Late-phase bronchial vascular responses in allergic sheep.

Sheep were classified on the basis of their airway response to Ascaris suum antigen aerosols as allergic or nonsensitive. Allergic sheep were classed as acute or dual responders. Acute responders had only an immediate increase in mean airflow resistance after antigen, whereas dual responders had an immediate and late-phase (6-8 h after antigen challenge) increase in mean airflow resistance; nonsensitive sheep had minimal airway responses to antigen (less than 30% increase from base line). The sheep were anesthetized 2 wk later and, after a left thoracotomy, were challenged with antigen to determine bronchial vascular responses; bronchial artery blood flow was measured with an electromagnetic flow probe. Airway responses to antigen aerosol challenge were similar in the anesthetized and conscious animals. The mean fall in bronchial vascular resistance (BVR) immediately after antigen challenge was similar in acute and dual responders (41 +/- 7 and 47 +/- 9% of base line, respectively). In dual responders, late-phase airway responses were preceded by a significant increase from base line in Qbr and a fall in bronchovascular resistance (BVR). The mean fall in BVR 6-8 h after antigen challenge in documented dual responders was significantly different from bronchial vascular responses in acute responders (59 +/- 3 vs. 89 +/- 10%, respectively). Sheep without airway responses to A. suum had no significant changes in bronchial hemodynamics or airways mechanics. Late-phase-associated changes in BVR are a specific response to antigen challenge and may be a sensitive index of mediators being released.(ABSTRACT TRUNCATED AT 250 WORDS)

Removal of excessive bronchial secretions by asymmetric high-frequency oscillations.

The present study evaluated whether high-frequency oscillations (HFO) with biased flow profiles applied at the airway opening are capable of altering mucus clearance. In eight anesthetized sheep, artificial mucus (100 P) was infused continuously (1 ml/min) into the left main bronchus via a cannula inserted through the dorsal wall of the left main bronchus after thoracotomy. Outcoming mucus was collected every 10 min from the end of a cuffed orotracheal tube. Animals were ventilated with a Harvard respirator at a low frequency with superimposed HFO at 14 Hz with asymmetrical waveforms generated by a digitally controlled electromagnetic piston pump (expiratory bias: peak expiratory flow 3.8 l/s, peak inspiratory flow 1.3 l/s; inspiratory bias: reverse of expiratory bias). The influence of posture and of HFO airflow bias on mucus clearance was determined. In the horizontal position, mucus clearance with expiratory biased HFO was 3.5 +/- 2 (SD) ml/10 min. Head-down tilt produced a clearance of 3.1 +/- 3 ml/10 min; addition of HFO with expiratory bias increased clearance to 11.0 +/- 2.0 ml/10 min (P less than 0.05). No clearance occurred with inspiratory biased HFO during head-down tilt. These results indicate that expiratory biased HFO at the airway opening can clear excessive airway secretions and augment clearance by postural drainage.

Measurement of airway mucosal perfusion and water volume with an inert soluble gas.

The purpose of this study was to develop and validate a new in vivo technique for the measurement of tracheal mucosal blood flow (Qtr) and tissue water volume (VH2O) with an inert soluble gas. The technique was based on the notion that the uptake of dimethyl ether (VDME) from an isolated tracheal segment is governed by VH2O (transient state) and Qtr (steady state). In lightly anesthetized sheep, an endotracheal tube with two cuffs placed 14.5-16.5 cm apart was placed to create a chamber into which dimethyl ether was introduced and from which VDMME into the mucosa was determined with a sensitive pneumotachograph. Mean Qtr was 1.20 ml/min (range 0.87-1.73), and mean VH2O was 1.67 ml (range 1.27-2.26). Qtr correlated with cardiac output but not with body weight or tracheal mucosal surface area determined by He dilution. VH2O did not show a correlation with any of these parameters. The response to selected pharmacological agents suggested that the measurements of Qtr and VH2O are independent of each other and from changes in tracheal diameter. Mean Qtr was 80% of mean tracheal mucosal blood flow measured with radiolabeled microspheres. We concluded that the inert soluble gas method is capable of measuring in vivo the perfusion and a water compartment of the intact tracheal mucosa.

Independent control of mucosal and total airway blood flow during hypoxemia.

