Effect of positive airway pressure on capillary transit time in rabbit lung

We used fluorescence videomicroscopy to measure the passage of fluorescent dye through the subpleural microcirculation of the lung. With the rabbit in the left lateral decubitus position, the subpleural microcirculation was viewed either through a transparent parietal pleural window located in the superior part of the chest or directly with the chest open. There was no physical contact with the chest or lung. The rabbit was anesthetized, paralyzed, and mechanically ventilated with 100% O2. The dye was injected into the right ventricle during a 2-min apneic period to eliminate lung movement due to ventilation. The video signal of the passage of the dye was analyzed frame by frame by use of digital image processing to compensate for cardiogenic oscillations of the lung surface. Gray scale levels of an arteriole and adjacent venule were measured every 1/30 s. Capillary transit time was determined from the difference between the concentration-weighted mean time values of the arteriolar and venular dye dilution curves. We studied the effect of airway pressure (0-20 cmH2O) on transit time. Cardiac output was measured at different airway pressures by the thermal dilution technique. Capillary transit time averaged 0.60 s at functional residual capacity. Right ventricular-to-arteriolar transit time was four times as large as the capillary transit time. An increase in airway pressure from 0-5 to 20 cmH2O resulted in a fourfold increase in both capillary and arterial transit times and a threefold decrease in cardiac output.     

Effects of lung inflation and blood flow on capillary transit time in isolated rabbit lungs.

In a previous study, direct measurements of pulmonary capillary transit time by fluorescence video microscopy in anesthetized rabbits showed that chest inflation increased capillary transit time and decreased cardiac output. In isolated perfused rabbit lungs we measured the effect of lung volume, left atrial pressure (Pla), and blood flow on capillary transit time. At constant blood flow and constant transpulmonary pressure, a bolus of fluorescent dye was injected into the pulmonary artery and the passage of the dye through the subpleural microcirculation was recorded via the video microscope on videotape. During playback of the video signals, the light emitted from an arteriole and adjacent venule was measured using a video photoanalyzer. Capillary transit time was the difference between the mean time values of the arteriolar and venular dye dilution curves. We measured capillary transit time in three groups of lungs. In group 1, with airway pressure (Paw) at 5 cmH2O, transit time was measured at blood flow of approximately 80, approximately 40, and approximately 20 ml.min-1.kg-1. At each blood flow level, Pla was varied from 0 (Pla less than Paw, zone 2) to 11 cmH2O (Pla greater than Paw, zone 3). In group 2, at constant Paw of 15 cmH2O, Pla was varied from 0 (zone 2) to 22 cmH2O (zone 3) at the same three blood flow levels. In group 3, at each of the three blood flow levels, Paw was varied from 5 to 15 cmH2O while Pla was maintained at 0 cmH2O (zone 2).(ABSTRACT TRUNCATED AT 250 WORDS)     

Moments of physiological transit time distributions and the time course of drug disposition in the body

Characterizing the tissue distribution kinetics of drugs by physiological and physico-chemical parameters and using a circulatory model the time course of blood concentration after intravenous injection is predicted for linear pharmacokinetic systems. The interrelationships between the first three (zero to second) moments of the distribution functions of organ transfer times, circulation times and residence times of drug molecules in the body are described. Utilizing literature data the model is applied to the analysis of lidocain kinetics in humans.     

Maternal separation and gastrointestinal transit time in neonate rats

Gastrointestinal transit times (GItts) were compared in separate litters of 10- and 15-day-old Sprague Dawley rats using barium sulphate. By tracking the leading front of the bolus on radiographs, the gastrocaecal transit times in pups were estimated. To measure the total GItt, the duration from orogastric gavage until an observable defecation of barium sulphate was recorded. The gastrocaecal times for 10-day-old pups maintained with their dam (n = 5) ranged from 4-5 h and those removed from the dam ranged from 2.5-5 h. For 15-day-old pups with their dam (n = 6) and without dam (n = 5), gastrocaecal times ranged from 4-6 h and 3.5-5 h, respectively. Ten-day-old pups that remained with the dam had a GItt of 13.8 ± 0.9 h and those kept in the absence of the dam had a time of 9.3 ± 0.7 h. This decrease (P < 0.05) in GItt in the absence of the dam was age-dependent in 10-day-old pups, and was not observed (P > 0.05) in 15-day-old pups. The results provide a basis, for the design of future studies involving neonate rat metabolism, to include maternal presence.     

