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Biomedical subjects

P A Henricks

Publications and source records attributed to P A Henricks.

At least 55 records · Page 3Linked to original sources

Pasteurella haemolytica leukotoxin enhances production of leukotriene B4 and 5-hydroxyeicosatetraenoic acid by bovine polymorphonuclear leukocytes.

The influence of the leukotoxin of Pasteurella haemolytica on the generation of arachidonic acid metabolites by bovine polymorphonuclear leukocytes (PMNs) was investigated. PMNs released 5-, 12-, and 15-hydroxyeicosatetraenoic acids (5-, 12-, and 15-HETE) and leukotriene B4 (LTB4) upon stimulation with arachidonic acid. The leukotoxin preparations dose dependently enhanced the release of the 5-lipoxygenase products 5-HETE and LTB4 in arachidonic acid-stimulated PMNs, whereas the release of 12- and 15-HETE was not affected. The enhanced release of LTB4 and 5-HETE was not due to a decreased cellular retention of the 5-lipoxygenase products. In addition, leukotoxin preparations by themselves were also able to induce LTB4 and 5-HETE production in the absence of exogenous arachidonic acid. Generation of 5-lipoxygenase products by PMNs stimulated by leukotoxin may represent an important cellular event that occurs during infections with P. haemolytica.

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9- and 13-hydroxy-linoleic acid possess chemotactic activity for bovine and human polymorphonuclear leukocytes.

The presence of polymorphonuclear leukocytes (PMNs) within the airways is a characteristic feature of a variety of lung diseases. Pulmonary alveolar macrophages (PAMs) and epithelial cells release many different factors which contribute to the recruitment of inflammatory cells into infected airways. PAMs and tracheal epithelial cells are able to produce linoleic acid metabolites (9-HODE and 13-HODE) besides arachidonic acid metabolites. The objective of the present study was to determine whether 9-HODE and 13-HODE possess chemotactic activity for isolated PMNs. It was found that 9-HODE and 13-HODE induced a chemotactic response of both human and bovine PMNs in vitro. The HODEs evoked chemotaxis with a linear dose response from 10(-10) to 10(-6) M to the same extent as the arachidonic acid metabolite 15-HETE. At 10(-8) M, 9-HODE and 13-HODE were approximately half as potent in inducing chemotaxis as compared to LTB4.

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Modulation of guinea pig airway reactivity by the linoleic acid metabolite 13-hydroxy-octadecadienoic acid (13-HODE).

The influence of the linoleic acid metabolite 13-hydroxy-octadecadienoic acid (13-HODE) on guinea pig tracheal reactivity to both contractile and relaxant agonists was investigated in vitro. 13-HODE induced an increased contraction of tracheal rings to histamine, whereas methacholine responsiveness was not significantly affected. Relaxant responses to isoprenaline and prostaglandin E2 were not influenced either.

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Epithelium-derived linoleic acid metabolites modulate airway smooth muscle function.

Cultured epithelial cells obtained from guinea pig tracheal preparations metabolized arachidonic acid into 5- and 15-hydroxy-eicosatetraenoic acid and into the prostaglandins E2 and F2 alpha. Linoleic acid was converted by the epithelial cells into 9-hydroxy-octadecadienoic acid (9-HODE) and smaller amounts of 13-HODE. It was further investigated whether linoleic acid metabolites are of importance for the regulation of airway smooth muscle function. 13-HODE caused an increase of maximal contraction of tracheal rings to histamine, while 9-HODE had no effect.

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Production of arachidonic and linoleic acid metabolites by guinea pig tracheal epithelial cells.

Pulmonary epithelial cells may be responsible for regulating airway smooth muscle function, in part by release of fatty acid-derived mediators. Incubation of isolated guinea pig tracheal epithelial cells with radiolabeled arachidonic acid (AA) leads to the production of 5- and 15-hydroxyeicosatetraenoic acid (5- and 15-HETE) and smaller amounts of leukotriene (LT) B4 and C4 and 12-hydroxyheptadecatrienoic acid (HHT). Epithelial cells also are able to release linoleic acid (LA) metabolites. Incubation with radiolabeled linoleic acid leads to the formation of 9- and 13-hydroxyoctadecadienoic acid (9- and 13-HODE). The biological significance of these mediators produced by epithelial cells is discussed.

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The effect of Pasteurella haemolytica cytotoxin on bovine polymorphonuclear leukocytes can be attenuated by beta-adrenoceptor antagonists.

