Red Cell Spacing Research Articles

A monolithic fluid-structure interaction framework applied to red blood cells.

A parallel fully coupled (monolithic) fluid-structure interaction (FSI) algorithm has been applied to the deformation of red blood cells (RBCs) in capillaries, where cell deformability has significant effects on blood rheology. In the present FSI algorithm, fluid domain is discretized using the side-centered unstructured finite volume method based on the Arbitrary Lagrangian-Eulerian (ALE) formulation; meanwhile, solid domain is discretized with the classical Galerkin finite element formulation for the Saint Venant-Kirchhoff material in a Lagrangian frame. In addition, the compatible kinematic boundary condition is enforced at the fluid-solid interface in order to conserve the mass of cytoplasmic fluid within the red cell at machine precision. In order to solve the resulting large-scale algebraic linear systems in a fully coupled manner, a new matrix factorization is introduced similar to that of the projection method, and the parallel algebraic multigrid solver BoomerAMG is used for the scaled discrete Laplacian provided by the HYPRE library, which we access through the PETSc library. Three important physical parameters for the blood flow are simulated and analyzed: (1) the effect of capillary diameter, (2) the effect of red cell membrane thickness, and (3) the effect of red cell spacing (hematocrit). The numerical calculations initially indicate a shape deformation in which biconcave discoid shape changes to a parachute-like shape. Furthermore, the parachute-like cell shape in small capillaries undergoes a cupcake-shaped buckling instability, which has not been observed in the literature. The instability forms thin riblike features, and the red cell deformation is not axisymmetric but three-dimensional.

CHLORIDE CONCENTRATION, NOT SODIUM AND POTASSIUM, IS SELECTIVELY HIGHER IN HEART FAILURE-ASSOCIATED PLEURAL EFFUSION

SESSION TITLE: Pleural Disease SESSION TYPE: Original Investigations PRESENTED ON: 10/07/2018 02:15 pm - 03:15 pm PURPOSE: My recent studies have shown that serum chloride is a key electrolyte for the regulation of the fluid distribution or shift between the intravascular and cellular (red blood cell) spaces in the heart failure (HF) pathophysiology. Electrolyte chloride may have an important role for accumulation of pleural fluid in acutely worsening HF. The present study evaluated the hypothesis that a differential chloride concentration exists between the pleural fluid and blood serum in patients with worsening HF. METHODS: Seventeen consecutive patients that presented to Nishida hospital with pleural effusion and underwent thoracentesis during the study period from May 2017 to December 2017 were retrospectively analyzed. Worsening HF was diagnosed based on clinical criteria (presentation, echocardiography, serum b-type natriuretic peptide, and response to therapy). Biochemical measurements were performed on samples of non-heparinized pleural fluid and peripheral venous blood obtained within 2 h from each other. Pleural effusion was classified as transudates or exudates according to the classic Light’s criteria. Namely, the effusion was classified as exudates when any of the following criteria were met: 1) pleural to serum protein ratio >0.5, 2) pleural fluid LDH >200 IU, and 3) pleural fluid-to-serum LDH ratio >0.6. The effusion-serum electrolyte gradient was determined as [(pleural fluid electrolyte) minus (serum electrolyte)]. Statistical analysis was performed by one-way analysis of variance with Tukey’s post-hoc test. RESULTS: Fifteen patients (53% men; 85±11 years) were diagnosed with HF-associated pleural effusion; based on the Light’s criteria, 10 of these patients were classified as having transudative effusion and the other 5 patients were classified as having exudative effusion [fulfilling only one (n=4) or both (n=1) of the LDH criteria]. The effusion-serum electrolyte gradient was significantly higher in chloride (7.4 ± 2.6 mEq/L; range 4 – 14 mEq/L) than in sodium (0.87 ± 1.36 mEq/L; range -1 – 4 mEq/L) and potassium (-0.12 ± 0.254 mEq/L; range -0.8 – 0.2 mEq/L; P < 0.001 for each). CONCLUSIONS: In acutely worsening HF patients with associated pleural effusion, concentration of chloride in the pleural effusion is greater than that in the serum, indicating that chloride may have an important and active role in the formation and retention of body fluid in the pleural space, and possibly in the interstitial space. CLINICAL IMPLICATIONS: The finding of higher chloride concentration in HF-associated pleural fluid as shown in this study should raise innovative idea for investigating the pleural fluid pathophysiology. According to the “revised Starling equation and the glycocalyx model”, anionic electrolyte chloride may penetrate the endothelial glycocalyx layer, diffuse into the pleural space, and hold pleural fluid by its tonicity under condition of insufficient drainage via venous and/or lymphatic channels. DISCLOSURES: No relevant relationships by Hajime Kataoka, source=Web Response

