Ultrasonic evaluation of erythrocyte aggregation dynamics

The dynamics of aggregation and disaggregation of blood of varying hematocrit in oscillatory flow in a distensible horizontal tube was determined by measuring the developing echo intensity of the blood samples with a 10 MHz B-mode ultrasonic scanner. Early aggregation could be detected within 10 sec. of stoppage of flow. The rate of echo intensity buildup and thus, presumably the rate of aggregation when flow was stopped was inversely related to hematocrit, as was the rate of echo intensity reduction when flow was resumed. Polycythemic blood of 60% hematocrit showed no echo intensity increase over 5 min. Increasing the shear stress when flow was resumed resulted in rapid decreases in aggregation. In all cases, disaggregation following flow resumption was faster than aggregation following flow stoppage.     

Effect of multiple arterial haemorrhage on the coronary circulation

In the open-chest anesthetized dog, multiple arterial haemorrhage induces a reduction of the mean coronary flow, an absote or relative increase of the phasic flow during the early ventricular ejection phase and a decrease during the remaining phases of the cardiac cycle. When blood pressure falls sharply during the haemorrhage, coronary vascular resistance increases, whereas it does not change or decreases when blood pressure falls slowly. Moreover, coronary vascular resistance decreases transiently after the stoppage of the haemorrhage.     

Influence of gravity on flow distribution of red blood cells in microcirculation

The influence of the gravity on flow distribution of erythrocytes in microcirculation was examined. We developed a new centrifuge system with a rotation disc. An observation system of blood flow in a micro-flow channel was arranged on the disc. Erythrocyte flow in the micro-flow tube was displaced under the gravity. This study suggests that the gravity affects the transfer of substances from blood vessels to tissues.     

The flow rate of blood in an environment of weightlessness

To discuss the decrease in the flow rate of blood experienced by astronauts, a theory of blood flow is presented taking account of the effect of gravity. The theory of two-dimensional Poiseuille flow is adopted. It is assumed that the flow is horizontal and that the width of the upper marginal layer filled with plasma thickens as gravity increases. The parameter xi which mainly indicates the effect of thickening of the upper marginal layer is introduced. The extent of decrease in the flow rate of blood in the environment of weightlessness compared to that in the gravitational field is calculated for various values of xi. The decrease is more remarkable in the flow rate of the cell fraction than that of whole blood for the same value of xi.     

Development of a whole blood staining device for use during space shuttle flights.

BACKGROUND: Exposure to microgravity during space flight results in profound physiologic changes. Numerous studies have shown changes in circulating populations of peripheral blood immune cells immediately after space flight. It is currently unknown if these changes result from exposure to microgravity or are caused by the stress of reentry and readaptation to gravity. METHODS: We have developed the whole blood staining device (WBSD) as a system for the staining of whole blood collected during space flight for subsequent flow cytometric analysis. This device contains all liquids to address safety issues concerned with space flight and also moves the cells through the staining, lyse/fixation, and dilution steps. RESULTS: Data from flow cytometric analysis of samples stained in the WBSD was found to be comparable to data from samples stained by the conventional methods. Cells stained with the WBSD remain stable in the device for up to 14 days. The necessary manipulations required to use the device were tested on the KC-135 aircraft during the reduced gravity segment of parabolic flight. CONCLUSIONS: With the WBSD immunophenotype analysis can be performed at various time points for the duration of an entire Shuttle flight. In addition, this device has significant terrestrial applications for rapid and easy immunofluorescence labeling of whole blood in remote and isolated locations where immediate access to specialized equipment and skilled laboratory personnel may not be available. The WBSD provides a simple mechanism to design specific immunophenotyping tests for use by nontechnical personnel at bedside or in field locations. Cytometry 37:74-80, 1999. Published 1999 Wiley-Liss, Inc.     

The venous circulation: a piscine perspective.