It was investigated whether beta-adrenoceptor antagonists could disturb the interaction between cytotoxin preparations isolated from Pasteurella haemolytica and bovine polymorphonuclear leukocytes (PMNs). The toxicity of the cytotoxin preparation was evaluated by measuring the chemiluminescence response and the viability of the cells after incubation with the cytotoxin. No effect on cell viability was detected when PMNs were incubated with 63 micrograms cytotoxin per ml while the chemiluminescence response was diminished by approximately 30%. The beta-adrenoceptor antagonists alprenolol (10(-5) M) and propranolol (5 X 10(-7) - 5 X 10(-6) M) were able to attenuate this effect of cytotoxin on the chemiluminescence response of PMNs. It seemed unlikely that propranolol and alprenolol diminished the effect of cytotoxin on the chemiluminescence response of PMNs by their beta-adrenoceptor blocking potency because other beta-adrenoceptor antagonists used were without effect. Also, the membrane stabilizing characteristics of the beta-adrenoceptor antagonists used were probably not responsible for the diminished interaction between PMNs and the cytotoxin. Whether beta-adrenoceptor antagonists could be used in vivo to prevent or treat P. haemolytica infections in bovines remains to be examined.

Adrenergic beta-Antagonists↗

The effect of stress on host defense system and on lung damage in calves experimentally infected with Pasteurella haemolytica type A1.

This paper describes the influence of stress, immediately before a massive inoculation of a log-phase P. haemolytica A1 culture in brain-heart-infusion broth by fiberoptic bronchoscopy in 5 months old conventional calves. Differences in phagocytic cell function and numbers, lung function and lung tissue damage between stressed and non-stressed, infected calves were studied. Compared with the non-stressed calves before infection, the superoxide generation by blood polymorphonuclear leucocytes of the stressed animals was markedly reduced immediately after a 2 hours stress period, but it recovered within 3 days. The total numbers of phagocytic cells, harvested from lung lavage fluid 3 days after inoculation were twice as high in the non-stressed calves than in the stressed calves. In contrast, no differences in pulmonary lesions and functions were found between stressed or non-stressed calves. The role of several stress factors on infiltration and metabolic response of polymorphonuclear leucocytes in P. haemolytica infections and the effects of bacterial endotoxin and leukotoxin on alveolar and vascular injury are discussed.

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Pulmonary lesions induced by a Pasteurella haemolytica cytotoxin preparation in calves.

Intrabronchial instillation of a Pasteurella haemolytica type A1 crude cytotoxin preparation in calves resulted in pulmonary gross and microscopic lesions comparable to spontaneous and experimental pasteurellosis. In the acute stage of the lesion electronmicroscopy revealed intravascular accumulation, degeneration and fragmentation of leukocytes in the interalveolar septa. Secondary thrombus formation and increased vascular permeability resulted in alveolar flooding, fibrin deposition, extravasation of erythrocytes and loss of alveolar epithelium. No cytotoxicity was observed for the tracheal (in vitro) and bronchial epithelium (in vivo). The pathogenesis of the vascular lesions and their significance for the development of the typical lesions of pneumonic pasteurellosis is discussed.

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The influence of mononuclear leukocytes on the chemotactic responsiveness of polymorphonuclear leukocytes of bovines.

Polymorphonuclear leukocytes (PMN) isolated from the blood of bovines which do not react with an influx of cells into the milk upon intramammary administration of low doses of endotoxin (Group II), show a diminished chemotactic activity in vitro as compared to PMN of bovines which do respond to intramammary endotoxin administration (Group I; 8). The purpose of the present study was to determine whether other immune cells affected the chemotactic activity of PMN of Group I and II bovines. Mononuclear leukocyte suspensions of Group I animals had no effect on the migration of PMN of Group I animals. In contrast, the distance migrated by Group II PMN was diminished in the presence of Group II mononuclear leukocytes. As a consequence, this phenomenon resulted in a much larger difference in chemotactic activity of PMN in vitro between Group I and Group II bovines. It can be concluded that the chemotactic activity of PMN of animals which are more susceptible for mastitis as indicated by non-responsiveness to intramammary endotoxin-infusions, was further down-regulated by mononuclear leukocytes.

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Receptors in airway disease. Beta-adrenoceptors in lung inflammation.