Effects of freshwater and saltwater adaptation and dietary salt on fluid compartments, blood pressure, and venous capacitance in trout

Trout are of interest in defining the relationship between fluid and salt balance on cardiovascular function because they thrive in freshwater (FW; volume loading, salt depleting), saltwater (SW; volume depleting, salt loading), and FW while fed a high-salt diet (FW-HS; volume and salt loading). The effects of chronic (>2 wk) adaptation to these three protocols on blood volume (51Cr red cell space), extracellular fluid volume (99mTc-diethylene triaminepenta-acetic acid space), arterial (dorsal aortic; P(DA)) and venous (ductus Cuvier; Pven) blood pressure, mean circulatory filling pressure (zero-flow Pven), and vascular capacitance were examined in the present study on unanesthetized rainbow trout. Blood volume, extracellular fluid volume, P(DA), Pven, and mean circulatory filling pressure progressively increased in the order SW < FW < FW-HS. Vascular capacitance in SW fish appeared to be continuous with the capacitance curve of FW fish and reflect a passive volume-dependent unloading of the venous system of FW fish. Vascular capacitance curves for FW-HS fish were displaced upward and parallel to those of FW fish, indicative of an active increase in unstressed blood volume without any change in vascular compliance. These studies are the first in any vertebrate to measure the relationship between fluid compartments and cardiovascular function during independent manipulation of volume and salt balance, and they show that volume, but not salt, balance is the primary determinant of blood pressure in trout. They also present a new paradigm with which to investigate the relative contributions of water and salt balance in cardiovascular homeostasis.

Blood and extracellular fluid volume in whole body and tissues of the Pacific hagfish, Eptatretus stouti.

Whole-body and 20 individual-tissue (51)Cr-RBC (red cell space; RCS) and (99)Tc-diethylenetriaminepentaacetic acid (extracellular space; ECS) spaces were measured in seven unanesthetized Pacific hagfish (Eptatretus stouti). Volume indicators were administered via a dorsal aortic cannula implanted the previous day. Blood samples were collected at 6, 12, 18, and 24 h after injection. Tissues were removed at 24 h and radioactivity was measured; tissue water content (percent of wet weight) was determined by desiccation at 95 degrees C for 48 h. Mixing rates of both indicators were identical and were essentially complete by 12 h, indicating that blood convection is the rate-limiting process. At 24 h, the whole-body RCS was 19.3+/-2.1 mL kg(-1) body weight, and the ECS was 338.5+/-15.2 mL kg(-1) body weight. Blood volume estimated from the 24-h RCS and the mean central hematocrit (14%) was 137.9 mL kg(-1) body weight. Liver RCS (118.6+/-30.5 microL g(-1) tissue weight) was twice that of any other tissue and was also the most variable, ranging from 59 to 263 microL g(-1), whereas liver ECS (406.0+/-34.3 microL g(-1)) was in the range of other tissues, and water content (66.9%+/-3.5%) was low. Gill RCS (55.9+/-5.7 microL g(-1)), ECS (415.3+/-37.7 microL g(-1)), and percent water (83.1%+/-0.8%) were higher than most other tissues. RCS, ECS, and percent water were consistently lowest in ovum (1.1+/-0.02 microL g(-1), 111.1+/-4.3 microL g(-1), 51.3%+/-3.5%, respectively). Tongue, notocord, and myotome had generally lower RCS (2.1+/-0.4, 2.2+/-0.5, 7.1+/-0.1 microL g(-1), respectively) and ECS (121.2+/-7.0, 246.3+/-17.4, 185.3+/-16.7 microL g(-1), respectively), although their water content was in the midrange (74.7+/-0.5, 81.2+/-1.6, 74.4%+/-0.6%, respectively). Skin had a low RCS (6.8+/-1.1) and midrange ECS (387.5+/-28.0) but very low water content (61.2%+/-2.1%). These findings confirm that hagfish blood volume is at least twice as large as other fish, whereas our estimate of extracellular fluid volume is larger than previously reported and more in line with the predicted interstitial volume. RCS, ECS, and water content vary, often independently, between tissues, which may perhaps be indicative of specific tissue needs or functions. A distinct spleen is lacking in hagfish, and the liver appears to serve this function by sequestering red cells. To our knowledge, this is the first report of tissue ECS in Myxiniformes.

