hiPSC and hiPSC-cardiomyocytes are alternative EV biofactories for cardiac regeneration

In cardiac regenerative medicine, there is a growing interest in using EV as cell-mimetic therapeutics due to their potential superior efficacy and overall advantages over cell transplantation: i) absence of oncogenic risk, ii) low immunogenicity [1], iii) easier large-scale manufacturing and iv) consistent product profile. However, most studies have focused particularly on the potential of either mesenchymal or cardiac progenitor cell-derived EV to promote cardiac repair [2]. Here, we study the potential of human induced pluripotent stem cells (hiPSC) and hiPSC-derived cardiomyocytes (hiPSC-CM) as alternative cell factories for the production of a high yield of therapeutic EV for cardiac regeneration. Due to their high self-renewal ability and capacity to differentiate into functional cardiomyocytes, these cells can provide an unlimited source of EV for application in cardiac regeneration. We generated and characterized EV derived from key stages of hiPSC-CM differentiation and maturation, i.e. from hiPSC (hiPSC-EV), cardiac progenitors (CPC-EV), immature (CMi-EV) and mature (CMm-EV) cardiomyocytes, with the goal of studying their potential role as therapeutics, and whether their yield and function was influenced by the state of their parent cell. Two hiPSC lines were differentiated into hiPSC-CM and cultured as 3D spheroids in a fatty acid supplemented medium to improve CM maturation [3,4]. EV isolation was performed based on density separation on an iodixanol discontinuous gradient, and EV were characterized in terms of particle size and particle size distribution, presence of EV-specific markers, and imaging through transmission electron microscopy. Functional studies were performed using human umbilical vein endothelial cells (HUVECs) to evaluate EV-uptake, migration and angiogenesis. EV yield varied along CM differentiation stages, with a minimum for CPC, for both cell lines. Bioactivity assays with HUVECs showed that uptake of PKH26-labelled EV could be blocked by dynasore, an inhibitor of dynamin-2, a GTPase that plays a crucial role in clathrin and caveolin-dependent endocytosis. Increased migration was observed in HUVECs treated with hiPSC, CPC and CM-derived EV (92.25 ± 14.69% wound closure at 24h for hiPSC, 77.13 ± 13.64 % for CPC, 74.71 ± 19.86% for CMi, 69.2 ± 19.12% for CMm versus 45.65 ± 7.26% for control), but angiogenic properties were found only for hiPSC-EV (fold change of 11.2 ± 4.59 in total segment length vs. control, p\u3c0.001). Current efforts towards the characterization of EV small RNA cargo aim at understanding the correlation between cargo composition and in vitro activity, to identify the optimal cell factory for scalable therapeutic EV production. Funding: FCT PhD fellowship PD/BD/139078/2018; IC&TD Projects MetaCardio” (PTDC/BTM-SAL/32566/2017) and NETDIAMOND (SAICTPAC/0047/2015), and iNOVA4Health Research Unit (UIDB/04462/2020). 1. Zhu, X., et al. Comprehensive toxicity and immunogenicity studies reveal minimal effects in mice following sustained dosing of extracellular vesicles derived from HEK293T cells. Journal of Extracellular Vesicles 6, 1, 2017. 2. El Harane, et al. Acellular therapeutic approach for heart failure: in vitro production of extracellular vesicles from human cardiovascular progenitors. European Heart Journal 39, 20, 2018. 3. Correia, C., et al. Distinct carbon sources affect structural and functional maturation of cardiomyocytes derived from human pluripotent stem cells. Scientific reports 7, 1, 2017. 4. Correia, C., et al. 3D aggregate culture improves metabolic maturation of human pluripotent stem cell derived cardiomyocytes. Biotechnology and Bioengineering 115, 3, 2018

Global transcriptome characterization of Candida glabrata biofilms in response to acetate and fluconazole

