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D Negrini

Publications and source records attributed to D Negrini.

At least 37 records · Page 2Linked to original sources

Model of pleural fluid turnover.

A model of pleural fluid turnover, based on mass conservation law, was developed from experimental evidence that 1) pleural fluid filters through the parietal pleura and is drained by parietal lymphatics and 2) lymph flow increases after an increase in pleural liquid volume, attaining a maximum value 10 times greater than control. From the differential equation describing the time evolution of pleural liquid pressure, we obtained the equation for the steady-state condition ("set point") of pleural liquid pressure: Pss = (KfPi*+KlPzf)/Kf+Kl), where Kf is parietal pleura filtration coefficient, Kl is initial lymphatic conductance, Pzf is lymphatic potential absorption pressure, and Pi* is a factor accounting for the protein reflection coefficient of parietal mesothelium and hydraulic and colloid osmotic pressure of parietal interstitium and pleural liquid. Lymphatics act as a passive negative-feedback control tending to offset increases in pleural liquid volume. Some features of this control are summarized here: 1) lymphatics exert a tight control on pleural liquid volume or pressure so that the set point is maintained close to the potential absorption pressure of lymphatics; 2) a 10-fold increase in Kf would cause only a 2- and 5-fold increase in pleural liquid volume with normal (1.8 g/dl) and increased (3.4 g/dl) protein concentration of the pleural fluid, respectively; and 3) the reduction in maximum lymph flow greatly reduces the range of operation of the control with increased filtration and/or protein concentration of pleural fluid.

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Microvascular pressure profile in intact in situ lung.

We measured the microvascular pressure profile in lungs physiologically expanded in the pleural space at functional residual capacity. In 29 anesthetized rabbits a caudal intercostal space was cleared of its external and internal muscles. A small area of endothoracic fascia was surgically thinned, exposing the parietal pleura through which pulmonary vessels were clearly detectable under stereomicroscopic view. Pulmonary microvascular pressure was measured with glass micropipettes connected to a servo-null system. During the pressure measurements the animal was kept apneic and 50% humidified oxygen was delivered in the trachea. Pulmonary arterial and left atrial pressures were 22.3 +/- 1.5 and 1.6 +/- 1.5 (SD) cmH2O, respectively. The segmental pulmonary vascular pressure drop expressed as a percentage of the pulmonary arterial to left atrial pressure was approximately 33% from pulmonary artery to approximately 130-microns-diam arterioles, 4.5% from approximately 130- to approximately 60-microns-diam arterioles, approximately 46% from approximately 60-microns-diam arterioles to approximately 30-microns-diam venules, approximately 9.5% from 30- to 150-microns-diam venules, and approximately 7% for the remaining venous segment. Pulmonary capillary pressure was estimated at approximately 9 cmH2O.

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Distribution of diaphragmatic lymphatic lacunae.

The morphology of the submesothelial lymphatic lacunae on the pleural and peritoneal surface over the tendinous and muscular portion of the diaphragm was studied in 10 anesthetized rabbits. The lymphatic network was evidenced by injecting 1 ml of colloidal carbon solution in the pleural (n = 5) or the peritoneal (n = 5) space. After 1 h of spontaneous breathing, the animal was killed and the diaphragm was fixed in situ by injection of approximately 5 ml of fixative in pleural and peritoneal spaces. Then both cavities were opened and the diaphragm was excised and pinned to a support. According to which cavity had received the injection, the peritoneal or the pleural side of the diaphragm was scanned by sequential imaging of the whole surface by use of a video camera connected to a stereomicroscope and to a video monitor. The anatomic design appeared as a network of lacunae running either parallel or perpendicular to the major axis of the tendinous or muscular fibers. The lacunae were more densely distributed on the tendinous peritoneal area than on the pleural one. Scanty lacunae were seen on the muscular regions of both diaphragmatic sides, characterized by large areas without lacunae. The average density of lacunae on tendinous and muscular regions was 6 and 1.7/cm2 for the pleural side and 25 and 3.4/cm2 for the peritoneal side, respectively. The average width of lacunae was 137.9 +/- 1.6 and 108.8 +/- 1.7 microns on the tendinous pleural and the peritoneal side, respectively, and 163 +/- 1.8 microns on the muscular portion of the pleural and peritoneal surfaces.

