Endotoxin induces leukocyte transmigration and changes in permeability of the airway epithelium via protein-kinase C and extracellular regulated kinase activation.

Lipopolysaccharide (LPS) endotoxin of Gram-negative bacteria compromises the integrity of the airway epithelial barrier and initiates migration of leukocytes across the epithelium. The goal of the present study was to identify the role of extracellular regulated kinase (ERK1/2) transduction pathways in these processes. The first aim was to determine whether LPS induces ERK1/2 activation and changes in epithelial permeability in epithelial cells alone or only in the presence of immune cells. The second aim was to determine whether the changes in the epithelial permeability were diminished by ERK1/2 blockade. The third aim was to investigate the role of protein kinase C (PKC) activation as an upstream event in activation of ERK1/2. In vitro 20 microg/ml LPS challenge reduced epithelial barrier function, and induced ERK1/2 phosphorylation in primary cultures of bovine tracheal epithelium and in the transformed human airway epithelial cell line, Calu-3. LPS initiated migration of neutrophil-like and monocyte-like transformed HL-60 cell across sheets of Calu-3 cells. The migration rate and the associated changes in the electrical resistance, permeability to albumin, and ERK1/2 phosphorylation were all blocked by calphostin C, the specific blocker of PKC and by PD98059 (2'-amino-3'-methoxyflavone), a selective cell-permeable inhibitor of MAP kinase kinase. In rats, in vivo perfusion of the lumen of an isolated segment of trachea with LPS (0.1 mg/ml) initiated migration of neutrophils and increased the permeability to albumin. Again, these effects were markedly inhibited by PD98059 and calphostin C (by > 50%). We conclude that epithelial ERK1/2 is activated by endotoxin via PKC and is an important pathway in regulation of epithelial permeability.

Effects of the ventilation pattern and pulmonary blood flow on lung heat transfer.

To investigate the relative role of pulmonary perfusion compared to ventilation on lung heat exchange, we determined the effects of blood flow, tidal volume and frequency of ventilation on the rate of lung heat transfer. In anesthetized dogs and isolated, perfused lungs, we investigated the dependence of the overall lung heat transfer coefficient (HTC) and lung thermal capacitance upon ventilation and pulmonary blood flow. The relationship between the HTC and pulmonary blood flow was strongly dependent on ventilation parameters. A distributed model of non-steady-state heat exchange adequately described these observations and demonstrated that changes in pulmonary blood flow may be considered as changes in the effective conductivity of the bronchial walls as 0.4 (0.1) J s(-1)m(-1) K(-1) per (l/min(-1)) of pulmonary blood flow. Our model describes the complex relationship between HTC, ventilation pattern, and effective thermal conductivity of the bronchial walls, all of which present limitations for the use of lung heat transfer to determine pulmonary blood flow.

Effects of pneumoperitoneum on intraoperative pulmonary mechanics and gas exchange during laparoscopic gastric bypass.

BACKGROUND: Hypercarbia and elevated intraabdominal pressure resulting from carbon dioxide (CO2) pneumoperitoneum can adversely affect respiratory mechanics. This study examined the changes in mechanical ventilation, CO2 homeostasis, and pulmonary gas exchange in morbidly obese patients undergoing a laparoscopic or open gastric bypass (GBP) procedure. METHODS: In this study, 58 patients with a body mass index (BMI) of 40 to 60 kg/m2 were randomly allocated to laparoscopic ( n = 31) or open ( n = 27) GBP. Minute ventilation was adjusted to maintain a low normal arterial partial pressure of CO2 (PaCO2), low normal end-tidal partial pressure of CO2 (ETCO2), and low airway pressure. Respiratory compliance, ETCO2, peak inspiratory pressure (PIP), total exhaled CO2 per minute (VCO2), and pulse oximetry (SO2) were measured at 30-min intervals. The acid-base balance was determined by arterial blood gas analysis at 1-h intervals. The pulmonary gas exchange was evaluated by calculation of the alveolar dead space-to-tidal volume ratio (V(Dalv)/V(T)) and alveolar-arterial oxygen gradient (PAO2-PaO2). RESULTS: The two groups were similar in age, gender, and BMI. As compared with open GBP, laparoscopic GBP resulted in higher ETCO2, PIP, and VCO2, and a lower respiratory compliance. Arterial blood gas analysis demonstrated higher PaCO2 and lower pH during laparoscopic GBP than during open GBP ( p < 0.05). The V(Dalv)/V(T) ratio and PAO2-PaO2 gradient did not change significantly during laparoscopic GBP. Intraoperative oxygen desaturation (SO2 < 90%) did not develop in any of the patients in either group. CONCLUSIONS: Laparoscopic GBP alters intraoperative pulmonary mechanics and acid-base balance but does not significantly affect pulmonary oxygen exchange. Changes in pulmonary mechanics are well tolerated in morbidly obese patients when proper ventilator adjustments are maintained.