In the larger airways, the blood circulation forms a subepithelial (mucosal) and outer (peribronchial) microvascular network. This raises the possibility that blood flow in these two networks is regulated independently. We used hypoxemia as a stimulus to induce changes in tracheal mucosal blood flow normalized for systemic arterial pressure (Qtr n) measured with an inert soluble gas technique and total bronchial blood flow (Qbr) and normalized Qbr (Qbrn) measured with an electromagnetic flow probe in anesthetized sheep. Fifteen minutes of hypoxemia [PO2 40 +/- 7 (SD) Torr] decreased mean Qtr n from 1.1 +/- 0.4 to 0.8 +/- 0.4 ml.min-1.mmHg-1.10(2) (-27%; P less than 0.05; n = 7) and increased mean Qbr n from 12.1 +/- 3.2 to 17.1 +/- 5.4 ml.min-1.mmHg-1.10(2) (+41%; P less than 0.05; n = 6). The rise in Qbr correlated with cardiac output (r = 0.68; P less than 0.05). Phentolamine pretreatment (0.1 mg/kg iv) blunted the hypoxemia-related decrease of mean Qtr n (-8%; P = NS). Tyramine (2.5 mg) applied locally to the trachea decreased mean Qtr n significantly after 30 and 45 min by 31 and 19%, respectively (P less than 0.05). 6-Hydroxydopamine (0.2 mg 4 times for 1 h locally applied) prevented the hypoxemia-induced as well as local tyramine-induced decrease in mean Qtr n (0 and 0%).(ABSTRACT TRUNCATED AT 250 WORDS)

Modification of bronchial blood flow during allergic airway responses.

Ascaris suum antigen effects on mean airflow resistance (RL) and bronchial arterial blood flow (Qbr) were studied in allergic anesthetized sheep with documented airway responses. Qbr was measured with electromagnetic flow probes, and supplemental O2 prevented antigen-induced hypoxemia. Aerosol challenge with this specific antigen increased RL and Qbr significantly. Cromolyn sodium aerosol pretreatment prevented antigen-induced increases in RL but not in Qbr. Intravenous cromolyn, however, prevented increases in Qbr and RL, suggesting a role for mast cell degranulation in both bronchomotor and bronchovascular responses to antigen. Antigen-induced increases in Qbr were not solely attributable to histamine release. Indomethacin pretreatment attenuated the antigen-induced increase in Qbr, thus suggesting that vasodilator cyclooxygenase products contribute to the vascular response. Antigen challenge significantly decreased Qbr after indomethacin and metiamide pretreatment, which suggests that vasoconstrictor substances released after antigen exposure also modulate Qbr; however, released vasodilators overshadow vasoconstrictor effects. Thus antigen challenge affects Qbr by locally releasing histamine and vasodilator prostaglandins as well as vasoconstrictor substances. These effects were independent of antigen-induced changes in systemic and pulmonary hemodynamics.

Effects of histamine on bronchial artery blood flow and bronchomotor tone.

The effects of aerosolized 5% histamine (10 breaths) on bronchial artery blood flow (Qbr), airflow resistance (RL), and pulmonary and systemic hemodynamics were studied in mechanically ventilated sheep anesthetized with pentobarbital sodium. Histamine increased mean Qbr and RL to 252 +/- 45 and 337 +/- 53% of base line, respectively. This effect was significantly different from base line for 30 min after challenge. The histamine-induced increase in RL was blocked by pretreatment with the histamine H1 receptor antagonist, chlorpheniramine, whereas the histamine-induced elevation in Qbr was prevented by the H2 antagonist, metiamide. Both responses were blocked only when both antagonists were present. Changes in Qbr were not directly associated with alterations in systemic and pulmonary hemodynamics or arterial blood gas composition. In vitro histamine caused a dose-dependent contraction of ovine bronchial artery strips that was prevented by H1 antagonist. The H2 agonist, impromidine, caused relaxation of precontracted arterial strips and was more potent and efficacious than histamine, whereas H1 agonists failed to elicit a relaxant response. Thus these findings indicate that histamine aerosol induces a vasodilation in the bronchial vascular bed; histamine has a direct effect on Qbr that is independent of alterations in RL, systemic and pulmonary hemodynamics, or arterial blood gas composition; and, histamine-induced bronchoconstriction is mediated predominantly by H1-receptors, whereas increased Qbr is controlled predominantly by H2-receptors, probably located in resistance vessels. This local effect of histamine on Qbr may have important implications in the pathophysiology of bronchial asthma and pulmonary edema.

Environmental radon daughters reveal pathognomonic changes in the brain proteins and lipids in patients with Alzheimer's disease and Parkinson's disease, and cigarette smokers.

