Exploring the chemotherapeutic potential of Brassicaceae extracts in colorectal cancer cell spheroids

Colorectal cancer is the third most common cause of mortality worldwide. Given the growth and increasing life expectancy of the world’s population, as well as the acquisition of unhealthy lifestyle habits, the global burden of colorectal cancer is estimated to increase in the next years. Despite the efforts made so far, its treatment is still very challenging due to cancer recurrence usually associated with prevalence of cancer stem cells (CSCs) after treatment. Hence, it is imperative to seek new therapeutic strategies that target colorectal CSCs. Epidemiological data have reported a positive correlation between cruciferous vegetables intake and decreased risk of colorectal cancer. Their chemo-preventive effect is mainly due to their high content in glucosinolates, the precursors of isothiocyanates (ITCs) that are known to modulate and target several aspects of carcinogenesis. Hence, by recurring to a green and sustainable high pressure extraction process to recover ITCs from cruciferous vegetables, namely watercress and broccoli, we intended to explore the anticancer mechanisms of Brassicaceae vegetables and respective ITCs in a tri-dimensional (3D) cell model of colorectal cancer (i.e. in cell spheroids), since this approach resembles best with the tumor microenvironment in comparison with the conventional two-dimensional (2D) cells models. Our results revealed that Brassicaceae extracts and ITCs have the potential to prevent cell proliferation and chemo-resistance, to induce apoptosis, and to target colorectal CSC population and its self-renewal ability. Therefore, our research provides new insights on colorectal cancer therapy using nutraceuticals derived from cruciferous vegetables

O ENSINO DE FUNÇÃO AFIM POR MEIO DA RESOLUÇÃO DE PROBLEMAS

The main objective of this work was to verify the contributions of the teaching of affine function through Problem Solving in Mathematics classes in classes of the 1st year of High School. The research was carried out in two classes of the 1st year of High School, in a State School located in the irrigated area of the municipality of Petrolina - PE, with a total of 41 participating students. In order to carry out this qualitative work, was used class diary to record all reports, doubts and progress of each student during the intervention, which following the ten steps of Problem Solving, as suggested by Onuchic and Allevato (2011). After analyzing the data obtained, the results indicate that teaching, through this methodology, linked to problem situations present in the students' daily lives, provided a change in the behavior of those involved, by arousing interest to learn more, both in the students and in the teacher-researcher. Problem solving proved to be an effective strategy, contributing positively to student learning.Este trabalho teve como objetivo principal verificar as contribuições do ensino de função afim por meio da Resolução de Problemas nas aulas de Matemática em turmas do 1° ano do Ensino Médio. A pesquisa foi realizada em duas turmas do 1° ano do Ensino Médio, em uma Escola Estadual localizada na área irrigada do município de Petrolina – PE, com um total de 41 alunos participantes. Para a realização deste trabalho de caráter qualitativo, utilizou-se de diário de campo para anotar todos os relatos, dúvidas e avanços de cada aluno durante a intervenção, a qual seguiu as dez etapas da Resolução de Problemas, conforme sugerida por Onuchic e Allevato (2011). Após a análise dos dados obtidos, os resultados apontaram que o ensino, por meio desta metodologia, atrelada a situações-problema presentes no cotidiano dos alunos, proporcionou uma mudança no comportamento dos envolvidos, ao despertar interesse para aprender mais, tanto nos alunos, quanto na professora-pesquisadora. A Resolução de Problemas se mostrou como uma estratégia eficaz, contribuindo, de maneira positiva, na aprendizagem dos estudantes

Leisure as a mechanism to democratic reinforce of the public spaces: a study of leisure activities at Estação das Docas (Docks), Belém (Pará, Brazil)