Tyloxapol attenuates the pathologic effects of endotoxin in rabbits and mortality following cecal ligation and puncture in rats by blockade of endotoxin receptor-ligand interactions.

We have previously demonstrated that the detergent Tyloxapol is effective in preventing reactions to endotoxin. We studied the effects of Tyloxapol on the morbidity and mortality from endotoxemia in rabbits and on the mortality in rats with sepsis. The effects of Tyloxapol on endotoxin binding and macrophage activation were studied in the macrophage cell line RAW264.7 and CHO cells expressing CD14. Isolated human leukocytes were used to study the effects of Tyloxapol on immune reactions, leukocyte motility, and phagocytosis. Intravenous Tyloxapol (200 mg/kg), given prior to or at the time of endotoxin infusion protected rabbits from developing shock. In rats with peritoneal sepsis, a lipid-rich diet and Tyloxapol given at the time of induction of peritonitis protected them from septic death. In vitro, Tyloxapol blocked the binding of endotoxin to murine macrophages and CHO cells expressing CD14, activation of macrophages, and also some antigen-antibody immune reactions (mediated by CD2, CD4, CD22, HLA-DR). Tyloxapol may prevent the reaction to endotoxin by desensitizing endotoxin-recognizing receptors.

Effect of heated and humidified carbon dioxide gas on core temperature and postoperative pain: a randomized trial.

BACKGROUND: Intraoperative hypothermia is a common event during laparoscopic operations. An external warming blanket has been shown to be effective in preventing hypothermia. It has now been proposed that using heated and humidified insufflation gas can prevent hypothermia and decrease postoperative pain. Therefore, we examined the extent of intraoperative hypothermia in patients undergoing laparoscopic Nissen fundoplication using an upper body warming blanket. We also attempted to determine whether using heated and humidified insufflation gas in addition to an external warming blanket would help to maintain intraoperative core temperature or decrease postoperative pain. METHODS: Twenty patients were randomized to receive either standard carbon dioxide (CO2) gas (control, n = 10) or heated and humidified gas (heated and humidified, n = 10). After the induction of anesthesia, an external warming blanket was placed on all patients in both groups. Intraoperative core temperature and intraabdominal temperature were measured at 15-min intervals. Postoperative pain intensity was assessed using a visual analogue pain scale, and the amount of analgesic consumption was recorded. Volume of gas delivered, number of lens-fogging episodes, intraoperative urine output, and hemodynamic data were also recorded. RESULTS: There was no significant difference between the two groups in age, length of operation, or volume of CO2 gas delivered. Compared with baseline value, mean core temperature increased by 0.4 degrees C in the heated and humidified group and by 0.3 degrees C in the control group at 1.5 h after surgical incision. Intraabdominal temperature increased by 0.2 degrees C in the heated and humidified group but decreased by 0.5 degrees C in the control group at 1.5 h after abdominal insufflation. There was no significant difference between the two groups in visual analog pain scale (5.4 +/- 1.6 control vs 4.5 +/- 2.8 heated and humidified), morphine consumed (27 +/- 26 mg control vs 32 +/- 19 mg heated and humidified), urine output, lens-fogging episodes, or hemodynamic parameters. CONCLUSION: Heated and humidified gas, when used in addition to an external warming blanket, minimized the reduction of intraabdominal temperature but did not alter core temperature or reduce postoperative pain.

Cardiac function during laparoscopic vs open gastric bypass.

