Lysosomes in Type Ii Glycogenosis

Short papers submitted expressly for this section, reporting original and significant findings of immediate interest and judged to be acceptable without major revision, will be published within approximately three months. See inside back cover for details. Type II glycogenosis is a fatal disease of infants (1) characterized by increased concentration of tissue glycogen and deficient activity of lysosomal acid a glucosidase (a glucosidase) (2, 3). In hepatic electron photomicrographs, Baudhuin et al. observed vacuoles filled with glycogen which they interpreted as abnormal lysosomes, i.e., as the morphological correlate of the biochemical deficiency (4). They injected intramuscularly an extract of the fungus Aspergillus niger which Pazur and Ando had shown to contain enzymes with a amylase, amyloglucosidase, and maltase activity (5). The treatment did not produce appreciable changes of the abnormal lysosomes. In our patient such changes were observed after prolonged, intravenous administrations of an identically prepared fungal extract. PATIENT AND METHODS An alcohol precipitate of Aspergillus niger served as the starting material. 20 g of the powder were extracted with 600 cc of distilled water filtered through a Buchner funnel, lyophilized, dissolved in 60 cc of 0.9% NaCI, sterilized with passages through Milli-pore filters, and kept in the refrigerator until used. The extract had a protein concentration of 25 mg/cc. It catalyzed the essentially complete conversion of glycogen to glucose at the rate of 22 moles of glucose formed per milligram of protein per minute at pH 4 and 37 0 C. The liver was biopsied with the Menghini needle (6). The concentration of glycogen and the activity of a glucosidase in the biopsy specimens were determined according to standard procedures (2) and as reported previously (7). For ultrastructural examination the tissue specimens were immersed immediately after the biopsy in cold (0-5 0 C), buffered 3% glutaraldehyde (pH 7.3; 0.1 M phosphate buffer), cut into small pieces, kept in the refrigerator for 24 hr, and washed overnight in cold buffer. Postfixation was in cold (0-5°C), buf-fered 1% OsO4 (pH 7.3; 0.1 M phosphate buffer) for 90 min. After dehydration in graded ethanol, the tissue was embedded in Epon 812. Thin sections were cut with glass knives on a Reichert OMU2 ultramicrotome, were stained on uncoated grids with uranyl acetate (5 min) followed by lead citrate (5 min), and were examined in a Zeiss EM 9 electron microscope. The patient was a 3 month old negro girl with the typical features …