmente IFN-g e TNF-a, que contribuem para a ativação de células efetoras capazes de eliminar patógenos. As células T CD8+ foram recentemente classificadas em subpopulações, de acordo com as citocinas que produzem, a saber IFN-g (Tc1), IL-4 (Tc2), IL-10 (Tc10), IL-17 (Tc17), IL-21 (Tc21) e IL-22 (Tc22). Com o objetivo de avaliar a participação das subpopulações de células T CD8+ na paracoccidioidomicose (PCM) humana determinamos a frequência e o perfil destas células em pacientes com as formas aguda (FA) e crônica (FC) da doença. Além disso, verificamos a capacidade de leveduras de P. brasiliensis em induzir a diferenciação de subpopulações de células T CD8+ in vitro. Foi realizada a coleta do sangue periférico de pacientes com PCM e controles saudáveis, seguida do isolamento das células mononucleares, estimulação in vitro para ativação/expansão dos linfócitos e posterior análise fenotípica em citômetro de fluxo. O plasma dos doadores foi utilizado para dosagem de citocinas pela técnica de ELISA. Para os experimentos in vitro realizamos ensaios de cocultura com células dendríticas obtidas a partir de monócitos do sangue periférico de doadores saudáveis, pulsadas in vitro com leveduras de P. brasiliensis e posteriormente cocultivadas com linfócitos T autólogos. A avaliação da polarização das células T CD8+ foi realizada por citometria de fluxo e as citocinas no sobrenadante foram quantificadas por ELISA. As análises realizadas mostraram uma maior frequência de células Tc1 e Tc10 no sangue periférico de pacientes com a FA da PCM. Por outro lado, em pacientes com a FC encontramos uma maior frequência de células T CD8+IFN-g+ (Tc1) e CD8+IL-21+(Tc21). Não foram observadas diferenças quanto a expressão de grânulos citotóxicos entre pacientes e controles. A quantificação das citocinas no plasma, mostrou maiores concentrações de TNF-a e IL-10 em pacientes com a FA em comparação aos controles e pacientes com a FC. Por outro lado, os pacientes com a FC apresentaram menores níveis de IL-22. Nos experimentos de polarização in vitro verificamos que células dendríticas pulsadas com P. brasiliensis foram capazes de induzir subpopulações de células T CD8+ produtoras de IL-17 e IL-22. Além disso, a análise das citocinas no sobrenadante mostrou um aumento na concentração de IFN-g acompanhada de uma diminuição de IL-4 em resposta a estimulação com o fungo. Em conjunto os resultados obtidos indicam que na paracoccidioidomicose, a maior contribuição das T CD8+ fica no âmbito da produção de citocinas, com um perfil misto Tc1/Tc10 na FA e Tc1/Tc21 na FC. Embora componham uma população minoritária em relação às células T CD4+, as células T CD8+ com fenótipo Tc10 poderiam ter, juntamente com as células Th2, uma participação na inibição da resposta celular observada nos pacientes com a forma aguda da PCM. Por outro lado, o perfil Tc1/Tc21 também está de acordo com o padrão predominante da resposta Th1/Th17 em pacientes com a forma crônica da PCM, que representam mais de 90% dos casos da doença. Na mesma linha, os ensaios in vitro evidenciaram que leveduras de P. brasiliensis são capazes de induzir a polarização de subpopulações Tc17/Tc22. Desta forma, podemos presumir que a disponibilidade de células T CD8+ capazes de responder prontamente a estímulos antigênicos com a produção de citocinas contribui para resposta mediada por células T CD4+, além de conferir ao hospedeiro um sistema de defesa alternativo para situações de depleção ou incapacidade das células T CD4+ como ocorre na AIDS e outros quadros de imunodeficiênciaAbstract: CD8+ T cells are characterized by their cytotoxic capacity, mediated by the release of cytotoxic granules or by the binding of FasL, which results in target cell apoptosis. In addition to cytotoxic function, these cells also secrete cytokines, mainly IFN?g and TNF-a, which contribute to the activation of effector cells capable of eliminating pathogens. CD8+ T cells have recently been classified into subpopulations, according to the cytokines they produce, as IFN-g (Tc1), IL-4 (Tc2), IL-10 (Tc10), IL-17 (Tc17), IL-21 (Tc21) and IL-22 (Tc22). Our work aimed to determine the frequency and the profile of CD8+ T cells subpopulations in patients with acute form (AF) and chronic form (CF) of paracoccidioidomycosis (PCM), Furthermore, we intended to verify the capacity of P. brasiliensis yeasts cells to induce the polarization of CD8+ T cells subpopulation in vitro. For that, we collected peripheral blood from patients and healthy donors, isolated the mononuclear cells, stimulated them to activate and expand the lymphocytes and subsequently assessed their phenotype by flow cytometry. The plasma from patients and healthy donor¿s was used for cytokine determination by ELISA. For in vitro experiments we performed a coculture assay with monocytes-derived dendritic cells from healthy donors, pulsed in vitro with yeasts of P. brasiliensis and later cocultivated with autologous T lymphocytes. The polarization of the cells stimulated in vitro was analyzed by flow cytometry and the cytokine production in the supernatant was quantified by ELISA. Our data showed a higher frequency of Tc1 and Tc10 cells in the peripheral blood of patients with the AF of PCM. On the other hand, in patients with the CF we found a higher frequency of CD8+ T cells IFN-a+ (Tc1) and CD8+ IL-21+ (Tc21). No differences were found between patients and healthy donors in relation to the expression of cytotoxic granules. The quantification of cytokines in plasma showed an increase of TNF-a and IL-10 in patients with AF and a decrease in the concentration of IL-22 in patients with CF. The in vitro polarization experiments showed that dendritic cells pulsed with P. brasiliensis were able to induce the polarization of the CD8+ T cells producing IL-17 and IL-22. We also detected higher concentrations of IFN-? in addition to lower concentration of IL-4 in the supernatants of the cultures. All together our results showed that in paracoccidioidomycosis the CD8+ T cells have a major contribution in cytokines production, with mixed profile Tc1/Tc10 in AF and Tc1/Tc21 in CF. In despite of being a small population when compared to CD4+ T cells, CD8+ T cells with Tc10 phenotype could, along with Th2 cells, have a role in inhibiting the cellular response observed in patients with the acute form of PCM. Contrarily, the Tc1 / Tc21 profile are in accordance with the predominant pattern Th1/Th17 seen in patients with the chronic form of PCM, which represent more than 90% of the cases of the disease. Similarly, the in vitro assays showed that P. brasiliensis yeasts are able to induce the polarization of Tc17/Tc22 subpopulations. Based on the results we may assume that the availability of CD8+ T cells capable of promptly respond to antigenic stimuli with cytokine production contributes to a CD4+ T cell-mediated response. Moreover these cells may confer to the host an alternative defense system for situations of depletion or incapacity of CD4+ T cells as in AIDS and other immunodeficiency conditionsMestradoPatologia ClinicaMestra em Ciências165822/2017-1CNP

