Biomedical subjects

D F Kern

Publications and source records attributed to D F Kern.

11 recordsLinked to original sources

PubMed1994

Characterization of pulmonary endothelial charge barrier.

To clarify the role of charge in protein movement across the pulmonary endothelial barrier, we simultaneously measured the permeability-surface area product (PS) for native [isoelectric point (pI) 4.4-5.1] and cationic (pI 7.2-8.0) albumin in isolated rabbit lungs perfused with and without protamine sulfate. We focused our measurement on the initial (endothelial) barrier by using a technique that is based on the very rapid (3 min) uptake of tracer. This allowed us to distinguish the charge properties of the endothelium separate from other barriers. In control studies, PS was greater for cationic than for native albumin (8.67 +/- 0.93 vs. 2.55 +/- 0.20 x 10(-2) ml.min-1.g dry lung-1). In the presence of 1 mg/ml protamine sulfate, cationic albumin permeability was not different from control (7.34 +/- 0.49 x 10(-2) ml.min-1.g dry lung-1), whereas PS for anionic albumin increased to 8.82 +/- 1.32 x 10(-2) ml.min-1.g dry lung-1. Thus the protamine sulfate eliminated the difference between native and cationic albumin PS. This selective increase in anionic albumin permeability is presumably due to the cation, protamine sulfate, binding to the anionic charges on the endothelium and reducing the anionic charge-charge repulsion. If protamine sulfate had produced a general endothelial injury, the PS for both albumins would have increased. Our results suggest that the normal pulmonary endothelium is an anionic charge barrier restricting the transcapillary movement of negatively charged molecules.

Animals↗

PubMed1993

HEPES buffer perfusate alters rabbit lung endothelial permeability.

N-2-hydroxyethylpiperazine-N'-2-ethanesulfonic acid (HEPES) has been shown to cause changes in cultured endothelial cells and smooth muscle function at concentrations from 5 to 25 mM. To determine whether HEPES also affects vascular permeability, the effects of two buffers, HEPES and phosphate, were compared in isolated perfused rabbit lungs. Hemodynamic parameters and vascular protein permeability-surface area products (PS) were measured after perfusion with the buffers. Endothelial permeability was measured for an anionic and a cationic albumin to assess the charge effects of the zwitterion buffer. With HEPES, there were no changes in vascular pressure or resistance but permeability was affected. Cationic albumin permeability increased with 12 mM HEPES (8.7phosphate-->30(12) mM HEPES x ml.min-1.g dry lung-1 x 10(-2)) as did the anionic albumin PS (2.7phosphate-->3.52(12) mM HEPES). The cationic PS returned to baseline (8.1(60) mM HEPES) at 60 mM HEPES, but the anionic PS did not change from the 12 mM HEPES (4.01(60) mM HEPES). In summary, we find that HEPES is not innocuous. Although hemodynamic parameters did not change, endothelial permeability was increased when HEPES was used at normal concentrations. Therefore, HEPES should be used with caution as a physiological buffer in perfused organ systems.

Animals↗

PubMed1993

Effect of common vasodilators on lung microvascular permeability.

The effect of papaverine on the albumin permeability-surface area product (PS), reflection coefficient (sigma), and capillary filtration coefficient (Kf) was examined in isolated rabbit lungs. Because PS and Kf are functions of vascular surface area and permeability, we also compared papaverine with two other means of maximizing lung surface area: isoproterenol (1 x 10(-7) M) and a mild increase in vascular pressure. Only lungs perfused with 0.1 mg/ml papaverine were significantly different from control. PS increased from control (2.80 +/- 0.16 to 5.53 +/- 0.20 ml.min-1.g dry lung-1 x 10(-2), whereas sigma decreased from control (0.92 +/- 0.01 to 0.78 +/- 0.03). Kf after papaverine was significantly lower than baseline predrug Kf (5.60 +/- 0.78 to 4.56 +/- 0.53 ml.s-1.cmH2O-1.g dry lung-1 x 10(-3). However, this group's predrug Kf was higher than that of any other group. Our results indicate that papaverine increases albumin permeability and decreases endothelial selectivity. The isolated perfused lung appears fully recruited, because Kf and PS did not increase with isoproterenol or increased vascular pressure. Papaverine should be used with caution in the Ringer-perfused lung.