Beta-adrenoceptor dysfunction and increase in airway reactivity can be induced by administration of gram-negative bacteria, endotoxin, viruses, and allergens in laboratory animals. However, the deterioration of lung beta-adrenoceptor function is not invariably associated with lung inflammation. Severe asthmatics, but not asthmatics per se, show a diminished beta-adrenoceptor function of airway smooth muscle. These changes are probably a consequence of the active disease state rather than an intrinsic component of asthma. Mediators released from inflammatory cells such as reactive oxygen species and fatty acid metabolites may directly or indirectly induce beta-adrenoceptor dysfunction. Beta-adrenoceptor function of leukocytes from asthmatic patients can be decreased as well and it is suggested that lymphokines like interleukin-2 and interferon-gamma may affect beta-adrenoceptor function. A disturbed beta-adrenoceptor function on inflammatory cells themselves may have consequences for their immune function, mediator release, and effect on surrounding tissues.

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Preinfection functions of blood polymorphonuclear leukocytes and the outcome of experimental Escherichia coli mastitis in the cow.

The relationship between preinfection functions of blood neutrophils and outcome of experimental Escherichia coli mastitis was studied in 11 cows. Random migration, chemotaxis, phagocytosis, and chemiluminescence by neutrophils were determined in white blood cell suspensions, and in purified neutrophil suspensions. The course of E. coli mastitis (10(4) E. coli 0:157 in rear quarters) was monitored using clinical parameters, counts of E. coli in mastitic secretion, and milk production. Regressions were calculated for areas under curves of these parameters and preinfection activities of neutrophils. Chemiluminescence by nonstimulated neutrophils in white blood cell suspensions was negatively correlated with counts of E. coli in secretion and with losses in milk production. The chemotactic differential in white blood cell suspensions minus the chemotactic differential in purified suspensions of neutrophils referred to as delta varied from -.66 to +.50, indicating, respectively, inhibition and stimulation of chemotactic activity of neutrophils in white blood cell suspensions. Delta correlated negatively with counts of E. coli in mastitis secretion, inhibition of the amplitude of rumen contractions, and losses in milk production. We hypothesize that a factor in white blood cell suspensions may be involved in the down-regulation of the migratory response of neutrophils during E. coli mastitis.

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Polymorphonuclear leucocyte function: relationship between induced migration into the bovine mammary gland and in vitro cell activity.

Low doses of 10(-7) mg Escherichia coli endotoxin applied as intramammary infusion into single bovine quarters induced a rise in milk cell count without other inflammatory signs. Significantly fewer quarters responded in early lactation than in mid lactation. Maximum cell count was also somewhat later and less pronounced in early lactation. The rise in milk cell count after infusion of E. coli endotoxin was related to in vitro chemotactic activity of blood polymorphonuclear leucocytes (PMN). PMN isolated from cows which did not respond with a rise in milk cell count upon endotoxin infusion showed a diminished chemotactic activity in vitro as compared to PMN isolated from animals which did respond to an intramammary endotoxin infusion with a rise in milk cell count. No differences in phagocytic and metabolic activity were observed in vitro between the PMN isolated from the two groups of animals.

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Modulatory activity of 9-hydroxy- and 9-hydroperoxy-octadecadienoic acid towards reactive oxygen species from guinea-pig pulmonary macrophages.

As guinea-pig pulmonary macrophages (PM) synthesize the linoleic acid metabolite 9-hydroxy-octadecadienoic acid (9-OH-Lin) under non-stimulated conditions in relatively large quantities, we investigated whether this product has an effect on the macrophage's own phagocytic cell function. 9-OH-Lin, and also its hydroperoxy precursor 9-hydroperoxy-octadecadienoic acid (9-OOH-Lin), influenced the generation of PM chemiluminescence, a measure of the production of reactive oxygen species. The generation of lucigenin-enhanced chemiluminescence by stimulated and non-stimulated PM was inhibited concentration-dependently. Inhibition was observed at concentrations as low as 10 nM. Since 9-OH-Lin and 9-OOH-Lin also inhibited the generation of chemiluminescence by a cell-free enzyme system, i.e. xanthine/xanthine oxidase, the inhibitory effects might represent a scavenging activity towards reactive oxygen species. 9-OH-Lin and 9-OOH-Lin did not influence other phagocytic cell functions, e.g. PM phagocytic capacity, the aggregatory response to the calcium ionophore A23187, or the release of lysosomal enzymes. The effects of 9-OH-Lin and 9-OOH-Lin could be ascribed to the hydroxy and hydroxyperoxy moiety, respectively, as evidenced by lack of effect of the native fatty acid linoleic acid. We conclude that the formation of 9-OH-Lin and 9-OOH-Lin by PM may represent a regulatory mechanism towards the cell's own activity by modulating reactive oxygen species.

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