Vascular capacitance describes the pressure-volume relationship of the circulatory system. The venous vasculature, which is the main capacitive region in the circulation, is actively controlled by various neurohumoral systems. In terrestrial animals, vascular capacitance control is crucial to prevent orthostatic blood pooling in dependent limbs, while in aquatic animals like fish, the effects of gravity are cancelled out by hydrostatic forces making orthostatic blood pooling an unlikely concern for these animals. Nevertheless, changes in venous capacitance have important implications on cardiovascular homeostasis in fish since it affects venous return and cardiac filling pressure (i.e. central venous blood pressure), which in turn may affect cardiac output. The mean circulatory filling pressure is used to estimate vascular capacitance. In unanaesthetized animals, it is measured as the central venous plateau pressure during a transient stoppage of cardiac output. So far, most studies of venous function in fish have addressed the situation in teleosts (notably the rainbow trout, Oncorhynchus mykiss), while any information on elasmobranchs, cyclostomes and air-breathing fishes is more limited. This review describes venous haemodynamic concepts and neurohumoral control systems in fish. Particular emphasis is placed on venous responses to natural cardiovascular challenges such as exercise, environmental hypoxia and temperature changes.     

The interdependent contributions of gravitational and structural features to perfusion distribution in a multiscale model of the pulmonary circulation

Recent experimental and imaging studies suggest that the influence of gravity on the measured distribution of blood flow in the lung is largely through deformation of the parenchymal tissue. To study the contribution of hydrostatic effects to regional perfusion in the presence of tissue deformation, we have developed an anatomically structured computational model of the pulmonary circulation (arteries, capillaries, veins), coupled to a continuum model of tissue deformation, and including scale-appropriate fluid dynamics for blood flow in each vessel type. The model demonstrates that both structural and the multiple effects of gravity on the pulmonary circulation make a distinct contribution to the distribution of blood. It shows that postural differences in perfusion gradients can be explained by the combined effect of tissue deformation and extra-acinar blood vessel resistance to flow in the dependent tissue. However, gravitational perfusion gradients persist when the effect of tissue deformation is eliminated, highlighting the importance of the hydrostatic effects of gravity on blood distribution in the pulmonary circulation. Coupling of large- and small-scale models reveals variation in microcirculatory driving pressures within isogravitational planes due to extra-acinar vessel resistance. Variation in driving pressures is due to heterogeneous large-vessel resistance as a consequence of geometric asymmetry in the vascular trees and is amplified by the complex balance of pressures, distension, and flow at the microcirculatory level.     

The venous circulation: a piscine perspective

Vascular capacitance describes the pressure–volume relationship of the circulatory system. The venous vasculature, which is the main capacitive region in the circulation, is actively controlled by various neurohumoral systems. In terrestrial animals, vascular capacitance control is crucial to prevent orthostatic blood pooling in dependent limbs, while in aquatic animals like fish, the effects of gravity are cancelled out by hydrostatic forces making orthostatic blood pooling an unlikely concern for these animals. Nevertheless, changes in venous capacitance have important implications on cardiovascular homeostasis in fish since it affects venous return and cardiac filling pressure (i.e. central venous blood pressure), which in turn may affect cardiac output. The mean circulatory filling pressure is used to estimate vascular capacitance. In unanaesthetized animals, it is measured as the central venous plateau pressure during a transient stoppage of cardiac output. So far, most studies of venous function in fish have addressed the situation in teleosts (notably the rainbow trout, Oncorhynchus mykiss), while any information on elasmobranchs, cyclostomes and air-breathing fishes is more limited. This review describes venous haemodynamic concepts and neurohumoral control systems in fish. Particular emphasis is placed on venous responses to natural cardiovascular challenges such as exercise, environmental hypoxia and temperature changes.     

Selected contribution: redistribution of pulmonary perfusion during weightlessness and increased gravity.

To compare the relative contributions of gravity and vascular structure to the distribution of pulmonary blood flow, we flew with pigs on the National Aeronautics and Space Administration KC-135 aircraft. A series of parabolas created alternating weightlessness and 1.8-G conditions. Fluorescent microspheres of varying colors were injected into the pulmonary circulation to mark regional blood flow during different postural and gravitational conditions. The lungs were subsequently removed, air dried, and sectioned into approximately 2 cm(3) pieces. Flow to each piece was determined for the different conditions. Perfusion heterogeneity did not change significantly during weightlessness compared with normal and increased gravitational forces. Regional blood flow to each lung piece changed little despite alterations in posture and gravitational forces. With the use of multiple stepwise linear regression, the contributions of gravity and vascular structure to regional perfusion were separated. We conclude that both gravity and the geometry of the pulmonary vascular tree influence regional pulmonary blood flow. However, the structure of the vascular tree is the primary determinant of regional perfusion in these animals.     

