ABSTRACT
OBJECTIVES: To evaluate the cytotoxic activity of the chloroform fraction of the Piper aduncum methanolic extract (PAMoCl) and its effect on the cell cycle in two gastric cancer cell lines: AGS and KATO III. MATERIALS AND METHODS: The cytotoxic effect of PAMoCl was evaluated in cell lines AGS and KATO III. The following PAMoCl concentrations were tested, 1.25, 2.5, 5, 10, 20, 40, 80 and 160 µg/mL. Resazurine was used to evaluate cell viability. In the cell cycle assay, the cells were treated with 19.62 µg/mL and 39.23 µg/mL of PAMoCl for AGS as well as 87.49 µg/mL and 160 µg/mL for KATO III. After 24 hours both cell lines were analyzed by flow cytometry. RESULTS: PAMoCl showed cytotoxic activity, inhibiting cell growth by 50%. It presented a (IC50) of 39.23 µg/mL and 87.49 µg/mL at 24 hours and a (IC50) of 49.47 µg/mL and 64.68 µg/mL at 48 hours against AGS and KATO III cell lines, respectively. In addition, it was observed that PAMoCl has an effect on the cell cycle, it causes an accumulation of cells in the G2/M phase. CONCLUSIONS: PAMoCl contains secondary metabolites with cytotoxic activity that have an effect on the G2/M phase of the cell cycle, in two gastric cancer cell lines, both primary and metastatic. The results of this study will allow us to deepen the search for more effective active ingredients found in PAMoCl for eliminating gastric cancer cells, but with less toxicity for healthy cells.
OBJETIVOS: Evaluar la actividad citotóxica de la fracción clorofórmica del extracto metanólico de Piper aduncum (PAMoCl) y su efecto en el ciclo celular en dos líneas celulares de cáncer gástrico: AGS y KATO III. MATERIALES Y MÉTODOS: El efecto citotóxico de PAMoCl se evaluó en las líneas celulares: AGS y KATO III. Se probaron concentraciones de PAMoCl: 1,25; 2,5; 5; 10; 20; 40; 80 y 160 µg/mL. Para evaluar la viabilidad celular se usó el reactivo resazurina. En el ensayo de ciclo celular las células fueron tratadas con 19,62 µg/mL y 39,23 µg/mL de PAMoCl para AGS, así como 87,49 µg/mL y 160 µg/mL para KATO III. Después de 24 horas ambas líneas celulares fueron analizadas por citometría de flujo. RESULTADOS: PAMoCl mostró actividad citotóxica con una inhibición del crecimiento celular en un 50% (IC50) de 39,23 µg/mL y 87,49 µg/mL a las 24 horas y un IC50 de 49,47 µg/mL y 64,68 µg/mL a las 48 horas frente a las líneas celulares AGS y KATO III, respectivamente. Además, se observó que PAMoCl tiene efecto a nivel del ciclo celular: provoca una acumulación de células en la fase G2/M. CONCLUSIONES: PAMoCl contiene metabolitos secundarios con actividad citotóxica que tienen efecto en la fase G2/M del ciclo celular, en dos líneas celulares de cáncer gástrico tanto primario como metastásico. Los resultados de este estudio permitirán profundizar en la búsqueda de principios activos presentes en PAMoCl que tengan mayor eficacia en la eliminación de células de cáncer gástrico, pero con menor toxicidad en células sanas.
