Peter L. Lefferts

Effects of platelet depletion on the unanesthetized sheep's pulmonary response to endotoxemia.

The effect of platelet depletion on the unanesthetized sheeps pulmonary response to endotoxemia was studied in eight unanesthetized sheep. Platelets were depleted with rabbit anti-sheep platelet antibodies (APA). Bolus injections of APA alone caused marked pulmonary hypertension (PPA increased from 21 +/- 2 to 62 +/- 5 cm H2O +/- SE) and alterations in lung mechanics (dynamic compliance of the lung [Cdyn] decreased to 38.5 +/- 4.6% and resistance to air flow across the lung [RL] increased to 705 +/- 162% +/- SE of control), which were attenuated by pretreatment with meclofenamate. It was possible to deplete platelets before endotoxemia through a slow continuous infusion of APA without altering base-line values of the measured variables. Platelet depletion did not significantly attenuate the alterations in pulmonary hemodynamics, lung mechanics, lung fluid and solute exchange, or the normal increase in lung lymph concentrations of thromboxane B2 or 6-keto-PGF1 alpha observed following endotoxemia in the sheep. We conclude that normal circulating platelet counts are not required for the full expression of the sheeps response to endotoxemia.

Effects of thromboxane synthase and cyclooxygenase inhibition on PAF-induced changes in lung function and arachidonic acid metabolism

PAF was administered as an intravenous bolus (0.1 micrograms/kg) to eight chronically instrumented awake sheep. The effects of pretreatment with an inhibitor of cyclooxygenase (meclofenamate) on PAF-induced changes in lung function were compared to those observed with a specific inhibitor of thromboxane synthase (DP1904). Each animal was studied four times in varied order: PAF alone, PAF + DP1904, PAF + meclofenamate, and DP1904 alone. Saline alone (control), DP1904 alone, and meclofenamate alone did not cause changes in any of the measured variables. DP1904 and meclofenamate significantly attenuated the PAF-induced fall in lung compliance, elevation in peak pulmonary artery pressure, and increased lung lymph flow. Both drugs abolished the PAF-induced increases in lung lymph thromboxane B2 concentrations. Meclofenamate, but not DP1904, blocked the rise in lymph 6-keto-PGF1 alpha. Although meclofenamate blocked the rise in lymph PGE2, DP1904 resulted in levels 2.7 times higher than PAF alone. We conclude that: (1) inhibition of thromboxane synthase is as effective as inhibition of cyclooxygenase in attenuating PAF-induced changes in lung function, and (2) thromboxane synthase inhibition results in augmented production of PGE2 following PAF administration in vivo.

Phorbol myristate acetate lung injury and airway responsiveness to aerosol histamine in awake sheep

To test the hypothesis that phorbol myristate acetate (PMA), a potent granulocyte activator, would increase airway responsiveness to aerosol histamine, we quantitated airway responsiveness to aerosol histamine in nine awake sheep before and 4 1/2 hours after intravenous PMA, 5 micrograms/kg. A dose-response curve to aerosol histamine was constructed by administering histamine aerosols of 0.1, 0.3, 1.0, 3.0, 10.0, and 30.0 mg/ml sequentially until dynamic lung compliance (Cdyn) decreased to less than or equal to 65% baseline. The dose of histamine that caused a reduction of Cdyn to 65% of baseline (ED65Cdyn) was determined by linear interpolation. We then administered PMA, defined a new baseline Cdyn 4 1/2 hours later, and administered aerosol histamine as before. Control experiments were performed on another day in eight of the nine sheep by administering aerosol histamine 4 1/2 hours apart without infusing PMA. Histamine responsiveness did not change significantly during 4 1/2 hours in the control experiments. Control (preaerosol histamine) Cdyn decreased from 0.090 +/- 0.010 L/cm H2O before PMA to 0.048 +/- 0.004 L/cm H2 4 1/2 hours after PMA. The mean pre-PMA ED65Cdyn for the nine sheep was 3.80 +/- 0.87 mg/ml, and the mean post-PMA ED65Cdyn was 2.13 +/- 0.98 mg/ml (p less than 0.05). When individual dose-response curves are examined, the small statistical increase in aerosol histamine responsiveness does not appear important, especially when the increase is compared to the changes previously reported after endotoxemia in unanesthetized sheep.

Effect of Pulmonary Edema on Tracheal Diameter

Background: Though it is well known that cardiogenic and noncardiogenic pulmonary edema can cause changes in lung mechanics, actual alterations in tracheal diameter have not been described. Objective: To evaluate the effects of pulmonary edema induced by increased left atrial pressure (cardiogenic) and Perilla ketone (PK; noncardiogenic) on tracheal diameter in chronically instrumented awake sheep. Methods: We investigated the effects of two mechanistically distinct types of pulmonary edema on tracheal diameter in chronically instrumented awake sheep. Cardiogenic pulmonary edema (analogous to congestive heart failure in humans) was induced by increasing left atrial pressure (↑PLA) by inflating the balloon on a Foley catheter positioned in the mitral valve annulus to cause partial obstruction to flow across the valve (n = 18). Noncardiogenic pulmonary edema (increased pulmonary microvascular permeability pulmonary edema analogous to the acute respiratory distress syndrome in humans) was produced by the intravenous administration of PK (n = 11). Lateral chest radiographs (CXRs) were scored by a standardized 5-point scoring system for the severity of pulmonary edema, and tracheal diameter was measured at a fixed location in the carina. Three radiologists, blinded to sheep identification number and experimental protocol, evaluated the radiographs independently at different points in time for edema severity and tracheal diameter. The sheep were sacrificed immediately after the final CXR, and wet/dry lung weight ratio (W/D ratio) was determined. Results: Both ↑PLA and PK were associated with statistically significant tracheal narrowing (↑PLA: 20.3 ± 0.6 to 15.1 ± 0.9 mm; PK: 20.2 ± 0.6 to 14.1 ± 1.4 mm). Tracheal narrowing correlated with the severity of the pulmonary edema determined radiographically (↑PLA: r = –0.69, p < 0.01; PK: r = –0.62, p < 0.01) and by W/D ratio (↑PLA: r = –0.64, p < 0.05; PK: r = –0.54, p < 0.05). Conclusions: We conclude that tracheal narrowing occurs in sheep models of both cardiogenic and noncardiogenic pulmonary edema and that the degree of narrowing correlates with the severity of the edema.

Meclofenamate blocks the pulmonary arterial vasopressor effects of leukotriene B4 in awake sheep

This study examined the hemodynamic effects of leukotriene B4 (LTB4) in chronically instrumented awake sheep, and the role of cyclooxygenase products in the sheeps response to LTB4. LTB4 (25 micrograms) was given as a bolus into the pulmonary artery. Six sheep were studied with LTB4, both before and after pretreatment with meclofenamate (5 mg/kg load, 3 mg/kg/hr maintenance infusion). LTB4 alone caused a rapid rise in pulmonary arterial pressure from 15 +/- 1 to 42 +/- 11 cm H2O. LTB4 had no effect on pulmonary arterial pressure following pretreatment with meclofenamate. LTB4 alone caused an increase in serum thromboxane B2 (TxB2) from 130 +/- 35 to 320 +/- 17 pg/ml 3 minutes after dosing but did not increase TxB2 following pre-treatment with meclofenamate. LTB4 caused a slight decrease in mean systemic arterial pressure and a transient fall in circulating white blood cells, both of which were unaffected by meclofenamate pre-treatment. The vasoactive effects of LTB4 in the pulmonary circulation appear to be mediated indirectly through the production of cyclooxygenase metabolites of arachidonic acid.