重力对锥形血管中血流压力的影响——基于考虑体位改变的三维流固耦合数学模型

重力是体位改变过程中最基本的生物力学刺激因素．血流压力是表征心血管功能状态的一个基本指标．目前,体位改变影响心血管系统的确切内部机制尚不清楚．为此,采用在流体和固体方程中分别引入体力项的方法,建立一个基于血流动力学概念的三维流固耦合数学模型,用以研究体位改变,确切量化重力对血流压力的影响．通过数值计算,得到以下结果．水平卧位条件下：a．单一血管中血流压力由无重力影响的轴对称二维分布变为重力影响下的三维不对称分布;b．随着进出口压差由小变大,重力对压力分布和极值的影响由大变小,当压差值分别达到10 665.6 Pa(80 mmHg)和2 666.4 Pa(20 mmHg)时,重力的影响就不再随进出口压差增大而变化;对三维单一流体,重力影响的总体趋势类似．对正、倒直立位,压力均为二维轴对称分布,其重力影响强度约为水平卧位的2倍以上．结果表明：基于血流动力学概念,引入体力项,建立三维流固耦合模型为研究体位改变提供了一种新思路,重力对单一血管中血流压力分布和大小的影响因体位不同而不同,并与进出口压差密切相关,提示,若血管进出口压差较小,忽略重力影响,不考虑体位改变,以二维轴对称模型来研究血管中血流状态,须谨慎解释所得结果．     

Hemodynamics for medical students

The flow of blood through the cardiovascular system depends on basic principles of liquid flow in tubes elucidated by Bernoulli and Poiseuille. The elementary equations are described involving pressures related to velocity, acceleration/deceleration, gravity, and viscous resistance to flow (Bernoulli-Poiseuille equation). The roles of vascular diameter and number of branches are emphasized. In the closed vascular system, the importance of gravity is deemphasized, and the occurrence of turbulence in large vessels is pointed out.     

Effect of decreased O2 supply on skeletal muscle oxygenation and O2 consumption during sepsis: role of heterogeneous capillary spacing and blood flow

One of the main aspects of the initial phase of the septic inflammatory response to a bacterial infection is abnormal microvascular perfusion, including decreased functional capillary density (FCD) and increased blood flow heterogeneity. On the other hand, one of the most important phenomena observed in the later stages of sepsis is an increased dependence of tissue O(2) utilization on the convective O(2) supply. This "pathological supply dependency" is associated with organ failure and poor clinical outcomes. Here, a detailed theoretical model of capillary-to-tissue O(2) transport during sepsis is used to examine the origins of abnormal supply dependency. With use of three-dimensional arrays of capillaries with heterogeneous spacing and blood flow, steady-state O(2) transport is simulated numerically during reductions in the O(2) supply. Increased supply dependency is shown to occur in sepsis for hypoxic (decreased hemoglobin O(2) saturation) and stagnant (decreased blood flow) hypoxia. For stagnant hypoxia, a reduction in FCD with decreasing blood flow is necessary to obtain the observed increase in supply dependency. Our results imply that supply dependency observed under normal conditions does not have its origin at the level of individual capillaries. In sepsis, however, diffusion limitation and shunting of O(2) by individual capillaries occur to a degree that is dependent on the heterogeneity of septic injury and the arrangement of capillary networks. Thus heterogeneous stoppage of individual capillaries is a likely factor in pathological supply dependency.     

Paradoxical redistribution of pulmonary blood flow in prone and supine humans exposed to hypergravity.

We hypothesized that exposure to hypergravity in the supine and prone postures causes a redistribution of pulmonary blood flow to dependent lung regions. Four normal subjects were exposed to hypergravity by use of a human centrifuge. Regional lung perfusion was estimated by single-photon-emission computed tomography (SPECT) after administration of (99m)Tc-labeled albumin macroaggregates during normal and three times normal gravity conditions in the supine and prone postures. All images were obtained during normal gravity. Exposure to hypergravity caused a redistribution of blood flow from dependent to nondependent lung regions in all subjects in both postures. We speculate that this unexpected and paradoxical redistribution is a consequence of airway closure in dependent lung regions causing alveolar hypoxia and hypoxic vasoconstriction. Alternatively, increased vascular resistance in dependent lung regions is caused by distortion of lung parenchyma. The redistribution of blood flow is likely to attenuate rather than contribute to the arterial desaturation caused by hypergravity.     