Capillary electrophoresis of glutamate and aspartate in rat brain dialysate Improvements in detection and analysis time using cyclodextrins

Addition of cyclodextrins (CDs) to the electrolyte buffer in the capillary zone electrophoresis (CZE) separation of derivatized amino acids was evaluated in terms of fluorescence signal enhancement, resolution, and migration time effects. Maximum fluorescence signal enhancement was observed with separation buffers containing 4M β-cyclodextrin or 10 mM hydroxypropyl β-cyclodextrin. Resolution values decreased as the CD concentrations increased. Migration times were dependent on CD concentration. Inclusion complex formation constants calculated using changes in migration time showed slight agreement with those calculated by the steady-state fluorescence enhancement technique. Analysis of 20 μl of rat brain microdialysate by CZE using 4 mM β-cyclodextrin in borate buffer resulted in baseline resolution of glutamate and aspartate in 3.6 min. The results of this work indicate that, when used as separation buffer additives, cyclodextrins are capable of increasing the fluorescence signal and decreasing the migration times of NDA-derivatized acidic amino acids.     

Prolonged small-intestinal transit time in cystic fibrosis.

A lactulose hydrogen breath test was performed on 10 patients with cystic fibrosis and 15 control subjects matched for age and sex. All normal subjects had a fasting breath hydrogen concentration of less than 20 ppm. In contrast, seven of the patients with cystic fibrosis had high concentrations (25-170 ppm), which fell to 20 ppm or below on prolonged fasting (14-23 hours). Two patients showed no rise in breath hydrogen concentrations after lactulose, and in one patient the breath hydrogen concentration rose at 15 minutes, suggesting bacterial colonisation of the small bowel. Seven of the patients had prolonged small-bowel transit times (160-390 minutes) compared with those in the control group (50-150 minutes).     

Changes in the rat sperm head during epididymal transit

Morphological changes in sperm are one aspect of a maturation process during epididymal transit in marnrnals. The literature mentions only, for different strains of rats, a remodeling and decrease in size of the acrosome. In the present work, the sperm were obtained from caput, corpus, and cauda epididymis of the albino rat. Samples were processed for scanning electron microscopy, with routine techniques, and for light microscopy and video microscopy. It appeared, with these techniques, that the acrosomal curvature and the whole head surface area of the rat sperm decrease during the epididymal transit. To measure these changes, a geometrical method was designed, and surface measurements were made using a computer program. It was found that the caput sperm head has the greatest surface area and a sharper acrosome bend than the cauda sperm. In an attempt to explain the above-mentioned changes, the suggestion is offered that some compactation of the nucleus and acrosomal material could be related to the decrease of the surface area.     

Lung water measurements with the mean transit time approach

The potential usefulness and limitations of the double-indicator mean transit time approach for measuring lung water are evaluated from both theoretical and empirical points of view. It is concluded that poor tissue perfusion is the most serious factor that can compromise the reliability of this approach. Replacement of the conventional water isotopes with a thermal signal enhances indicator delivery to ischemic areas but the diffusion of heat is not sufficiently rapid to permit measurements of water in macroscopic collections of fluid which remain unperfused. The frequency of pulmonary vascular obstruction in patients with pulmonary edema related to lung injury suggests that interpretation of transit time data will be complicated by uncertainties concerning perfusion. Thermal-dye measurements of lung water may prove more helpful in situations where pulmonary blood flow remains relatively uniform.     

