Histological and Biochemical Study on Acid Glycosaminoglycans from a Case of Benign Localized Pleural Mesothelioma

Abstract

The fraction of acid glycosaminoglycans was isolated from the tissue of a benign localized pleural mesothelioma and was analyzed by histological and biochemical means. The weakly PAS-positive substances in the interstitium was digested almost completely with hyaluronidase “Amano”. Disappearance of alcianophilia of the specimen after methylation and the results of staining with alcian blue in graded concentrations of MgCl² indicated the presence of a large amount of hyaluronic acid in the tissue. In the next experiment the tissue from the above pleural mesothelioma was digested exhaustively with pronase. The crude glycosaminoglycan was isolated from the resulting digest by means of fractionation with ethanol and precipitated with cetylpyridinium chloride. The crude glycosaminoglycan was analyzed by column chromatography using Dowex-1. Elution was effected with graded concentratons of NaCl. The 0.5 M-NaCl eluate from the column showed the same mobility as the hyaluronic acid from the human umbilical cord on the membrane of cellulose acetate. Only glucosamine was found in the acid hydrolysate of the 0.5 M-NaCl eluate. It was digested with hyaluronidase “Amano” just as the hyaluronic acid from human umbilical cord. In addition the hyaluronic acid from meothelioma (0.5 M-NaCl eluate) and that from the human umbilical cord had the same infrared spectrum pattern. However the former migrated more slowly than the latter in electrophoresis on paper. The mobility of the former remained unchanged even after repeated digestion with pronase. In gel filtration chromatography using Sepharose 4B it was found that the hyaluronic acid from mesothelioma was eluted prior to the hyaluronic acid from human umbilical cord.
Thus it was suggested that hyaluronic acid from pleural mesothelioma had a larger molecular size than that from the human umbilical cord.