Lena Mårtensson

Nitric oxide inhalation decreases pulmonary platelet and neutrophil sequestration during extracorporeal circulation in the pig

OBJECTIVE The inhibiting effect of nitric oxide on the aggregation and adhesion of neutrophils and platelets has been well documented in vitro. In vivo evidence, however, is more scant. In this study, we studied the effects of inhaled nitric oxide on pulmonary cellular sequestration in our sham hemodialysis model. Accumulation of neutrophils and platelets in the lungs has been shown to be an early event in this model. DESIGN Prospective, randomized, controlled study. SETTING Animal laboratory at a university medical center. SUBJECTS Twenty-six anesthetized, mechanically ventilated pigs. INTERVENTIONS 111Indium-oxine was used to selectively label neutrophils or platelets and the activity over the lungs was followed dynamically with a gamma camera. Sham hemodialysis, using a cuprophan hollow-fiber dialyzer, was instituted via catheters in the femoral vessels. The animals were divided into two main groups: a) the nitric oxide recipient group (n = 12, with platelets labeled in seven animals and neutrophils labeled in five animals); and b) the control group (n = 14, with platelets labeled in seven animals and neutrophils labeled in seven animals). The animals in the former group were given 50 parts per million of nitric oxide in the inspiratory gas from the beginning of dialysis and for 30 mins onward. MEASUREMENTS AND MAIN RESULTS Inhalation of nitric oxide attenuated the increase in activity over the lungs in both the neutrophil and platelet groups during sham hemodialysis. In addition, an inhibiting effect on the increase in pulmonary pressure was noted. CONCLUSION Apart from the effects of nitric oxide on central hemodynamics in this model, the scintigraphic findings indicate an in vivo effect of nitric oxide on the accumulation of platelets and neutrophils in the lungs, probably due to inhibition of the adhesion and/or aggregation of these cells.

Polymorphonuclear leucocyte sequestration in the lungs and liver following soft-tissue trauma: an in vivo study

Neutrophils are thought to sequestrate in the lungs and the liver in association with shock. Indications for this have previously been demonstrated in different in vitro studies. In this experiment an in vivo technique for dynamic studies of pulmonary and liver neutrophil sequestration (PNT and LNT, respectively) is described. Autologous neutrophils from ten pigs were labelled with 111Indium-oxine. The pigs were placed under a scintillation camera for continuous recording of the activity distribution in the pigs during 105 minutes. Following a steady-state period of 15 minutes seven pigs were subjected to a standardized soft-tissue trauma. Three pigs were used as controls and not traumatized. Within 1-3 minutes after trauma the radioactivity over the lungs increased dramatically, indicating PNT. This was followed by a fast decrease but 90 minutes after trauma PNT levels were still significantly elevated. LNT showed a similar pattern, although the immediate increase was less dramatic. This study shows that PNT and LNT occur immediately after soft-tissue trauma and can be studied dynamically in vivo.

Changes in Plasma Levels of Vasoactive Peptides during Sequential Bicarbonate Hemodialysis

The hemodynamic response to isolated ultrafiltration (IUF) is characterized by a vasoconstriction, while there is no significant change in peripheral vascular resistance during isovolemic bicarbonate hemodialysis (IVHD). The present investigation was designed to study the plasma levels of vasoactive regulatory peptides together with noradrenaline (NA) and plasma renin activity (PRA) in 11 patients during sequential hemodialysis (SQHD) - IUF for 60 min, followed by IVHD for 210 min. During IUF, the vasoconstrictors arginine vasopressin (AVP), gamma 2-melanocyte-stimulating hormone (gamma 2-MSH), neuropeptide Y (NPY), NA and PRA increased. During IVHD, NPY and PRA remained unchanged on a higher level. A decrease in AVP below the baseline and in gamma 2-MSH and NA to the baseline levels occurred during IVHD. In the case of vasodilators, there were no changes in calcitonin gene-related peptide or motilin during SQHD. An increase in beta-endorphin (beta-END) occurred during IUF, followed by a decrease during IVHD. Substance P and vasoactive intestinal peptide were unchanged during IUF but decreased during IVHD. We conclude that SQHD is characterized by an increase in all the measured vasoconstrictors during IUF in response to loss of fluid, and by a decrease in some vasoconstrictors (AVP, gamma 2-MSH, NA) during IVHD. With the exception of beta-END, there were no changes or only minor ones in vasodilators during SQHD. There are changes in plasma levels of vasoactive substances during SQHD but the importance of these changes for the hemodynamic adaptation to ultrafiltration and dialysis needs to be studied further.

Circulatory and ventilatory effects of intermittent nitric oxide inhalation during porcine endotoxemia.