This paper presents an investigation of the retention of environmental radon daughters, 210Po (alpha particle emitting radio-nuclide) and 210Bi (beta particle emitting radio-nuclide), in lipid and protein fractions of the cortical grey and subcortical white matter from the frontal and temporal brain lobes of patients who had suffered from Alzheimer's disease or Parkinson's disease, of cigarette smokers, and of control subjects. 210Po and 210Bi radioactivity increased tenfold in the cortical grey and subcortical white protein fraction in patients with Alzheimer's disease and smokers, and tenfold in the cortical grey and subcortical white lipid fraction in patients with Parkinson's disease. Free radicals generated by radon daughters may add to the severity of the radio-chemical injury to the brain astrocytes. The pathognomonic distribution of radon daughters to lipids in patients with Parkinson's disease and to proteins in patients with Alzheimer's disease was attributed to high chlorine affinity of radon daughters. The changes in the membrane protein pores, channels, and gates in patients with Alzheimer's disease and in the lipid bilayer in patients with Parkinson's disease are at the core of what the authors think are two systemic brain diseases.

Contribution of viable and nonviable heart myocytes to substrate oxidation.

An approach is described that allows metabolic data obtained from a mixture of vital dye-excluding (T-) and nonexcluding (T+) myocytes to be extrapolated to a homogeneous cell population. Myocytes from adult dog hearts were dispersed by enzymatic treatment and separated into two fractions: one containing predominantly T-, and the other containing predominately T+ cells. Measuring the oxidation rate and viability of each fraction allows the determination of the rate of oxidation of a homogeneous cell population when palmitate, glucose, or lactate is the oxidizable substrate. The calculated rate of oxidation of these substrates by 100% T- cells was: 0.15, 0.46, and 2.99 nmol . mg protein-1 . min-1, respectively. Oxidation of palmitate and lactate by T+ cells was one-fifth of the T-cell rate. Glucose oxidation of T+ cells was not significantly different from zero. Use of this procedure will permit study of myocardial metabolism when experimental procedures may cause altered cell viability.

Influence of Escherichia coli endotoxin on palmitate, glucose, and lactate utilization by isolated dog heart myocytes.

The in vitro effects of E. coli 0127:B8 endotoxin on the oxidation of palmitate, glucose, and lactate by isolated, beating, adult dog heart myocytes were studied. The result were correlated with the in vitro effect of endotoxin on the transport of palmitate and glucose by using their respective analog, hexadecanol and 3-O-methyl-D-glucose. Endotoxin decreased palmitate but increased glucose and lactate oxidation in a dose-dependent manner. The changes in palmitate and lactate oxidation induced by endotoxin could not be corrected by addition of L-carnitine to the reaction mixture. The transport of hexadecanol or of 3-O-methyl-D-glucose was not affected by endotoxin. These results suggest that the endotoxin-induced alteration in palmitate and glucose oxidation by the heart is not the result of an impairment in metabolite transport, but rather a defect in the subsequent intracellular oxidative processes.

Bronchial circulation in asthma.

Whereas the anatomical changes of the bronchial circulation in response to a wide variety of congenital and acquired cardiopulmonary diseases have been well described, little is known about its functional response. There is growing evidence that the bronchial circulation plays a major role in the pathophysiology of hyperreactive airway disease. The bronchial vascular system appears to be involved in mediator transport to and from target tissues in the airway wall, in the development of airway wall edema which may contribute to airflow obstruction, and in heat and water exchange in the tracheobronchial tree. Although our current understanding of these functions is rather sketchy, enough is known to outline the contributions of the bronchial, i.e. the systemic circulation to the mechanisms of bronchial asthma.

Mobilization of mucus by airway oscillations.

The effects of high frequency asymmetric airway oscillations on mucus clearance were evaluated in excised tracheas of sheep, in an animal model of excessive mucus production, and in patients with bronchiectasis. Asymmetric high frequency ventilation (15 Hz) with expiratory biased flow profiles (expiratory peak-flow greater than inspiratory peak-flow) could move mucus droplets towards the pharynx in rigid and flexible tracheas by gas-liquid interaction. In rigid tracheas the mucus was transported towards the periphery of the model lung if the oscillations were inspiratory biased. In very collapsible tracheas, however, even inspiratory biased oscillations moved the mucus cephalad. Parameters influencing direction and speed of mucus are airflow profile, peak-flow, airway compliance and lung resistance. Gamma-camera studies showed that in anesthetized dogs radiolabeled artificial mucus followed the direction of the bias during high frequency ventilation. In five human volunteers with bronchiectasis and excessive secretions the asymmetric airway oscillations were superimposed during spontaneous breathing using a mouthpiece. Airway wall vibrations following the pressure swings of the oscillator could be observed. During forced expiration inward bulging of the posterior membranes of trachea and bronchi occurred at the negative pressure phase of the oscillations. This event was associated with increased appearance of sputum in the central airways. We conclude that high frequency ventilation with asymmetric flow profiles applied via tube or mouthpiece might be an effective future treatment of mucostasis.

