BACKGROUND: Multidrug-resistant infections due to Mycobacterium abscessus often require complex and prolonged regimens for treatment. Here, we report the evaluation of a new ex vivo antimicrobial susceptibility testing model using organotypic cultures of murine precision-cut lung slices, an experimental model in which metabolic activity, and all the usual cell types of the organ are found while the tissue architecture and the interactions between the different cells are maintained. METHODS: Precision cut lung slices (PCLS) were prepared from the lungs of wild type BALB/c mice using the Krumdieck® tissue slicer. Lung tissue slices were ex vivo infected with the virulent M. abscessus strain L948. Then, we tested the antimicrobial activity of two drugs: imipenem (4, 16 and 64 µg/mL) and tigecycline (0.25, 1 and 4 µg/mL), at 12, 24 and 48 h. Afterwards, CFUs were determined plating on blood agar to measure the surviving intracellular bacteria. The viability of PCLS was assessed by Alamar Blue assay and corroborated using histopathological analysis. RESULTS: PCLS were successfully infected with a virulent strain of M. abscessus as demonstrated by CFUs and detailed histopathological analysis. The time-course infection, including tissue damage, parallels in vivo findings reported in genetically modified murine models for M. abscessus infection. Tigecycline showed a bactericidal effect at 48 h that achieved a reduction of > 4log10 CFU/mL against the intracellular mycobacteria, while imipenem showed a bacteriostatic effect. CONCLUSIONS: The use of this new organotypic ex vivo model provides the opportunity to test new drugs against M. abscessus, decreasing the use of costly and tedious animal models.
Anti-Bacterial Agents/administration & dosage , Lung/microbiology , Mycobacterium Infections, Nontuberculous/drug therapy , Mycobacterium abscessus/drug effects , Animals , Humans , In Vitro Techniques , Male , Mice , Mice, Inbred BALB C , Microbial Sensitivity Tests , Models, Biological , Mycobacterium Infections, Nontuberculous/microbiology , Mycobacterium abscessus/physiology
Infections caused by Trichomonas vaginalis in humans are one of the main public health problems caused by sexually transmitted diseases. Objective of this study was to evaluate potential biological activity of the medicinal plant Argemone mexicana (Mexican poppy) on T. vaginalis. Methanolic extracts of the stems and leaves of A. mexicana, and different fractions were prepared with solvents of different polarities. The extracts and functional groups were detected containing sterols, triterpenes, quinones, flavonoids and, alkaloids. Extracts from both the stems and leaves of A. mexicana inhibited the growth of T. vaginalis with half-maximal inhibitory concentration value of 70.6 and 67.2 µg/ml, respectively. In the active fractions, the most abundant compounds were berberine and jatrorrhizine, with presumed antiparasitic activity.
Plant Extracts/pharmacology , Trichomonas vaginalis/drug effects , Trichomonas vaginalis/growth & development , Antineoplastic Combined Chemotherapy Protocols , Bacterial Vaccines , Cyclophosphamide , Depression, Chemical , Dose-Response Relationship, Drug , Doxorubicin , Fluorouracil , In Vitro Techniques , Leucovorin , Methanol , Plant Extracts/chemistry , Plant Leaves/chemistry , Plant Stems/chemistry , Quinones , Sterols , Triterpenes
Due to the biological activities of Syzygium aromaticum essential oil, its incorporation in methacrylate polymeric (Eudragit E100) nanoparticles (NP), physical characterization, and antimicrobial essays were evaluated. The clove bears great potential for applications in dentistry. The oil was obtained by hydrodistillation and oil loaded NP using the nanoprecipitation method. Particle size and polydispersity index were determined by photon correlation spectroscopy, and physical morphology by electron microscopy. Loading capacity and in vitro eugenol release were evaluated by gas mass chromatography, and the antimicrobial activity of oil loaded-NP was calculated against Streptococcus mutans. Different chemical ingredients were characterized, and eugenol was the principal compound with 51.55%. Polymer content was directly related to NP homogenous size, which was around 150 nm with spherical morphology. A 73.2% loading capacity of eugenol was obtained. Oil loaded NP presented a fickian-type release mechanism of eugenol. Antimicrobial activity to 300 µg/mL was obtained after 24 h.
