RESUMEN
An estimated one-third of tuberculosis (TB) cases go undiagnosed or unreported. Sputum samples, widely used for TB diagnosis, are inefficient at detecting infection in children and paucibacillary patients. Indeed, developing point-of-care biomarker-based diagnostics that are not sputum-based is a major priority for the WHO. Here, in a proof-of-concept study, we tested whether pulmonary TB can be detected by analyzing patient exhaled breath condensate (EBC) samples. We find that the presence of Mycobacterium tuberculosis (Mtb)-specific lipids, lipoarabinomannan lipoglycan, and proteins in EBCs can efficiently differentiate baseline TB patients from controls. We used EBCs to track the longitudinal effects of antibiotic treatment in pediatric TB patients. In addition, Mtb lipoarabinomannan and lipids were structurally distinct in EBCs compared to ex vivo cultured bacteria, revealing specific metabolic and biochemical states of Mtb in the human lung. This provides essential information for the rational development or improvement of diagnostic antibodies, vaccines and therapeutic drugs. Our data collectively indicate that EBC analysis can potentially facilitate clinical diagnosis of TB across patient populations and monitor treatment efficacy. This affordable, rapid and non-invasive approach seems superior to sputum assays and has the potential to be implemented at point-of-care.
Asunto(s)
Líquidos Corporales , Mycobacterium tuberculosis , Tuberculosis Pulmonar , Tuberculosis , Humanos , Niño , Tuberculosis/diagnóstico , Tuberculosis/microbiología , Tuberculosis Pulmonar/diagnóstico , Tuberculosis Pulmonar/microbiología , Esputo/microbiología , Sensibilidad y EspecificidadRESUMEN
Histoplasma capsulatum is a dimorphic, thermal, and nutritional fungus. In the environment and at an average temperature of 28 °C, it develops as a mold that is composed of infecting particles. Once in the host or in cultures at 37 °C, it undergoes a transition into the parasitic form. In the present work, we performed chemical extraction and characterization using chromatography techniques of the associated lipid composition of the external surface of the cell wall of the mycelial phase of two isolates of the H. capsulatum: one clinical and one environmental. Several differences were evidenced in the fatty acids in the phospholipid composition. Surface pressure-area isotherms and compression module curves of the Amphotericin B and lipid extract monolayers, as well as (AmB)-lipid extract mixed monolayers were recorded. Results show a high affinity of AmB towards lipid extracts. The most stable monolayers were formed by AmB + environmental with a mass ratio of 1:3 and AmB + clinical with a mass ratio of 1:2. Knowledge of the AmB aggregation processes at a molecular level and the characterization of the lipid extracts allows the possibility to understand the interaction between the AmB and the lipid fractions of H. capsulatum.
RESUMEN
The anti-mycobacterial immune response in adults and children with tuberculosis (TB), as well as the response in bacteriologically positive and negative patients, is different. However, knowledge of the immunological events occurring in the lungs in these clinical situations remains scarce. Exhaled breath condensate (EBC) samples may be useful for studying the inflammatory environment of the lower airways in TB patients. The fatty acid, cytokine, and chemokine profiles in EBC from healthy adults; smear-positive and smear-negative adult patients; and healthy, asthmatic, and TB children were determined using gas chromatography and LUMINEX, respectively. Unsaturated fatty acids, particularly oleate, were increased in TB adults and children compared with healthy individuals. Elevated levels of IL-17 were characteristic of paucibacillary patients (adults and children), whereas elevated MCP-1 (monocyte chemotactic protein-1) levels were characteristic of adult patients (smear-positive and smear-negative). The levels of all of the molecules were comparable to the controls after anti-TB treatment, suggesting that changes in the levels of the molecules detected in the EBC samples were the result of the active pulmonary TB. EBC samples may be an important tool for the detection of potential early biomarkers in the different clinical manifestations of pulmonary TB and a useful tool for the diagnosis of TB, particularly in children.