Dynamics of lung collapse and recruitment during prolonged breathing in porcine lung injury

Oleic acid (OA)injection, lung lavage, and endotoxin infusion are three commonly usedmethods to induce experimental lung injury. The dynamics of lungcollapse and recruitment in these models have not been studied,although knowledge of this is desirable to establish ventilatorytechniques that keep the lungs open. We measured lung density bycomputed tomography during breath-holding procedures. Lung injury wasinduced with OA, lung lavage, or endotoxin in groups of sixmechanically ventilated pigs. After a stabilization period, repetitivecomputed tomography scans of the same slice were obtained duringprolonged expirations with and without positive end-expiratory pressureand during prolonged inspirations after 5 and 30 s of expiration. Lungcollapse and recruitment occurred mainly within the first 4 s ofbreath-holding procedures in all three lung injury models, and somecollapse and recruitment occurred even within 0.6 s. OA-injured lungswere significantly more unstable than lungs injured by bronchoalveolarlavage or endotoxin infusion. In this experimental setting, expirationtimes <0.6 s are required to avoid cyclic alveolar collapse duringmechanical ventilation without extrinsic positive end-expiratorypressure.

     

A micro-CT analysis of murine lung recruitment in bleomycin-induced lung injury.

The effects of lung injury on pulmonary recruitment are incompletely understood. X-ray computed tomography (CT) has been a valuable tool in assessing changes in recruitment during lung injury. With the development of preclinical CT scanners designed for thoracic imaging in rodents, it is possible to acquire high-resolution images during the evolution of a pulmonary injury in living mice. We quantitatively assessed changes in recruitment caused by intratracheal bleomycin at 1 and 3 wk after administration using micro-CT in 129S6/SvEvTac mice. Twenty female mice were administered 2.5 U of bleomycin or saline and imaged with micro-CT at end inspiration and end expiration. Mice were extubated and allowed to recover from anesthesia and then reevaluated in vivo for quasi-static compliance measurements, followed by harvesting of the lungs for collagen analysis and histology. CT images were converted to histograms and analyzed for mean lung attenuation (MLA). MLA was significantly greater for bleomycin-exposed mice at week 1 for both inspiration (P<0.0047) and exhalation (P<0.0377) but was not significantly different for week 3 bleomycin-exposed mice. However, week 3 bleomycin-exposed mice did display significant increases in MLA shift from expiration to inspiration compared with either group of control mice (P<0.005), suggesting increased lung recruitment at this time point. Week 1 bleomycin-exposed mice displayed normal shifts in MLA with inspiration, suggesting normal lung recruitment despite significant radiographic and histological changes. Lung alveolar recruitment is preserved in a mouse model of bleomycin-induced parenchymal injury despite significant changes in radiographic and physiological parameters.     

Biological half time of deposited nickel oxide aerosol in rat lung by inhalation

Wistar male rats were exposed to nickel oxide (NiO) aerosols (mass median aerodynamic diameter, 1.2 and 4.0 μm). The average exposure concentration was controlled from a low level of 0.6 mg/m³ to a high level of 70 mg/m³ and total exposure time was 140 h. Some rats were sacrificed just after the exposure, whereas others were exposed for 1 mo and kept for 12 and 20 mo clearance periods before sacrifice. There were no differences in body weight gain between NiO exposure groups and controls. Nickel concentrations in lungs of exposure groups were much higher than those of controls and decreased with the passing of the clearance time. No apparent deposition of nickel was observed in the liver, kidney, spleen, and blood immediately after the exposure, but in the case of the high exposure groups, the nickel concentration in the liver, spleen, and blood slightly increased with the increasing time of clearance. The biological half time of NiO deposited in the lungs was estimated by the assumption that the amount of the clearance is proportional to the amount of the NiO deposited. This resulted in a biological half time of 11.5 and 21 mo for 1.2 and 4.0 μm, respectively.     