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Intrapleural fluid movements described by a porous flow model.

We injected technetium-labeled albumin (at a concentration similar to that of the pleural fluid) in the costal region of anesthetized dogs (n = 13) either breathing spontaneously or apneic. The decay rate of labeled activity at the injection site was studied with a gamma camera placed either in the anteroposterior (AP) or laterolateral (LL) projection. In breathing animals (respiratory frequency approximately 10 cycles/min), 10 min after the injection the activity decreased by approximately 50% on AP and approximately 20% on LL imaging; in apneic animals the corresponding decrease in activity was reduced to approximately 15 and approximately 3%, respectively. We considered label translocation from AP and LL imaging as a result of bulk flows of liquid along the costomediastinal and gravity-dependent direction, respectively. We related intrapleural flows to the hydraulic pressure gradients existing along these two directions and to the geometry of the pleural space. The pleural space was considered as a porous medium partially occupied by the mesh of microvilli protruding from mesothelial cells. Solution of the Kozeny-Carman equation for the observed flow velocities and pressure gradients yielded a mean hydraulic radius of the pathways followed by the liquid ranging from 2 to 4 microns. The hydraulic resistivity of the pleural space was estimated at approximately 8.5 x 10(5) dyn.s.cm-4, five orders of magnitude lower than that of interstitial tissue.

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Regional blood flow to canine parietal pleura and internal intercostal muscle.

Transcapillary Starling forces in the parietal pleura and the underlying interstitium may potentially contribute to the exchange of fluid across this barrier. However, the extent of blood flow to the parietal pleura has not been measured. Thus, using standard microsphere techniques, we compared blood flow to the parietal pleura, including the subpleural interstitium, with blood flow to the adjacent internal intercostal muscle, as well as with flows to other serous tissues, including mediastinal pleura, pericardium, and parietal peritoneum, in anesthetized dogs that were either breathing spontaneously (n = 9) or ventilated to control arterial PCO2 (n = 5). Blood flow (ml.min-1.g-1) was measured after 20 min of equilibration in four successive body positions: right lateral decubitus, supine, left lateral decubitus, and prone. Overall, flow to parietal pleura was not different in spontaneous [1.07 +/- 0.14 (SE)] and mechanically ventilated animals (0.74 +/- 0.11). Flow to the internal intercostal muscle was significantly less than pleural blood flow, averaging 0.24 +/- 0.03 and 0.16 +/- 0.03 in the same groups, although again there was no effect of ventilation mode. Blood flow to other serous tissues in the thoracic cavity, specifically the mediastinal pleura (0.67 +/- 0.14) and pericardium (0.88 +/- 0.22), was similar to parietal pleural flow, whereas that to the parietal peritoneum was an order of magnitude lower (0.09 +/- 0.02, P less than 0.05). Changing body position had no effect on blood flow to any of the sampled tissues. Blood flow to the dorsal aspect of the chest wall muscle in spontaneously breathing animals tended to be greater than that to lateral or ventral portions of the chest wall.(ABSTRACT TRUNCATED AT 250 WORDS)

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Distribution of diaphragmatic lymphatic stomata.

In seven anesthetized rabbits we measured the size, shape, and density of lymphatic stomata on the peritoneal and pleural sides of the diaphragm. The diaphragm was fixed in situ and processed for scanning electron microscopy. Results are from 2,902 peritoneal and 3,086 pleural fields (each 1,620 microns 2) randomly chosen from the various specimens. Stomata were seen in 9% of the fields examined, and in 30% of the cases they appeared grouped in clusters with 2-14 stomata/field. Stoma density was 250 +/- 242 and 72 +/- 57 (SD) stomata/mm2 on peritoneal and pleural sides, respectively, and it was similar over the muscular and tendinous portion of the two surfaces. The maximum diameter ranged from less than 1 to approximately 30 microns, with an average value of 1.2 +/- 3.1 micron. The ratio of the maximum to the minimum diameter and the surface area averaged 2 +/- 1.4 and 0.7 +/- 2.4 micron 2, respectively. The maximum and minimum diameter and surface area values followed a lognormal frequency distribution, suggesting that stomata geometry is affected by diaphragmatic tension.

