Medicinal pastures. Seed germinability acessment of Bellis perennis and Bellis sylvestris accessions from Alentejo and Spanish Extremadura

Com o objectivo de contribuir para o conhecimento da biologia reprodutiva da margarida (Bellis perennis e Bellis sylvestris), fez-se um estudo preliminar da capacidade germinativa de acessions silvestres destes taxa colhidos no Alentejo e Extremadura espanhola. O tratamento para quebra de dormência das sementes foi a pré-refrigeração a 5ºC durante 7 dias e um control. A temperatura de incubação foi de 20 ºC e testou-se também o efeito da duração do fotoperiodo na germinação das sementes (0 horas-escuro e 12 h-luz). As sementes testadas foram colhidas em prados naturais durante o ano de 2009; uma acession de B. perennis foi colhida simultaneamente em 2009 e 2010. Os melhores resultados foram obtidos com fotoperíodo de 12 h de luz e sem pré-tratamento de quebra de dormência; no entanto, na modalidade de 0 h de fotoperíodo (escuro), houve germinação de um maior número de acessions. O melhor resultado (48,5 %) foi obtido para a acession de B. perennis colhida em Marvão. Com estes resultados preliminares, conclui-se que existe uma enorme biodiversidade relativa à biologia reprodutiva das acessions estudadas. A germinação obtida para as espécies estudadas foi muito baixa para fins comerciais. Estudos futuros devem ser efectuados com outras acessions regionais, outros tratamentos de quebra de dormência e condições de acondicionamento da sement

Innate immune evasion revealed in a colorectal zebrafish xenograft model

Funding Information: We thank Champalimaud Foundation, LPCC-Terry Fox prize 2018, Congento (LISBOA-01-0145-FEDER-022170, co-financed by FCT/Lisboa2020) for funding. FCT fellowships for V.P. (SFRH/BD/118252/2016), M.M.L. (PD/BD/138203/2018), A.R.G. (CEECIND/ 02699/2017), and D.S. (PTDC/MED-ONC/28660/2017). We are grateful for M.G. Fer-reira’s initial support of this project; M.G. Carvalho for co-supervising MD bioinformatic analysis; all members of the Fior Lab for critical discussions; R. Mendes, M. Negrão, and B. Costa for sharing data; the Surgery Digestive Units from Champalimaud Clinical Center (CCC, Dr. N. Figueiredo) and Hospital Professor Fernando Fonseca (HPFF, Dr. V. Nunes and Dr. A. Gomes); The Histopathology Units of CCC (Dr. A. Beltran) and HPFF (Dr. A. Alves); Champalimaud Foundation (CF) Imaging (D. Accardi) and Flow Cytometry Platform (A. Vieira and R. Colaço) for support. We also thank Dr. M. Castillo-Martin, Director of the CF Biobank (CFB), and all the CFB team for human specimens procurement and advice; the CF Fish Platform (C. Certal, J. Monteiro et al.) and Vivarium Rodent Platform (I. Campos) for excellent animal care. We thank M.F. Moraes-Fontes, P. Moura Alves, and D. Henrique for the critical reading of the manuscript; Single Cell Discoveries (M. Muraro) for help with the scRNAseq initial data analysis and the generosity of the zebrafish community for sharing fish (S. Renshaw, F. Djouad, and Z. Wen). Finally, we would like to specially thank Professor Maria de Sousa (1939–2020) for her mentoring, support, enthusiasm, and critical reading of the manuscript.Cancer immunoediting is a dynamic process of crosstalk between tumor cells and the immune system. Herein, we explore the fast zebrafish xenograft model to investigate the innate immune contribution to this process. Using multiple breast and colorectal cancer cell lines and zAvatars, we find that some are cleared (regressors) while others engraft (progressors) in zebrafish xenografts. We focus on two human colorectal cancer cells derived from the same patient that show contrasting engraftment/clearance profiles. Using polyclonal xenografts to mimic intra-tumor heterogeneity, we demonstrate that SW620_progressors can block clearance of SW480_regressors. SW480_regressors recruit macrophages and neutrophils more efficiently than SW620_progressors; SW620_progressors however, modulate macrophages towards a pro-tumoral phenotype. Genetic and chemical suppression of myeloid cells indicates that macrophages and neutrophils play a crucial role in clearance. Single-cell-transcriptome analysis shows a fast subclonal selection, with clearance of regressor subclones associated with IFN/Notch signaling and escaper-expanded subclones with enrichment of IL10 pathway. Overall, our work opens the possibility of using zebrafish xenografts as living biomarkers of the tumor microenvironment.publishersversionpublishe

Do HACCP à cadeia de valor : estudo de caso aplicado à Vinícola Castelar, Lda.

