RESUMEN
Achyrocline satureioides is widely consumed as infusion or aperitif and shows important therapeutic properties. Previously, we reported absence of genotoxicity of cold aqueous extract (CAE) of A. satureioides by Allium test. However, one test cannot predict the genotoxic effects of a substance. Thus, the aim of this work was to investigate cytotoxicity, genotoxicity and apoptotic ability of CAE of A. satureioides. In addition, CAE was chemically characterized. The cytotoxicity was evaluated by Trypan blue and MTT assays. The apoptotic capacity was evaluated by Hoechst staining and DNA fragmentation-analysis. The genotoxicity was studied by comet assay (CA) and micronucleus test. The identification and quantification of flavonoids were performed by HPLC-ESI-MS/MS. The cytotoxicity studies indicated low toxicity of CAE. In addition, CAE did not induce apoptotic effects on human PBMCs. CAE did not show genotoxicity in vitro against Vero cells, at 10-50 µg/mL. CAE did not induce in vivo genotoxic effects, but it showed at high concentrations cytotoxicity by micronucleus assay. CAE presented flavonoids such as quercetin, 3-O-methylquercetin and luteolin. In conclusion, A. satureioides at popularly concentrations used, in aperitif or infusion, can be consumed safely because did not show any cytotoxic or genotoxic effects.
Asunto(s)
Achyrocline/química , Apoptosis/efectos de los fármacos , Daño del ADN/efectos de los fármacos , Extractos Vegetales/farmacología , Animales , Chlorocebus aethiops , Cromatografía Líquida de Alta Presión , Ensayo Cometa , Fragmentación del ADN , Evaluación Preclínica de Medicamentos , Femenino , Humanos , Dosificación Letal Mediana , Leucocitos Mononucleares/efectos de los fármacos , Luteolina/análisis , Luteolina/farmacología , Masculino , Ratones , Ratones Endogámicos BALB C , Pruebas de Micronúcleos , Extractos Vegetales/análisis , Quercetina/análogos & derivados , Quercetina/análisis , Quercetina/farmacología , Espectrometría de Masas en Tándem , Células VeroRESUMEN
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Asunto(s)
Adulto , Masculino , Humanos , Transporte de Pacientes/métodos , Traumatismos Craneocerebrales/terapia , Servicios Médicos de Urgencia/métodos , Escala de Coma de Glasgow , Traumatismos Craneocerebrales/clasificaciónRESUMEN
Mitogenic and antigenic lymphocyte stimulation was examined in pigs that had been immunized with 2 inactivated vaccines which had been prepared with Aujesky's disease virus (ADV), strain RC/79. One vaccine was partially purified by ultra-centrifugation (Vaccine A) the other one was not (Vaccine B). A second dose of vaccine had no influence over the blastogenic response when the lymphocytes were stimulated with phytohemoagglutinin (PHA). Lymphocyte response to the ADV antigen in the immunized pigs was significantly higher at day 30 post inoculation than at day 0 indicating that it was highly specific. Cellular antigens contained in the viral cultures produced a slight non-specific response as shown by a low increase in the levels of lymphocyte blastic transformation (LBT) in the control group at day 30 p.i., this group only received a non infected Vero cell suspension. This was the case in pigs that received vaccine A as well as in those that were vaccinated with vaccine B. Vaccine B contains a greater quantity of contaminating cellular antigens, since it is an impure vaccine. Such antigens could act as non-specific immunomodulators, potentiating cell-mediated immunity (CMI). This assay demonstrated that inactivated vaccines produced with VPR-RC/79, partially purified and unpurified are capable of inducing a humoral immune response. The blastogenic reaction of the peripheral blood lymphocytes to antigens of ADV strain RC/79, indicated that the employed immunogens also induced the CMI. Results indicate that the analyzed immunogens could be considered for the possible implementation of epidemiological measures, which imply the use of vaccines to prevent pseudo-rabies in Argentina.
Asunto(s)
Herpesvirus Suido 1/inmunología , Activación de Linfocitos , Seudorrabia/prevención & control , Sus scrofa/inmunología , Enfermedades de los Porcinos/prevención & control , Vacunas Virales/inmunología , Adyuvantes Inmunológicos , Animales , Formación de Anticuerpos , Antígenos de Superficie/inmunología , Antígenos Virales/inmunología , Chlorocebus aethiops , Femenino , Inmunidad Celular , Activación de Linfocitos/efectos de los fármacos , Linfocitos/efectos de los fármacos , Linfocitos/inmunología , Masculino , Fitohemaglutininas/farmacología , Vacunas de Productos Inactivados/inmunología , Vacunas de Productos Inactivados/aislamiento & purificación , Células Vero/inmunología , Vacunas Virales/aislamiento & purificaciónRESUMEN
The effect produced on Vero cell monolayers by toxins derived from Staphylococcus strains was characterized. 210 milk samples taken from dairy cows suffering from sub-clinical mastitis were analyzed. Strains belonging to the Staphylococcus genus were isolated from 73 of these milk samples. The production of toxins was then stimulated from these strains when they were cultured in Dolman's medium. The study of cell cultures showed that 53 toxin samples induced marked and irreversible cellular changes. This is compared to 42 samples (57.5%) which were strongly cytotoxic. The remaining 11 samples were shown to be slowly cytotoxic. 16% of the total toxins did not induce cell damage and 11% of the toxins produced cellular damage that was reversible in less than 24 hrs, and were designated as cytotonic. Haemolytic actively in vitro, using sheep red blood cells, was assessed using toxins that caused alteration in the monolayers. The results indicate that 46.51% of the toxins showed beta haemolytic activity, 2.32% alpha haemolytic activity, and 51.16% showed neither alpha nor beta haemolytic activity. The later type of activity did however cause damage to cultured cells, which suggests that the causative agent could be delta toxin. This study reveals a strong predominance of beta haemolytic strains in the dairy farm studied. These strains induced in vitro cell damage, and it is possible to speculate that mammary gland tissue damage is similarly produced, which may be attributed to both beta and/or delta haemolytic toxins.