Effect of anemia on cardiac function, microvascular structure, and capillary hematocrit in rat hearts.

The effect of anemia on the coronary microcirculation was studied in young male rats. Chronic anemia resulted in increased left ventricular end-diastolic pressure and decreased functional reserve. Cardiac mass in anemic animals increased by 25%. Capillary and arteriolar densities in these hearts remained unchanged, indicating angiogenesis in this experimental situation (estimated aggregate capillary length in the left ventricle of anemic hearts was 3.06 km compared with 2.35 km in control hearts). Capillary hematocrit was decreased in chronic anemia less than systemic hematocrit: from 25 to 18% in anemia versus 45 to 28% in controls. Capillary hematocrit and red blood cell spacing were also studied after acute blood withdrawal. Here, capillary hematocrit was preserved even more: 22 versus 24% in systemic hematocrit. Finally, the same was studied in isolated hearts perfused with solutions of various hematocrits. After perfusion with low-hematocrit solution (14%), the capillary hematocrit (24%) was even higher than the perfusate hematocrit! In conclusion, we found evidence of angiogenesis in cardiomegaly induced by chronic anemia. Microvascular growth was accompanied by advantageous regulation of red blood cell spacing within these vessels. This was even more pronounced during acute hemodilution and in isolated perfused hearts.

Tissue and whole-body extracellular, red blood cell and albumin spaces in the rainbow trout as a function of time: a reappraisal of the volume of the secondary circulation

[58Co]EDTA, [51Cr]RBC and [125I]albumin spaces in the whole body and 28 tissue samples were examined at timed intervals over 16 h in rainbow trout Oncorhynchus mykiss. [58Co]EDTA space (which approximates extracellular fluid volume; ECF) in fins, skin, gallbladder and eye are reported for the first time. After a 16 h equilibration, ECF volume was large (376-726 microl g-1 wet tissue mass) in kidney, swimbladder, skin and fins, moderate (219-313 microl g-1 wet tissue mass) in stomach, skull, spleen, liver, intestine, gills, eye and cecum, and small (53-181 microl g-1 wet tissue mass) in red muscle, fat, brain, gallbladder and white muscle. Whole-body ECF was 387+/-10.6 microl g-1 (mean +/- s.e.m.; N=11). [51Cr]RBC space relative to [58Co]EDTA space was large in spleen, liver, intestine and gill, and low in skin, fins, stomach and skull. Whole-body [51Cr]RBC space was 9.9+/-0.6 microl g-1 body mass (N=17). Blood volume calculated from [51Cr]RBC space at 16 h and a dorsal aortic hematocrit of 24.5 % was 40.4 microl g-1 body mass. Whole-body [125I]albumin space at 16 h was 118.0+/-7.4 microl g-1 body mass (N=6), which resulted in an estimated blood volume of 156. 6 microl g-1 body mass, nearly four times that estimated from the [51Cr]RBC space. Tissue hematocrits, calculated from [125I]albumin and [51Cr]RBC spaces, were significantly lower than dorsal aortic hematocrit in all tissues except spleen, kidney and liver. [58Co]EDTA and [51Cr]RBC spaces reached equilibrium in nearly all tissues within 1 h, whereas [125I]albumin continued to accumulate in many tissues up 24 h. The disparity between [125I]albumin distribution kinetics compared with the kinetics of [58Co]EDTA and [51Cr]RBC distribution, as well as the accumulation of [125I]albumin in tissues not known to have a secondary circulation, indicates that [125I]albumin is a poor marker of plasma volume in trout and that previous studies based on [125I]albumin clearance from the plasma have overestimated both the volume and the turnover rate of the secondary system. Revised estimates of secondary circulation volume, based on [58Co]EDTA distribution rate, indicate that it is no more than 10-20 % of the volume of the primary circulation. <P>

Blood volume, plasma volume and circulation time in a high-energy-demand teleost, the yellowfin tuna (Thunnus albacares)

