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detection of β-Glucuronidase Activity within Actinomadura madurae Grains of Human Actinomycetoma

Advances in Microbiology 2014年第6期4卷 317-323页

作者： Alejandro Palma-Ramos Samantha Reyes-Mayén laura Estela Castrillón-Rivera Silvia Elena Fernández-López Carmen Padilla-desgarennes María Elisa Vega-Memije Roberto Arenas-Guzmán María Elisa drago-Serrano Teresita Sainz-Espunes department of Biological Systems Universidad Autónoma Metropolitana-XochimilcoMexico CityMéxico dermatology center“dr.ladislao de la pascua” Mycology ServiceMexico City Health SecretaryMexico CityMéxico General Hospital“Manuel Gea González” Dermatology ServiceMexico City Health SecretaryMexico CityMéxico General Hospital“Manuel Gea González” Mycology ServiceMexico City Health SecretaryMexico CityMéxico

Actinomycetoma syndrome by Actinomadura (A.) madurae is characterized by a subcutaneous chronic lesion that affects fascia, muscle and bone. A. madurae produces colonies that form grains of less than 1 mm in diameter. Grains are surrounded and infiltrated by neutrophils involved in the grain disruption by enzymes like β-glucuronidase released after the neutrophil degranulation. The aim of this work was to evaluate the polysaccharide degradation of grains treated with β-glucuronidase and to detect the presence and activity of β-glucuronidase within the A. madurae grains. Actinomadura madura grains from patients infected were processed to quantify the total content of polysaccharide with the phenol-sulfuric acid reaction. Grains were treated with β-glucuronidase at different conditions to evaluate the optimal polysaccharide degradation. Grains were analyzed to detect the enzyme by using anti-human β-glucuronidase antibody while enzymatic activity was assessed by evaluating the release of reduced sugars and by in situ enzymatic activity. Optimal degradation of polysaccharide in the grains treated with β-glucuronidase was found with 300 units/ml of enzyme and 24 hr of incubation at 37°C. Presence and activity of β-glucuronidase enzyme within the grains were detected. Results suggested that β-glucuronidase present within A. madurae grain resulted from degranulated neutrophils surrounding and/or infiltrated within the grain.

关键词： Actinomycetoma A. madurae β-Glucuronidase

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