BACKGROUND: Hypercarbia and increased intraabdominal pressure during prolonged pneumoperitoneum can adversely affect cardiac function. This study compared the intraoperative hemodynamics of morbidly obese patients during laparoscopic and open gastric bypass (GBP). METHODS: Fifty-one patients with a body mass index (BMI) of 40-60 kg/m2 were randomly allocated to undergo laparoscopic (n = 25) or open (n = 26) GBP. Cardiac output (CO), mean pulmonary artery pressure (MPAP), pulmonary artery wedge pressure (PAWP), central venous pressure (CVP), heart rate (HR), and mean arterial pressure (MAP) were recorded at baseline, intraoperatively at 30-min intervals, and in the recovery room. Systemic vascular resistance (SVR) and stroke volume (SV) were also calculated. RESULTS: The two groups were similar in terms of age, weight, and BMI. Operative time was longer in the laparoscopic than in the open group (p < 0.05). The HR and MAP increased significantly from baseline intraoperatively, but there was no significant difference between the two groups. In the laparoscopic group, CO was unchanged after insufflation, but it increased by 5.3% at 2.5 h compared to baseline and by 43% compared to baseline in the recovery room. In contrast, during open GBP, CO increased significantly by 25% after surgical incision and remained elevated throughout the operation. CO was higher during open GBP than during laparoscopic GBP at 0.5 h and at 1 h after surgical incision (p < 0.05). During laparoscopic GBP, CVP, MPAP, and SVR increased transiently and PAWP remained unchanged. During open GBP, CVP, MPAP, and PAWP decreased transiently and SVR remained unchanged. There was no significant difference in the amount of intraoperative fluid administered during laparoscopic (5.5 +/- 1.6 L) and open (5.6 +/- 1.7 L) GBP. CONCLUSION: Prolonged pneumoperitoneum during laparoscopic gastric bypass does not impair cardiac function and is well tolerated by morbidly obese patients.

Pulmonary and bronchial circulations: contributions to heat and water exchange in isolated lungs.

The relative contribution of the pulmonary and bronchial circulatory systems to heat and water exchange in normal lungs was evaluated in 20 isolated, in situ perfused dog lungs and in four patients undergoing elective cardiopulmonary bypass. In isolated dog lungs, if the pulmonary artery was perfused at a nominal flow rate (0.5 l/min), bronchial artery perfusion (up to 70 ml/min) did not significantly affect the expired gas temperature. When the lungs were not perfused through either system, 8 min of ventilation with cool, dry gas decreased the temperature of the expired gas by 6.2 +/- 1.4 degrees C. Selective perfusion of bronchial arteries at 68 +/- 10 mmHg resulted in a mean flow rate of 28 +/- 16 ml/min and increased the average temperature of the expired gas by 0.6 degrees C. An increase in the rate of bronchial arterial perfusion to 55 +/- 14 ml/min increased the average temperature of the expired gas by 1.3 degrees C. The time constant for equilibration of tritiated water between the perfusate and the lung parenchyma was 130 +/- 33 min for pulmonary arterial perfusion and 35 +/- 13 min for combined bronchial and pulmonary perfusion, which indicated that filtration of water from high-pressure bronchial vessels facilitated water exchange in the lung interstitium. The rate of tracer equilibration was similar between the perfusate and gas in both variants of perfusion, but the ratios of tracer gas to perfusate were different (0.42 +/- 0.06 for pulmonary, 0.98 +/- 0.07 for combined), which indicates that bronchial vessels contribute mainly to the hydration of the bronchial mucosa. In humans, the bronchial blood flow was capable of maintaining heat supply after the initiation of cardiopulmonary bypass. Before bypass, when both pulmonary and bronchial blood flow were present, the mean time constant of the temperature decay after a switch to ventilation with cool, dry gas was 35 +/- 12 s. The average temperature difference between the blood and expired gas was 2.4 +/- 0.50 degrees C. After 5 min of dry gas ventilation, the temperature difference between the expired gas and initial blood temperature decreased an average of 3.8 +/- 0.06 degrees C (P < 0.05). The time constant of temperature decay increased to 56 +/- 14 s (P < 0.05). We conclude that bronchial perfusion has a less important role in the temperature balance of the respiratory tract compared with pulmonary arterial perfusion because heat flux is "flow limited" but is important in providing water for hydration of the mucosal surface and interstitial compartments of peribronchial tissues.