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Parenchymal stress affects interstitial and pleural pressures in in situ lung.

After resecting the intercostal muscles and thinning the endothoracic fascia, we micropunctured the lung tissue through the intact pleural space at functional residual capacity (FRC) and at volumes above FRC to evaluate the effect of increasing parenchymal stresses on pulmonary interstitial pressure (Pip). Pip was measured at a depth of approximately 230 microns from the pleural surface, at 50% lung height, in 12 anesthetized paralyzed rabbits oxygenated via a tracheal tube with 50% humidified O2. Pip was -10 +/- 1.5 cmH2O at FRC. At alveolar pressure of 5 and 10 cmH2O, lung volume increased by 8.5 and 19 ml and Pip decreased to -12.4 +/- 1.6 and -12.3 +/- 5 cmH2O, respectively. For the same lung volumes held by decreasing pleural surface pressure to about -5 and -8.5 cmH2O, Pip decreased to -17.4 +/- 1.6 and -23.8 +/- 5 cmH2O, respectively. Because Pip is more negative than pleural pressure, the data suggest that in intact pulmonary interstitium the pressure of the liquid phase is primarily set by the mechanisms controlling interstitial fluid turnover.

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Permeability-surface area product and reflection coefficient of the parietal pleura in dogs.

The parameters describing the permeability of the parietal pleura to liquid and total plasma proteins were measured in five anesthetized adult dogs. Small areas of parietal pleura (approximately 1 cm2) and the underlying endothoracic fascia were exposed through resection of the skin and the intercostal muscles. The portion of the thorax containing the pleural windows was removed from the chest and fixed over a bath of whole autologous plasma, the inner parietal pleural surface facing the bath. Small hemispheric Perspex capsules (surface area 0.28 cm2) connected to a pressure manometer were glued to the pleural windows; a subatmospheric pressure was set into the capsule chamber to create step hydraulic transpleural pressure gradients (delta P) ranging from 5 to 60 cmH2O. Transpleural liquid flows (Jv) and protein concentration of the capsular filtrate (Cfilt) and of the plasma bath were measured at each delta P. The transpleural protein flux (Js) at each delta P was calculated by multiplying Jv by the corresponding Cfilt. The hydraulic conductivity (Lp) of the parietal pleura was obtained from the slope of the Jv vs. delta P linear regression. The average Lp from 14 capsules was 9.06 +/- 4.06 (SD) microliters.h-1.cmH2O-1.cm-2. The mathematical treatment of the Js vs. Jv relationship allowed calculation of the unique Peclet number at the maximal diffusional protein flux and a corresponding osmotic permeability coefficient for plasma protein of 1 x 10(-5) +/- 0.97 x 10(-5) cm/s. The reflection coefficient calculated from the slope of the linear phase of the Js vs. Jv relationship was 0.11 +/- 0.05.(ABSTRACT TRUNCATED AT 250 WORDS)

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Hydraulic conductivity of canine parietal pleura in vivo.

The hydraulic conductivity (Lp) of the parietal pleura was measured in vivo in spontaneously breathing anesthetized dogs in either the supine (n = 8) or the prone (n = 7) position and in an excised portion of the chest wall in which the pleura and its adjacent tissue were intact (n = 3). A capsule was glued to the exposed parietal pleura after the intercostal muscles were removed. The capsule was filled with either autologous plasma or isotonic saline. Transpleural fluid flow (V) was measured at several transpleural hydrostatic pressures (delta P) from the rate of meniscus movement within a graduated pipette connected to the capsule. Delta P was defined as the measured difference between capsule and pleural liquid pressures. The Lp of the parietal pleura was calculated from the slope of the line relating V to delta P by use of linear regression analysis. Lp in vivo averaged 1.36 X 10(-3) +/- 0.45 X 10(-3) (SD) ml.h-1.cmH2O-1.cm-2, regardless of whether the capsule was filled with plasma or saline and irrespective of body position. This value was not significantly different from that measured in the excised chest wall preparation (1.43 X 10(-3) +/- 1.1 X 10(-3) ml.h-1.cmH2O-1.cm-2). The parietal pleura offers little resistance to transpleural protein movement, because there was no observed difference between plasma and saline. We conclude that because the Lp for intact parietal pleura and extrapleural interstitium is approximately 100 times smaller than that previously measured in isolated stripped pleural preparations, removal of parietal pleural results in a damaged preparation.