Animals↗

PubMed1986

Pulmonary vascular effects of a fibrin(ogen)-derived vasoactive peptide.

We examined the effects of the 5 amino acid fibrin(ogen)-degradation product Ala-Arg-Pro-Ala-Lys (Peptide 6A) on pulmonary hemodynamics and vascular permeability in the isolated-perfused guinea pig lung. Infusion of this peptide (100 mumoles) resulted in a small increase in pulmonary vascular resistance. The pulmonary vasoconstrictor responses was not mediated by histamine or cyclooxygenase metabolites since neither H1 blockers nor meclofenamate inhibited the response. The capillary filtration coefficient (a measure of water transvascular permeability) did not change after Peptide 6A infusion. Small molecular weight fibrin(ogen)-degradation products may contribute to increased pulmonary vascular resistance associated with pulmonary intravascular coagulation, but not to the increase in lung hydraulic conductivity.

Animals↗

PubMed1986

Pulmonary microvascular responses to arachidonic acid in isolated perfused guinea pig lung.

We examined the effects of arachidonic acid (AA) on pulmonary hemodynamics and fluid balance in Ringer- and blood-perfused guinea pig lungs during constant-flow conditions. Mean pulmonary arterial (Ppa), venous (Pv), and capillary pressures (Pcap, estimated by the double-occlusion method) were measured, and arterial (Ra) and venous resistances (Rv) were calculated. Bolus AA injection (500 micrograms) caused transient increases (peak response 1 min post-AA) in Ppa, Pcap, and Rv without affecting Ra in both Ringer- and blood-perfused lungs. The response was sustained in blood-perfused lungs. AA had no effect on the capillary filtration coefficient in either Ringer- or blood-perfused lungs. AA stimulated the release of thromboxane B2 and 6-ketoprostaglandin F1 alpha in both Ringer- and blood-perfused lungs, but the responses were sustained only in the blood-perfused lungs. Meclofenamate (1.5 X 10(-4) M), a cyclooxygenase inhibitor, abolished the AA-induced pulmonary hemodynamic responses in both Ringer- and blood-perfused lungs, whereas U-60257 (10 microM), a lipoxygenase inhibitor, attenuated the response only in the blood-perfused lungs. In conclusion, AA does not alter pulmonary vascular permeability to water in either Ringer- or blood-perfused lungs. AA mediates pulmonary venoconstriction and thus contributes to the rise in Pcap. The venoconstriction results from the generation of cyclooxygenase-derived metabolites from lung parenchymal cells and blood-formed elements. Lipoxygenase metabolites may also contribute to the vasoconstriction in the blood-perfused lungs.

6-Ketoprostaglandin F1 alpha↗

PubMed1986

Mechanism of peptidoleukotriene-induced increases in pulmonary transvascular fluid filtration.

We examined the effects of leukotrienes C4 (LTC4) and D4 (LTD4) (1 microgram) on the pulmonary vascular filtration coefficient, a measure of vessel wall conductivity to water, and the alterations in pulmonary vascular resistance (PVR) in isolated-perfused guinea pig lungs. We also assessed whether LTC4 and LTD4 increased the permeability to albumin in cultured monolayers of pulmonary artery endothelial cells. In Ringer-perfused and blood-perfused lungs, LTC4 resulted in increases in pulmonary arterial pressure (Ppa) and the pulmonary capillary pressure (Pcap) measured as the equilibration pressure after simultaneous pulmonary arterial and venous occlusions. Pulmonary venous resistance (Rv) increased to a greater extent than arterial resistance (Ra) in both Ringer-perfused and blood-perused lungs challenged with LTC4. The greater increase in PVR in blood-perfused lungs corresponded with a greater elevation of lung effluent thromboxane B2 (TxB2) concentration. The LTC4-stimulated increase in PVR was prevented by pretreatment with meclofenamate (10(-4) M). LTD4 also induced rapid increases in Ppa and Pcap in both Ringer-perfused and blood-perfused lungs; however, Ppa decreased before stabilizing at a pressure higher than base line. The increases in Rv with LTD4 were greater than Ra. The LTD4-stimulated increases in Ra and Rv also paralleled the elevation in TxB2 concentration. As with LTC4, the increases in Ppa, Pcap, PVR, and TxB2 concentration were greater in blood-perfused than in Ringer-perfused lungs. Pretreatment with meclofenamate reduced the magnitude of the initial increase in Ppa, but did not prevent the response.(ABSTRACT TRUNCATED AT 250 WORDS)

6-Ketoprostaglandin F1 alpha↗

PubMed1985

Pulmonary microcirculatory responses to leukotrienes B4, C4 and D4 in sheep.

