1,721,113 research outputs found
Purification and partial characterization of elastase activity from rat alveolar and peritoneal macrophages
No full textMacrophage elastase was purified from conditioned media from alveolar and thioglycollate-elicited peritoneal macrophages. The enzyme was purified to apparent electrophoretic homogeneity by preparative isoelectric focusing after a purification step consisting of low ionic strength dialysis and sequential batch fractionation on DEAE-Sephadex A-50. The proteinase activities isolated from alveolar and peritoneal macrophages showed the same physical and biochemical properties. This fact suggests that the same enzyme activity is present in rat macrophages of two different anatomical sites. The molecular weight and isoelectric point of the enzyme were estimated to be 22,500 and 8.3, respectively. The enzyme, characterized as a metallo proteinase, had elastolytic activity, as well as activity toward Suc-(Ala)3-NA. It is inhibited by o-phenanthroline, chicken ovoinhibitor, and EDTA, but not by phenylmethylsulfonyl fluoride or soybean trypsin inhibitor. The macrophage enzyme possesses biochemical and biophysical properties different from the rat pancreatic and granulocyte elastases (which are serine proteinases), and from the metallo proteinase with elastolytic activity isolated from rat platelets
Isolation and partial characterization of rat elastolytic enzymes from various cells and tissues
No full textDifferent elastolytic enzymes were isolated from rat aorta and platelets, as well as from granulocyte and pancreatic extracts. The active fractions were purified to electrophoretic apparent homogeneity by precipitation with ammonium sulfate, sequential batch fractionation on DEAE-Sephadex A-50, and finally by isoelectric focusing (IF) on Sephadex G-75 Superfine. The molecular weight and the isoelectric point of the isolated enzymes were estimated by sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) and by analytical IF, respectively. All the enzymes have elastolytic activity as well as activity toward Suc-(Ala)3-NA. The inhibition profile of the different isolated enzymes toward various inhibitors indicates that aortic, pancreatic, and granulocyte enzymes all belong to the group of serine proteinases, unlike the platelet elastase which is a metalloproteinase
Detection of elastase activity with a zymogram method after isoelectric focusing in polyacrylamide gel
No full textA zymogram method for detecting elastase activity following isoelectric focusing in polyacrylamide gel is described. After enzyme activity has been visualized, the gel itself is available for protein staining and for analysis in sodium dodecyl sulfate-polyacrylamide gel electrophoresis in second dimension. The zymogram method is suitable for detecting microgram amounts of elastase and has one step only. It can be used with the purified enzyme as well as with crude extracts of tissue containing elastases showing activity toward succinyl-(Ala)3-p-nitroanilide. By this method a major component of elastase in both porcine and rat pancreas was detected. In addition, two forms of elastase with isoelectric points of 8.2 and 8.8, respectively, were identified in rat leukocyte extracts
Correlation between biochemical and morphological repair in rabbit lungs after elastase injury
No full textThe structural and elastic properties of the lung depend primarily on the concentration in connective tissue of proteins, collagen and elastin. The relationships between biochemical and morphological aspects of pulmonary damage were studied in 50 male New Zealand rabbit's, randomly divided into two groups of 25 animals, instilled with 245 IU of pig pancreatic elastase solution or physiological saline. Animals were killed at 8, 15, 30, 60 and 90 days after treatment. The changes in pulmonary morphology and in collagen and elastin content have been studied chemically and by light microscopy. The left lungs were used for biochemical determinations of insoluble elastin and total, insoluble and salt-extractable collagens. Morphometric measurements (mean linear intercepts or Lm) and histologic evaluations were performed on right lungs. Three months after instillation, the elastase treated lungs showed a minimal degree of alveolar wall rupture, and no significant difference in Lm from the control group. Quantitative restoration of normal biochemical parameters preceded connective tissue repair. Statistical evaluations showed significative correlations among Lm, total collagen (p < 0.01) and insoluble elastin/dry weight (p < 0.01). No significant differences between Lm and insoluble elastin, salt-extractable collagen or total collagen/dry weight were found. © 1980 Springer-Verlag
Isolamento di attività elastolitiche nell'aorta, nel pancreas, nei granulociti e nelle piastrine di ratto. Confronto tra alcune delle loro proprietà chimico -fisiche ed enzimatiche
No full textIsolamento di attività elastolitiche nell'aorta, nel pancreas, nei granulociti e nelle piastrine di ratto.Confronto tra alcune delle loro proprietà chimico-fisiche ed enzimatiche
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Pulmonary changes induced in rabbits by long-term exposure to n-hexane
No full textNew Zealand male rabbits were exposed in an inhalation chamber to 3,000 ppm of n-hexane 8 h/day, 5 days/week for 24 weeks, resulting in a total of 120 exposures. After a further 120 days in clean air, morphologic examination of lungs by light and transmission electron microscopy (TEM) revealed three important exposure-related lesions. These consisted of air space enlargement centered on respiratory bronchioles and alveolar ducts (centriacinar emphysema), scattered foci of pulmonary fibrosis, and papillary tumors of non-ciliated bronchiolar epithelial cells. At TEM examination these tumors were rather homogeneous and were found to consist of cells showing both small and large electron dense ovoid granules in their cytoplasmic matrix, a feature that has been described for Clara cells. The implications of these findings are discussed
Response of lung enzyme activities in rabbits following short-term exposure to n-hexane: correlation between morphological and biochemical changes
No full textThe activity of lactate dehydrogenase, beta-glucuronidase, glucose-6-phosphate dehydrogenase, acid and alkaline phosphatase was studied in lung homogenate from New Zealand rabbits exposed to 3000 p.p.m. of n-hexane 8 h per day for 8 days of filtered air. In hydrocarbon-treated animals all enzymes examined, except alkaline phosphatase, were markedly increased. The biochemical changes correlated well with the morphological changes and the results of cytological evaluation of bronchopulmonary lavage. It is suggested that high values in lung lysosomal enzymes from treated rabbits reflect the acute inflammation whilst the increase in lung glucose-6-phosphate dehydrogenase may depend upon reparative process subsequent to n-hexane-induced lung damage
Ultrastructural abnormalities in respiratory cilia and sperm tails in a patient with Kartagener's syndrome
No full textUltrastructural abnormalities of spermatozoa and respiratory cilia have been reported in a male patient with Kartagener's syndrome and infertility. In this patient both respiratory cilia and sperm tails showed defects in radial spokes and dynein arms. Such defects are heretofore undescribed in the same subject with immotile cilia syndrome. Absence of both inner and outer dynein arms and absence of the inner dynein arms only were detected in spermatozoa and in respiratory tract cilia, respectively. Moreover, total absence of axoneme was seen in several sperm tails from this patient. The possibility that the features described are of genetic origin is discussed
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