We measured red cell space with 51Cr-labeled red blood cells, and dextran space with 500 kDa fluorescein-isothiocyanate-labeled dextran (FITC-dextran), in two groups of yellowfin tuna (Thunnus albacares). Red cell space was 13.8+/-0.7 ml kg-1 (mean +/- s.e.m.) Assuming a whole-body hematocrit equal to the hematocrit measured at the ventral aortic sampling site and no significant sequestering of 51Cr-labeled red blood cells by the spleen, blood volume was 46. 7+/-2.2 ml kg-1. This is within the range reported for most other teleosts (30-70 ml kg-1), but well below that previously reported for albacore (Thunnus alalunga, 82-197 ml kg-1). Plasma volume within the primary circulatory system (calculated from the 51Cr-labeled red blood cell data) was 32.9+/-2.3 ml kg-1. Dextran space was 37.0+/-3.7 ml kg-1. Because 500 kDa FITC-dextran appeared to remain within the vascular space, these data imply that the volume of the secondary circulatory system of yellowfin tuna is small, and its exact volume is not measurable by our methods. Although blood volume is not exceptional, circulation time (blood volume/cardiac output) is clearly shorter in yellowfin tuna than in other active teleosts. In a 1 kg yellowfin tuna, circulation time is approximately 0.4 min (47 ml kg-1/115 ml min-1 kg-1) compared with 1. 3 min (46 ml kg-1/35 ml min-1 kg-1) in yellowtail (Seriola quinqueradiata) and 1.9 min (35 ml kg-1/18 ml min-1 kg-1) in rainbow trout (Oncorhynchus mykiss). In air-breathing vertebrates, high metabolic rates are necessarily correlated with short circulation times. Our data are the first to imply that a similar relationship occurs in fishes.

A method to determine red blood cell spacing in capillaries of rat heart.

Researchers have traditionally focused on blood capillaries as the primary site of diffusive oxygen transfer to tissue. Many theoretical studies assume that capillary blood is continuous and homogeneous with respect to its oxygen supply. However, this “continuum assumption” ignores the particulate nature of blood, whereby erythrocytes are discrete O2 sources.

Red Blood Cell Spacing in Rat Coronary Capillaries during the Cardiac Cycle

Theoretical studies have demonstrated a pronounced effect of red blood cell (RBC) spacing on tissue oxygen supply. Our objective was to collect data regarding RBC spacing and related linear capillary hematocrit (Hct) in rat coronary capillaries, from two distinct locations within the capillary bed (proximal and distal portions), in the subendo- and midmyocardium, during systole and diastole. Hearts were rapidly frozenin situ,and tissue sections were stained in order to distinguish capillaries in proximal and distal portions of the capillary bed, as well as the RBCs within them. Morphometric data were compared according to capillary type, cardiac region, and cardiac phase. Distal portions of the capillary bed had a significantly greater capillary Hct (P< 0.001) and significantly lower RBC spacing values (P< 0.01), compared to proximal portions, irrespective of cardiac phase or region. There was a greater frequency of RBC spacing values equal to 0 μm in systole compared to diastole (P< 0.001) and a greater frequency of RBC spacing values greater than 40 μm in proximal portions of the capillary bed compared to distal portions (P< 0.001). These results suggest that intracapillary resistance to O2transport is reduced, and O2carrying capacity of the blood increased, in distal portions of the capillary bed, where PO2in the blood is lower.

Effect of [Hb] on blood flow distribution and O2 transport in maximally working skeletal muscle.