Asunto(s)
Pruebas Respiratorias/métodos , Quimiocinas/análisis , Espiración , Ácidos Grasos/análisis , Tuberculosis/diagnóstico , Adolescente , Adulto , Anciano , Biomarcadores/análisis , Estudios de Casos y Controles , Niño , Preescolar , Diagnóstico Diferencial , Análisis Discriminante , Ésteres/análisis , Espiración/fisiología , Femenino , Estudios de Seguimiento , Humanos , Masculino , Persona de Mediana Edad , Reproducibilidad de los Resultados , Tuberculosis/tratamiento farmacológico , Adulto JovenRESUMEN
The fatty acid composition of monocytes changes substantially during differentiation into macrophages, increasing the proportion of saturated fatty acids. These changes prompted us to investigate whether fatty acid accumulation in the extracellular milieu could affect the differentiation of bystander mononuclear phagocytes. An esterified fatty acid derivative, stearate, was the only fatty acid that significantly increased in macrophage supernatants, and there were higher levels when cells differentiated in the presence of Mycobacterium tuberculosis H37Rv or purified protein derivative (PPD). Exogenous stearic acid enhanced the expression of HLA-DR and CD64; there was also accumulation of IL-12, TNF-α, IL-6, MIP-1 α and ß and a reduction in MCP-1 and the bacterial load. These results suggested that during differentiation, a derivative of stearic acid, which promotes the process as well as the effector mechanisms of phagocytes against the mycobacterium, accumulates in the cell supernatants.
Asunto(s)
Leucocitos Mononucleares/efectos de los fármacos , Macrófagos/efectos de los fármacos , Mycobacterium tuberculosis/fisiología , Ácidos Esteáricos/farmacología , Tuberculosis/metabolismo , Adolescente , Adulto , Carga Bacteriana/efectos de los fármacos , Diferenciación Celular/efectos de los fármacos , Células Cultivadas , Citocinas/metabolismo , Antígenos HLA-DR/genética , Antígenos HLA-DR/metabolismo , Humanos , Leucocitos Mononucleares/fisiología , Macrófagos/fisiología , Persona de Mediana Edad , Fagocitosis/efectos de los fármacos , Receptores de IgG/genética , Receptores de IgG/metabolismo , Ácidos Esteáricos/química , Tuberculosis/tratamiento farmacológico , Adulto JovenRESUMEN
Introducción: la búsqueda de metabolitos de origen natural, con actividad biológica promisoria, particularmente la actividad insecticida, es un blanco interesante en las investigaciones sobre productos naturales. Objetivos: evaluar la bioactividad de extractos de diferente polaridad de semillas de Crotalaria pallida Aiton sobre el modelo biológico Drosophila melanogaster. Métodos: la bioactividad de los extractos de diferente polaridad de semillas secas de C. pallida se evalúo por ingestión en el modelo biológico; permitiendo purificar y determinar la estructura química del principio activo usando RMN. Resultados: la bioactividad expresada resultó de dos tipos; uno causó la inhibición de los estados larvarios, evidenciada con la disminución del número de pupas de los tratamientos con respecto a los controles no tratados, la relación dosis-respuesta permitió calcular una CI 50 de 156,47 ppm; el otro efecto inhibió el paso pupa-adulto, disminuyendo el número de adultos de los tratamientos frente a los controles, estableciéndose una CI50 de 7,95 ppm. Con el uso de diferentes ensayos de RMN se determinó el alcaloide usaramina como responsable de esta actividad biológica. Conclusiones : la bioactividad de los extractos de polaridad media/baja permitió el aislamiento de un metabolito con actividad insecticida promisoria, manifestada con la inhibición del normal desarrollo del ciclo de vida de D. melanogaster, el extracto no exhibe actividad sobre la oviposición, en el intervalo de concentraciones evaluado; a bajas concentraciones inhibe la eclosión de pupas y a altas concentraciones afecta el desarrollo de las larvas; actividades que se mantienen al probar el metabolito purificado.