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Transperitoneal fluid dynamics in rabbit liver.

The peritoneal cavity of 18 anesthetized spontaneously breathing supine rabbits was opened through a midline section. One or two hollow capsules (surface area 0.8 cm2) were glued to the exposed liver surface, filled with whole or 25% diluted plasma, and connected to a transducer and a graduated pipette. Various hydraulic pressures (Pcap) were set in the capsule; at each Pcap the liquid flow per unit surface area (V/S) between the Disse's interstitial space and the capsule was measured from the rate of liquid displacement in the pipette. The slope of the V/S vs. Pcap linear regression was utilized to estimate the hydraulic conductivity of the Glissonian-peritoneal membrane and averaged 5.1 x 10(-3) +/- 4.7 x 10(-3) (SD) ml.h-1.cmH2O-1.cm-2 (n = 25). Hydraulic pressure in the Disse's space (Pd) was measured by closing the capsule against the transducer disconnected from the pipette. At portal and hepatic venous pressures of 7.6 +/- 2.9 and 2.6 +/- 1 cmH2O, respectively, Pd was 2.05 +/- 2 cmH2O. Physiologically, Starling pressure gradients cause fluid transfer from the sinusoids to the Disse's space; transperitoneal fluid filtration only occurs through the liver surface that faces the diaphragm, which corresponds to one-fifth of the total hepatic surface.

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Direct measurement of interstitial pulmonary pressure in in situ lung with intact pleural space.

We developed an experimental approach to measure the pulmonary interstitial pressure with the micropuncture technique in in situ lungs with an intact pleural space. Experiments were done in anesthetized paralyzed rabbits that were oxygenated via an endotracheal tube with 50% humidified oxygen and kept in either the supine or the lateral position. A small area of an intercostal space was cleared of the intercostal muscles down to the endothoracic fascia. Subsequently a "pleural window" was opened by stripping the endothoracic fascia over a 0.2-cm2 surface and leaving the parietal pleura (approximately 10 microns thick). Direct micropuncture through the pleural window was performed with 2- to 3-microns-tip pipettes connected to a servo-null pressure-measuring system. We recorded pleural liquid pressure and, after inserting the pipette tip into the lung, we recorded interstitial pressure from subpleural lung tissue. Depth of recording for interstitial pressure averaged 263 +/- 122 (SD) microns. We report data gathered at 26, 53, and 84% lung height (relative to the most dependent portion of the lung). For the three heights, interstitial pressure was -9.8 +/- 3, -10.1 +/- 1.6, and -12.5 +/- 3.7 cmH2O, respectively, whereas the corresponding pleural liquid pressure was -3.4 +/- 0.5, -4.4 +/- 1, and -5.2 +/- 0.3 cmH2O, respectively.

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Liquid drainage through the peritoneal diaphragmatic surface.

In 14 spontaneously breathing anesthetized rabbits, we used cyanoacrylate to glue a hollow capsule, at end expiration or at end inspiration, to the peritoneal surface of the tendinous portion of the diaphragm. The capsule was connected to a pressure transducer and a pipette calibrated in microliters. We filled the system with fluid and measured flow into the diaphragmatic surface facing the capsule (Fcap, microliter/cm2), from liquid displacement in the pipette at different hydraulic pressures in the system (Pcap). Pleural liquid pressure was simultaneously measured in the supraphrenic region (Psup). Fcap was positively correlated to transdiaphragmatic pressure gradient (Psup-Pcap) and breathing frequency but was unaffected by protein concentration of capsular fluid. For a breathing frequency of 30 cycles/min and a Psup - Pcap = -2 cmH2O, Fcap was 0.54 microliter.min-1.cm-2 for capsules applied at end expiration and 10-fold greater for capsules applied at end inspiration. Data indicate that the diaphragmatic tendinous portion in rabbits is a draining site for peritoneal fluid and that the conductance of the draining pathways (lymphatic stomata) is related to diaphragmatic tension. In the intact rabbit the average peritoneal fluid drainage through the tendinous portion of the diaphragm (approximately 16 cm2) was estimated at 43 microliters/min.