We investigated whether the reduction in calculated muscle diffusion capacity for O2 (DmO2) previously shown to occur with lowered hemoglobin concentration ([Hb]) perfusion of maximally working muscle is related to changes in the blood flow distribution. If blood flow distribution is altered during low [Hb] conditions, the reduction in the calculated DmO2 may in fact be due to increasing heterogeneity and not to some other hemoglobin-related factor. Color-stained (15-microns-diam) microspheres were injected into the artery supplying maximally working isolated in situ dog gastrocnemius muscle (n = 6) while it was being perfused (flow controlled by pump perfusion) with whole blood at three different levels of [Hb] (14.1 +/- 0.5, 8.9 +/- 0.4, and 5.7 +/- 0.4 (SE) g/100 ml] in a blocked-order design. Muscle blood flow and arterial PO2 were not changed as [Hb] was altered. Maximal O2 uptake (11.8 +/- 1.3, 8.2 +/- 0.8, and 6.0 +/- 0.9 ml.100 g-1 min-1 for those [Hb] values, respectively) and the associated estimate of DmO2 (0.25 +/- 0.03, 0.18 +/- 0.03, and 0.15 +/- 0.03 ml.100 g-1.min-1.Torr-1) declined significantly (P < 0.05) with [Hb]. However, the dispersion of the blood flow distribution did not change significantly and, if anything, indicated less heterogeneity at lower [Hb] (coefficient of variation - 0.52 +/- 0.06, 0.46 +/- 0.05, and 0.43 +/- 0.03). These results suggest that in maximally working canine muscle in situ, when O2 delivery is reduced by lowering [Hb] (at constant blood flow), changes in blood flow distribution play no significant role in the reduction of maximal O2 uptake and calculated DmO2. The apparent increase in the resistance to O2 diffusion (i.e., reduction in the DmO2) during anemia may therefore be a result of increased red blood cell spacing in the capillary, slow chemical off-loading kinetics of O2 from Hb, or some other effect that remains to be determined.

Effect of reduced hemoglobin concentration on leg oxygen uptake during maximal exercise in humans

Maximum oxygen uptake (VO2max) is affected by hemoglobin concentration ([Hb]). Whether this is simply due to altered convection of O2 into the muscle microcirculation or also to [Hb]-dependent diffusive transport of O2 out of the muscle capillary is unknown in humans. To examine this, seven healthy volunteers performed four maximal cycle exercise bouts at sea level immediately after 8 wk at altitude (3,801 m, barometric pressure 485 Torr), a sojourn designed to increase [Hb]. The first two bouts were at ambient [Hb] of 15.9 +/- 0.7 g/100 ml breathing 21 or 12% O2 in random order. [Hb] was then decreased to a prealtitude level of 13.8 +/- 0.6 g/100 ml by venesection and isovolemic replacement with 5% albumin in 0.9% saline, and the exercise bouts were repeated. At whole body VO2max, PO2, PCO2, pH, and O2 saturation were measured in radial arterial and femoral venous blood. Femoral venous thermodilution blood flow was determined for calculation of leg VO2. Mean muscle capillary PO2 and muscle diffusing capacity (DO2) were computed by Bohr integration between measured arterial and femoral venous PO2. Averaged over both fractional concentrations of inspired O2, leg VO2 at maximum decreased by 17.7 +/- 4.3% as [Hb] was lowered while leg O2 delivery decreased by 17.5 +/- 2.6% and DO2 decreased by 10.7 +/- 2.7% (all P < 0.05). The relative contributions of decreases in leg O2 delivery and DO2 to the decrease in VO2max were computed to be 64 and 36%, respectively. These findings suggest that [Hb] is an important determinant of O2 diffusion rates into working muscle in humans. Possible mechanisms include 1) dependence of DO2 on intracapillary red blood cell spacing, 2) changes in the total rate of dissociation of O2 from [Hb], and 3) increased red blood cell flow heterogeneity as [Hb] is reduced.

Effects of chronic hypoxia on capillary flow and hematocrit in rat skeletal muscle.

The cremasteric microcirculation was studied in rats exposed to chronic hypoxia. Control male weanling Sprague-Dawley rats (n = 8) were raised for 42-49 days at 752 mmHg. Hypoxic rats (n = 9) were reared for 3 days at 551 mmHg, 4 days at 461 mmHg, 3 days at 371 mmHg, and 31-38 days at 311 mmHg (6,000 m). Red blood cells labeled with fluorescein isothiocyanate were injected. The exposed cremaster was observed using fluorescence microscopy. Rats acutely breathed 10, 21, and 30% O2 spontaneously in random order. Hypoxia-adapted animals had greater (P less than 0.01) red cell flux (10.6 +/- 1.0 vs. 5.7 +/- 0.4/s), capillary hematocrits, capillary-to-systemic hematocrit ratios (0.42 +/- 0.02 vs. 0.33 +/- 0.02), and reduced red cell spacing (11.4 +/- 1.3 vs. 22.3 +/- 2.0 microns) than controls under 21% O2. Chronically hypoxic rats also demonstrated significantly (P less than 0.05) larger capillary diameters (6.52 +/- 0.04 vs. 6.15 +/- 0.06 microns) and greater perfused (135 +/- 5 vs. 94 +/- 3 mm/mm3) and anatomic (182 +/- 5 vs. 151 +/- 8 mm/mm3) microvessel length-densities at 21% O2. Results were generally similar for 10 and 30% O2. Bulk capillary blood flow was significantly (P less than 0.01) greater in controls (2.75 +/- 0.32 vs. 1.87 +/- 0.12 pl/s) only under 30% O2. Our experiments demonstrate that numerous physiological, in addition to anatomic, alterations can occur in the cremasteric microcirculation in response to chronic hypoxia.