Association between IL-27 and Tr1 cells in severe form of paracoccidioidomycosis

Interleukin-27, a cytokine of the IL-12 family, is secreted by antigen-presenting cells such as macrophages and dendritic cells (DCs). Recent studies suggest an anti-inflammatory role for IL-27 by inducing IL-10 producing Tr1 cells capable of inhibiting Th1 and Th17 type responses. Our study aimed to investigate the involvement of IL-27 and Tr1 cells in the immunomodulation of paracoccidioidomycosis (PCM), the most prevalent systemic mycosis in Brazil. The presence of IL-27 was evaluated in serum and biopsies of patients with PCM by ELISA, immunohistochemistry, and immunofluorescence. The presence of Tr1 in peripheral blood was analyzed by flow cytometry. In vitro assays were performed to verify the ability of P. brasiliensis yeast to induce IL-27 production by DCs and macrophages, as well as the polarization of lymphocytes to the Tr1 phenotype. Patients with the acute form and severe chronic form, the most severe and disseminated forms of PCM, presented higher serum concentrations of IL-27 and higher percentage of Tr1 cells compared to patients with mild chronic form. IL-27 was also detected in lesions of patients with PCM and associated with DCs and macrophages. P. brasiliensis Pb18 yeasts were able to induce IL-27 production by both DCs and macrophages. We found that DCs pulsed with Pb18 were able to induce Tr1 lymphocytes in vitro. Our data suggest that IL-27 and Tr1 cells could contribute to the deficient immune response to P. brasiliensis that leads to severe and disseminated forms of the disease12