Evaluation of core temperature during laparoscopic and open gastric bypass.

BACKGROUND: Intraoperative hypothermia is a common event during open and laparoscopic abdominal surgery. The aim of this study was to compare changes in core temperature between laparoscopic and open gastric bypass (GBP). METHODS: 101 patients with a body mass index (BMI) of 40-60 kg/m2 were randomly assigned to open (n = 50) or laparoscopic (n = 51) GBP. Anesthetic technique was similar for both groups. An external warming blanket and passive airway humidification were used intraoperatively. Core temperature was recorded at preanesthesia, at baseline (after induction) and at 30-min intervals; intra-abdominal temperature was additionally measured at 30-min intervals in a subset of 30 laparoscopic GBP patients. The number of patients who developed intraoperative and postoperative hypothermia (< 36 degrees C) was recorded. Length of operation for both groups and the amount of CO2 gas delivered during laparoscopic operations were also recorded. RESULTS: There was no significant difference between groups with respect to age, gender, mean BMI, and amount of intravenous fluid administered. After induction of anesthesia, core temperature significantly decreased in both groups; 36% of patients in the open group and 37% of patients in the laparoscopic group developed hypothermia. This percentage increased to 46% in the open group and 41% in the laparoscopic group during the operation, and then decreased to 6% in the open group and 8% in the laparoscopic group in the recovery-room. Core temperature increased during the operative procedure to reach 36.5 +/- 0.6 degrees C in the open group and 36.3 +/- 0.5 degrees C in the laparoscopic group at 2.5 hours after surgical incision. Intra-abdominal temperature during laparoscopic GBP was significantly lower than core temperature at all measurement points (p < 0.05). Operative time was longer in the laparoscopic group than in the open group (232 +/- 43 vs 201 +/- 38 min, p < 0.01). Mean volume of gas delivered during laparoscopic GBP was 650 +/- 220 liters. CONCLUSION: Perioperative hypothermia was a common event during both laparoscopic and open GBP. Despite a longer operative time, laparoscopic GBP did not increase the rate of intraoperative hypothermia when efforts were made to minimize intraoperative heat loss.

Comparison of the intubation conditions provided by rapacuronium (ORG 9487) or succinylcholine in humans during anesthesia with fentanyl and propofol.

BACKGROUND: Currently, the only approved muscle relaxant with a rapid onset and short duration of action is succinylcholine, a drug with some undesirable effects. Rapacuronium is an investigational nondepolarizing relaxant that also has a rapid onset and short duration and consequently should be compared with succinylcholine in its ability to facilitate rapid tracheal intubation. METHODS: This prospective, randomized clinical trial involved 336 patients. Anesthesia was induced with fentanyl and propofol and either 1.5 mg/kg rapacuronium or 1.0 mg/kg succinylcholine. The goal was to accomplish tracheal intubation by 60 s after administration of the neuromuscular blocking drug. Endotracheal intubation was performed, and conditions were graded by a blinded investigator. Recovery of neuromuscular function was assessed by electromyography. RESULTS: Intubation conditions were evaluated in 236 patients. Intubation by 60 s after drug administration occurred in 100% of patients with rapacuronium and in 98% with succinylcholine. Intubation conditions were excellent or good in 87% of patients with rapacuronium and in 95% with succinylcholine (P < 0.05). The time (median and range) to the first recovery of the train-of-four response was 8.0 (2.8-20.0) min with rapacuronium and 5.7 (1.8-17.7) min with succinylcholine (P < 0.05). The overall incidence of adverse effects was similar with both drugs. CONCLUSIONS: A 1.5-mg/kg dose of rapacuronium effectively facilitates rapid tracheal intubation. It can be considered a valid alternative to 1.0 mg/kg succinylcholine for this purpose.

Airway thermal volume in humans and its relation to body size.