Na cidade de Belém (Pará, Brasil), percebe-se que alguns espaços muitas vezes limitam os moradores, através de seus muros e gestão excludente. Nesta perspectiva, o objetivo principal do presente artigo foi analisar as diferentes práticas de lazer vivenciadas na Estação das Docas, além de buscar identificar e analisar se havia barreiras para a vivência do lazer, e quais seriam essas barreiras. Neste estudo, os procedimentos metodológicos incluíram a pesquisa de campo, através da observação direta e pesquisa bibliográfica, além de entrevistas semiestruturadas junto a quarenta frequentadores da Estação. Concluiu-se que as práticas de lazer na Estação eram das mais diversas, constituindo-se como um lugar para passear, namorar, apreciar o pôr-do-sol e conversar nos barzinhos e restaurantes. No entanto, havia diversas barreiras para a apropriação de algumas atividades promovidas pela Estação, como cinema, teatro e outras apresentações artísticas, que eram pouco conhecidas pelos frequentadores.In Belém (Pará, Brazil) it is noticed that some places imply limits to their neighborhood with walls or exclusionary management. In this sense, the aim of this paper was analyze different ways of leisure offered by Estação das Docas and thereby seek to identify and analyze if there were any barriers for the people to have leisure activities and what were these barriers. The methodology included field research, using direct observation and bibliographic research and besides, the study applied forty interviews to the users of the place. It was concluded that there were a great variety of leisure practices, being a place to walk, date, watch the sunset or talk in restaurants and pubs. Otherwise it was identified some barriers at that place, as movies, theater and another cultural activities which were little known by users

Gellan gum-based hydrogels support the recreation of the dermal papilla microenvironment

The dermal papilla (DP), a specialized compartment within the hair follicle, regulates hair growth. However, human DP cells rapidly lose their inductivity in 2D-culture given the loss of positional and microenvironmental cues. Spheroids have been capable of recreating the 3D intercellular organization of DP cells, however, DP cell-matrix interactions are poorly represented. Considering the specific nature of the DP's extracellular matrix (ECM), we functionalized gellan gum (GG) with collagen IV-(HepIII) or fibronectin-(cRGDfC) derived peptide sequences to generate a 3D environment in which the phenotype and physiological functions of DP cells are restored. We further tuned the stiffness of the microenvironments by varying GG amount. Biomimetic peptides in stiffer hydrogels promoted the adhesion of DP cells, while each peptide and amount of polymer independently influenced the type and quantity of ECM proteins deposited. Furthermore, although peptides did not seem to have an influence, stiffer hydrogels improved the inductive capacity of DP cells after short term culture. Interestingly, independently of the peptide, these hydrogels supported the recapitulation of basic hair morphogenesis-like events when incorporated in an organotypic human skin in vitro model. Our work demonstrates that tailored GG hydrogels support the generation of a microenvironment in which both cell-ECM and cell-cell interactions positively influence DP cells towards the creation of an artificial DP.Authors would like to acknowledge the financial support from the Consolidator Grant “ECM_INK” (ERC-2016-COG-726061) and from FCT/MCTES (Fundação para a Ciência e a Tecnologia/Ministério da Ciência, Tecnologia e Ensino Superior) through the PD/59/2013, PD/BD/113800/2015 (Carla Abreu), PD/169/2013 (Manuela Lago) and IF/00945/2014 (Alexandra Marques) grants. The authors also appreciate the support given by the FSE/POCH (Fundo Social Europeu through the Programa Operacional do Capital Humano) under the scope of the NORTE-08-5369-FSE-000037 grant (Manuela Lago)