Absorption↗

Size-related differences in parietal extrapleural and pleural liquid pressure distribution.

The hydraulic pressure in the extrapleural parietal interstitium (Pepl) and in the pleural space over the costal side (Pliq) was measured in anesthetized spontaneously breathing supine adult mammals of increasing size (rats, dogs, and sheep) using saline-filled catheters and cannulas, respectively. From the Pliq and Pepl vs. lung height regressions it appears that in all species Pliq was significantly more subatmospheric than Pepl simultaneously measured at the same lung height. The vertical pleural liquid pressure gradient increased with size, amounting to -1, -0.69, and -0.44 cmH2O/cm in rats, dogs, and sheep, respectively. The vertical extrapleural liquid pressure gradient also increased with size, being -0.6, -0.52, and -0.33 cmH2O/cm in rats, dogs, and sheep, respectively. With increasing body size, the transpleural hydraulic pressure gradient (Ptp = Pepl - Pliq) at the level of the right atrium increased from 1.45 to 5.6 cmH2O going from rats to sheep. In all species Ptp increased, with lung height being greatest in the less dependent part of the pleural space.

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Intrapleural liquid flow down a gravity-dependent hydraulic pressure gradient.

We studied the vertical movement of 2 mg technetium-labeled albumin injected intrapleurally in 0.5 ml saline (15% of pleural liquid volume) in eight spontaneously breathing anesthetized dogs subject to a sudden change in posture (prone to supine or vice versa). The albumin movements were evaluated through a large field gamma camera placed laterally to the animal and detecting total (AT) and regional activities from two superimposed equal areas (At and Ab, top and bottom, respectively). The At/Ab ratio decreased from 2.1 to 1.3 in four animals up to 20 min from the change in posture and from 0.9 to 0.5 in four more animals studied from 50 to 90 min from turning maneuver. The rate of change in At and Ab was similar in the two groups of animals and unaffected by the acquisition posture. AT decreased by 7.7 and 3.5% for the two groups, respectively, reflecting albumin clearance from the pleural space. The opposite time course of regional activities and the independence of their rate of change of the At/Ab ratio and of the animal posture suggest a top-to-bottom albumin transfer occurring through a bulk flow of liquid estimated at 0.006 ml.kg-1.h-1. The data are consistent with a measured vertical pleural liquid pressure gradient that does not reflect a hydrostatic condition.

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Pleural and extrapleural interstitial liquid pressure measured by cannulas and micropipettes.

In 15 anesthetized apneic, oxygenated rabbits we simultaneously measured pleural liquid and interstitial extrapleural parietal pressures by using catheters and/or cannulas and micropipettes connected to a servonull system. With the animal in lateral posture, at an average recording height of 4.4 +/- 0.9 (SD) cm from the most dependent part of the cavity, the extrapleural catheter and the pleural cannula yielded -2.5 +/- 0.6 and -5.5 +/- 0.2 cmH2O; the corresponding values for micropipette readings in the two compartments were -2.4 +/- 0.6 and -5.4 +/- 0.4 cmH2O, respectively (not significantly different from those measured with catheters and cannulas). In the supine animal, interstitial extrapleural catheter pressure data obtained at recording heights ranging from 15 to 80% of pleural cavity lay on the identity line when plotted vs. the micropipette pressure values simultaneously gathered from the same tissues. We conclude that 1) micropipettes and catheters-cannulas yield similar results when recording from the same compartment and 2) the hydraulic pressure in the parietal extrapleural interstitium is less negative than that in the pleural space.

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Gravity-dependent distribution of parietal subpleural interstitial pressure.

