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131


Separation of equine bronchopulmonary lavage cells by density gradient centrifugation and expression of procoagulant activity in unpurified cells and cell subpopulations

Grunig, G; Hulliger, C; Hermann, M; Winder, C; von Fellenberg, R
Bronchopulmonary lavage was performed in 10 healthy horses and in 39 horses with chronic pulmonary disease. The predominant cell types were macrophages in healthy horses and neutrophils in severely diseased horses. Procoagulant activity (PCA) was detected in all 32 cell-free supernatants examined and in all 49 unpurified cell suspensions. Cells were separated by centrifugation on discontinuous gradients prepared either with Percoll or with Metrizamide. Macrophages were enriched in subpopulations of low density. Neutrophils could not be purified by density gradient centrifugation using either gradient medium. PCAs of cell subpopulations were plotted against their respective macrophage, neutrophil, and lymphocyte content. PCA was positively correlated with macrophage content (P less than 0.001) and negatively correlated with neutrophil (P less than 0.02) and lymphocyte (P less than 0.001) content. Therefore, PCA of equine lung cells most likely originates from macrophages as shown in other species. The density shift of lung neutrophils requires further investigation
PMID: 2382054
ISSN: 0034-5288
CID: 117374

Fibrin/fibrinogen in lungs and respiratory secretions of horses with chronic pulmonary disease

Winder, N C; Grunig, G; Hermann, M; von Fellenberg, R
The concentration of soluble fibrinogen derivatives (SFD) and protease and procoagulant activities were determined in cell-free supernatants of equine respiratory secretions obtained from horses with chronic pulmonary disease. The concentration of neutrophils was estimated from direct smears of the secretions. Lung specimens and smears of the secretions were evaluated for the presence of fibrin or fibrinogen by use of immunohistochemical methods. Thirty-five of 80 specimens tested contained SFD. Respiratory secretions from horses with moderate or severe chronic pulmonary disease contained SFD more frequently than did secretions from mildly affected horses (P less than 0.05). Respiratory secretions with vast numbers of neutrophils had significantly (P less than 0.05) higher SFD concentrations than respiratory secretions with fewer neutrophils. Protease and procoagulant activities in respiratory secretion specimens were positively correlated with neutrophil content, clinical diagnosis, and SFD concentration. Immunohistochemically, macrophages that stained for fibrin or fibrinogen were observed in direct smears of respiratory secretions from horses with moderate and severe chronic small airway disease, but not in smears from mildly affected horses. Fibrin or fibrinogen was detected in a few thickened alveolar septa from 10 horses with moderate or severe chronic small airway disease, but not in lungs from horses with mild or no evidence of chronic small airway disease. Fibrin or fibrinogen was detected in alveolar septa, granulomas, and on alveolar macrophages in lungs of all horses with chronic granulomatous and chronic bronchointerstitial pneumonia. The presence of SFD in equine respiratory secretions may be an indicator of pulmonary inflammation
PMID: 2114807
ISSN: 0002-9645
CID: 117372

Comparison of bronchoalveolar lavage and respiratory secretion cytology in horses with clinically diagnosed chronic pulmonary disease

Winder, N C; Hermann, M; Grunig, G; Hulliger, C; Von Fellenberg, R
Thirty-nine horses and 3 ponies underwent a thorough respiratory examination and were grouped as follows: healthy (4 horses and 1 pony); mild chronic pulmonary disease (CPD 11 horses); moderate CPD (16 horses and 1 pony); and severe CPD (8 horses and 1 pony). Bronchoalveolar lavage (BAL) fluid collected from all animals and respiratory secretions (RS) obtained from 39 of these animals were evaluated cytologically and the results were compared. It was concluded that cytological examination of either BAL fluid or RS was useful in diagnosing various equine pulmonary diseases. The only advantage that BAL offered over RS sampling was in cases in which there was no RS available in the trachea. In addition, the severity of the CPD did not always correlate with either RS or BAL cytology
PMID: 2267569
ISSN: 0036-7281
CID: 117373

Partial divergence between airway inflammation and clinical signs in equine chronic pulmonary disease

Grunig, G; Hermann, M; Howald, B; Winder, C; von Fellenberg, R
PMID: 2707235
ISSN: 0425-1644
CID: 117375

[Determination of complement in serum and tracheobronchial secretions of horses with chronic lung diseases]

Grunig, G; Lerch, C; Hermann, M; von Fellenberg, R
PMID: 3194744
ISSN: 0036-7281
CID: 117378

Procoagulant activity in respiratory tract secretions from horses with chronic pulmonary disease