Stroma-free hemoglobin: its presence in plasma does not improve oxygen supply to the resting hindlimb vascular bed of hemodiluted dogs

In hemodilution, red cell spacing in the microcirculation is increased, flow distribution may become more heterogeneous, and, as a result, oxygen supply to tissues may suffer. We tested the hypothesis that oxygen extraction from diluted blood may be enhanced by the presence of hemoglobin in the plasma phase in relatively low concentrations. In anesthetized dogs, the hindlimb vascular bed was isolated and perfused with the animal's own blood by a roller pump. One group of dogs (n = 6) was hemodiluted (hematocrit = 15.0 +/- 1.0%) with a 6% solution of dextran. A second group of dogs (n = 6) was similarly hemodiluted (hematocrit = 16.0 +/- 0.4%) with dextran containing stroma-free hemoglobin solution whereby plasma-phase hemoglobin concentration was raised to 1.1 +/- 0.1 g.dL-1. Systemic hemodynamic observations were made repeatedly over the subsequent 2.5 h, while blood flow to the hindlimb was progressively reduced in stepwise decrements. The hemoglobin-hemodiluted group showed increased systemic arterial blood pressure and total peripheral resistance when compared with the control (dextran diluted) group. The isolated hindlimb also showed evidence of increased vascular resistance in the hemoglobin-treated group. In each individual animal, critical oxygen delivery and extraction were determined by finding the intercept of the supply-independent and supply-dependent portions of the oxygen uptake/oxygen delivery relationship. Neither the critical oxygen delivery rates (5.75 +/- 0.83 vs. 6.41 +/- 0.53 mL.kg-1.min-1) nor critical oxygen extraction ratios (0.75 +/- 0.03 vs. 0.76 +/- 0.04) were found to be significantly different in the two groups.(ABSTRACT TRUNCATED AT 250 WORDS)

Response of rainbow trout to constant-pressure and constant-volume hemorrhage.

Plasma oncotic pressure (OC), protein concentration (P), osmolality (Osm), and hematocrit (Hct) were measured in conscious rainbow trout subjected to 120 min of constant-pressure (CPH) or constant-volume (CVH) hemorrhage. Blood volume, tissue red cell, and plasma space were determined in control and CPH trout with 51Cr-labeled red cells and 125I-albumin. At the onset of CPH, blood loss exceeded 1.5 ml.min-1.kg-1 but quickly dropped to a constant rate of 0.1 ml.min-1.kg-1 by 15 min. After 2 h, 73% of the initial blood volume was lost, and Hct, OC, and P were reduced by 55 +/- 3, 39 +/- 4, and 19 +/- 9%, respectively; Osm did not change. Calculated blood volume was reduced by 39% (51Cr method) or by 56% (125I method). Red cell space was reduced in all tissues except brain. Plasma space decreased in stomach, increased in liver and kidney, and was unchanged in other tissues. CVH had similar effects on Hct, OC, and P, while Osm and plasma sodium increased. The time required for arterial pressure to recover to within 90% of control increased with successive bleeding from 39 to greater than 160 min. Twenty-four hours after CVH, Hct remained depressed, while all other parameters were returning toward control levels. Trout incorporate a variety of mechanisms to respond to hemorrhage, including mobilization of stored blood and reabsorption of fluid from interstitial and probably intracellular sources. High permeability of trout capillaries to protein may facilitate the restoration of protein and fluid and allow fish to survive extensive hypovolemia.

Effects of red cell spacing and red cell movement upon oxygen release under conditions of maximally working skeletal muscle.