TECNOLOGIAS DE INFORMAÇÃO E COMUNICAÇÃO COMO INSTRUMENTO DE INCLUSÃO EM CASOS DE DIFICULDADES DE APRENDIZAGEM

The current information and communication technology (ict) is an importanttool in the learning consolidation process of people with a profile of school failure derivedfrom cognitive disabilities and neurological dysfunction. This is the type of problem thathappens to individuals with dyslexia – who display specific difficulties in reading and writing,and those with adhd, two of the major types of neuropsychological learning difficulties,conditions that cause low self-esteem and lack of social adjustment. such individuals have amedium-superior to a superior type of intelligence, but their brain activities differ from themajority of the population. This causes serious disturbances in their private and group life.this article aims at discussing the characteristics of those two types of disabilities and presentboth the use of technological tools for neurological research and the role of ict - informationand communication technology, as a medium for social inclusion through education.As atuais tecnologias de informação e comunicação (tic) são ferramentasimportantes na consolidação da aprendizagem de pessoas com perfil de fracasso escolarresultante de desabilidades cognitivas e disfunções neurológicas. Tais problemas ocorrem noscasos de indivíduos disléxicos – que apresentam dificuldade específica em leitura e escrita, enaqueles com déficit de atenção e hiperatividade, dois dos mais prevalentes tipos dedificuldades de aprendizagem neuropsicológicas, ambos causadores de baixa auto-estima edesajuste social. tais indivíduos têm inteligência média superior ou superior, mas umfuncionamento cerebral diverso do da maioria da população, o que causa sérios transtornosem sua vida pessoal e em grupo. O artigo pretende apresentar as características desses doistipos de distúrbio e comentar o estado da arte da utilização de ferramentas tecnológicasempregadas em pesquisas sobre essas diferenças cerebrais e o papel da tecnologia deinformação e comunicação usada como mediadora da inclusão social através da educação

Pre-selection of fibroblasts subsets prompt prevascularization of tissue engineered skin analogues

The papillary and reticular dermis harbors phenotypically distinct fibroblasts, whose functions such as maintenance of skin's microvasculature are also distinct. Thus, we hypothesized that pre-selection of the subpopulations of fibroblasts would benefit the generation of skin tissue engineered (TE) constructs, promoting their prevascularization in vitro. We first isolated papillary and reticular fibroblasts using fluorescence-activated cell sorting and studied the effect of their secretome and extracellular matrix (ECM) on human dermal microvascular endothelial cell (hDMEC) organization. Subsequently, we developed a bilayered 3D polymeric structure with distinct layer-associated features to house the subpopulations of fibroblasts, to generate a skin analogue. Both papillary and reticular fibroblasts were able to stimulate capillary-like network formation in a Matrigel assay. However, the secretome of the two subpopulations was substantially different, being enriched in VEGF, IGF-1, and Angio-1 in the case of papillary fibroblasts and in HGF and FGF-2 for the reticular subset. In addition, the fibroblast subpopulations deposited varied levels of ECM proteins, more collagen I and laminin was produced by the reticular subset, but these differences did not impact hDMEC organization. Vessel-like structures with lumens were observed earlier in the 3D skin analogue prepared with the sorted fibroblasts, although ECM deposition was not affected by the cell's pre-selection. Moreover, a more differentiated epidermal layer was obtained in the skin analogue formed by the sorted fibroblasts, confirming that its whole structure was not affected. Overall, we provide evidence that pre-selection of papillary and reticular fibroblasts is relevant for promoting the in vitro prevascularization of skin TE constructs.The authors would like to acknowledge the financial support from the Consolidator Grant “ECM_INK” (ERC-2016- COG-726061), to the FSE/POCH (Fundo Social Europeu através do Programa Operacional do Capital Humano) under the scope of the PD/169/2013, NORTE-08-5369-FSE-000037 (H.R. M.)

Neurotensin Decreases the Proinflammatory Status of Human Skin Fibroblasts and Increases Epidermal Growth Factor Expression

