Asunto(s)
Tornillos Óseos , Falanges de los Dedos de la Mano/lesiones , Fijación Interna de Fracturas/métodos , Fracturas Óseas/cirugía , Adolescente , Adulto , Anciano , Femenino , Fijación Interna de Fracturas/instrumentación , Fracturas Óseas/diagnóstico por imagen , Humanos , Masculino , Persona de Mediana Edad , Radiografía , Adulto JovenRESUMEN
We devised an improved technique of thin layer chromatography, which permitted the high resolution of urinary neutral oligosaccharides and the qualitative determination of blood group related oligosaccharides as well as oligosaccharides pathologically secreted in lysosomal storage diseases. This procedure can be used inscreening for disorders associated with abnormal excretion of oligosaccharides, as well as in the purification of oligosaccharides.
Asunto(s)
Antígenos de Grupos Sanguíneos/inmunología , Errores Innatos del Metabolismo/prevención & control , Oligosacáridos/orina , Sistema del Grupo Sanguíneo ABO/inmunología , Adolescente , Adulto , Antígenos/orina , Secuencia de Carbohidratos , Cromatografía en Capa Delgada/métodos , Humanos , Antígenos del Grupo Sanguíneo de Lewis/inmunología , Tamizaje Masivo , Errores Innatos del Metabolismo/orina , Datos de Secuencia Molecular , Oligosacáridos/química , Oligosacáridos/inmunologíaRESUMEN
We describe the partial characterization and some properties of leucocyte alpha-glucosidase towards disaccharides with the alpha-1,4 (maltose) and alpha-1,6-glucosidic linkage (isomaltose) and tetrasaccharides with the alpha-1,4 (maltotetraose) and alpha-1,6-glucosidic linkage (tetrasaccharide, Glc alpha 1----6Glc alpha 1----4Glc alpha 1----4Glc, which was isolated from the urine of a patient with glycogenosis type II). Leucocyte alpha-glucosidase showed optimal activity towards all four oligosaccharides under two conditions, acidic (pH 4.0-4.5) and neutral (pH 6.0-6.5) regions. Our comparative studies on enzyme kinetics showed that leucocyte alpha-glucosidase was able to hydrolyze both the 1----4 isomers and the 1---6 isomers at acidic and neutral pH. Acid alpha-glucosidase could hydrolyze maltose about 10 times faster than isomaltose, and maltotetraose about 5 times faster than tetrasaccharide isolated from urine. In leucocytes of the patient with late onset glycogenosis type II, acid alpha-glucosidase activities towards maltose, isomaltose, maltotetraose and tetrasaccharide isolated from urine showed 75.3%, 67.4%, 76.5% and 41.4% of normal control values, respectively. Neutral alpha-glucosidase activities towards these four oligosaccharides were normal. Tetrasaccharide with alpha-1,6-glucosidic linkage might be accumulated by the impaired hydrolysis in the circulation as well as the leakage of undegraded glycogen to the circulation from the affected muscle.
Asunto(s)
Enfermedad del Almacenamiento de Glucógeno Tipo II/metabolismo , Oligosacáridos/metabolismo , alfa-Glucosidasas/sangre , Adulto , Conformación de Carbohidratos , Secuencia de Carbohidratos , Humanos , Concentración de Iones de Hidrógeno , Cinética , Leucocitos/enzimología , Masculino , Datos de Secuencia Molecular , Oligosacáridos/químicaRESUMEN
We describe the biochemical characterization of lymphocyte alpha-glucosidase in a 23-year-old man with intermediate clinical features between the childhood and adult forms of glycogenosis type II (Pompe's disease). Acid alpha-glucosidase activity was markedly reduced, but immunologic cross-reactive material against human liver acid alpha-glucosidase protein could be detected, and its amount was normal. In this patient, the disorder was induced by the catalytically inactive enzyme with a normal amount of enzyme protein.
Asunto(s)
Enfermedad del Almacenamiento de Glucógeno Tipo II/enzimología , Linfocitos/enzimología , alfa-Glucosidasas/metabolismo , Adulto , Humanos , Técnicas para Inmunoenzimas , Técnicas Inmunológicas , Linfocitos/metabolismo , Masculino , Coloración y EtiquetadoRESUMEN
We describe partial characterization and properties of leucocyte alpha-glucosidase from a patient with clinical features intermediate of juvenile and adult onset forms of glycogenosis type II. Acid and neutral alpha-glucosidase activities toward 4-methylumbelliferyl glucopyranoside as substrate were studied in total leucocytes, and separately in lymphocytes and granulocytes. Lymphocytes, which showed markedly reduced activities of acid alpha-glucosidase in the patient, are the most reliable peripheral blood cells for the diagnosis of glycogenosis type II. Moreover, the ratio of acid/neutral alpha-glucosidase activities, especially in lymphocytes, is a useful parameter for the diagnosis. In lymphocytes, the Km values of both acid and neutral alpha-glucosidases were essentially the same between the patient and normal controls; the Vmax value of acid alpha-glucosidase from the patient was markedly reduced, and the Vmax value of neutral alpha-glucosidase from the patient was reduced by 36% as compared with that from normal controls. Heat-inactivation experiments revealed that acid alpha-glucosidase activities of lymphocytes were relatively heat-stable, while both acid and neutral alpha-glucosidases of granulocytes were heat-labile. No differences in these properties, however, could be detected between the patient and normal controls.
Asunto(s)
Enfermedad del Almacenamiento de Glucógeno Tipo II/enzimología , Leucocitos/enzimología , alfa-Glucosidasas/metabolismo , Adulto , Calor , Humanos , MasculinoRESUMEN
Neutral oligosaccharides were isolated from urine of an adult patient with glycogen storage disease type II, a deficiency of lysosomal acid alpha-glucosidase, by chromatography on columns of activated charcoal, Dowex 50 X 2 and Dowex 1 X 2. Total neutral oligosaccharides in the urine of the patient were increased about 5-fold as compared with those in normal controls. The most accumulated oligosaccharide was separated by Bio-Gel P-2 column chromatography, and finally purified by paper chromatography. Based on various studies, including carbohydrate analysis, chemical ionization mass spectrometry, fast atom bombardment mass spectrometry, degradation by glucoamylase and isopullulanase, and methylation analysis, the structure of this oligosaccharide was deduced to be Glc alpha 1----6Glc alpha 1----4Glc alpha 1----4Glc. This oligosaccharide appears to be accumulated in urine of the patient with acid alpha-glucosidase deficiency as an end product of the hydrolysis of glycogen.