Effects of insulin on cardiac lysosomes and protein degradation.

Hearts perfused in the absence of added insulin had 1) accelerated rates of protein degradation, as assessed by release of phenylalanine and tyrosine; 2) increased rates of release of seven other amino acids; 3) decreased lysosomal latency and sedimentable lysosomal enzyme activity; 4) increased numbers of autophagic vacuoles in cardiac muscle cells; and 5) decreased activity of beta-N-acetylglucosaminidase in dense lysosomes (1.06-1.09 g/ml), as compared to hearts perfused in the presence of the hormone. After 3 h of perfusion in the absence of insulin, the changes that developed in protein degradation, lysosomal latency, and sedimentability, and in enzyme activity in dense lysosomes, were reversed by insulin addition during 90 min of subsequent perfusion. These studies suggest a role for insulin in controlling the activity of the lysosomal system and the involvement of this system in protein degradation, particularly in insulin-deprived tissue.

Effects of diabetes on cardiac lysosomes and protein degradation.

Rates of proteolysis by hearts obtained from alloxan diabetic rats and perfused as working preparations with buffer simulating control sera were accelerated 30% above identically perfused control hearts. The total homogenate activities of cathepsin D and beta-N-acetylglucosaminidase, assayed in the presence of Triton X-100, decreased 15-35% in diabetic heart, but the activities of these lysosomal enzymes assayable in the absence of detergent were unchanged in the diabetic tissue. The effects of diabetes were examined further by centrifugation of particulate fractions from subtilisin-treated hearts of control and diabetic rats on polyvinylpyrrolidone-coated colloidal silica (Percoll) gradients. Two species of lysosomes were resolved on the basis of their densities. Both dense and buoyant lysosomes accumulated radioactivity when hearts were exposed to [14C]phenylalanine methyl ester. Dense lysosomes (1.06-1.09 g/ml) sedimented with mitochondria while buoyant lysosomes banded with Golgi and sarcolemmal particles (1.02-1.03 g/ml). When particulate fractions of hearts from diabetic animals were layered on the Percoll gradients, total activities of beta-N-acetylglucosaminidase and cathepsin D were decreased from control in buoyant lysosomes, but unchanged in dense lysosomes. These results demonstrated that the increase in proteolysis in the diabetic heart was associated with decreased total activity and latency of cathepsin D and beta-N-acetylglucosaminidase and an increased proportion of dense lysosomes in the particulate fraction.

Effects of amino acid methyl esters on cardiac lysosomes and protein degradation.

Perfusion of rat hearts as Langendorff preparations for 20 min with buffer containing 15 mM glucose and either 10 mM methionine methyl ester or 10 mM leucine methyl ester decreased 1) latency of beta-N-acetylglucosaminidase, 2) buoyant density of cardiac lysosomes, 3) the rate of proteolysis, and 4) heart rate and coronary flow. A significant inhibitory effect on proteolysis was not observed during this brief exposure of insulin-treated hearts to the methyl esters. The effects of the methyl esters were reversible when they were washed away. Heart rate and coronary flow returned to control values within 5 min. After 90 min of recovery, lysosomes distributed on Percoll gradients in dense and buoyant bands that were indistinguishable from control hearts. The reversibility of these effects correlated with the net release of either methionine or leucine from hearts exposed to the corresponding methyl ester. During recovery, rates of proteolysis returned toward those observed in control hearts. In insulin-treated hearts, methionine methyl ester inhibited proteolysis during the recovery period, but leucine methyl ester had no effect. These results indicate that exposure to amino acid methyl esters led to swelling of cardiac lysosomes and inhibition of protein degradation. These effects appeared to be related to the amount of free amino acid that was retained.

The effects of high-dose corticosteroids in patients with septic shock. A prospective, controlled study.