Fibroblasts colonization into injured areas during wound healing (WH) is responsible for skin remodelling and is also involved in the modulation of inflammation, as fibroblasts are immunologically active. Herein, we aimed to determine neurotensin effect on the immunomodulatory profile of fibroblasts, both in homeostatic and inflammatory conditions. Neurotensin mediated responses occurred through NTR1 or NTR3 receptors, while under inflammatory conditions NTR1 expression increase seemed to modulate neurotensin responses. Among different immunomodulatory genes, CCL11, IL-8, and IL-6 were the most expressed genes, while CCL4 and EGF were the less expressed genes. After neurotensin exposure, IL-8 mRNA expression was increased while CCL11 was decreased, suggesting a proinflammatory upregulation and chemoattractant ability downregulation of fibroblasts. Under inflammatory conditions, gene expression was significantly increased. After neurotensin exposure, CCL4 and IL-6 mRNA expression were decreased while CCL11 was increased, suggesting again a decrease in the chemoattractant capacity of fibroblasts and in their proinflammatory status. Furthermore, the expression of EGF, a crucial growth factor for skin cells proliferation and WH, was increased in all conditions. Overall, neurotensin, released by nerve fibers or skin cells, may be involved in the decrease of the chemotaxis and the proinflammatory status in the proliferation and remodelling phases of WH

Production of fermented cheese whey-based beverage using kefir grains as starter culture : evaluation of morphological and microbial variations

Whey valorization concerns have led to recent interest on the production of whey beverage simulating kefir. In this study, the structure and microbiota of Brazilian kefir grains and beverages obtained from milk and whole/deproteinised whey was characterized using microscopy and molecular techniques. The aim was to evaluate its stability and possible shift of probiotic bacteria to the beverages. Fluorescence staining in combination with Confocal Laser Scanning Microscopy showed distribution of yeasts in macro-clusters among the grain’s matrix essentially composed of polysaccharides (kefiran) and bacteria. Denaturing gradient gel electrophoresis displayed communities included yeast affiliated to Kluyveromyces marxianus, Saccharomyces cerevisiae, Kazachatania unispora, bacteria affiliated to Lactobacillus kefiranofaciens subsp. Kefirgranum, Lactobacillus kefiranofaciens subsp. Kefiranofaciens and an uncultured bacterium also related to the genus Lactobacillus. A steady structure and dominant microbiota, including probiotic bacteria, was detected in the analyzed kefir beverages and grains. This robustness is determinant for future implementation of whey-based kefir beverages.Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES), CAPES-GRICES

Stem cell-containing hyaluronic acid-based spongy hydrogels for integrated diabetic wound healing

The detailed pathophysiology of diabetic foot ulcers is yet to be established and improved treatments are still required. We propose a strategy that directs inflammation, neovascularization, and neoinnervation of diabetic wounds. Aiming to potentiate a relevant secretome for nerve regeneration, stem cells were precultured in hyaluronic acid-based spongy hydrogels under neurogenic/standard media before transplantation into diabetic mice full-thickness wounds. Acellular spongy hydrogels and empty wounds were used as controls. Reepithelialization was attained 4 weeks after transplantation independently of the test groups, whereas a thicker and more differentiated epidermis was observed for the cellular spongy hydrogels. A switch from the inflammatory to the proliferative phase of wound healing was revealed for all the experimental groups 2 weeks after injury, but a significantly higher M2(CD163 þ )/M1(CD86 þ ) subtype ratio was observed in the neurogenic preconditioned group that also failed to promote neoinnervation. A higher number of intraepidermal nerve fibers were observed for the unconditioned group probably due to a more controlled transition from the inflammatory to the proliferative phase. Overall, stem cell-containing spongy hydrogels represent a promising approach to enhance diabetic wound healing by positively impacting re-epithelialization and by modulating the inflammatory response to promote a successful neoinnervation.The authors would like to acknowledge Gene2Skin Project (H2020-TWINN2015-692221) and Fundac¸a˜o para a Cieˆncia e Tecnologia for SFRH/BD/ 78025/2011 (LPdS), SFRH/BPD/96611/2013 (MTC), SFRH/BPD/101886/2014 (RPP), SFRH/BPD/101952/2014 (TCS) grants. Moreover, the authors would also like to acknowledge Teresa Oliveira for histology support, Andreia Carvalho for hASCs supply, Luca Gasperini for cell profiler analysis, and Manuela E. L. Lago and Carla M. Abreu for intraepidermal nerve fiber quantification.info:eu-repo/semantics/publishedVersio