RBC spacing in capillaries plays an important role in that it determines the total number of RBCs contained in a capillary and, therefore, the total O2 flux out of the capillary. The detrimental effects of increased RBC spacing upon capillary O2 release are in part compensated for by enhanced O2 release out of single RBCs due to improved diffusion geometry and RBC movement. Non-uniformity of O2 flux brought about by the particulate nature of blood is considerably smaller than calculations which do not consider RBC movement indicate. It creates oscillations in the O2 supply to the tissue, the periodicity of which is fast, however, compared to the time constant of the PO2 decay in a temporarily unsupplied tissue. We conclude that non-uniformity of O2 flux out of capillaries due to large inter-erythrocytic plasma gaps does not play an important role for tissue O2 supply as long as average RBC spacing is sufficiently small to guarantee an appropriate overall capillary O2 flux.

Microvessel hematocrit: measurement and implications for capillary oxygen transport.

Microvascular preparations of the hamster cheek pouch and cremaster muscle were used to establish the relationship among three measures of red cell distribution: 1) the systemic hematocrit defined as the volume percentage of red cells sampled from a toe clip; 2) the tube hematocrit defined as the instantaneous volume fraction of red cells in a microvessel segment of measured length and diameter; and 3) the discharge hematocrit defined as the volume percentage of red cells that flowed into micropipettes inserted into microvessels. The results show the tube hematocrit is low and highly variable in the presence of a nearly constant systemic hematocrit. The discharge hematocrit, in contrast, consistently approximated systemic values in blood taken from arterioles or venules with diameters of 6-98 micron. Indeed, the mean ratio of the discharge to the systemic hematocrit did not differ from unity (P greater than 0.25). The observed similarity between the discharge and systemic hematocrit indicates that red cells are distributed uniformly across capillary networks. Differences between the absolute value and variance of the tube hematocrit compared with the discharge hematocrit imply that the anatomical volume of a microvessel can differ from the volume available to cells and plasma within a microvessel. Thus moment-to-moment variation in the tube hematocrit or capillary red cell spacing may be indicative of a change in the effective capillary flow cross section and not solely on the inflow hematocrit.

Blood volume and red cell space in tissues of the rainbow trout, Salmo gairdneri

1. 1. Whole body blood volume and red cell space of 22 tissues were measured in unanesthetized rainbow trout at 4, 12, 30, 60, 150 and 240 min after dorsal aortic injection of 51Cr-labeled red blood cells. 2. 2. Apparent blood volume decreased during the initial 30 min after injection and increased thereafter. At 240 min the blood volume was 33.5 ± 3.1 ml/kg body wt. 3. 3. Tissue red cell space varied as a function of the interval between labeled red cell injection and tissue collection. Red cell space was highest in spleen followed by heart, kidney and liver. Lowest red cell spaces were found in stomach and red and white skeletal muscle. 4. 4. Variability in blood volume and tissue red cell space over time suggests that caution should be exercised in the design of experiments that employ indicator dilution measurements to measure vascular volumes.

An examination of the measurement of flow heterogeneity in striated muscle.

This review leads us to a number of conclusions and suggestions for further study. First, we find wide differences in the meaning of flow heterogeneity, arising as a result of the different methods used. These differences will have to be reconciled to form a comprehensive view of the role of heterogeneity in determining vascular function. Second, in the future, the meaning of heterogeneity must be clearly defined and related to a particular microvascular component, and it is imperative that the differences in scale of heterogeneity be appreciated when comparing data from various laboratories. These heterogeneities have different implications for function, and failure to distinguish among them leads to confusion. Third, the degree to which perfusion heterogeneity is regulated in the microcirculation remains in doubt. Reports of variations in flow heterogeneity in response to physiological stimuli are for the most part based on highly questionable indirect methods. Fourth, the heterogeneity that can be demonstrated at the capillary level within striated muscle does not appear to be large relative to the capacity for the microcirculation to exchange most diffusible solutes. Thus, the inferences regarding heterogeneity, as evidenced by diffusible indicators, are likely to be the result of different preparations, damage to the preparations, or perhaps large-scale heterogeneities in the tissue. An alternate possibility would be that the heterogeneity occurs at the microvascular level but reflects some other aspect of microcirculatory function, such as length or hematocrit heterogeneities, but not flow heterogeneities. Fifth, flow heterogeneity within microvessels implies important consequences for capillary exchange and tissue oxygenation. Heterogeneities of velocity of a magnitude comparable to those observed by direct visualization of microcirculation can clearly produce reductions in oxygen supply to small tissue regions of a degree that may limit oxygen delivery, and thereby, tissue function. Sixth, flow heterogeneity may also influence capillary hematocrit and/or red cell spacing by producing cell separation at bifurcations and a resultant reduction in mean capillary tube hematocrit. There is as yet no agreement on why and how these hematocrits influence tissue oxygenation and function. Although several hypotheses are advanced to explain the distribution of blood flow and red cells within microcirculation, each lacks a critical experimental test at present.(ABSTRACT TRUNCATED AT 400 WORDS)