Introduction: the search for metabolites of natural origin with promising biological -particularly insecticidal- activity, is an interesting target for research about natural products. Objectives: evaluate the bioactivity of extracts of varying polarity from Crotalaria pallida Aiton seeds using the Drosophila melanogaster biological model. Methods: an evaluation was conducted of the bioactivity of extracts of varying polarity from dry C. pallida seeds by ingestion into the biological model, with the purpose of purifying and determining the chemical structure of the active principle through NMR spectroscopy. Results: two types of bioactivity were expressed. One caused inhibition of larval stages, evidenced in a smaller number of pupae in treatment controls with respect to non-treatment controls. The dose-response relationship allowed estimation of a CI50 of 156.47 ppm. The other effect inhibited progress from pupa to adult, reducing the number of adults in the treatment vs. control groups, with an CI50 of 7.95 ppm. With the use of various NMR assays, it was determined that the alkaloid usaramine was responsible for this biological activity. Conclusions: the bioactivity of extracts of medium / low polarity permitted the isolation of a metabolite with promising insecticidal activity, manifested in the inhibition of the normal development of the life cycle of D. melanogaster. The extract does not show any activity on oviposition in the concentration range studied. At low concentrations it inhibits the eclosion of pupae, whereas at high concentrations it affects larval development. Both activities remained when the purified metabolite was tested.
RESUMEN
RESUMEN Objetivo Realizar la caracterización físico-química, microbiológica y parasitológica en biosólidos generados en la planta de tratamiento de agua residual San Fernando, Itagüí (Antioquia, Colombia). Métodos Se analizaron 12 muestras de biosólido tomadas cada mes, de enero a diciembre de 2010 a las cuales se les realizó un análisis físico-químico y microbiológico de acuerdo a lo establecido en la Norma Técnica Colombiana 5167. Para la determinación y viabilidad de huevos de helmintos se siguió el protocolo descrito en la Norma Oficial Mexicana 004 con modificaciones. Resultados En las doce muestras analizadas, se encontró una concentración de huevos de Ascarislumbricoides en un rango entre 4 a 22 huevos viables/2g ST. Salmonella estuvo presente en todas las muestras y las Enterobacterias estuvieron en una concentración mínima de 3000 UFC/g. La concentración de los metales pesados estuvo dentro de los valores recomendados por la norma. Los macro y micronutrientes cumplieron con los parámetros establecidos para los materiales orgánicos usados como abonos. No hubo asociación entre la precipitación y el contenido de patógenos en los biosólidos. Conclusión Los biosólidos generados en la planta San Fernando tienen un gran potencial para ser usados como abono orgánico, no obstante, deben ser sometidos a procesos de sanitización con el propósito de cumplir con los parámetros indicados en la Norma NTC 5167.
ABSTRACT Objective This study was aimed at evaluating pertinent physicochemical and microbiological (bacteria and parasites) parameters regarding the biosolids produced by the San Fernando wastewater treatment plant (WWTP) in Itagui, Antioquia, Colombia. Methods Twelve samples were collected and evaluated every month from January to December during 2010. The chemical, physical and microbiological tests followed the protocol described in Colombian technical guideline 5167. The protocol described in Mexican official Norm 004 (with some modifications) was used for identifying helminth ova and assessing their viability. Results All samples proved positive for Ascarislumbricoides, viable ova count ranging from 4 to 22 eggs/2gTS. Both Salmonella and Enterobacteriawere detected in all samples evaluated, the latter having 3,000 colony forming unit (CFU)/g minimum concentration. Biosolid sample values met the heavy metal concentration requirement established by national guidelines. There was no statistical association between rainfall and the pathogen's presence in the biosolids. Conclusion Our results suggested that the biosolids being produced by the San Fernando wastewater treatment plant (WWTP) could be used as organic fertilizer; however they should be treated/sanitized to meet the stipulations in Colombian technical guideline 5167.
