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Humoral aspects are being discussed in development of arthrosis. Punctuate of 144 traumatized knee joints has been investigated. Proteolytic enzymes elastase and cathepsin D were assessed. Furthermore, protease inhibitors alpha 1-antitrypsin and alpha 2-macroglobulin were determined. Activity of chondrolytic enzymes elastase and cathepsin D clearly correlates with the clinical severity of injuries. At the same time, primary inhibitory mechanisms for compensation seem to be non-sufficient, however.

A 59-year-old female patient who developed fever, headache, arthralgia and painful, raised red cutaneous plaques located predominantly on the upper trunk is presented. On the basis of the clinical and histological findings acute febrile neutrophilic dermatosis (Sweet's syndrome) was diagnosed. In affected skin we applied immunohistochemical methods to examine the topographical organization of neutrophil-associated proteinase/proteinase inhibitor systems, i.e. the PMN-elastase/alpha 1-proteinase inhibitor and the cathepsin G/alpha 1-antichymotrypsin systems. Cathepsin G, PMN-elastase and alpha 1-antichymotrypsin were found in association with the polymorphonuclear infiltrating cells, whereas alpha 1-proteinase inhibitor was localized exclusively in the subepidermal oedema. In conclusion, our immunohistological findings support the view that free PMN-elastase is involved in the pathogenesis of cutaneous lesions in Sweet's syndrome.

Pulmonary emphysema is thought to be due to an elastase-antielastase imbalance which leads to the destruction of alveolar walls. It is generally agreed that cigarette smoking is the major cause of acquired emphysema although many smokers fail to develop overt disease. Cigarette smoke inactivates alpha 1-proteinase inhibitor (alpha 1PI) which is believed to be the major antielastase in the lower respiratory tract. There is, however, controversy regarding the activity of alpha 1PI in smokers compared with nonsmokers. We, therefore, investigated the trypsin inhibitory capacity (TIC), the pancreatic elastase inhibitory capacity (PEIC), and the amount of immunoreactive alpha 1PI in serum and bronchoalveolar lavage fluid (BALF) of 24 individuals (15 smokers, 5 former smokers, 4 non-smokers) with clinical signs of emphysema and 32 persons (15 smokers, 6 former smokers, 11 non-smokers) without emphysema. Pancreatic elastase is known to be inhibited only by non-oxidised alpha 1PI whereas trypsin is inhibited by both native and oxidised alpha 1PI. Serum values of TIC/alpha 1PI and PEIC/alpha 1PI did not differ between the groups of subjects with emphysema (TIC/alpha 1PI: 0.71 +/- 0.12; PEIC/alpha 1PI: 0.44 +/- 0.06) and without emphysema (TIC/alpha 1PI: 0.68 +/- 0.13; PEIC/alpha 1PI: 0.41 +/- 0.07). Both serum-inhibitory capacities were found to be smaller than unit (= 1) indicating inactivation of alpha 1PI. BALF values of TIC/alpha 1PI showed a wide variation. TIC/alpha 1PI was greater than unity in 84% of subjects with emphysema (2.06 +/- 1.28) compared to 61% without emphysema (1.14 +/- 1.07) providing evidence for the presence of additional inhibitor(s) in those lavage fluids.(ABSTRACT TRUNCATED AT 250 WORDS)

In a pilot study paraffin-embedded sections of open skin wounds (stab and slash wounds, lacerations) were investigated to determine the presence of a vital reaction. Granulocytes were detected by naphthol AS-D chloroacetate esterase, the enzyme "lysozyme", and eight proteinase inhibitors by the indirect immunoperoxidase method. The tissue specimens were taken from consecutive autopsy material. The survival time could be determined in 14 cases (10-165 min) and was unknown in 12 other cases of sudden death due to injury of the major vessels or heart. The controls were cases with injuries inflicted after and cases of sudden death due to massive blunt trauma served death. In vital injuries, accumulations of proteinase inhibitors, particularly alpha-2-macroglobulin and alpha-1-antichymotrypsin, were demonstrable in the corium parallel to the wound surface. In comparison, the reaction of proteinase inhibitors that neutralize only enzymes participating in blood coagulation or complement activation (C1-esterase inhibitor and protein C) was absent or weak. Protein accumulation was observed only sporadically in cases of sudden death and never in cases with wounds inflicted after death. No relationship could be established between semiquantitatively estimated staining and survival time. Granulocytes and lysozyme were first observed in the corium after a survival time of more than 60 min.

The complete amino-acid sequence of the alpha A- and the beta-chains of the major component (HbA) and the alpha D- and the beta-chains of the minor component (HbD) of Andean Condor (Vultur gryphus) is presented. The minor component with the alpha D-chains is present in smaller amounts (17%) than in other birds (25%). The comparison with the corresponding chains of Greylag Goose (Anser anser) shows 17 different amino acids (17 nucleotides, only one-point mutations) in the alpha A-chains and 8 (8 nucleotides) in the beta-chains. The alpha D-chains differ from those of the pheasant (Phasanius cholchicus cholchicus) in 24 amino acids (27 nucl., 3 two-point mutations). Seven alpha 1 beta 1-, one alpha 1 beta 2-, three alpha 1 alpha 1-contacts and one beta 1 beta 1-contact are exchanged. The systematy of Cathartiformes, Ciconiiformes and Phoenicopteriformes is discussed, based on the amino-acid exchanges of all known adult hemoglobins of birds.