Gravity-independent inequality in pulmonary blood flow in humans

Single-photon emission computerized tomography of the lung with 99mTc-labeled human albumin macroaggregates (99mTc-MAA) was used in six healthy subjects to study the three-dimensional distribution of pulmonary blood flow. 99mTc-MAA was injected while the subjects were resting in the supine position and holding their lung volume at normal end expiration. Tomography was performed on each subject from 120 projections of radioactivity in the lungs acquired with a rotating gamma camera. To minimize lung motion artifacts, the subjects were asked to hold their breath at end expiration during the 10-s duration of data acquisition in each projectional angle. Perfusion images of lung slices (11 mm thick) were reconstructed, and the radioactivity within each slice was expressed per unit lung volume of 3.7 X 3.7 X 11 mm. Perfusion images of a midcoronal slice from each subject manifested a concentric pattern of radioactivity that decreased significantly from the center to the periphery, suggesting that blood flow rate per unit lung volume was up to 10 times larger near the central region. This gradient in activity between the center and the periphery of the coronary slices was gravity independent as the subjects were supine. Images of sagittal slices from the middle of the right lung also manifested a similar pattern of concentric gradient in activity, with the vertical distribution (gravity related) almost comparable with the horizontal distribution (gravity independent). These results indicate that pulmonary blood flow in resting supine humans is spatially stratified with a marked central-to-peripheral gradient in all directions. It appears that zone 4 (reduced blood flow) is not a phenomenon limited to the dependent region of the lung as commonly thought but rather is a manifestation of this spatial distribution whereby blood flow is lowest in all peripheral regions of the lung.     

Impulse activity of the nodose ganglion neurons in myocardial ischemia

Standard microelectrode techniques were used to study the impulse activity of different types of nodosal ganglion neurons in the course of the development of myocardial ischemia. The cardiopulmonary late inspiratory and inspiratory-expiratory neuronal responses were estimated upon ligation of the coronary artery during the first respiratory cycle after blood flow stoppage. Spontaneous activity of cardiopulmonary neurons was not dependent on coronary circulation disturbances at the moment of coronary artery ligation. Later on, however, during the development of myocardial ischemia, there occurred substantial changes in all the types of nodosal ganglion neuronal activity, excluding real inspiratory neurons.     

Pulmonary blood flow distribution in anesthetized ponies.

Results of recent investigations in humans and dogs indicate that gravity-independent factors may be important in determining the distribution of pulmonary blood flow. To further evaluate the role of gravity-independent factors, pulmonary blood flow distribution was examined using 15-microns radionuclide-labeled microspheres in five prone ponies over 5 h of pentobarbital sodium anesthesia. The ponies were killed, and the lungs were excised and dried by air inflation (pressure 45 cmH2O). The dry lungs were cut into transverse slices 1-2 cm thick along the dorsal-ventral axis, parallel to gravity. Radioactivity of pieces cut from alternate slices was measured with a gamma well counter. The main finding was a preferential distribution of pulmonary blood flow to dorsal-caudal regions and higher flow in the center of each lung slice when compared with the slice periphery. Flow was lowest in cranial and ventral areas. Differences of +/- 2 SD were observed between core and peripheral blood flow. No medial-lateral differences were found. Pulmonary blood flow distribution did not change over 5 h of anesthesia, and the basic flow pattern was not different in the left vs. right lung. These results suggest that in the intact prone mechanically ventilated pony (inspired O2 fraction greater than or equal to 0.95) factors other than gravity are primary determinants of pulmonary blood flow.     

