Extracellular Vesicles Derived-LAT1 mRNA as a Powerful Inducer of Colorectal Cancer Aggressive Phenotype.

Colorectal cancer (CRC) is the third most common cancer in the world and represents the third most deadly tumor worldwide. About 15-25% of patients present metastasis in the moment of diagnosis, the liver being the most common site of metastization. Therefore, the development of new therapeutic agents is needed, to improve the patients' prognosis. Amino acids transporters, LAT1 and ASCT2, are described as upregulated in CRC, being associated with a poor prognosis. Extracellular vesicles have emerged as key players in cell-to-cell communication due to their ability to transfer biomolecules between cells, with a phenotypic impact on the recipient cells. Thus, this study analyzes the presence of LAT1 and ASCT2 mRNAs in CRC-EVs and evaluates their role in phenotype modulation in a panel of four recipient cell lines (HCA-7, HEPG-2, SK-HEP-1, HKC-8). We found that HCT 116-EVs carry LAT1, ASCT2 and other oncogenic mRNAs being taken up by recipient cells. Moreover, the HCT 116-EVs' internalization was associated with the increase of LAT1 mRNA in SK-HEP-1 cells. We also observed that HCT 116-EVs induce a higher cell migration capacity and proliferation of SK-HEP-1 and HKC-8 cells. The present study supports the LAT1-EVs' mRNA involvement in cell phenotype modulation, conferring advantages in cell migration and proliferation.

Direito Penal e COVID-19 no espaço lusófono / Criminal Law and COVID-19 in the lusophone world / Derecho Penal y COVID-19 en el espacio lusofono

Objetivo: convocar as normas penais relevantes para o contexto pandémico dos ordenamentos jurídicos tomados como referência (Portugal, Brasil, Angola e Região Administrativa Especial de Macau) a fim de aferir das vantagens e desvantagens dos respetivos modelos. Metodologia: através de uma análise comparativa dos diferentes modelos, foi utilizado o método dedutivo para asserção da possível vantagem ou desvantagem, tendo em conta a função do Direito Penal. Resultados: as normas penais mais relevantes, em especial, o crime de desobediência, comum a todos os ordenamentos, acabam por assumir a natureza de normas penais em branco. Conclusão: tendo o contexto pandémico evidenciado a importância de uma efetiva tutela penal da saúde pública, a mesma deve ser feita através de um modelo que melhor cumpra a função de garantia do direito penal.

Objective: to identify the relevant criminal law norms for the pandemic context of the legal systems taken as reference (Portugal, Brazil, Angola and Macao Special Administrative Region) in order to assess the advantages and disadvantages of the respective models. Methods:through a comparative analysis of the different models, the deductive method was used to assert the possible advantage or disadvantage, taking into account the function of Criminal Law. Results: the most relevant criminal norms, in particular, the crime of disobedience, common to all ordinances, end up assuming the nature of blank criminal norms. Conclusion: having the pandemic context highlighted the importance of an effective criminal protection of public health, it should be done through a model that better fulfills the function of guarantee of criminal law.

Objetivo: identificar las leyes penales relevantes para el contexto pandémico de los sistemas jurídicos tomados como referencia (Portugal, Brasil, Angola y la Región Administrativa Especial de Macao) para evaluar las ventajas y desventajas de los respectivos modelos. Metodología: através de un análisis comparativo de los diferentes modelos, se utilizó e método deductivo para afirmar la posible ventaja o desventaja, teniendo en cuenta la función del derecho penal. Resultados: las normas penales más relevantes, en particular, el delito de desobediencia, común a todas las ordenanzas, acaban asumiendo la naturaleza de normas penales en blanco. Conclusión: habiendo evidenciado el contexto pandémico la importancia de una efectiva tutela penal de la salud pública, la misma debe hacerse a través de un modelo que cumpla mejor la función de garantía del derecho penal.

Loss of hierarchical imprinting regulation at the Prader-Willi/Angelman syndrome locus in human iPSCs.

The human chr15q11-q13 imprinted cluster is linked to several disorders, including Prader-Willi (PWS) and Angelman (AS) syndromes. Recently, disease modeling approaches based on induced pluripotent stem cells (iPSCs) have been used to study these syndromes. A concern regarding the use of these cells for imprinted disease modeling is the numerous imprinting defects found in many iPSCs. Here, by reprogramming skin fibroblasts from a control and AS individuals, we generated several iPSC lines and addressed the stability of imprinting status across the PWS/AS domain. We focused on three important regulatory DNA elements which are all differentially methylated regions (DMRs), methylated on the maternal allele: the PWS imprinting center (PWS-IC), which is a germline DMR and the somatic NDN and MKRN3 DMRs, hierarchically controlled by PWS-IC. Normal PWS-IC methylation pattern was maintained in most iPSC lines; however, loss of maternal methylation in one out of five control iPSC lines resulted in a monoallelic to biallelic switch for many imprinted genes in this domain. Surprisingly, MKRN3 DMR was found aberrantly hypermethylated in all control and AS iPSCs, regardless of the methylation status of the PWS-IC master regulator. This suggests a loss of hierarchical control of imprinting at PWS/AS region. We confirmed these results in established iPSC lines derived using different reprogramming procedures. Overall, we show that hierarchy of imprinting control in donor cells might not apply to iPSCs, accounting for their spectrum of imprinting alterations. Such differences in imprinting regulation should be taken into consideration for the use of iPSCs in disease modeling.

Flow cytometry CD45-negative B-NHL: a case report of a diffuse large B-cell lymphoma without extranodal involvement.

BACKGROUND: The loss of CD45, the leukocyte-common antigen, has been described in rare cases of large B-cell lymphoma (LBCL) subtypes with extranodal involvement by immunohistochemical methods. Here we report a case of a patient with LBCL, with no extranodal lesions, which is CD45 negative by flow cytometry (FC) immunophenotyping. METHODS: Immunophenotyping and DNA content analysis was performed by multiparametric FC on lymph node and bone marrow aspirate obtained from a 65 year old male patient. RESULTS: Malignant B-lymphocytes were CD5-, CD10+/++, CD11c-, CD19+, CD20+/++, CD23-, CD34-, CD38-/+, CD45-, CD79b++/+++, BCL2 overexpressed, FMC7++, IgM++/+++, TdT- with Lambda light chain restriction. This pathological cellular population showed near-diploid DNA content, with a high proliferate rate. CONCLUSIONS: To our knowledge, we describe the first case of a CD19+ B-cell non-Hodgkin lymphoma without expression of CD45 detected by FC, and the first case without extranodal involvement presentation. This case is reported not only because it is a rare one but also to raise awareness of FC users of its correct diagnosis.