The pulmonary microvascular responses to leukotrienes B4, C4, and D4 (total dosage of 4 micrograms/kg i.v.) were examined in acutely-prepared halothane anesthetized and awake sheep prepared with lung lymph fistulas. In anesthetized as well as unanesthetized sheep, LTB4 caused a marked and transient decrease in the circulating leukocyte count. Pulmonary transvascular protein clearance (pulmonary lymph flow X lymph-to-plasma protein concentration ratio) increased transiently in awake sheep, suggesting a small increase in pulmonary vascular permeability. The mean pulmonary artery pressure (Ppa) also increased. In the acutely-prepared sheep, the LTB4-induced pulmonary hemodynamic and lymph flow responses were damped. Leukotriene C4 increased Ppa to a greater extent in awake sheep than in anesthetized sheep, but did not significantly affect the pulmonary lymph flow rate (Qlym) and lymph-to-plasma protein concentration (L/P) ratio in either group. LTD4 increased Ppa and Qlym in both acute and awake sheep; Qlym increased without a significant change in the L/P ratio. The LTD4-induced rise in Ppa occurred in association with an increase in plasma thromboxane B2 (TxB2) concentration. The relatively small increase in Qlym with LTD4 suggests that the increase in the transvascular fluid filtration rate is the result of a rise in the pulmonary capillary hydrostatic pressure. In conclusion, LTB4 induces a marked neutropenia, pulmonary hypertension, and may transiently increase lung vascular permeability. Both LTC4 and LTD4 cause a similar degree of pulmonary hypertension in awake sheep, but had different lymph flow responses which may be due to pulmonary vasoconstriction at different sites, i.e. greater precapillary constriction with LTC4 because Qlym did not change and greater postcapillary constriction with LTD4 because Qlym increased with the same rise in Ppa.

Animals↗

PubMed1985

Microvascular albumin permeability in isolated perfused lung: effects of EDTA.

We examined the effects of decreases in perfusate concentrations of calcium and magnesium on the pulmonary vascular permeability in the isolated perfused rabbit lung. The albumin permeability-surface area product (PS) and the albumin reflection coefficient (sigma) were determined in the same lung using 125I- and 131I-labeled albumin tracers. Decreases in vascular Ca2+ and Mg2+ concentrations were induced by adding ethylenediaminetetraacetic acid (EDTA) to the perfusate. Decreases in the concentration of these cations resulted in an increase in the PS from a control value of 1.18 +/- 0.13 X 10(-3) to 7.69 +/- 0.75 X 10(-3) cm3 X min-1 X g wet lung wt-1 and a decrease in the sigma from 0.96 +/- 0.01 to 0.74 +/- 0.02. The decrease in sigma suggests an increase in the calculated equivalent pore radius from 44 to 63 A. The results indicate that Ca2+ and Mg2+ play a role in the maintenance of normal pulmonary vascular permeability to proteins.

Animals↗

PubMed1984

Pulmonary vascular response to thrombin: effects of thromboxane synthetase inhibition with OKY-046 and OKY-1581.