Cellular recruitment and cytokine generation in a rat model of allergic lung inflammation are differentially modulated by progesterone and estradiol

We evaluated the role of estradiol and progesterone in allergic lung inflammation. Rats were ovariectomized (Ovx) and, 7 days later, were sensitized with ovalbumin (OA) and challenged after 2 wk with inhaled OA; experiments were performed 1 day thereafter. Ovx-allergic rats showed reduced cell recruitment into the bronchoalveolar lavage (BAL) fluid relative to sham-Ovx allergic rats, as was observed in intact allergic rats treated with ICI-182,780. Estradiol increased the number of cells in the BAL of Ovx-allergic rats, whereas progesterone induced an additional reduction. Cells of BAL and bone marrow (BM) of Ovx-allergic rats released elevated amounts of IL-10 and reduced IL-1 and TNF-. BM cells of Ovx-allergic rats released increased amounts of IL-10 and lower amounts of IL-4. Estradiol treatment of Ovx-allergic rats decreased the release of IL-10 but increased that of IL-4 by BM cells. Estradiol also caused an increased release of IL-1 and TNF- by BAL cells. Progesterone significantly increased the release of IL-10, IL-1, and TNF- by BAL cells and augmented that of IL-4 by BM cells. Degranulation of bronchial mast cells from Ovx rats was reduced after in vitro challenge, an effect reverted by estradiol but not by progesterone. We suggest that the serum estradiol-to-progesterone ratio might drive cellular recruitment, modulating the pulmonary allergy and profile of release of anti-inflammatory or inflammatory cytokines. The existence of such dual hormonal effects suggests that the hormone therapy of asthmatic postmenopausal women and of those suffering of premenstrual asthma should take into account the possibility of worsening the pulmonary conditions. asthma; eosinophils     

The diffusion transit time; A simple derivation

The mean first passage time for free diffusion can be derived directly by solving a simple analogue steady state problem. In this problem the diffusion starting region is considered as a time independent source of diffusing particles and the diffusion target assumes the behaviour of a perfectly absorbing sink. It is shown here that the transit time between the source and the sink, which in this particular problem is equal to the ratio between the holdup of the system and the total flux, is identical to the Brownian movement concept of the mean first passage time for free diffusion. This established identity considerably facilitates the derivation and investigation of the timing of diffusion in complicated structures such as those commonly found in living organisms.     

Colonization resistance of the digestive tract and gastrointestinal transit time in SPF mice.

The gastro-intestinal colonization resistance to Escherichia coli was assessed in individual CBA/Rij, C3H/StZ and Swiss/Cpb:SE mice. Gastro-intestinal transit time was determined by feeding small steel balls and x-ray examination of sequentially collected faeces. No correlation was found between transit times measured on 3 subsequent days, nor between them and colonization resistance.     

Regulation of lung neutrophil recruitment by VE-cadherin

Lung inflammatory disease is characterized by increased polymorphonuclear leukocyte (PMN) infiltration and vascular permeability. PMN infiltration into tissue involves signaling between endothelial cells and migrating PMNs, which leads to alterations in the organization of adherens junctions (AJs). We addressed the possible role of the protein constituents of AJs, endothelium-specific vascular-endothelial (VE)-cadherin, in the migration of PMNs. Studies were made using VE-cadherin mutant constructs lacking the extracellular domain (DeltaEXD) or, additionally, lacking the COOH-terminus beta-catenin-binding domain (DeltaEXDDeltabeta). Either construct was transduced in pulmonary microvessel endothelia of mice using cationic liposome-encapuslated cDNA constructs injected intravenously. Optimal expression of constructs was seen by Western blot analysis within 24 h. Vessel wall liquid permeability measured as the lung microvessel capillary filtration coefficient increased threefold in DeltaEXD-transduced lungs, indicating patency of interendothelial junctions, whereas the control DeltaEXDDeltabeta construct was ineffective. To study lung tissue PMN recruitment, we challenged mice intraperitoneally with LPS (3 mg/kg) for 6 h and measured PMN numbers by bronchoalveolar lavage and their accumulation morphometrically in lung tissue. DeltaEXD expression markedly reduced the PMN sequestration and migration seen in nontransfected (control wild type) or DeltaEXDDeltabeta-transfected (negative control) mice challenged with LPS. In addition, DeltaEXD transfection suppressed LPS-induced activation of NF-kappaB and consequent ICAM-1 expression. These results suggest that disassembly of VE-cadherin junctions serves as a negative signal for limiting transendothelial PMN migration secondary to decreased ICAM-1 expression in the mouse model of LPS-induced sepsis.