The objective of this study was to investigate the influence of volume ventilation (VE) and cardiac output (Q) on the temperature of the expired gas at the distal end of the endotracheal tube in anesthetized humans. In 63 mechanically ventilated adults, we used a step decrease in the humidity of inspired gas to cool the lungs. After change from humid to dry gas ventilation, the temperature of the expired gas decreased. We evaluated the relationship between the inverse monoexponential time constant of the temperature fall (1/tau) and either VE or Q. When VE was increased from 5.67 +/- 1.28 to 7.14 +/- 1.60 (SD) l/min (P = 0. 02), 1/tau did not change significantly [from 1.25 +/- 0.38 to 1.21 +/- 0.51 min-1, P = 0.81]. In the 11 patients in whom Q changed during the study period (from 5.07 +/- 1.81 to 7.38 +/- 2.45 l/min, P = 0.02), 1/tau increased correspondingly from 0.89 +/- 0.22 to 1. 52 +/- 0.44 min-1 (P = 0.003). We calculated the airway thermal volume (ATV) as the ratio of the measured values Q to 1/tau and related it to the body height (BH): ATV (liters) = 0.086 BH (cm) - 9. 55 (r = 0.90).

Neuromuscular blocking action of suxamethonium after antagonism of vecuronium by edrophonium, pyridostigmine or neostigmine.

The reported effects of edrophonium on a subsequent dose of suxamethonium are variable and the effects of pyridostigmine have not been evaluated extensively. We have studied this interaction in patients anaesthetized with propofol and sufentanil. After recovery from an initial bolus (1 mg kg-1) of suxamethonium, vecuronium was infused to produce 75% block. After 30 min, the infusion was discontinued and saline 5 ml, edrophonium 0.75 mg kg-1, pyridostigmine 0.24 mg kg-1 or neostigmine 0.05 mg kg-1 was given. Fifteen minutes later the mean durations of a second bolus of suxamethonium were: 10.5 (SD 3.9) min (saline), 10.9 (3.7) min (edrophonium), 18.7 (5.4) min (pyridostigmine) and 23.8 (7.4) min (neostigmine). Corresponding plasma cholinesterase activities (percentage of baseline) were: 91 (18), 87 (9), 21 (10) and 52 (26). When both treatment groups and individual patients were compared, the changes in duration of action did not correlate with changes in cholinesterase activity. These data suggest that other mechanisms in addition to cholinesterase inhibition may contribute to this drug interaction.

Cholinesterase inhibitors do not prolong neuromuscular block produced by mivacurium.

Cholinesterase inhibitors antagonize neuromuscular block produced by mivacurium, but some may also decrease its metabolism by inhibiting pseudocholinesterase. These opposing interactions were examined in rats anaesthetized with pentobarbitone. After spontaneous recovery from an initial bolus dose of 0.03 mg kg-1, mivacurium was infused to produce 80-90% block of gastrocnemius muscle twitch. After 15 min, the infusion was discontinued and saline, edrophonium, pyridostigmine or neostigmine was administered. Fifteen minutes later, a second bolus dose of mivacurium was given. Edrophonium, pyridostigmine and neostigmine reduced the subsequent maximum block, compared with the change in saline control, by 3%, 19% and 35%, respectively. Correspondingly, the time to recovery of T1 to 50% was decreased by 20%, 58% and 62%. In rats, acetylcholinesterase-mediated antagonism of neuromuscular block predominated over decreased pseudocholinesterase-mediated metabolism, such that prior administration of a cholinesterase inhibitor did not prolong the neuromuscular blocking effects of mivacurium.

Effects of cholinesterase inhibitors on the neuromuscular blocking action of suxamethonium.

Prolonged neuromuscular block occurs when suxamethonium is given after neostigmine or pyridostigmine; however, studies of edrophonium and suxamethonium have yielded conflicting results. We have studied, therefore, interactions between suxamethonium and all three anticholinesterases in rats anaesthetized with pentobarbitone. After recovery from an initial bolus of suxamethonium, saline, edrophonium, pyridostigmine or neostigmine was administered and a second dose of suxamethonium was then given. All three anticholinesterases prolonged the duration of neuromuscular block (90% suppression to 50% twitch recovery) to 127 (SEM 9)%, 127(10)% and 138 (11)% of baseline for edrophonium, pyridostigmine and neostigmine, respectively. Recovery index (25% to 75% twitch recovery) was increased also to 125 (9)%; 149 (10%) and 185 (15)% of baseline, respectively for the three drugs.

Acute effects of calcitonin gene-related peptide on the mechanical and electrical responses of the rat hemidiaphragm.