Asunto(s)
Residuos Sólidos , Aguas Residuales , Colombia , Aguas Residuales/análisis , Aguas Residuales/microbiología , Aguas Residuales/parasitologíaRESUMEN
Introducción: el aceite esencial extraído de especies del género Eucalyptus, entre estas Eucalyptus tereticornis, ofrece un importante potencial para el control de hongos patógenos e insectos plaga y de importancia médica. Objetivo: evaluar la actividad anti-insecto (aguda, crónica y repelente) de una emulsión aceite/agua del aceite esencial de Eucalyptus tereticornis, en función de su composición química y fraccionamiento. Métodos: se prepararon emulsiones aceite/agua con aceite esencial previamente extraído, separado y caracterizado, y 2 de sus fracciones (A y B). Se evaluó su actividad anti-insecto mediante bioensayos con Drosophila melanogaster. Se determinó la fitotoxicidad foliar sobre plántulas de Phaseolus vulgaris. Resultados: el aceite completo mostró considerable actividad repelente e insecticida aguda que guarda relación con su composición química. La actividad insecticida aguda de las fracciones disminuyó de manera notable con respecto a la mostrada por el aceite completo, sin embargo, no se observaron efectos significativos sobre la actividad repelente. En ningún caso se presentó fitotoxicidad foliar sobre frijol. Conclusión: el aceite esencial de esta especie de eucalipto posee actividad anti-insecto considerable y podría formularse directamente como insecticida para el control de insectos de interés agrícola y médico.
Introduction: the essential oil extracted from Eucalyptus species, including Eucalyptus tereticornis, offers significant potential for control of fungal pathogens and insect pests and medical importance. Objectives: to evaluate the anti-insect activity (acute, chronic and repellent) of an oil/water emulsion of Eucalyptus tereticornis essential oil in terms of their chemical composition. Methods: oil/water emulsion was prepared with essential oil previously extracted, separated and characterized, and two of its fractions (A and B). Their anti-insect activity was evaluated through bioassays on Drosophila melanogaster. Foliar phytotoxicity was evaluated on seedlings of Phaseolus vulgaris. Results: the complete oil showed considerable acute repellent and insecticide activity related to its chemical composition. The acute insecticidal activity of the fractions decreased significantly if compare with that of the whole oil , but neither significant effects on the repellent activity nor phytotoxicity on the bean leaves were observed. Conclusions: the essential oil of this eucalyptus species has considerable anti-insect properties and could be formulated as an insecticide for the control of insects of agricultural and medical significance.
RESUMEN
ABSTRACT Objective This study was aimed at evaluating pertinent physicochemical and microbiological (bacteria and parasites) parameters regarding the biosolids produced by the San Fernando wastewater treatment plant (WWTP) in Itagui, Antioquia, Colombia. Methods Twelve samples were collected and evaluated every month from January to December during 2010. The chemical, physical and microbiological tests followed the protocol described in Colombian technical guideline 5167. The protocol described in Mexican official Norm 004 (with some modifications) was used for identifying helminth ova and assessing their viability. Results All samples proved positive for Ascarislumbricoides, viable ova count ranging from 4 to 22 eggs/2gTS. Both Salmonella and Enterobacteriawere detected in all samples evaluated, the latter having 3,000 colony forming unit (CFU)/g minimum concentration. Biosolid sample values met the heavy metal concentration requirement established by national guidelines. There was no statistical association between rainfall and the pathogen's presence in the biosolids. Conclusion Our results suggested that the biosolids being produced by the San Fernando wastewater treatment plant (WWTP) could be used as organic fertilizer; however they should be treated/sanitized to meet the stipulations in Colombian technical guideline 5167.
