Antitumoral Activity of the Universal Methyl Donor S-Adenosylmethionine in Glioblastoma Cells.

Glioblastoma (GBM), the most frequent and lethal brain cancer in adults, is characterized by short survival times and high mortality rates. Due to the resistance of GBM cells to conventional therapeutic treatments, scientific interest is focusing on the search for alternative and efficient adjuvant treatments. S-Adenosylmethionine (AdoMet), the well-studied physiological methyl donor, has emerged as a promising anticancer compound and a modulator of multiple cancer-related signaling pathways. We report here for the first time that AdoMet selectively inhibited the viability and proliferation of U87MG, U343MG, and U251MG GBM cells. In these cell lines, AdoMet induced S and G2/M cell cycle arrest and apoptosis and downregulated the expression and activation of proteins involved in homologous recombination DNA repair, including RAD51, BRCA1, and Chk1. Furthermore, AdoMet was able to maintain DNA in a damaged state, as indicated by the increased γH2AX/H2AX ratio. AdoMet promoted mitotic catastrophe through inhibiting Aurora B kinase expression, phosphorylation, and localization causing GBM cells to undergo mitotic catastrophe-induced death. Finally, AdoMet inhibited DNA repair and induced cell cycle arrest, apoptosis, and mitotic catastrophe in patient-derived GBM cells. In light of these results, AdoMet could be considered a potential adjuvant in GBM therapy.

S-Adenosylmethionine Inhibits Colorectal Cancer Cell Migration through Mirna-Mediated Targeting of Notch Signaling Pathway.

Metastasis is a leading cause of mortality and poor prognosis in colorectal cancer (CRC). Thus, the identification of new compounds targeting cell migration represents a major clinical challenge. Recent findings evidenced a central role for dysregulated Notch in CRC and a correlation between Notch overexpression and tumor metastasis. MicroRNAs (miRNAs) have been reported to cross-talk with Notch for its regulation. Therefore, restoring underexpressed miRNAs targeting Notch could represent an encouraging therapeutic approach against CRC. In this context, S-adenosyl-L-methionine (AdoMet), the universal biological methyl donor, being able to modulate the expression of oncogenic miRNAs could act as a potential antimetastatic agent. Here, we showed that AdoMet upregulated the onco-suppressor miRNAs-34a/-34c/-449a and inhibited HCT-116 and Caco-2 CRC cell migration. This effect was associated with reduced expression of migration-/EMT-related protein markers. We also found that, in colorectal and triple-negative breast cancer cells, AdoMet inhibited the expression of Notch gene, which, by luciferase assay, resulted the direct target of miRNAs-34a/-34c/-449a. Gain- and loss-of-function experiments with miRNAs mimics and inhibitors demonstrated that AdoMet exerted its inhibitory effects by upregulating miRNAs-34a/-34c/-449a. Overall, these data highlighted AdoMet as a novel Notch inhibitor and suggested that the antimetastatic effects of AdoMet involve the miRNA-mediated targeting of Notch signaling pathway.

A rapid and inexpensive genotyping method using dried blood spots for mutational analysis in a mutant mouse model: an update.

BACKGROUND: Dried blood spot (DBS) testing is a well-known method of bio-sampling by which blood samples are blotted and dried on filter paper. The dried samples can then be analyzed by several techniques such as DNA amplification and HPLC. We have developed a non-invasive sampling followed by an alternative protocol for genomic DNA extraction from a drop of blood adsorbed on paper support. This protocol consists of two separate steps: (1) organic DNA extraction from the DBS, followed by (2) DNA amplification by polymerase chain reaction (PCR). The PCR-restriction fragment length polymorphism (PCR-RFLP) is an advantageous and simple approach to detect single nucleotide polymorphisms (SNPs). RESULTS: We have evaluated the efficiency of our method for the extraction of genomic DNA from DBS by testing its performance in genotyping mouse models of obesity and herein discuss the specificity and feasibility of this novel procedure. CONCLUSIONS: Our protocol is easy to perform, fast and inexpensive and allows the isolation of pure DNA from a tiny amount of sample.

Climbing plants of Porto Ferreira State Park, southeastern Brazil / Trepadeiras do Parque Estadual de Porto Ferreira, Brasil

Abstract A floristic survey of climbing plants was carried out in an ecotone area of seasonal semideciduous forest (SSF) and forested savanna (CER), in Porto Ferreira State Park (PFSP), Southeastern Brazil. We sampled the reproductive specimens every month during two periods, March 2010 to September 2011 and April and July 2015. The surveys were performed by the walking method, and the sampled individuals were classified by habit, climbing mechanism and dispersal mode. Overall, 109 species, belonging to 67 genera and 29 families, were recorded; 49 species occurred in both, 29 and 31 were exclusive to SSF and CER, respectively. Bignoniaceae and Malpighiaceae were the richest families, with 17 species, followed by Sapindaceae (12 species), Asteraceae and Apocynaceae (8 species each) and Fabaceae (6). The majority of climbers were lianas, twining and anemochoric species, corresponding to 70%, 47% and 66% of all samples, respectively. In this work, we added one new family and 14 species to the Cerrado's list of climbing plants from São Paulo state, and 10 species to the Brazilian seasonal semideciduous forest's list. Therefore, we contributed to the understanding of diversity of climbing plants in vegetation types poorly studied for this plant group, mainly in the Cerradão, wherein we found new records for several species.

Resumo O levantamento florístico das trepadeiras foi realizado em um ecótono de Floresta Estacional Semidecidual (FES) e Cerradão (CER), no Parque Estadual de Porto Ferreira, Sudeste do Brasil. Realizamos coletas mensais dos espécimes reprodutivos ao longo de dois períodos, março 2010 a setembro 2011, abril e julho 2015. Os levantamentos foram realizados por meio do método de caminhada e os indivíduos amostrados foram classificados quanto ao hábito, mecanismo de ascensão e síndrome de dispersão. No geral, foram registradas 109 espécies, pertencentes a 67 gêneros e 29 famílias. Dentre essas, 49 espécies ocorrem nos dois tipos de vegetação, sendo que 29 e 31 espécies são exclusivas de FES e CER, respectivamente. Bignoniaceae e Malpighiaceae foram as famílias mais ricas com 17 espécies, seguidas por Sapindaceae (12 espécies), Asteraceae e Apocynaceae (8 espécies cada) e Fabaceae (6). A maioria das espécies de trepadeiras são lianas, volúveis e anemocóricas, correspondendo a 70%, 47% e 66% de toda a amostra, respectivamente. Neste trabalho, acrescentamos uma nova família e 14 espécies para a lista de trepadeiras do Cerrado paulista e 10 espécies para a lista brasileira de trepadeiras em Floresta Estacional Semidecidual. Portanto, contribuímos para o conhecimento da diversidade de trepadeiras em tipos vegetacionais pouco estudados para este grupo de planta, destacando o Cerradão, no qual encontramos novos registros para um grande número de espécies.