Replacement of α-galactosidase A in Fabry disease: effect on fibroblast cultures compared with biopsied tissues of treated patients

The function and intracellular delivery of enzyme therapeutics for Fabry disease were studied in cultured fibroblasts and in the biopsied tissues of two male patients to show diversity of affected cells in response to treatment. In the mutant fibroblasts cultures, the final cellular level of endocytosed recombinant α-galactosidases A (agalsidases, FabrazymeTM, and ReplagalTM) exceeded, by several fold, the amount in control fibroblasts and led to efficient direct intra-lysosomal hydrolysis of (3H)Gb3Cer. In contrast, in the samples from the heart and some other tissues biopsied after several months of enzyme replacement therapy (ERT) with FabrazymeTM, only the endothelial cells were free of storage. Persistent Gb3Cer storage was found in cardiocytes (accompanied by increase of lipopigment), smooth muscle cells, fibroblasts, sweat glands, and skeletal muscle. Immunohistochemistry of cardiocytes demonstrated, for the first time, the presence of a considerable amount of the active enzyme in intimate contact with the storage compartment. Factors responsible for the limited ERT effectiveness are discussed, namely post-mitotic status of storage cells preventing their replacement by enzyme supplied precursors, modification of the lysosomal system by longstanding storage, and possible relative lack of Sap B. These observations support the strategy of early treatment for prevention of lysosomal storage

Hydrothermal decomposition of various crystalline celluloses as treated by semi-flow hot-compressed water

Various types of crystalline celluloses I, II, IIII, IIIII, IVI and IVII that have been adjusted for their degree of polymerization were treated by semi-flow hot-compressed water (HCW) at 230–270 °C/10 MPa/15 min to study their hydrothermal decomposition. The treatments resulted in either partial or complete decomposition of the celluloses and the decomposed products were primarily recovered as hydrolyzed, dehydrated and fragmented ones as well as organic acids in the water-soluble (WS) portions. Their results of hydrothermal decomposition and its kinetics revealed that the celluloses decomposition is dependent on the types of crystalline celluloses as well as temperature of the HCW treatment. The outcome from the WS portions at 270 °C/10 MPa/15 min showed that the degree of difficulty for decomposition is lower in group II (cell II, cell IIIII, cell IVII) than group I (cell I, cell IIII, cell IVI), indicating that group II is less resistant to decomposition by HCW treatment. Therefore, the decomposition behaviors of the cellulose are due to the inherent differences in the crystalline structures