Calcitonin gene-related peptide (CGRP) acutely augments the contractile response of skeletal muscle to both direct and indirect stimulation. However, studies in whole muscle tissues have produced extremely variable results. To determine if differences in stimulation parameters are the source of this variation, the effects of increasing stimulus duration were evaluated in the isolated perfused rat hemidiaphragm. The effect of CGRP on the maximum force of contraction (Fc) was dependent upon the stimulus duration. CGRP (10(-6) M) produced an 11% increase in Fc with a 0.1-msec duration stimulus and a 32% increase with a 1.0-msec stimulus. In contrast, CGRP decreased the time for twitch relaxation and this effect was independent of stimulus duration. Additional studies demonstrated that CGRP shortened the relative refractory period. Compound muscle action potential recordings revealed that stimulus durations greater than 0.5 msec produced a second peak of electrical activity with an associated increased Fc. The effects of CGRP on Fc corresponded to the effects of CGRP on this second peak of compound muscle action potential activity. We conclude that, by decreasing the relative refractory period of the muscle tissue, CGRP enhances the potential for repetitive stimulation with commonly used experimental parameters.

Effects of succinylcholine on the pharmacodynamics of pipecuronium and pancuronium.

To study the effects of succinylcholine on subsequent pharmacodynamics of nondepolarizing muscle relaxants, a comparative pharmacodynamic study was carried out in patients having balanced anesthesia (thiopental, fentanyl, nitrous oxide/oxygen) in whom equipotent doses of pipecuronium (80 micrograms/kg) and pancuronium (100 micrograms/kg) were given with or without prior administration of succinylcholine (1 mg/kg). Fifty-two patients were randomly assigned to one of the following four groups: 1, pancuronium (100 micrograms/kg); 2, pipecuronium (80 micrograms/kg); 3, succinylcholine (1 mg/kg) plus pancuronium (100 micrograms/kg); and 4, succinylcholine (1 mg/kg) plus pipecuronium (80 micrograms/kg). In groups 3 and 4, the nondepolarizing relaxant was given after succinylcholine when the twitch height recovered to 75% of its control value. For maintenance of neuromuscular blockade, additional increments of pancuronium (20 micrograms/kg) or pipecuronium (15 micrograms/kg) were given. Neuromuscular function was monitored throughout induction, maintenance, spontaneous recovery, and pharmacologic reversal of the neuromuscular block. Mean onset times for pancuronium (group 1) and pipecuronium (group 2) given without succinylcholine were (mean +/- SEM) 2.5 +/- 0.3 and 2.8 +/- 0.2 min, respectively. Mean onset times (times to maximum twitch depression) of the two drugs given after succinylcholine (groups 3 and 4) were significantly shorter (1.4 +/- 0.4 and 1.6 +/- 0.1 min, respectively). Clinical durations (i.e., until 25% twitch recovery of pancuronium and pipecuronium) were not significantly different among the four groups, varying from 81.1 +/- 5.4 (group 4) to 107.0 +/- 17.0 (group 2) min.(ABSTRACT TRUNCATED AT 250 WORDS)

Mechanisms of respiratory failure produced by neostigmine and diisopropyl fluorophosphate.

Acetylcholinesterase inhibitors produce diverse physiologic effects, but lethal exposure consistently produces respiratory failure due to neuromuscular paralysis or depression of respiratory control centers in the medulla. Simultaneous measurement of gastrocnemius muscle contraction and efferent phrenic nerve activity was used to determine the primary cause of respiratory failure produced by neostigmine and diisopropyl fluorophosphate (DFP) in anesthetized cats. Both neostigmine and DFP abolished phrenic nerve activity prior to producing neuromuscular blockade. Furthermore, neostigmine did not alter brain acetylcholinesterase activity and pretreatment with either atropine methylbromide or atropine increased the dose of neostigmine required to abolish phrenic nerve activity. In contrast, DFP abolished brain cholinesterase activity and only atropine inhibited its respiratory effects. Despite the loss of efferent phrenic nerve activity, there is no evidence of a direct effect of neostigmine on respiratory control centers. Neostigmine may instead alter afferent inputs which modulate respiration to produce a reflex respiratory failure.