Using liquid-filled catheters, we recorded, in 30 anesthetized, spontaneously breathing supine rabbits, the hydraulic pressure from the parietal subpleural interstitial space (Pspl). Through a small exposed area of parietal pleura a plastic catheter (1 mm ED), with a closed and smooth tip and several holes on the last centimeter, was carefully advanced between the muscular layer and the parietal pleura, tangentially to the pleural surface to reach the submesothelial layer. Simultaneous measurements of pleural liquid pressure (Pliq) were obtained from intrapleurally placed cannulas. End-expiratory Pspl decreased (became more negative) with increasing height (LH) according to the following: Pspl (cmH2O) = -1 - 0.4 LH (cm), the corresponding equation for Pliq being Pliq (cmH2O) = -1.5 - 0.7 LH (cm). Thus at end expiration a transpleural hydraulic pressure difference (Pliq-Pspl) developed at any height, increasing from the bottom to the top of the cavity as Pliq - Pspl (cmH2O) = -0.5 - 0.3 LH (cm). The Pliq-Pspl difference increased during inspiration due to the much smaller tidal change in Pspl than in Pliq. By considering the gravity-dependent distribution of the functional hydrostatic pressure in the systemic capillaries of the pleura (Pc) and the Pspl and Pliq values integrated over the respiratory cycle we estimated that on the average, the Pc-Pspl difference is sevenfold larger than the Pspl-Pliq difference.

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Kinetics of the intrapleural distribution of a radioactive bolus.

Through the use of a gamma-camera placed horizontally over the chest of 5 supine, anesthetized and spontaneously breathing dogs we studied the kinetics of the intrapleural spreading of 1 ml saline containing 2 mg of 99mTc labelled albumin (250 microCi/kg) injected at various points on the costal side. From the postinjection sequence of gamma-camera images, we defined, along the intrapleural spreading pathways, some regions of interest (ROI) displaying a preferential label accumulation. In all the animals the ROI roughly corresponded to the apical, mediastinal and latero-diaphragmatic regions. The kinetics of the intrapleural label distribution, estimated from the time course of the activity under the ROI, displayed an initial rapid phase, lasting a few minutes, followed by a marked slowing down of the exchange processes. At 30 min regional counts reached an apparent steady state, the activity values greatly varying (even by 10 fold) among the animals, for the ROI corresponding to the same anatomical region, independently on the site of injection. The mathematical modelling of the regional activity curves required the assumption of a progressive decrease in the exchange coefficients to account for a marked slowing down of the exchange processes with increasing time.

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Contribution of Starling and lymphatic flows to pleural liquid exchanges in anesthetized rabbits.

We studied the time course of volume and protein reabsorption of a 2-ml hydrothorax using whole (WP) or diluted (DP) homologous plasma injected into the right pleural cavity in anesthetized spontaneously breathing supine rabbits. Animals were killed at 5 (WP, n = 4; DP, n = 3), 36 (WP, n = 3; DP, n = 4), 55 (WP, n = 4), 90 (WP, n = 8; DP, n = 4), and 150 (WP, n = 4; DP, n = 5) min after the injection. The volume and protein content of the pleural liquid in control conditions (n = 12) amounted to 0.35 +/- 0.015 (SE) ml/kg and 1.8 +/- 0.27 g/100 ml, respectively, which are not significantly different at 90 min (n = 7). Pleural liquid volume decreased at a similar rate during WP or DP reabsorption according to the equation V = 0.84 +/- 0.05 X e-0.02t, with net reabsorptive flow expressed as dV/dt. The globulin quantity (Q) of the pleural liquid for WP and DP, respectively, decreased according to the equations Qwp = 1 + 1.5 X e-0.04t and Qdp = 0.7 + 0.6 X e-0.03t. Assuming a major lymphatic globulin clearance and no filtration into the cavity, we obtained lymph flow using the equation VL = dQ/dt X l/C where dQ/dt is calculated from the equations for Qwp and Qdp and C represents globulin concentration. The Starling flow (Vs) was then calculated by the equation Vs = dV/dt-VL. With increasing time, lymph flow was found to decrease progressively and was not significantly different from net flow with DP, which implied a Starling flow value of zero. During WP reabsorption, lymph flow initially exceeded the net flow, with the difference disappearing at approximately 60 min; accordingly, Starling filtration flow decreased progressively, becoming zero at the same time.

Anesthesia↗