Circulatory Adaptations of Snakes to Gravity

Comparative investigations of diverse taxa of snakes demonstratenumerous adaptations for counteracting effects of gravity onthe circulation, including morphological, physiological andbehavioral specializations. Arboreal and terrestrial snakesthat are normally subjected to stresses from gravity are characterizedby relatively high arterial pressures and ability to regulatepressure by physiological adjustments of flow and flow resistance.The heart occupies an anterior position, and the arterial bloodcolumn between the heart and head is comparatively short. Terrestrialsnakes characteristically possess short vascular lungs whicheliminate risks of pulmonary edema due to gravity effects duringvertical posture. Problems of blood pooling in peripheral systemicvasculature are counteracted by relatively non-compliant bodytissue, vasomotor adjustments, and specific movements that facilitatethe venous cardiac return. Anatomical valves appear to be absentfrom major venous channels, but gravity, acting in concert withspecificfeatures of venous morphology, can create valving actions thatimpede shifts of blood volume to dependent segments of thesevessels. Nearly all of these characteristics are absent or deficientin several independent lineages of aquatic snakes that are farless subject to gravitational disturbance of hydrostatic pressures.Thus, snakes provide diverse and particularly useful modelsfor examining cardiovascular adaptations to gravity, includingmechanisms of function and the evolution of cardiovascular design.     

Optimisation of flow cytometric measurement of parasitaemia in plasmodium-infected mice

Mouse malaria is often used as a model for drug testing. The results of drug trials are monitored by tedious (and consequently, sometimes inaccurate) microscopic counting of blood smears, or by flow cytometry. We suggest an improved, accurate and time-saving flow cytometric method for determination of parasitaemias in mice infected with Plasmodium vinckei petteri or Plasmodium berghei. The method involves collection of drops of blood from the tail vein, fixation, storage, permeabilisation, staining and analysis with a visible range flow cytometer. Three nucleic acid dyes, YOYO-1, propidium iodide and acridine orange were compared. YOYO-1 was found to be the best stain for the discrimination of parasitised erythrocytes from non-infected ones. A good direct correlation was obtained between parasitaemia determined by conventional microscopy and parasitaemia measured by flow cytometry. Drug effects could be assessed by the cytometric method. For the detection of low level of parasitemia, parasitised cells were treated with RNAse to completely cancel RNA-derived signals originating from host reticulocytes. This procedure also revealed discrete peaks arising from red cells infected with multiple parasites or from parasites with different numbers of nuclei.     

Aggregation of red cells in disease: some deductions and speculations based on results of "ARC" experiment on the space shuttle "Discovery" STS 51-C

Experiment on STS 51-C in January 1985, carried out on blood samples obtained from patients with heart disease, diabetes, hyperlipidaemia and cancer showed that, under zero gravity, the morphology of red cell aggregates aggregates was normal, in contradistinction to the parallel and simultaneous observations under 1 g, which showed large and unorientated clumps of red cells. As such clumps could be considered of disadvantage in the microcirculation and tissue perfusion, the zero gravity observations were significant in a number of ways. In particular, a preliminary deduction (subject to further zero g experimentation) was that cell-cell interaction and adhesion are affected by zero gravity, and that most likely the microarchitecture of the cell membrane is modified; and that probably the receptors, their position and/or activity, are affected by zero gravity. Of particular interest could be a possible change in the properties of the discrete surface areas which respond preferentially to specific macromolecules (or ligands). There is a dissonance between these in vitro results and theoretical deductions on flow in the microcirculations by Oka, and as well of deductions on space sickness by Dintenfass, both assuming a disabling effect of zero g on the in vivo microcirculation. This dissonance should be explored, as effect of zero g might be different on blood flow in vivo and in vitro. However, the data available from the in vitro experiment suggest that studies in immunology and oncology might be enriched by zero gravity findings; and that studies under zero gravity might open a new avenue of research in these important fields.     

Differences in regional vascular conductances in isolated dog lungs

The distribution of pulmonary blood flow is influenced by gravity, regional lung expansion, and hypoxic pulmonary vasoconstriction. However, these factors cannot completely explain the three-dimensional distribution of blood flow in the lung. The present study was designed to see whether anatomically related factors could contribute. Regional blood pressure vs. flow curves were determined in 100-230 small parenchymal samples (0.3-0.4 ml) from 12 isolated perfused dog lungs held at constant inflation pressure. In each region four blood flows were measured using radioactively labeled microspheres, and the four corresponding regional perfusion pressures were determined by correcting the measured perfusion pressure for hydrostatic effects. There were considerable differences in the slopes of the pressure vs. flow curves among lung regions. Dorso-caudal regions of the lung had higher vascular conductances than ventrocephalad regions, independent of the vertical orientation of the lung or the inflation volume during injections of microspheres. Thus the distributions of regional vascular conductances were related to the anatomic location and were not related to gravity, nor were they caused by nonuniformities in regional lung expansion or by hypoxic vasoconstriction or edema.