Thyroid tissues containing lymphoid secondary follicles were studied immunohistochemically for the presence of acid cysteine proteinase inhibitor (ACPI). Reticulum cells in the lymphoid secondary follicles, mainly dendritic reticulum cells, exhibited a positive reaction. In addition, in 2 cases a positive reaction could be detected in some of the thyroid epithelial cells, adjacent to the lymphoid secondary follicles. The possible function of ACPI generally and its role in the function of those cells containing it, is discussed.

2 low-molecular cysteine-proteinase inhibitors were purified from human tonsillar tissues: an acid cysteine proteinase inhibitor (ACPI), and a neutral cysteine proteinase inhibitor (NCPI). Their biochemical and immunological characteristics appeared to be identical to those inhibitors which we have identified in other human tissues in previous studies (epidermis and spleen). An immunohistological analysis revealed in tonsillar squamous epithelium a strong and consistent immunoreactivity for both inhibitors. In the tonsillar lymphatic tissue, ACPI-immunoreactivity appeared to be a characteristic mainly of dendritic reticulum cells whereas a prominent NCPI-immunoreactivity was confined mostly for the histiocytic reticulum cells in lymphoid secondary follicles. We also compared the distribution of the 2 immunoreactive inhibitors with the immunohistology as revealed by antisera raised against keratin and muramidase. Limitations of the immunohistochemistry of cysteine proteinase inhibitors are discussed in the light of an extensive trial of various fixation procedures. We deem that proposal of Barrett (1984) for the nomenclature of cysteine proteinase inhibitors is appropriate. According to it the ACPI is the cystatin A, and the NCPI is the cystatin B.

Using a specific antiserum, it was possible to demonstrate (with the Ouchterlony-test, immunofluorescence and peroxidase-antiperoxidase reaction) that the epidermal thiolprotease inhibitor was present in all the squamous epithelia tested by us, e.g. oesophagus, vagina and portio. Immunological methods further showed that epidermoid carcinoma of the skin, squamoepithelial carcinomas of the portion and oesophagus, and squamoepithelial carcinoma of the lung contained an immunoreactive protein reminiscent of the epidermal protease. The immunoreactive protein typical of squamous epithelium was also visible in part of the anaplastic lung carcinomas, which means that the determination of the inhibitor may be of significance in the classification of lung tumors.

In this work, we have studied the occurrence of acid and neutral cysteine proteinase inhibitors in 3 histologically different lung cancers. The acid cysteine proteinase inhibitor ( ACPI ) seems to be associated to squamous differentiation as our previous observations have shown. As a new observation we could show that the neutral cysteine proteinase inhibitor ( NCPI ) was localized in alveolar macrophages.

Investigation of vitality was done in paraffin-embedded tissue sections of open skin wounds by demonstration of a neutrophilic reaction marking the cells by naphthol AS-D chloroacetate esterase, by demonstration of enzyme activity by application of lysozyme antibodies and by demonstration of concentration of proteinase inhibitors by application of inhibitor antibodies using the indirect immunohistochemical peroxidase method. Survival time of the wounds could be determined in 10 cases (15-165 mins) and was unknown in 9 other cases of sudden death due to injury of the major vessels or heart. Postmortally inflicted injuries and cases of sudden death due to massive blunt trauma served as controls. In vital injuries, accumulation of proteinase inhibitors, particularly alpha-1-antichymotrypsin and alpha-2-macroglobulin, were demonstrable in the corium parallel to the wound surface. Protein accumulation was observed only sporadically in cases of sudden death and never in cases with postmortally inflicted wounds. Granulocytes and lysozyme were first observed in the corium after a survival time of more than 60 minutes.

Two apparent endothelial cell growth factors were isolated and characterized from serum free cell culture medium of hepatoma cells by McKeehan et al. The factors were identified as proteinase inhibitors with known primary structure. They are the pancreatic secretory trypsin inhibitor (HPSTI) and the double headed inhibitor HI-30, the inhibitory active part of the inter-alpha-trypsin inhibitor complex. We were able to isolate acid resistant inhibitory active material from serum free culture medium of 4 out of 11 tumor cell lines, which we have analyzed. The cell lines were not derived from liver cells. The inhibitory active material was identified as the inhibitor HI-30 by N-terminal amino acid sequence analysis. The results indicate that HI-30 is a real growth factor, since it is expressed as well in tumor cells which are not derived from liver cells.

The significance of proteinase inhibitors in lacrimal fluid is discussed. While an increase is diagnostically important, localized deficiencies may lead to serious symptoms. To our knowledge this is the first time that local antiproteinase treatment has been described.