We examined the effects of thromboxane synthetase inhibition with OKY-1581 and OKY-046 on pulmonary hemodynamics and lung fluid balance after thrombin-induced intravascular coagulation. Studies were made in anesthetized sheep prepared with lung lymph fistulas. Pulmonary intravascular coagulation was induced by i.v. infusion of alpha-thrombin over a 15 min period. Thrombin infusion in control sheep resulted in immediate increases in pulmonary artery pressure (Ppa) and pulmonary vascular resistance (PVR), which were associated with rapid 3-fold increase in pulmonary lymph flow (Qlym) and a delayed increase in lymph-to-plasma protein concentration (L/P) ratio, indicating an increase in the pulmonary microvascular permeability to proteins. Thrombin-induced intravascular coagulation also increased arterial thromboxane B2 (a metabolite of thromboxane A2) and 6-keto-PGF1 alpha concentrations (a metabolite of prostacyclin). Both OKY-1581 and OKY-046 prevented thromboxane B2 and 6-keto-PGF1 alpha generation. The initial increments in Ppa and PVR were attenuated in both treated groups. The increases in Qlym were gradual in the treated groups but attained the same levels as in control group. However, the increases in Qlym were associated with decreases in L/P ratio. In both treated groups, the leukocyte count decreased after thrombin infusion but then increased steadily above the baseline value, whereas the leukocyte count remained depressed in the control group after thrombin. These studies indicate that a part of the initial pulmonary vasoconstrictor response to thrombin-induced intravascular coagulation is mediated by thromboxane generation. In addition, thromboxane may also contribute to the increase in lung vascular permeability to proteins that occurs after intravascular coagulation and this effect may be mediated by a thromboxane-neutrophil interaction.

Acrylates↗

PubMed1984

Comparison of alpha- and gamma-thrombin on lung fluid balance in anesthetized sheep.

We examined the effects of varying dosages of thrombin on lung fluid balance in halothane-anesthetized sheep prepared with lung lymph fistulas. A 15-min iv infusion of sublethal doses of alpha-thrombin (2.5 clotting units/micrograms), the native enzyme, at 0.6 or 1.1 nmol active enzyme/kg body wt increased the mean pulmonary arterial pressure (Ppa) and pulmonary vascular resistance (PVR) two- to threefold. Neither parameter increased in a dose-dependent manner. Platelet counts decreased 50% with both dosages. Leukocyte counts decreased 35 and 75% from base line in the low- and high-dosage groups, respectively, and reached comparable levels of 50% below base line at 60-min postinfusion in both groups. Plasma fibrinogen concentrations decreased in a dose-dependent manner preceding dose-dependent increases in pulmonary lymph flow (Qlym) and lymph protein clearance (Clym). Fibrin deposition in pulmonary vessels was greater at 30 than at 180 min postinfusion. In contrast, a 15-min iv infusion of gamma-thrombin (0.002 clotting units/micrograms), which lacks the fibrinogen recognition site, at 1.2 nmol active enzyme/kg produced no significant increases in PVR, Ppa, Qlym, or Clym. The fibrinogen concentration did not change significantly, whereas platelet and leukocyte counts decreased 25% within 15 min. Fibrin microthrombi were less prominent in pulmonary vessels. Fibrin deposition associated with intravascular coagulation may be an important factor mediating thrombin-induced increases in pulmonary transvascular fluid and protein exchange.

6-Ketoprostaglandin F1 alpha↗

PubMed1983

Endothelial albumin permeability measured with a new technique in perfused rabbit lung.

This study describes two methods of measuring the albumin permeability-surface area product (PS) of the pulmonary microvascular endothelium in isolated, perfused rabbit lungs. Both involve direct sampling of lung tissue to measure how much labeled albumin has entered the extravascular space during a known exposure period. In one, the traditional "multiple-sample" method, extravascular albumin is measured for several different exposure times to determine the kinetics of transcapillary albumin movement, and PS is calculated from that data. We found single-exponential kinetics, with a half time of 77 min and a PS of 2.28 X 10(-3) cm3 . min-1 . g blood-free wet lung-1. The second, "single-sample" method, is a new modification that involves exposing the tissue to labeled albumin for only 3 min, followed by a 3-min vascular washout. Tissue is then analyzed for extravascular albumin and PS calculated assuming the interstitial albumin concentration is essentially zero. The PS obtained was 2.44 X 10(-3) cm3 . min-1 . g blood-free wet lung-1, not significantly different from the value obtained using the more accepted method. Possible sources of error in the new method are discussed, and we conclude that it is an easy, quick, reliable way of accurately measuring macromolecular PS in the lung and presumably other organs as well.

Animals↗