Subject(s)
Chloroform , Piper , Stomach Neoplasms , Cell Cycle/drug effects , Cell Line, Tumor/cytology , Cell Line, Tumor/drug effects , Chloroform/pharmacology , Cytotoxins , Humans , Stomach Neoplasms/drug therapyABSTRACT
RESUMEN Objetivos: Evaluar la actividad citotóxica de la fracción clorofórmica del extracto metanólico de Piper aduncum (PAMoCl) y su efecto en el ciclo celular en dos líneas celulares de cáncer gástrico: AGS y KATO III. Materiales y métodos: El efecto citotóxico de PAMoCl se evaluó en las líneas celulares: AGS y KATO III. Se probaron concentraciones de PAMoCl: 1,25; 2,5; 5; 10; 20; 40; 80 y 160 µg/mL. Para evaluar la viabilidad celular se usó el reactivo resazurina. En el ensayo de ciclo celular las células fueron tratadas con 19,62 µg/mL y 39,23 µg/mL de PAMoCl para AGS, así como 87,49 µg/mL y 160 µg/mL para KATO III. Después de 24 horas ambas líneas celulares fueron analizadas por citometría de flujo. Resultados: PAMoCl mostró actividad citotóxica con una inhibición del crecimiento celular en un 50% (IC50) de 39,23 µg/mL y 87,49 µg/mL a las 24 horas y un IC50 de 49,47 µg/mL y 64,68 µg/mL a las 48 horas frente a las líneas celulares AGS y KATO III, respectivamente. Además, se observó que PAMoCl tiene efecto a nivel del ciclo celular: provoca una acumulación de células en la fase G2/M. Conclusiones: PAMoCl contiene metabolitos secundarios con actividad citotóxica que tienen efecto en la fase G2/M del ciclo celular, en dos líneas celulares de cáncer gástrico tanto primario como metastásico. Los resultados de este estudio permitirán profundizar en la búsqueda de principios activos presentes en PAMoCl que tengan mayor eficacia en la eliminación de células de cáncer gástrico, pero con menor toxicidad en células sanas.
ABSTRACT Objectives: To evaluate the cytotoxic activity of the chloroform fraction of the Piper aduncum methanolic extract (PAMoCl) and its effect on the cell cycle in two gastric cancer cell lines: AGS and KATO III. Materials and methods: The cytotoxic effect of PAMoCl was evaluated in cell lines AGS and KATO III. The following PAMoCl concentrations were tested, 1.25, 2.5, 5, 10, 20, 40, 80 and 160 μg/mL. Resazurine was used to evaluate cell viability. In the cell cycle assay, the cells were treated with 19.62 μg/mL and 39.23 μg/mL of PAMoCl for AGS as well as 87.49 μg/mL and 160 μg/mL for KATO III. After 24 hours both cell lines were analyzed by flow cytometry. Results: PAMoCl showed cytotoxic activity, inhibiting cell growth by 50%. It presented a (IC50) of 39.23 μg/mL and 87.49 μg/mL at 24 hours and a (IC50) of 49.47 μg/mL and 64.68 μg/mL at 48 hours against AGS and KATO III cell lines, respectively. In addition, it was observed that PAMoCl has an effect on the cell cycle, it causes an accumulation of cells in the G2/M phase. Conclusions: PAMoCl contains secondary metabolites with cytotoxic activity that have an effect on the G2/M phase of the cell cycle, in two gastric cancer cell lines, both primary and metastatic. The results of this study will allow us to deepen the search for more effective active ingredients found in PAMoCl for eliminating gastric cancer cells, but with less toxicity for healthy cells.
Subject(s)
Stomach Neoplasms , Cell Cycle , Cell Line , Chloroform , Piper , Neoplasm MetastasisABSTRACT
BACKGROUND: Mycoplasma pneumoniae and Chlamydia pneumoniae are atypical pathogens responsible for pneumonia and a leading cause of morbidity and mortality in low income countries. The study objective is to determine the prevalence of this pathogens in Peruvian children with acute respiratory infections. METHODS: A consecutive cross-sectional study was conducted in Lima, Peru from May 2009 to September 2010. A total of 675 children admitted with clinical diagnoses of acute respiratory infections were tested for Mycoplasma pneumoniae and Chlamydia pneumoniae detection by polymerase chain reaction (PCR), and clinical symptoms were registered by the attending physician. RESULTS: Mycoplasma pneumonia was detected in 25.19% (170/675) of nasopharyngeal samples and Chlamydia pneumonia in 10.52% (71/675). The most common symptoms in patients with these atypical pathogens were rhinorrhea, cough and fever. A higher prevalence of Mycoplasma pneumoniae cases were registered in summer, between December 2009 and March 2010. CONCLUSIONS: Mycoplasma pneumoniae and Chlamydia pneumonia are a significant cause of morbidity in Peruvian children with acute respiratory infections (ARI). Further studies should evaluate the use of reliable techniques such as PCR in Peru in order to avoid underdiagnoses of these atypical pathogens.