Dissertação de mestrado em Sistemas de Prevenção e Controlo Alimentar, apresentada na Escola Superior Agrária de Santarém, Instituto Politécnico de SantarémVivemos hoje rodeados de produtos que trouxeram outra visibilidade e competitividade às empresas, independentemente da alavanca que inicia o processo de desenvolvimento e notoriedade. O desafio deste trabalho prende-se com a construção do posicionamento da cadeia de valor na empresa, Vinícola Castelar, Lda., tendo sido impulsionada por ferramentas com capacidade de criar valor, constituindo um estudo de caso. A implementação do HACCP foi uma das mais importantes etapas, assim como a aplicação da normativa ISO 22000:2005. A tomada de decisão pela gestão de introduzir a melhoria continua na organização partiu da avaliação de desempenho dos processos que integram a cadeia de valor do negócio. Esta opção resultou num aumento de vendas e no incremento da notoriedade da Vinícola Castelar culminando em valor acrescentado para o produto, para a empresa, para os seus clientes e fornecedores.Today we live surrounded by products that have brought another visibility and competitiveness to businesses, regardless of what triggers the process of development and awareness. The focus of this work relates to the construction of the positioning of the value chain in the company, Vinícola Castelar, Lda., which has been boosted by tools capable of creating value, constitutes a case study. The implementation of HACCP was one of the most important steps, as well as implementing the ISO 22000:2005 standards. The decision by the management to introduce continuous improvement in the organization originated from the performance assessment processes that integrate the business’s value chain. This option resulted in sales growth and increased the reputation of the Vinícola Castelar culminating in added value to the product, the company, its customers and suppliers

microRNAs regulate TAL1 expression in T-cell acute lymphoblastic leukemia.

The transcription factor TAL1 is a proto-oncogene whose aberrant expression in committed T-cell precursors is associated with the development of T-cell acute lymphoblastic leukemia (T-ALL). The mechanisms leading to aberrant activation of TAL1 in T-ALL patients who lack chromosomal rearrangements involving the TAL1 locus remain largely unknown. We hypothesized that TAL1 levels decrease during normal T-cell development at least in part due to miRNA-dependent silencing, in which case TAL1 over-expression in some T-ALL cases could be the consequence of deregulated miRNA expression. By performing computational prediction of miRNAs that bind to the human TAL1 mRNA we compiled a list of miRNAs that are candidates to regulate TAL1. Using a luciferase reporter system and mutagenesis assays we confirmed the miRNA-TAL1 mRNA interactions and selected candidate miRNAs: miR-101, miR-520d-5p, miR-140-5p, miR-448 and miR-485-5p. Over-expression of these microRNAs in different T-ALL cell lines consistently resulted in the down-regulation of TAL1 protein. In accordance, inhibition of miR-101 and miR-520d-5p promoted TAL1 protein expression. Importantly, we found that miR-101, miR-140-5p, miR-448 and miR-485-5p were down-regulated in T-ALL patient specimens and T-ALL cell lines. Our results show for the first time the existence of epigenetic regulation of TAL1 by specific miRNAs which may contribute, at least in part, to the ectopic expression of TAL1 in some T-ALL cases.These studies were financed by Liga Portuguesa Contra o Cancro (Terry Fox Award) and by Fundacao para a Ciencia e a Tecnologia (project PTDC/BIM-ONC/1548/2012). NCC received an FCT-SFRH PhD fellowship. JTB is supported by an FCT Consolidation Grant. The Cell Division and Cancer group of the CNIO is funded by the Spanish Ministry of Economy and Competitiveness (MINECO; SAF2012-38215: Consolider-Ingenio 2010 Programme SAF2014-57791-REDC; Red Tematica CellSYS BFU2014-52125-REDT), and the Comunidad de Madrid (OncoCycle Programme S2010/BMD-2470).S