Candida glabrata is considered a major opportunistic fungal pathogen of humans and has emerged as a leading cause of nosocomial fungal infections. The capacity of this yeast species to cause infections is dependent on the ability to grow within the human host environment and to assimilate the carbon sources available. Previous studies have suggested that Candida can encounter glucose-poor microenvironments during infection and that the ability to use alternative non-fermentable carbon sources, such as carboxylic acids, contributes to the virulence of these fungi. Our recent study (Mota et al., 2015) supported this view by demonstrating that the host metabolite acetate influences C. glabrata biofilm formation, antifungal drug resistance and phagocytosis; and suggesting a potential role of putative carboxylate transporters on these processes. In order to extend our studies and provide a comprehensive view of the C. glabrata biofilms' response to alternative carbon sources and antifungal treatment, we performed comparative transcriptomics analyses using RNA-sequencing. An overview of the most significant results will be presented. Our data support the view that adaptative responses of Candida cells to the different carbon sources present in host niches affects their virulence through multifarious mechanisms.info:eu-repo/semantics/publishedVersio

Development and Validation of a Social Capital Questionnaire for Adolescent Students (SCQ-AS)

Objectives: Social capital has been studied due to its contextual influence on health. However, no specific assessment tool has been developed and validated for the measurement of social capital among 12-year-old adolescent students. The aim of the present study was to develop and validate a quick, simple assessment tool to measure social capital among adolescent students. Methods: A questionnaire was developed based on a review of relevant literature. For such, searches were made of the Scientific Electronic Library Online, Latin American and Caribbean Health Sciences, The Cochrane Library, ISI Web of Knowledge, International Database for Medical Literature and PubMed Central bibliographical databases from September 2011 to January 2014 for papers addressing assessment tools for the evaluation of social capital. Focus groups were also formed by adolescent students as well as health, educational and social professionals. The final assessment tool was administered to a convenience sample from two public schools (79 students) and one private school (22 students), comprising a final sample of 101 students. Reliability and internal consistency were evaluated using the Kappa coefficient and Cronbach's alpha coefficient, respectively. Content validity was determined by expert consensus as well as exploratory and confirmatory factor analysis. Results: The final version of the questionnaire was made up of 12 items. The total scale demonstrated very good internal consistency (Cronbach's alpha: 0.71). Reproducibility was also very good, as the Kappa coefficient was higher than 0.72 for the majority of items (range: 0.63 to 0.97). Factor analysis grouped the 12 items into four subscales: School Social Cohesion, School Friendships, Neighborhood Social Cohesion and Trust (school and neighborhood). Conclusions: The present findings indicate the validity and reliability of the Social Capital Questionnaire for Adolescent Students

AVALIAÇÃO DO CLIMA ESCOLAR EM UMA ESCOLA PÚBLICA DO ENSINO FUNDAMENTAL II E ENSINO MÉDIO – AS POSSIBILIDADES DE DIAGNÓSTICO E INTERVENÇÃO NA DIMENSÃO DAS RELAÇÕES SOCIAIS NA ESCOLA

Vivemos tempos de violência, desrespeito e intolerância na sociedade. A escola pública não está isenta dessa triste realidade sendo permeável aos problemas do território que adentram seu cotidiano. Na escola ocorrem inúmeras situações de incivilidades, conflitos e violências, entre os seus atores. Objetivo: Essa pesquisa objetivou avaliar o clima relacional em uma escola pública estadual de Educação Fundamental e Médio, avaliando a dimensão das relações sociais que compõe o clima escolar, com ênfase nos conflitos e violência. Metodologia: Avaliou-se o clima escolar sob o ponto de vista dos estudantes, na instituição supracitada, de uma cidade do interior do estado de São Paulo. Participaram da pesquisa 59 estudantes, das séries do ensino fundamental II e 1º e 3º anos do ensino médio. Como instrumento utilizou-se o questionário de avaliação do clima escolar. Resultados: Após a avaliação, os dados foram organizados para apresentação e discussão junto aos docentes e gestores, identificando as necessidades de mudança. Pudermos detectar nas respostas dos estudantes em situações graves de violência, inclusive, a presença de situações de intimidação entre alunos e o porte de armas e tráfico de drogas na escola. Discussão final: Os resultados foram discutidos com os professores e gestores da escola participante, de modo a oferecer uma reflexão sobre as potencialidades e necessidades de melhorias no ambiente escolar. O intuito foi de buscar caminhos para superação de dificuldades e potencializar capacidades para contribuir assim com o aprimoramento na formação dos adolescentes das escolas públicas estaduais como pessoa humana, promovendo a formação ética, autonomia intelectual e pensamento crítico