The effects of intermittent inhalation of 57 ppm nitric oxide (NO) were studied in eight anesthetized, ventilated pigs given a continuous infusion of Escherichia coli endotoxin. Seven animals served as controls. By administering NO synchronized with inspiration and close to the orotracheal tube, measurable amounts of the toxic metabolite, NO2, in the inspiratory gas mixture were avoided. No direct systemic effects of NO inhalation were seen, but through counteracting pulmonary vasoconstriction, a fall in cardiac output was delayed. Nitric oxide effectively attenuated the initial peak rise in mean pulmonary artery pressure and resistance, both returning to control levels after cessation of NO. These effects were reproduced during later phases of endotoxemia, giving further proof to the role of gaseous NO as a selective pulmonary vasodilator. Nitric oxide diminished pulmonary shunting, but unimpaired oxygenation was preserved only during the first inhalation period. Leukocyte counts decreased drastically and platelet aggregation was enhanced, but after 1.5 hours of endotoxin infusion, platelet hyperaggregation was maintained in the NO group while it decreased in the control group.

Myoendothelial and placental blood flow responses to ritodrine infusion in the guinea pig.

The effect of ritodrine hydrochloride on uterine blood flow was investigated in near-term guinea pigs. The infusion of ritodrine in doses sufficient to inhibit uterine activity provoked tachycardia, and the cardiac output tended to rise. The percentage of cardiac output reaching the gravid uterus did not alter during the infusion of 12 micrograms per minute of ritodrine but decreased from 12% to 10% when the infusion rate was 120 micrograms per minute. There was an increase in the myoendothelial fraction of cardiac output in both instances, whereas the placental fraction decreased at the higher rate of infusion. Perfusion of the myoendothelial tissue improved during the infusion of 12 or 120 micrograms per minute of ritodrine, increasing by 25% and 18% respectively. No significant alteration occurred in the perfusion of the placental tissue.

Dialysis fluid temperature and vasoactive substances during routine hemodialysis

Blood pressure stability is better during cold hemodialysis (HD). This has mainly been attributed to a more pronounced sympathetic activation during cold than during warm HD. The authors studied the effect of dialysate temperature on vasoactive peptides, noradrenaline (NA), and renin (PRA). Ten hemodynamically stable patients were dialyzed for 240 min with each of two dialysate temperatures: 38.5 degrees C (warm HD = WHD) and 34.5 degrees C (cold HD = CHD). A decrease (P < 0.05) in blood pressure occurred during WHD; however, during CHD, blood pressure was stable. There were no differences in vasoconstrictors between the two regimens. There was a decrease in NA (P < 0.05), a tendency of PRA to increase (NS owing to a large statistical spread), while arginine vasopressin was unchanged. During CHD, there was a small increase in neuropeptide Y (NPY); however, during WHD, NPY only tended to increase. However, the relative NPY levels (percent of baseline levels) after WHD and CHD did not differ. The vasodilator response was similar during both treatments. Calcitonin gene related peptide was unaltered. Motilin tended to decrease initially, but then increased (P < 0.05) to baseline levels. An increase occurred in beta-endorphin (P < 0.05) and substance P(P < 0.01). There was an initial rise (P < 0.05) in vasoactive intestinal peptide (VIP), followed by a tendency to decrease during the remainder of treatment. The authors concluded that blood pressure stability was better during CHD. However, this was not reflected by differences in plasma levels of the vasoactive peptides, nor did they find any difference in the sympathetic drive between the two regimens.

Differential effects of nitric oxide synthase modulation on porcine systemic and pulmonary circulation in vivo.

OBJECTIVE To study and compare the effects of inhibiting endothelial nitric oxide synthase on systemic and pulmonary circulation in an in vivo model. DESIGN Prospective, randomized, controlled study. SETTING Laboratory for experimental surgery at a university medical center. SUBJECTS Seventeen anesthetized, mechanically ventilated pigs. INTERVENTIONS To produce a stable and continuous stimulation of endothelial nitric oxide synthase, an infusion of acetylcholine was given to one group of animals (n = 5) in a dose that decreased mean arterial pressure by 15%. After 45 mins, N(G)-monomethyl-L-arginine (L-NMMA) was given in a dose of 3 mg/kg for 5 mins in order to inhibit the enzyme. A second dose of 10 mg/kg was given 30 mins later. L-arginine was then given in a dose of 100 mg/kg to reverse the inhibition. One group of animals (n = 6) received a single dose of indomethacin (2.5 mg/kg) 15 mins after the start of acetylcholine infusion. L-NMMA and L-arginine were then given. In a control group (n = 5), the effects of L-NMMA and L-arginine were studied without acetylcholine. Circulatory parameters were monitored and resistance indices were calculated via arterial, central venous, and pulmonary artery catheters. MEASUREMENTS AND MAIN RESULTS In control animals, 3 and 10 mg/kg of L-NMMA induced an increase in mean arterial pressure of 14% and 25%, respectively, with similar increases in systemic vascular resistance. Mean pulmonary arterial pressure increased by 22% and 48%, respectively. Acetylcholine lowered mean arterial pressure by 15% and did not affect the relative changes induced by L-NMMA. Acetylcholine had no effect on pulmonary resting tone but enhanced the pulmonary hypertension and increase in resistance induced by L-NMMA. This enhancement was abolished by indomethacin, which produced systemic hypertension while no effect on pulmonary pressure was seen. CONCLUSIONS A basal release of nitric oxide contributes to the maintenance of normal vascular tone in the anesthetized pig. Stimulation of endothelial nitric oxide synthase by acetylcholine did not result in any further pulmonary vasodilation as was seen in the systemic circulation. Inhibition of nitric oxide synthase had a greater effect on pulmonary pressure than on systemic pressure. However, this difference was abolished by the administration of indomethacin. Increased nitric oxide release or acetylcholine itself seems to stimulate the production of a vasoconstricting prostanoid in the pulmonary circulation.