Asunto(s)
Residuos Sólidos , Aguas Residuales , Colombia , Aguas Residuales/análisis , Aguas Residuales/microbiología , Aguas Residuales/parasitologíaRESUMEN
Introducción: la piel o pericarpio del café que define su estado de madurez y calidad, es prácticamente lignocelulosa y no tiene ningún valor comercial, no obstante podría considerarse como una fuente importante de compuestos antioxidantes, sobre todo de antocianinas. Objetivos: comparar el potencial antioxidante del pericarpio del fruto de café en diferentes estados de maduración con respecto al contenido de antocianinas. Métodos: la piel del fruto del café liofilizada en cinco estados de maduración se sometió a extracciones sólido-líquido con metanol acidulado; se determinó el contenido de antocianinas por el método diferencial de pH y el potencial antioxidante se evaluó sobre la base de los métodos de DPPH (catión radical a-a-difenil-ß-picrilhidrazilo), ORAC (oxygen radical absorbance capacity) y FRAP (ferric reducing/antioxidant power). Resultados: la muestra de granos sobremaduros presentó un contenido de antocianinas 63,14 mg cianidin-3-glucósido/L. Este valor resulta de 2 a 18 veces mayor con respecto a los otros estados de maduración. No obstante, el estado "pintón" fue el que mostró una mayor capacidad antioxidante (³ 2-5 veces), en todos los métodos evaluados, lo cual sugiere una mayor presencia de otros componentes con actividad antioxidante durante esta etapa de maduración. Conclusiones: todos los extractos presentaron buena capacidad antioxidante, por lo tanto, aquellos frutos que no sean empleados para la industria de consumo de café pueden ser usados como fuente de antioxidantes, independiente de su estado de maduración.
Introduction: the maturity and quality of the coffee beans is defined by the color of its outer and often edible layer (pericarp), and this tissue is mainly lignocellulose without any commercial value; however it could be considered as a source of important antioxidant compounds, mostly anthocyanins. Objectives: to compare the antioxidant potential of the pericarp of the coffee bean at different states of maturation in terms of the content of anthocyanins. Methods: lyophilized and powered pericarp of Coffee arabica on different stages of maturation underwent solid-liquid extractions with acidulated methanol and total anthocyanins content was determined by pH differential method; the antioxidant potential was evaluated by using DPPH (radical cathion a-a-diphenyl-ß-picrylhydrazyl, ORAC (oxygen radical absorbance capacity) and FRAP (ferric reducing/antioxidant power) tests. Results: the sample from over-matured coffee beans showed a content of anthocyanins equal to 63.14 mg cyanidin-3-glucoside/L, which is 2 to 18 times higher than that of other maturation states. Nevertheless, the "pintón" stage demonstrated the highest antioxidant capacity (³ 2-5 times) in all the evaluated methods. This indicates the presence of additional components with antioxidant power during this maturation stage. Conclusions: all the extracts showed significant antioxidant capacity, therefore, the undesired fruits to coffee industry may be used as a source of antioxidant compounds regardless of their maturity stage.
RESUMEN
The objective of present paper was to determine the antifungal activity of the Eucalyptus tereticornis (Myrtaceae) essential oil and two fractions on the Fusarium oxysporum mushroom, a pathogen with clinical and agricultural significance. The total citronelal (44.8 percent) and geraniol (9.78 percent) essential oil had a fungicidal effect at a 3 g/L concentration and a fungicidal activity at small concentrations. The A and B fractions composed most of p-mentane-3,8-diol (18.95 percent) and geraniol acetate (24.34 percent), respectively were more active than the total extract. The observations at microscopic level showed damages and changes in hyphae and chlamydospores, as well as a decrease in the number of conidia. The observed fungicidal activity and the morphologic damages were dependent on the concentration.
El objetivo de este trabajo fue determinar la actividad antifúngica del aceite esencial de Eucalyptus tereticornis (Myrtaceae) y 2 fracciones sobre el hongo Fusarium oxysporum, patógeno de importancia tanto clínica como agrícola. El aceite esencial total, compuesto principalmente por citronelal (44,8 por ciento), citronelol (9,78 por ciento) presentó un efecto fungicida a una concentración de 3 g/L y actividad fungistática a concentraciones menores. La fracciones A y B compuestas en su mayoría por p-mentano-3,8-diol (18,95 por ciento) y acetato de citronelol (24,34 por ciento) respectivamente fueron más activas que el extracto total. Las observaciones a nivel microscópico mostraron daños y cambios en hifas y clamidosporas, así como disminución en el número de conidias. La actividad fungistática observada y los daños morfológicos fueron dependientes de la concentración.