Grunig, G; Hermann, M; Winder, C; Von Fellenberg, R
Cell-free supernatants (sol phases), obtained after centrifugation (50,000 x g for 45 minutes) of respiratory tract secretions from horses with chronic pulmonary disease, were assayed for procoagulant activity (PCA) in a one-stage clotting assay. Of the 103 specimens tested, 59% (61) contained PCA. Procoagulant activity was detected most often in respiratory tract secretions of severely affected horses and was correlated with the quantity of neutrophils in the respiratory tract secretions. In 12 of the 17 secretions tested, the clotting time was decreased in a dose-dependent manner. However, in the coagulation assay, some reversal of PCA or inhibition of coagulation was observed in 4 secretion specimens when greater volumes of sol phase were added. Procoagulant activity was characterized tentatively as tissue factor, because it was temperature stable and was inhibited by phospholipase C and by concanavalin A. Clotting was induced in factor VIII-deficient human plasma; however, with the exception of 1 respiratory secretion specimen, clotting was not enhanced in factor VII-deficient human plasma. Procoagulant activity is a useful indicator of airway inflammation
PMID: 3395015
ISSN: 0002-9645
CID: 117379

[Eosinophilic granulocytes in tracheobronchial secretions of horses: evidence of parasitic lung disease?]

Hermann, M; Grunig, G; Bracher, V; Howald, B; Winder, C; Hurlimann, J; von Fellenberg, R
PMID: 2964720
ISSN: 0036-7281
CID: 117376

Elastase-producing microorganisms in horse lungs: their possible role in the pathogenesis of chronic pulmonary disease in the horse

Grunig, G; Von Fellenberg, R; Maier, R; Corboz, L
Seventeen out of 21 horses had pulmonary microbial organisms which reached considerable numbers in seven cases. Elastase-producing microorganisms from the environment (Streptomyces species and to a lesser extent Bacillus species) constituted 22 per cent to 99 per cent (mean 79 per cent) of the total growth. There was a considerable number of microorganisms with in vitro-produced elastases which were not or only slightly affected by horse serum. There was no correlation between numbers of organisms and pulmonary histopathological findings thus the significance of these microorganisms in the pathogenesis of alveolar emphysema is unknown. The growth of a strain of Streptomyces collinus/diastatochromogus isolated from the lungs was suppressed by fresh horse serum but not by decomplemented horse serum. Complement activation in response to this organism could contribute to airway inflammation through the production of mediators
PMID: 3639820
ISSN: 0425-1644
CID: 117380

Comparison of neutrophil elastases and of neutrophil protease inhibitors in the horse and man

von Fellenberg, R; Kohler, L; Grunig, G; Pellegrini, A
Neutral neutrophil protease, elastase activities, and cytosol protease inhibitors of these enzymes of horses and man were compared. Human neutrophils had 5 times the elastase activity of equine neutrophils, and neutral protease activity was approximately 50% greater in human neutrophils than that in equine neutrophils. Cytosol inhibitors for elastase and neutral proteases were not found in human neutrophils, whereas large amounts were found in equine neutrophils. Using fibrinogen-agarose electrophoresis, 4 cytosol inhibitors of different enzyme specificities were detected. These cytosol inhibitors were differentiated on the basis of different electrophoretic migration and on the basis of differences in enzyme specificity
PMID: 3853453
ISSN: 0002-9645
CID: 117381

Proteinase inhibitors of horse seminal plasma. A high molecular mass, acid-soluble proteinase inhibitor

von Fellenberg, R; Zweifel, H R; Grunig, G; Pellegrini, A
Horse seminal plasma does not possess a proteinase inhibitor corresponding to human HUSI-I (human seminal plasma inhibitor). Instead a protein complex of high relative molecular mass (Mr) containing proteinase inhibitory activity was detected, which was called horse seminal plasma protein complex or HSPC. The compound had a broad enzyme-inhibiting spectrum. Its Mr was estimated to be 800 000 and it was composed of 7 different polypeptides with Mr values ranging from 11 000 to 30 000. Its carbohydrate content was between 3.5% and 5%. Despite the high molecular mass, the complex was soluble in diluted perchloric acid and did not lose its biological activity. The high recovery of seminal plasma protein (69%) after perchloric acid treatment, the unaltered immunoelectrophoretic precipitation pattern of the perchloric acid soluble part of seminal plasma, and the similarity of the polypeptide patterns of unfractionated seminal plasma and HSPC suggest that HSPC is one of the major components of horse seminal plasma. In addition to HSPC, horse seminal plasma contained a group of three electrophoretically distinguishable proteinase inhibitors, corresponding roughly to a Mr of 6500. They inhibited only trypsin. The similar Mr values and the identical narrow enzyme specificity suggest that they are isoinhibitors and may be analogues of human HUSI-II (human seminal plasma inhibitor). The lack of a HUSI-I analog in the horse is discussed in relation to a previously made observation that horse tracheobronchial fluid contains no detectable perchloric acid-soluble proteinase inhibitors
PMID: 2998413
ISSN: 0177-3593
CID: 117377