RESUMEN
Gastric cancer (GC) is the one of the most prevalent cancers and one of the leading causes of cancer-induced deaths. Previously, we found that the expression of purinergic P2Y2 receptor (P2Y2R) is increased in GC samples as compared to adjacent healthy mucosa taken from GC-diagnosed patients. In this work, we studied in detail purinergic signaling in the gastric adenocarcinoma-derived cell lines: AGS, MKN-45, and MKN-74, and compared them to a nontumoral epithelial cell line: GES-1. In GC-derived cells, we detected the expression of several purinergic receptors, and found important differences as compared to GES-1 cells. Functional studies revealed a strong contribution of P2Y2Rs in intracellular calcium increases, elicited by adenosine-triphosphate (ATP), uridine-triphosphate (UTP), and the P2Y2R agonist MRS2768. Responses were preserved in the absence of extracellular calcium and inhibited by P2Y2R antagonists. In GES-1 cells, ATP and UTP induced similar responses and the combination of P2X and P2Y receptor antagonists was able to block them. Proliferation studies showed that ATP regulates AGS and MKN-74 cells in a biphasic manner, increasing cell proliferation at 10-100 µM, but inhibiting at 300 µM ATP. On the other hand, 1-300 µM UTP, a P2Y2R agonist, increased concentration-dependent cell proliferation. The effects of UTP and ATP were prevented by both wide-range and specific purinergic antagonists. In contrast, in GES-1 cells ATP only decreased cell proliferation in a concentration-dependent manner, and UTP had no effect. Notably, the isolated application of purinergic antagonists was sufficient to change the basal proliferation of AGS cells, indicating that nucleotides released by the cells can act as paracrine/autocrine signals. Finally, in tumor-derived biopsies, we found an increase of P2Y2R and a decrease in P2X4R expression; however, we found high variability between seven different biopsies and their respective adjacent healthy gastric mucosa. Even so, we found a correlation between the expression levels of P2Y2R and P2X4R and survival rates of GC patients. Taken together, these results demonstrate the involvement of different purinergic receptors and signaling in GC, and the pattern of expression changes in tumoral cells, and this change likely directs ATP and nucleotide signaling from antiproliferative effects in healthy tissues to proliferative effects in cancer.
RESUMEN
An alternative method for determination of total trans fatty acids expressed as elaidic acid by capillary zone electrophoresis (CZE) under indirect UV detection at 224 nm within an analysis time of 7.5 min was developed. The optimized running electrolyte includes 15.0 mmol L(-1) KH(2)PO(4)/Na(2)HPO(4) buffer (pH approximately 7.0), 4.0 mmol L(-1) SDBS, 8.0 mmol L(-1) Brij35, 45%v/v ACN, 8% methanol, and 1.5% v/v n-octanol. Baseline separation of the critical pair C18-9cis/C18:1-9t with a resolution higher than 1.5 was achieved using C15:0 as the internal standard. The optimum capillary electrophoresis (CE) conditions for the background electrolyte were established with the aid of Raman spectroscopy and experiments of a 3(2) factorial design. After response factor (R(F)) calculations, the CE method was applied to total trans fatty acid (TTFA) analysis in a hydrogenated vegetable fat (HVF) sample, and compared with the American Oil Chemists' Society (AOCS) official method by gas chromatography (GC). The methods were compared with an independent sample t test, and no significant difference was found between CE and GC methods within the 95% confidence interval for six genuine replicates of TTFA analysis (p-value > 0.05). The CE method was applied to TTFA analysis in a spreadable cheese sample. Satisfactory results were obtained, indicating that the optimized methodology can be used for trans fatty acid determination for these samples.
Asunto(s)
Queso/análisis , Electroforesis Capilar/métodos , Ácidos Grasos trans/análisisRESUMEN
O uso de de derivados clorados de origem inorgânica, como gás cloro, hipoclorito de sódio, hipoclorito de cálcio e dos derivados clorados de origem orgânica, cujo principal representante é o dicloroisocianurato de sódio, tem contribuído para o controle das doenças de origem hídrica e alimentar, do processo de desinfecção de pisos, equipamentos e utensílios em áreas de industriais e de residências. Atualmente existe no mercado o dicloroisocianurato de sódio na forma comprimido efervescente, em diversos tamanhos, ou seja, o tamanho do comprimido a ser utilizado é em função do volume da solução sanificante a ser preparada e da concentração de cloro residual livre que se deseja, o que evita erros na dosagem do teor de matéria ativa e na perda do produto pelo consumo em excesso. O uso de derivados clorados de origem orgânica, principalmente o dicloroisocianurato de sódio, nos laticínios e mesmo no nosso dia a dia, no processo de desinfecção de água, equipamentos/utensílios, embalagens, ambientes, etc. ,em função da praticidade no manuseio, medição, transporte e armazenamento; maior solubilidade, maior período de validade, dosagem mais precisa, menor risco químico (corrosividade), modernidade do produto (tecnologia, embalagem e ainda efervescentes), a não formação de subprodutos da desinfecção, que associadas às necessidades atuais indicam que é uma questão de tempo a sua utilização em todos os setores. Pesquisa de opinião pública, realizada pela Data Kirsten por solicitação da Bayer Saúde Ambiental, avaliou a preferência entre dois agentes descontaminantes usados para água de consumo, o hipoclorito de sódio e o dicloroisocianurato de sódio, os resultados obtidos comprovam nossa afirmação anterior, onde o grau de preferência pelo dicloroisocianurato de sódio alcançou 69,6 por cento, contra 23,9 por cento do hipoclorito de sódio, sendo que 6,5 por cento dos entrevistados eram indiferentes.