WHY IS IT BETTER AT HOME? SERVICE USERS’ AND CAREGIVERS’ PERCEPTION OF HOME CARE

O objetivo deste estudo foi analisar as perspectivas dos usuários sobre os atendimentos de suas necessidades pelo serviço de atenção domiciliar. Trata-se de estudo qualitativo, com abordagem descritiva e exploratória. Participaram do estudo, usuários de serviços de atenção domiciliar de 16 municípios de Minas Gerais. Os dados foram obtidos de entrevistas com 20 usuários. Os resultados indicam que o perfil da população atendida pelos serviços remete a um público idoso. Os motivos de admissão são diversos e remetem à cronicidade. As equipes buscam realizar as atividades considerando as necessidades naturais e radicais de usuários. O atendimento no domicílio é apresentado como melhor, pelo conforto do lar, vínculo e pela superação quanto as barreiras de acesso a outros pontos da rede. Foi possível compreender que na perspectiva de usuários que o cuidar em casa apresenta novas relações que ampliam o acesso, a autonomia e a qualidade de vida do usuário. Estudio cuyo intuito fue analizar las perspectivas de los usuarios acerca de los atendimientos de sus necesidades por el servicio de Atención Domiciliar. Es un estudio cualitativo, de abordaje descriptivo y exploratorio, realizado entre febrero de 2014 y julio de 2015. Participaron 15 usuarios y 15 cuidadores de la Atención Domiciliar de ocho municipios de Minas Gerais. Los resultados apuntaron que el perfil de la población atendida es de ancianos en mayoría. Los motivos de esa opción son variables y se asocian a la cronicidad. Los equipos buscan realizar las actividades considerando las necesidades naturales y radicales de los usuarios. El atendimiento en domicilio es presentado como mejor opción a causa del conforto del hogar, vínculo con la familia y con el equipo, además de la superación de dificultades de acceso a otros puntos de la red de atención a la salud. Se constató que, en la perspectiva de los usuarios, cuidar en casa presenta nuevas relaciones que extienden el acceso, la autonomía y la cualidad de vida del usuario.The objective was to analyze service users’ perspectives regarding the attendance of their needs by the Home Care Service. This is a qualitative study, with a descriptive and exploratory approach, undertaken between February 2014 and July 2015. A total of 15 service users and 15 caregivers participated, from the Home Care Service of eight municipalities in the Brazilian state of Minas Gerais. The results indicated that the profile of the population attended is mainly older adults. The reasons for admission are diverse, and relate to the chronic nature of the health conditions. The team seeks to undertake the activities taking into account the service users’ natural and radical needs. Attendance in the home is presented as the best option due to the comfort of the home, the link with the family and with the team, and to the fact that this overcomes the barriers to accessing other points in the healthcare network. It is possible to understand that, in the service users’ perspective, care provided in the home presents new relationships which broaden access, autonomy and quality of life for the service user

ACIDENTES DE TRÂNSITO ENVOLVENDO ADOLESCENTES: UMA CARACTERIZAÇÃO

Problema: Os acidentes de trânsito são a segunda principal causa de mortalidade entre jovens de até 29 anos. No trânsito, em sua grande maioria, os jovens morrem ou ficam inválidos por motivos comuns que poderiam ser evitados. Objetivo: O estudo objetiva conhecer o perfil e o desfecho dos acidentes de trânsito envolvendo adolescentes em Montes Claros - MG. Método: Trata-se de uma pesquisa descritiva, retrospectiva e documental, com abordagem quantitativa. O projeto de pesquisa foi aprovado pelo Comitê de Ética e Pesquisa, com parecer consubstanciado nº 401.508. Resultados: A avaliação dos dados permitiu a constatação de que o sexo masculino lidera os acidentes de trânsito em um valor de 73,5% dos casos. No que tange a idade da vítima, 85% são de 15 a 19 anos de idade. Quando se faz a comparação entre tipo de veículo envolvido no acidente com localização da ocorrência, percebe-se que a motocicleta corresponde cerca de 70% dos números, com maior índice na região leste, com 26,4%. Conclusão: A atual condição desses adolescentes requer uma reorganização para a educação no trânsito