Silver Nimesulide Complex in Bacterial Cellulose Membranes as an Innovative Therapeutic Method for Topical Treatment of Skin Squamous Cell Carcinoma

Oxidative stress and inflammation act on skin squamous cell carcinoma (SSCC) development and progression. Curative therapy for SSCC patients is mainly based on surgical resection, which can cause various sequelae. Silver ions have in vitro activities over tumor cells, while nimesulide has antioxidant and anti-inflammatory activities. This study aimed to evaluate the effects of a silver(I) complex with nimesulide (AgNMS) incorporated in a sustained release device based on bacterial cellulose membrane, named AgNMS@BCM, on topic SSCC treatment. The antiproliferative effect of AgNMS complex was evaluated in the SCC4, SCC15 and FaDu SCC lines. AgNMS complex activity on exposure of phosphatidylserine (PS) residues and multicaspase activation were evaluated on FaDu cells by flow cytometry. The AgNMS@BCM effects were evaluated in a SSCC model induced by 7,12-dimethylbenzanthracene/12-o-tetradecanoyl-phorbol-13-acetate (DMBA/TPA) in mice. Toxicity and tumor size were evaluated throughout the study. AgNMS complex showed antiproliferative activity in SCC15 and FaDu lines in low to moderate concentrations (67.3 µM and 107.3 µM, respectively), and induced multicaspase activation on FaDu cells. The AgNMS@BCM did not induce toxicity and reduced tumor size up to 100%. Thus, the application of AgNMS@BCM was effective and safe in SSCC treatment in mice, and can be seen as a potential and safe agent for topic treatment of SSCC in humans

Detection of mutations in GATA1 gene using automated denaturing high-performance liquid chromatography and direct sequencing in children with Down syndrome

Denaturing high-performance liquid chromatography (dHPLC) was developed to screen DNA variations by separating heteroduplex and homoduplex DNA fragments by ion-pair reverse-phase liquid chromatography. in this study, we have evaluated the dHPLC screening method and direct sequencing for the detection of GATA1 mutations in peripheral blood and bone marrow aspirates samples from children with Down syndrome (DS). Cases were ascertained consecutively as part of an epidemiological study of DS and hematological disorders in Brazil. A total of 130 samples corresponding to 115 children with DS were analysed using dHPLC and direct sequencing methods to detect mutations in GATA1 exons 2, 3 and 4 gene sequences. the overall detection rate of sequencing and dHPLC screening methods was similar. Twenty mutations were detected in exon 2 and one mutation in exon 3 (c.231_232 dupGT) sequences of acute megakaryoblastic leukemia and transient leukemia samples. Four GATA1 mutations were newly described [c.155CG; c.156_178 del23 bp; c.29_30 del GG; c.182CA and c.151AT,c.153_162 del 10 bp). Out of four, three had single nucleotide change. in conclusion, our results indicate that dHPLC is an efficient and valuable tool for GATA1 mutational analysis.Inst Nacl Canc, Div Genet CPq, BR-20331050 Rio de Janeiro, BrazilHosp Darcy Vargas, Pediat Oncohematol Serv, São Paulo, BrazilSoc Oncol Bahia, Salvador, BrazilCtr Infantil Invest Hematol D Boldrini, São Paulo, BrazilHosp Santa Casa Misericordia, Pediat Oncohematol Serv, Itabuna, Bahia, BrazilHosp Martagao Gesteira, Pediat Oncohematol Serv, Salvador, BA, BrazilUniv Santa Maria Rio Grande Sul, Dept Hematol, Porto Alegre, RS, BrazilHosp Joana Gusmao Florianopolis, Pediat Oncohematol Serv, Santa Catarina, BrazilUniversidade Federal de São Paulo, Inst Oncol Pediat, São Paulo, BrazilHosp AC Camargo Fund Antonio Prudente, Pediat Oncohematol Serv, São Paulo, BrazilHosp Canc Cascavel, UOPECCAN, Cascavel, Parana, BrazilHosp Napoleao Laureano, Joao Pessoa, Paraiba, BrazilUFRJ, Inst Pediat & Puericultura Martagao Gesteira, Rio de Janeiro, Paraiba, BrazilHosp Serv Estado Rio de Janeiro, Dept Pediat, Rio de Janeiro, BrazilUniversidade Federal de São Paulo, Inst Oncol Pediat, São Paulo, BrazilWeb of Scienc