Object detection for automatic cancer cell counting in zebrafish xenografts

Cell counting is a frequent task in medical research studies. However, it is often performed manuallythus, it is time-consuming and prone to human error. Even so, cell counting auto- mation can be challenging to achieve, especially when dealing with crowded scenes and overlapping cells, assuming different shapes and sizes. In this paper, we introduce a deep learning-based cell detection and quantification methodology to automate the cell counting process in the zebrafish xenograft cancer model, an innovative technique for studying tumor biology and for personalizing medicine. First, we implemented a fine-tuned architecture based on the Faster R-CNN using the Inception ResNet V2 feature extractor. Second, we performed several adjustments to optimize the process, paying attention to constraints such as the presence of overlapped cells, the high number of objects to detect, the heterogeneity of the cells’ size and shape, and the small size of the data set. This method resulted in a median error of approximately 1% of the total number of cell units. These results demon- strate the potential of our novel approach for quantifying cells in poorly labeled images. Compared to traditional Faster R-CNN, our method improved the average precision from 71% to 85% on the studied data set

Adult B-cell acute lymphoblastic leukemia cells display decreased PTEN activity and constitutive hyperactivation of PI3K/Akt pathway despite high PTEN protein levels

© 2014 Ferrata Storti Foundation. This is an open-access paperAdult B-cell acute lymphoblastic leukemia remains a major therapeutic challenge, requiring a better characterization of the molecular determinants underlying disease progression and resistance to treatment. Here, using a phospho-flow cytometry approach we show that adult diagnostic B-cell acute lymphoblastic leukemia specimens display PI3K/Akt pathway hyperactivation, irrespective of their BCR-ABL status and despite paradoxically high basal expression of PTEN, the major negative regulator of the pathway. Protein kinase CK2 is known to phosphorylate PTEN thereby driving PTEN protein stabilization and concomitant PTEN functional inactivation. In agreement, we found that adult B-cell acute lymphoblastic leukemia samples show significantly higher CK2 kinase activity and lower PTEN lipid phosphatase activity than healthy controls. Moreover, the clinical-grade CK2 inhibitor CX-4945 (Silmitasertib) reversed PTEN levels in leukemia cells to those observed in healthy controls, and promoted leukemia cell death without significantly affecting normal bone marrow cells. Our studies indicate that CK2-mediated PTEN posttranslational inactivation, associated with PI3K/Akt pathway hyperactivation, are a common event in adult B-cell acute lymphoblastic leukemia and suggest that CK2 inhibition may constitute a valid, novel therapeutic tool in this malignancy.This study was supported by grant PTDC/IC/83023/2007, from Fundação para a Ciência e a Tecnologia (FCT). A.M.G. and V.P. received BI fellowships from FCT. L.R.M. and A.M. received a post-doctoral and a PhD fellowship, respectively, both from FCT.info:eu-repo/semantics/publishedVersio

The Core-Clock Gene NR1D1 Impacts Cell Motility In Vitro and Invasiveness in a Zebrafish Xenograft Colon Cancer Model

International audienceMalfunctions of circadian clock trigger abnormal cellular processes and influence tumorigenesis. Using an in vitro and in vivo xenograft model, we show that circadian clock disruption via the downregulation of the core-clock genes BMAL1, PER2, and NR1D1 impacts the circadian phenotype of MYC, WEE1, and TP53, and affects proliferation, apoptosis, and cell migration. In particular, both our in vitro and in vivo results suggest an impairment of cell motility and a reduction in micrometastasis formation upon knockdown of NR1D1, accompanied by altered expression levels of SNAI1 and CD44. Interestingly we show that differential proliferation and reduced tumour growth in vivo may be due to the additional influence of the host-clock and/or to the 3D tumour architecture. Our results raise new questions concerning host–tumour interaction and show that core-clock genes are involved in key cancer properties, including the regulation of cell migration and invasion by NR1D1 in zebrafish xenograft