RESUMEN
Fulvic acid (FA) was extracted and purified from Canadian Sphagnum peat (CP-FA) and characterized by using an element analysis meter, Fourier transform infrared (FT-IR) spectroscopy, electron spin resonance (ESR) spectroscopy, and (13)C-nuclear magnetic resonance ((13)C-NMR) spectroscopy. To investigate the antiallergic effect of CP-FA, we incubated rat basophilic leukemia (RBL-2H3) cells with 0.001-10.0 microg/ml of CP-FA and determined the beta-hexosaminidase release inhibition at different response stages. The intracellular calcium [Ca(2+)](i) level was also determined by using Fluo 3-AM, a calcium-specific fluorescent probe, and the cytotoxicity of CP-FA was determined by the 3-(4,5-dimethythiazol-2-yl)-2,5-diphenyl-tetrazolium bromide (MTT) assay. The results revealed that RBL-2H3 cells incubated for 48 h with 0.001-10.0 microg/ml of CP-FA did not show any decreased viability. CP-FA inhibited the beta-hexosaminidase release by IgE-sensitized, antigen-stimulated RBL-2H3 cells at the antigen-antibody binding stage and the antibody-receptor binding stage. CP-FA also inhibited histamine release from A23187 plus PMA- or compound 48/80-stimulated KU812 cells. Furthermore, there was a decrease in the intracellular [Ca(2+)](i) level in IgE-sensitized cells incubated with CP-FA and stimulated with antigen. Our results show that CP-FA may be useful for the treatment or prevention of allergic diseases.
Asunto(s)
Antialérgicos/farmacología , Basófilos/metabolismo , Benzopiranos/farmacología , Liberación de Histamina/efectos de los fármacos , Extractos Vegetales/farmacología , Sphagnopsida/química , Animales , Complejo Antígeno-Anticuerpo/inmunología , Antígenos/inmunología , Calcimicina/farmacología , Calcio/análisis , Línea Celular Tumoral , Relación Dosis-Respuesta a Droga , Colorantes Fluorescentes , Formazáns/metabolismo , Humanos , Inmunoglobulina E/farmacología , Ionóforos/farmacología , Leucemia Basofílica Aguda , Leucemia Experimental , Leucemia Mielógena Crónica BCR-ABL Positiva , Resonancia Magnética Nuclear Biomolecular , Ratas , Espectroscopía Infrarroja por Transformada de Fourier , Acetato de Tetradecanoilforbol/farmacología , Sales de Tetrazolio/metabolismo , beta-N-Acetilhexosaminidasas/antagonistas & inhibidores , p-Metoxi-N-metilfenetilamina/farmacologíaRESUMEN
Wastewater and surface water samples from three wastewater treatment plants (WWTPs) and three rivers in Tunisia were assayed for estrogenic activity using the E-screen assay and enzyme-linked immunosorbent assay (ELISA). Results showed that all the Tunisian raw wastewater samples as well as the Roriche river water sample induced a strong proliferative response in human MCF-7 breast cancer cells. Tunisian raw wastewater had an average 17beta-estradiol content of 2,705.4 pg/ml, whereas that of the Roriche river was 36.7 pg/ml, which is sufficient for inducing endocrine-mediated responses in aquatic organisms. Results further showed that the Mornag WWTP, which uses the activated-sludge treatment system, has a higher estrogen removal efficiency than the stabilization ponds of the Gammart and pilot WWTPs. This study, which is the first of such studies in Tunisia, and probably the first in the North African region, underscores the need to detect and monitor the estrogenic activity of water and wastewater, given the scarcity of water in Tunisia and the detrimental impact of endocrine-disrupting compounds on the physiology of both animals and humans.
Asunto(s)
Estrógenos/toxicidad , Contaminantes Químicos del Agua/toxicidad , Bioensayo , Biomarcadores/análisis , Biomarcadores/metabolismo , Biomarcadores de Tumor/análisis , Biomarcadores de Tumor/metabolismo , Células CACO-2 , División Celular , Línea Celular Transformada , Línea Celular Tumoral , Proteínas de Unión al ADN/análisis , Proteínas de Unión al ADN/metabolismo , Ensayo de Inmunoadsorción Enzimática , Estrógenos/análisis , Humanos , Factor 1 de Elongación Peptídica/análisis , Factor 1 de Elongación Peptídica/metabolismo , Fosfopiruvato Hidratasa/análisis , Fosfopiruvato Hidratasa/metabolismo , Proteómica , Proteínas Supresoras de Tumor/análisis , Proteínas Supresoras de Tumor/metabolismo , Túnez , Eliminación de Residuos Líquidos , Contaminantes Químicos del Agua/análisisRESUMEN
Organophosphorous pesticides are currently widely used in China to help boost agricultural production. However, these pesticides pose various threats to organisms, including humans, and are thus a cause of concern. Five organophosphorous pesticides, monocrotophos, omethoate, parathion-methyl, phoxim and dichlorvos, were examined for their effects on mammalian cell lines to determine their potential impact on physiological functions in vivo. Results show an increased proliferation of MCF-7 cells treated with 0.2 microM monocrotophos or 0.4 microM omethoate, suggesting that these compounds can induce breast cancer cell proliferation at relatively low concentrations. Murine primary spleen cells markedly decreased in number starting at a pesticide concentration of 0.01 microM; no cytotoxicity was observed below 0.001 microM. BALB/c3T3 murine fibroblasts treated with 0.25 microM monocrotophos showed enhanced DNA synthesis, while those treated with the other pesticides showed results similar to that of the control. The different pesticides reduced the acetylcholinesterase (AChE) activity of the rat neuronal cell line PC12 in a dose-dependent manner up to 100 microM. Parathion-methyl and phoxim showed acute toxicity at 0.01 microM. Finally, phoxim and parathion-methyl significantly reduced the transepithelial electrical resistance (TEER) of human intestinal Caco-2 cells, indicating that these pesticides can disrupt the tight-junction permeability of cell monolayers. These in vitro assays, which are rapid, reproducible, simple and inexpensive, clearly show the effects of organophosphorous pesticides on mammalian cells and suggest the potential impact of these pesticides on organisms in vivo.