Subject(s)
Acute Disease/epidemiology , Chlamydial Pneumonia/epidemiology , Pneumonia, Mycoplasma/epidemiology , Respiratory Tract Infections/epidemiology , Adolescent , Child , Child, Preschool , Chlamydial Pneumonia/microbiology , Chlamydophila pneumoniae/isolation & purification , Chlamydophila pneumoniae/pathogenicity , Female , Humans , Infant , Male , Mycoplasma pneumoniae/isolation & purification , Mycoplasma pneumoniae/pathogenicity , Peru , Pneumonia, Mycoplasma/microbiology , Respiratory Tract Infections/microbiologyABSTRACT
Introducción: La tuberculosis (TB) sigue siendo uno de las principales causas de muerte en el mundo. La TB meníngea como complicación requiere detección pronta e instalación inmediata de la terapia apropiada. Dada la baja sensibilidad de la baciloscopia y del cultivo del bacilo tuberculoso, se presenta el estudio de las isoenzimas adenosina deaminasa (ADA) como contribución al diagnóstico diferencial de la TB meníngea. Objetivo: Determinar la actividad de isoenzimas ADA en líquido cefalorraquídeo (LCR) de pacientes con TB meníngea. Diseño: Investigación descriptiva, con muestreo no probabilístico. Institución: Centro de Investigación de Bioquímica y Nutrición, Universidad Nacional Mayor de San Marcos. Material biológico: Muestras de LCR de pacientes con TB. Intervenciones: Se consideró muestras de LCR con actividad ADA mayor a 9 U/L, que fueron conservadas a -40 °C hasta la corrida electroforética en sistema vertical. El análisis estadístico se realizó aplicando Mann - Withney, con un nivel de significancia de 0,1 para comparar los valores ADA de TB meníngea frente a otras enfermedades del sistema nervioso central (SNC). Principales medidas de resultados: Isoenzimas ADA1m, ADA1cp y ADA2. Resultados: Las medianas de la actividad de ADA total en LCR de pacientes tuberculosos fueron mayores que en otras enfermedades parainfectivas del SNC. ADA1cp tuvo mayor contribución para ADA total en ambos casos, mostrando mayor incremento en tuberculosis meníngea. También, ADA2 aumentó en TB meníngea frente a otras enfermedades del SNC. Conclusiones: Las isoenzimas ADA en LCR expresan niveles elevados de ADA2 en tuberculosis meníngea, como consecuencia del incremento de la línea celular monocito-macrófago.
Introduction: Tuberculosis (TB) is a leading cause of death worldwide. Meningeal tuberculosis is a complication that requires early detection and immediate installation of appropriate therapy. Given the low sensitivity of sputum smear and culture of bacillus tuberculosis, other tools should be explored in the differential diagnosis of tuberculous meningitis such as adenosine deaminase (ADA) isoenzymes. Objectives: To determine ADA isoenzymes activity in cerebrospinal fluid (CSF) of patients with tuberculous meningitis. Design: Descriptive study, with non-probability sampling. Institution: Biochemistry and Nutrition Research Center, Universidad Nacional Mayor de San Marcos, Lima, Peru. Biological materials: Samples of CSF from patients with clinical symptoms of tuberculous meningitis. Interventions: Samples of CSF with ADA activity above 9 U / L were stored at -40°C until vertical electrophoresis was run. Mann - Whitney statistical analysis was used with 0.1 significance level to compare meningeal tuberculosis ADA values with other nervous system diseases. Main outcome measures: Isoenzymes ADA1m, ADA1cp, and ADA2. Results: Median total ADA enzyme activity in CSF in all patients with TB meningitis was higher than in other parainfective diseases of the central nervous system. In both cases ADA1cp had greater contribution to total ADA and showed larger increase in tuberculous meningitis. ADA2 increase was higher in meningeal tuberculosis compared with other CNS diseases. Conclusions: CSF electrophoresis distinguishes ADA isoenzymes, showing elevated levels of ADA2 in meningeal tuberculosis as a result of increased monocyte-macrophage cell line.