Debido a las actividades biológicas del aceite esencial de Syzygium aromaticum, se evaluó su incorporación en nanopartículas (NP) de metacrilato polimérico (Eudragit E100), su caracterización y ensayos antimicrobianos. El clavo tiene un gran potencial para aplicaciones en odontología. El aceite se obtuvo por hidrodestilación y las NP cargado de aceite utilizando el método de nanoprecipitación. El tamaño de partícula y el índice de polidispersidad se determinaron mediante espectroscopia de correlación fotónica y su morfología por microscopía electrónica. La capacidad de carga y la liberación de eugenol in vitro se evaluaron mediante cromatografía de gases en masa, y la actividad antimicrobiana se evaluó contra Streptococcus mutans. Se caracterizaron diferentes ingredientes químicos, siendo el eugenol el principal compuesto con 51.55%. El contenido de polímero se relacionó directamente con el tamaño homogéneo de NP, que fue de alrededor de 150 nm con morfología esférica. Se obtuvo un 73,2% de capacidad de carga de eugenol. El aceite cargado en NP presentó un mecanismo de liberación de eugenol de tipo fickiano. La actividad antimicrobiana a 300 µg/mL se obtuvo después de 24 h.
Polymers/chemistry , Oils, Volatile/administration & dosage , Syzygium/chemistry , Nanoparticles/chemistry , Anti-Bacterial Agents/administration & dosage , Streptococcus mutans/drug effects , Eugenol/pharmacology , Oils, Volatile/pharmacology , Administration, Oral , Chromatography, Thin Layer , Drug Delivery Systems , Gas Chromatography-Mass Spectrometry , Anti-Bacterial Agents/pharmacology
Maceration and Soxhlet methods were used to obtain methanol extracts from a Mexican grape (Ruby Cabernet) pomace and the biological activity and phenolic profiles were compared. The antioxidant capacity was used to evaluate the mechanism of action, using a physiological model (erythrocytes) of damage induced by AAPH-generated free radicals. The extract obtained by maceration presented a total phenolic content twice the one obtained using the Soxhlet method. It also contained the most potent antioxidants, reducing anisotropy in the presence of AAPH to the levels of untreated cells, restoring membrane fluidity, preventing the morphological changes, as demonstrated by scanning electron microscopy (SEM), and providing protection against protein oxidation at the higher concentration. Our work showed that both extracts presented significant antioxidant activity through positive interactions with the lipid bilayer.
Antioxidants/chemistry , Erythrocytes/drug effects , Phenols/chemistry , Vitis/chemistry , Free Radicals , Mexico , Oxidation-Reduction , Plant Extracts/chemistry
Cuatro reguladores comerciales (Biofol, Biozyme, Cytokin y Activol) y un extracto de algas (AlgaEnzims) se aplicaron en las dosis recomendadas, sobre un cultivo de papa (S.tuberosum L.var gigant) con el objeto de evaluar su objeto sobre el valor nutricional (materia seca, humedad, ceniza, grasa, proteína, carbohidratos y fibra dietética). Dichos parámetros fueron analizados de acuerdo al official methods of analysis (AOAC). El diseño experimental consistió en bloques al azar con 8 tratamientos y 4 repeticiones de 32 parcelas. El análisis de varianza muestra diferencias significativas entre los tratamientos para todas las variables bromatológicas. El mayor contenido de ceniza (6,20) se presentó con el tratamiento AlgaEnzims-suelo. Los tratamientos AlgaEnzims-suelo (9,30), AgaEnzims-follaje (8.90) y Cytokin (8.70) dieron valores más altos de proteína comparados con el testigo (6,20). el contenido de carbohidratos fue más alto con Biofol (88,21). La fibra dietética fue más alta en el tratamiento AlgaEnzims-suelo-follage (5,84) y más baja en Biofol (1,67). Con Biozyme y Cytokin se obtuvieron los mayores contenidos de grasa en tubérculos. Con activol se obtuvo el mejor peso de tubérculos y con AlgaEnzims-suelo el contenido de proteínas más alto
Eukaryota , Solanum tuberosum , Mexico , Nutritional Sciences