To determine whether corticosteroids are efficacious in severe septic shock, we conducted a prospective study of 59 patients randomly assigned to a methylprednisolone, dexamethasone, or control group. Patients were treated 17.5 +/- 5.4 hours (mean +/- S.E.M.) after the onset of shock, and 55 patients required vasopressor agents. Early in the hospital course, reversal of shock was more likely in patients who received corticosteroids than in those who did not. Four (19 per cent) of 21 methylprednisolone-treated, 7 (32 per cent) of 22 dexamethasone-treated, and none of 16 control patients had reversal of shock 24 hours after drug administration (corticosteroid groups vs. control group, P less than 0.05). Patients treated with corticosteroids within four hours after the onset of shock had a higher incidence of shock reversal (P less than 0.05). At 133 hours after drug administration, 17 (40 per cent) of 43 corticosteroid-treated patients had died, and 11 (69 per cent) of 16 control patients had died (P less than 0.05). However, these differences in reversal of shock and survival disappeared later in the course. Overall, 16 (76 per cent) of 21 patients receiving methylprednisolone, 17 (77 per cent) of 22 patients receiving dexamethasone, and 11 (69 per cent) of 16 controls in the hospital died. We conclude that corticosteroids do not improve the overall survival of patients with severe, late septic shock but may be helpful early in the course and in certain subgroups of patients.

Complement activation in septic shock patients.

To evaluate the status of the complement system and to determine the effects of corticosteroids on complement component levels in septic shock, C3, C4, and Factor B were measured in 42 patients with severe late septic shock. Serum levels of C4 and Factor B correlated with C3 levels (r = 0.48 and 0.64, respectively; p less than .01) in patients in shock for more than 4 h, but only Factor B correlated with C3 (r = 0.85; p less than .01) in patients in shock for 4 h or less. C3 and Factor B levels were significantly (p less than .05) lower in patients who died (12,174 +/- 1,524 CH50 U/ml and 14 +/- 1 mg/dl, respectively) than in patients who survived (18,418 +/- 2,833 CH50 U/ml and 21 +/- 2 mg/dl, respectively). Corticosteroids did not alter complement component levels. The alternative pathway appears to be activated early in septic shock, whereas the classical pathway is activated later. C3 and Factor B levels may predict survival of patients in septic shock. In this study, corticosteroids did not change the complement component levels of patients in late severe septic shock.

Distribution of proteins in pulmonary edema. The value of fractional concentrations.

Pulmonary edema fluid (PEF) and serum (S) were obtained from 14 patients with cardiac pulmonary edema (CPE) and 25 noncardiac pulmonary edema (NCPE) patients. The type of pulmonary edema was based on a clinical classification. The protein content of PEF was not significantly different between CPE (3.89 +/- 0.25 g/dl, mean +/- SEM) and NCPE (4.35 +/- 0.34 g/dl) patients, but the protein content of serum was different (7.18 +/- 0.27 versus 5.13 +/- 0.23, respectively, p less than 0.001). As expected then, the PEF/S ratio was greater in NCPE than in CPE (0.85 +/- 0.06 versus 0.54 +/- 0.03, respectively, p less than 0.05). Thus, differences in the PEF/S ratios in CPE as compared with those in NCPE are independent of mean edema fluid protein content and dependent on differences in serum protein content. PEF and S proteins were separated into 9 fractions of increasing molecular radius by electrophoresis, and fractional concentrations (percent of total protein content) were calculated. The fractional concentrations of these combined fractions of serum proteins were not different between CPE and NCPE. Fractional PEF/S ratios were significantly greater for combined fractions I-III (p less than 0.01) in CPE than in NCPE and significantly less in fractions VII-IX (p less than 0.01). In CPE, a higher percentage of proteins with smaller molecular radii (45 A) enter the airway compartment in part because of higher hydrostatic pressures, whereas in NCPE, larger proteins enter the airways consequent to increased permeability to proteins with molecular radii greater than 72 A.(ABSTRACT TRUNCATED AT 250 WORDS)

Environmental lead-210 and bismuth-210 accrue selectively in the brain proteins in Alzheimer disease and brain lipids in Parkinson disease.

We studied the occurrence of the environmental radon daughters, 210Po (alpha particles), and 210Bi (beta particles), in the protein and lipid fractions of cortical gray and subcortical white matter from the frontal and temporal lobes of human brains of persons with Alzheimer disease (AD), persons with Parkinson disease (PD), smokers, or persons with no previous evidence of clinical neurologic disease (controls). We found a 10-fold increase in 210Po and 210Pb radioactivity in the protein fraction from both the cortical gray and subcortical white matter in AD and smokers, and a similar increase in the lipid fraction in PD. The pathognomonic distribution of the radon daughters to the lipids in PD and to the proteins in AD was inferred to reflect the increase of local chlorine availability to which radon daughters bound selectively. Cigarette smoking strongly increases radon daughter retention in the central nervous system.