Estrategias para la evaluación de extractos de polifenoles en modelos in vitro de cáncer de vías digestivas / Strategies for Evaluation of Polyphenol Extracts on in vitro Models of Digestive Tract Cancer / Estratégias para a avaliação de extratos de polifenóis em modelos in vitro de câncer do sistema digestivo

Introducción: Los polifenoles son compuestos que se encuentran naturalmente en alimentos como frutas, verduras, té, vino y chocolates, a los que se les atribuyen beneficios a la salud humana por su capacidad antioxidante. El cáncer de las vías digestivas se encuentra entre la tercera y quinta causas de muerte para la población, por lo que se ha incrementado el interés por realizar los estudios dirigidos a encontrar compuestos polifenólicos que ayuden en su prevención o tratamiento. Objetivo: Identificar las estrategias disponibles para la evaluación de polifenoles en células de cáncer de vías digestivas. Metodología: Búsqueda de literatura en bases de datos como Ovid, Pubmed, Science Direct y Elsevier Journal. Se seleccionaron artículos en los cuales se reporta el efecto biológico de los polifenoles sobre líneas celulares de cáncer de vías digestivas publicados entre 2012 y 2022. Resultados: Varios estudios han reportado el uso de un buen número de líneas celulares como modelos in vitropara estudios de polifenoles en cáncer y han resaltado las líneas AGS y HT-29, además de técnicas para la caracterización de los polifenoles, como el ensayo 2,2-Difenil-I-Picril Hidrazilo (DPPH). Sin embargo, para evaluar el efecto biológico se identifican diversas pruebas que deben analizarse antes de su implementación. Conclusiones: En la literatura se identifica que existen varias alternativas y estrategias para la evaluación de extrac-tos vegetales en cultivos in vitro de cáncer de vías digestivas; no obstante, antes de pasar al diseño experimental, deben tenerse en cuenta una serie de consideraciones para garantizar la utilidad de los resultados.

Introduction: Polyphenols are compounds naturally found in foods such as fruits, vegetables, tea, wine and chocolates, and it was attributed with benefits to human health due to their antioxidant capacity. Cancer of the digestive tract is between the third and fifth cause of death for the population, increasing the interest in carrying out studies aimed at finding polyphenolic compounds that help in their prevention or treatment. Objective: Identify the available strategies for the evaluation of polyphenols in digestive tract cancer cells. Method: A literature search was performed in databases such Ovid, Pubmed, Science Direct and Elsevier Journal and selected articles reporting the biological effect of polyphenols on digestive tract cancer cell lines, published between 2012 and 2022. Results: Currently studies report the use of a good number of cell lines as in vitro models for poly-phenol studies in cancer highlighting the AGS and HT-29 lines, in addition to techniques for the characterization of polyphenols such as the α, α-diphenyl-ß-picrylhydrazyl DPPH assay, however, to evaluate the biological effect various tests are identified that should be analyzed before implemen-tation. Conclusions: The literature identifies that there are many alternatives and strategies for the evaluation of plant extracts in in vitro cultures of digestive tract cancer, however, before moving on to the experimental design, a number of considerations should be taken into account to ensure the usability of the results

Introdução: Os polifenóis são compostos encontrados naturalmente em alimentos como frutas, legumes, chá, vinho e chocolates, aos quais são atribuídos benefícios para a saúde humana devido à sua capacidade antioxidante. O câncer do sistema digestivo está entre a terceira e a quinta principais causas de morte na população, o que levou a um interesse crescente em estudos destinados a encon-trar compostos polifenólicos que ajudem a prevenir ou tratar esse tipo de câncer. Objetivo: Identificar as estratégias disponíveis para a avaliação dos polifenóis nas células cancerosas do sistema digestivo. Metodologia: Pesquisa bibliográfica em bases de dados como Ovid, Pubmed, Science Direct e Elsevier Journal. Foram selecionados artigos que relatam o efeito biológico dos polifenóis em linhas celulares de câncer do sistema digestivo, publicados entre 2012 e 2022. Resultados: Vários estudos relataram a utilização de várias linhas celulares como modelos in vitro para estudos de polifenóis no câncer destacando as linhas AGS e HT-29, bem como técnicas para a ca-racterização de polifenóis, como o ensaio 2,2-Difenil-I-Picril Hidrazil (DPPH). No entanto, para avaliar o efeito biológico, são identificados vários testes que devem ser analisados antes da sua aplicação. Conclusões: A literatura identifica que existem várias alternativas e estratégias para a avaliação de extratos de plantas em culturas in vitro de câncer do sistema digestivo; no entanto, antes de passar à concepção experimental, é necessário ter em conta uma série de considerações para garantir a uti-lidade dos resultados

Characterisation of iunH gene knockout strain from Mycobacterium tuberculosis

BACKGROUND Tuberculosis (TB) is an infectious disease caused mainly by the bacillus Mycobacterium tuberculosis. The better understanding of important metabolic pathways from M. tuberculosis can contribute to the development of novel therapeutic and prophylactic strategies to combat TB. Nucleoside hydrolase (MtIAGU-NH), encoded by iunH gene (Rv3393), is an enzyme from purine salvage pathway in M. tuberculosis. MtIAGU-NH accepts inosine, adenosine, guanosine, and uridine as substrates, which may point to a pivotal metabolic role. OBJECTIVES Our aim was to construct a M. tuberculosis knockout strain for iunH gene, to evaluate in vitro growth and the effect of iunH deletion in M. tuberculosis in non-activated and activated macrophages models of infection. METHODS A M. tuberculosis knockout strain for iunH gene was obtained by allelic replacement, using pPR27xylE plasmid. The complemented strain was constructed by the transformation of the knockout strain with pNIP40::iunH. MtIAGU-NH expression was analysed by Western blot and LC-MS/MS. In vitro growth was evaluated in Sauton’s medium. Bacterial load of non-activated and interferon-γ activated RAW 264.7 cells infected with knockout strain was compared with wild-type and complemented strains. FINDINGS Western blot and LC-MS/MS validated iunH deletion at protein level. The iunH knockout led to a delay in M. tuberculosis growth kinetics in Sauton’s medium during log phase, but did not affect bases and nucleosides pool in vitro. No significant difference in bacterial load of knockout strain was observed when compared with both wild-type and complemented strains after infection of non-activated and interferon-γ activated RAW 264.7 cells. MAIN CONCLUSION The disruption of iunH gene does not influence M. tuberculosis growth in both non-activated and activated RAW 264.7 cells, which show that iunH gene is not important for macrophage invasion and virulence. Our results indicated that MtIAGU-NH is not a target for drug development.