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

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 

机构地区：[1]Department of Biological Systems,Universidad Autónoma Metropolitana-Xochimilco,Mexico City,México [2]Dermatology Center“Dr.Ladislao de la Pascua”,Mycology Service,Mexico City Health Secretary,Mexico City,México [3]General Hospital“Manuel Gea González”,Dermatology Service,Mexico City Health Secretary,Mexico City,México [4]General Hospital“Manuel Gea González”,Mycology Service,Mexico City Health Secretary,Mexico City,México

出　　处：《Advances in Microbiology》2014年第6期317-323,共7页微生物学（英文）

摘　　要：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.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 

分 类 号：R73[医药卫生—肿瘤]