Theoretical analysis of oxygen supply to contracted skeletal muscle.

Honig and collaborators reported striking contradictions in current understanding of O2 supply to working skeletal muscle. Therefore we re-examined the problem by means of a new composite computer simulation. As inclusion of erythrocytic O2 desaturation and oxygen transport and consumption inside the muscle cell into a single model would entail immense numerical difficulties, we broke up the whole process into its several components: O2 desaturation of erythrocytes O2 transport and consumption in muscle fiber capillary transit time characterizing the period of contact between red cell and muscle fiber. "Erythrocyte model" as well as "muscle fiber model" both consist of a central core cylinder surrounded by a concentric diffusion layer representing the extracellular resistance to O2 diffusion (Fig. 1). Resistance layers in both models are to be conceived of as one and the same anatomical structure--even though in each model their shape is adapted to the respective geometry. By means of this overlap region a spatial connexion between both is given, whereas temporal coherence governing O2 fluxes and red cell spacing is derived from capillary transit time. Analysis of individual components is outlined as follows: Assuming axial symmetry of the problem a numerical algorithm was employed to solve the parabolic system of partial differential equations describing red cell O2 desaturation. Hb-O2 reaction kinetics, free and facilitated O2 diffusion in axial and radial directions, and red cell movement in capillary were considered. Resulting time courses of desaturation, which are considerably faster than the ones computed by Honig et al., are given in the following table (see also Fig. 3). (Formula: see text) Furthermore, we studied the respective importance of the several processes included in our model: Omission of longitudinal diffusion increased desaturation time by 15% to 23%, whereas effects of reaction kinetics and axial movement were 5% and 2% respectively. For time courses see Fig. 2. Nature and magnitude of extra-erythrocytic resistance to O2 diffusion playing a prominent part in O2 desaturation are scarcely explored. Calculated desaturation times based upon our new estimates (line 3 of above table) correspond well, however, with findings by Sinha, who observed 1.75 to 4-fold prolongation in omental and mesenteric capillaries compared to desaturation through equivalent plasma layers. The 3-dimensional elliptic system of partial differential equations describing stationary O2 transport through resistance layer and subsequent free and facilitated O2 diffusion and O2 consumption in muscle fiber was solved analytically.(ABSTRACT TRUNCATED AT 400 WORDS)

The end-points of the oxygen path: transport resistance in red cells and mitochondria.

The variation of oxygen pressure from red cell to mitochondrion is, for a given oxygen flux, determined by the distribution of transport resistance over the oxygen path. The total drop in oxygen pressure may be resolved into several component drops: (1) the red cell drop, from cell center to cell surface, (2) the transcapillary drop, from red cell surface to outer capillary wall, (3) the tissue drop, from the capillary wall to an interior point in the tissue, and (4) the mitochondrial drop, from tissue to mitchondrial interior. The frequently used Krogh model, as usually formulated, deals only with the tissue drop. The other components have received somewhat less attention in the literature. The transcapillary drop, which depends on hematocrit, blood flow rate, red cell geometry, capillary geometry, and other things, is by far the most complex. Some recent progress in the theory of this drop has been made — for example, the work of Federspiel and Sarelius (1984) on the effect of red cell spacing on oxygen delivery. Nevertheless, the transcapillary drop probably will be the last component to be understood quantitatively. The red cell drop and the mitochondrial drop, on the other hand, are readily amenable to analysis, and we summarize here our recent theoretical results on these parts of the oxygen path.KeywordsOxygen PressureOxygen TransportOxygen FluxOxygen GradientTransport ResistanceThese keywords were added by machine and not by the authors. This process is experimental and the keywords may be updated as the learning algorithm improves.