Dynamic Pulmonary Accumulation of Labelled Neutrophils by Blood Membrane Contact in the Pig

Nine anesthetized pigs were subjected to short (90 min) sham dialysis with blood membrane contact with the aim to select effects of the artificial surface during dialysis. The importance of the neutrophil (PMN) was investigated by the selective isotope labelling, dynamically followed by gamma-camera imaging and biochemical assays specifically oriented for PMN function. These assays included cell count, PMN aggregation, PMN luminescence, fibronectin and catalase activity. Additionally, pulmonary and systemic hemodynamics and acid base balance were monitored. Sham dialysis induced an accumulation of labelled PMN attaining a maximum between 15 and 17 min. This was coupled with a time-related neutropenia, pulmonary vasoconstriction, increased in vitro PMN aggregation and luminescence response. The neutrophil response abated by the end of dialysis. Cardiac output and arterial blood pressure declined to a steady level after 30 min of sham dialysis. There was an insignificant decrease in catalase activity. All other parameters remained unaltered. The results indicate that PMN accumulates in the pulmonary vessels, in association with neutropenia and activation. The transience of the event points to a protective mechanisms of humoral and/or cellular character.

Beneficial Effect of Cold Dialysate for the Prevention of Hemodialysis-lnduced Hypoxia

Hypoxia occurs frequently during routine hemodialysis (HD). In this study the effect of dialysate temperature on arterial blood gas parameters was investigated. Ten stable HD patients (2 smokers) were dialyzed for 240 min with each of three different dialysate temperatures: 36.5 degrees C (normal temperature HD; NHD), 38.5 degrees C (warm HD; WHD) and 34.5 degrees C (cold HD; CHD). A cuprophane plate dialyzer was used. The ultrafiltration volume was identical in each patient. Arterial blood gas samples were frequently (approximately 10 times/treatment) taken during the dialysis and immediately analyzed. The dialysate temperature significantly affected PaO2 (p < 0.001) but not PaCO2. We also compared the effect of NHD with that of WHD and CHD, respectively, as regards PaO2. NHD and WHD differed significantly p < 0.01), whereas NHD and CHD were not significantly different. However, the relative PaO2 value (% of the baseline value) at the end of CHD (105 +/- 5%) was significantly higher than after both NHD (96 +/- 4%, p < 0.01) and WHD (91 +/- 3%, p < 0.01). In the case of NHD and WHD the fraction of time during which the patients had a PaO2 value below 80 mm Hg was 62 and 64%, respectively. The corresponding figure for CHD was 44%. Arterial oxygen saturation (SaO2) increased during CHD from 95.2 +/- 0.6 to 96.7 +/- 0.6% (p < 0.05), while SaO2 was unchanged during NHD and WHD. The positive effect of CHD was evident in 7 patients. In 1 patient PaO2 was not affected by the dialysate temperature, while in the remaining 2 patients (smokers) a decrease in PaO2 was induced by WHD as well as CHD. A separate statistical analysis with the 2 smokers excluded was performed, which showed that the dialysate temperature significantly affected PaO2 (p < 0.001). A comparison between NHD and CHD showed a significant difference (p < 0.001), whereas NHD and WHD did not differ significantly. When the 2 smokers were excluded from the analysis the fraction of time with a PaO2 value below 80 mm Hg was 60% during NHD and 56% during WHD, but it was reduced to 31% during CHD. In conclusion, despite the existence of interindividual variations most patients seemed to benefit from cold dialysate for the prevention of dialysis-induced hypoxia.

Effect of terbutaline sulphate on ovarian, uterine and maternal placental blood flow in the anaesthetized guinea pig

The effect of terbutaline sulphage, a beta-adrenergic agonist, on uteroplacental blood flow was studied in the guinea pig during late pregnancy. Organ blood flows and cardiac output were assessed by the radioactive microsphere technique. Infusion of terbutaline at a rate of 2nmol min-1 kg-1 body weight caused a significant increase in cardiac output and in the perfusion of the uterus and ovaries, but there was no significant alteration in maternal placental blood flow. When the infusion rate was increased tenfold, there was also a significant rise in the blood supply to the bronchi, urinary bladder, kidneys, and adrenal glands. Maternal placental blood flow was not adversely affected even by this large dose of the drug, although there was a significant reduction in the placental share of the cardiac output.