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C G Ellis

Publications and source records attributed to C G Ellis.

At least 37 records · Page 2Linked to original sources

Heterogeneity of red blood cell perfusion in capillary networks supplied by a single arteriole in resting skeletal muscle.

Flow heterogeneity within capillary beds may have two sources: (1) unequal distribution of red blood cell (RBC) supply among arterioles and (2) unique properties of RBC flow in branching networks of capillaries. Our aim was to investigate the capillary network as a source of both spatial and temporal heterogeneity of RBC flow. Five networks, each supplied by a single arteriole, were studied in frog sartorius muscle (one network per frog) by intravital video microscopy. Simultaneous data on RBC velocity (millimeters per second), lineal density (RBCs per millimeter), and supply rate (RBCs per second) were measured continuously (10 samples per second) from video recordings in 5 to 10 capillary segments per network for 10 minutes by use of automated computer analysis. To quantify heterogeneity, mean values from successive 10-second intervals were tabulated for each flow parameter in each capillary segment (ie, portion of capillary between successive bifurcations), and percent coefficient of variation (SD/mean.100%) was calculated for (1) spatial heterogeneity among vessels (CVs) every 10 seconds and for the entire 10-minute sample and (2) temporal heterogeneity within vessels for every capillary segment and for the mean flow parameter. Analysis of these data indicates that (1) capillary networks are a significant source of both spatial and temporal flow heterogeneity, and (2) continuous redistributions of flow occur within networks, resulting in substantial temporal changes in CVs, although a persistent spatial heterogeneity of perfusion still exists on a 10-minute basis. In most networks, CVs decreased as supply rate within the network increased, thus indicating that rheology plays a significant role in determining the perfusion heterogeneity.

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Relationship between capillary and systemic venous PO2 during nonhypoxic and hypoxic ventilation.

We evaluated the relationship between end-capillary and systemic venous PO2 values in the retractor muscle of 14 anesthetized hamsters during both nonhypoxic and hypoxic ventilation to ascertain whether the level of tissue oxygenation could be reliably estimated from the systemic parameter. End-capillary PO2 was estimated from measurements of oxygen saturation in capillaries at the venular end of the network obtained using in vivo video microscopy and computer-aided image-analysis techniques at three different levels of inspired oxygen (0.3, 0.21, and 0.1). Measurements of systemic arterial and venous blood gases were made in conjunction with these capillary determinations. In addition, in a portion of the study we utilized an oxygen microelectrode to determine the PO2 in the first-order venule draining the portion of the muscle containing the capillaries under study. We found that only when the animals were made acutely hypoxic was there any correspondence between the systemic venous and end-capillary PO2 values. In addition, these data provide support for the presence of arteriovenous shunting of oxygen during nonhypoxic ventilation.

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Ischemia-reperfusion induced microvascular dysfunction in skeletal muscle: application of intravital video microscopy.

Video microscopy of red cell flow in capillaries at the surface of skeletal muscle provided the opportunity to quantitate ischemia-reperfusion (I-R) induced microcirculatory changes, in vivo. Extensor Digitorum Longus (EDL) muscles of 22 male Wistar rats (300-400 g), anesthetized with sodium pentobarbital (Somnotol, 65 mg kg,-1 IP), were used to measure the number of perfused capillaries (CDper: mm-1) crossing lines drawn perpendicular to the muscle axis, and red blood cell velocity (VRBC: mm/s) within individual capillaries from controls (n = 6), and after 2 hr (n = 4), 3 hr (n = 4), and 4 hr (n = 5) of no-flow ischemia with the muscle temperature maintained at its normal value of 32 degrees C. Ischemia was induced by tightening a tourniquet placed around the limb above the EDL muscle. Measurements were made after 30, 60, and 90 min of reperfusion. To test the usefulness of this skeletal muscle model for evaluating proposed interventions in I-R, the effect of hypothermia (24 degrees C) on the microcirculation following 4 hr ischemia (n = 3) was measured. Edema formation was estimated from the wet/dry weight ratio of the ischemic and contralateral control EDL muscles. Capillary perfusion at the surface of the control muscles was remarkably stable over the 5 hr period studied, while significant changes occurred following the ischemic periods. Significantly lower CDper was measured 30 min following all periods of normothermic ischemia. However, unlike the 2 and 4 hr ischemic periods 3 hr normothermic ischemia resulted in a progressive decline in CDper throughout the reperfusion period. VRBC showed evidence of a hyperemic response following 2 hr normothermic ischemia (control: 0.12 mm/s +/- 0.19 compared to 0.26 mm/s +/- 0.03 following 90 min reperfusion; mean +/- sem). However, no such hyperemia was measured following either 3 or 4 hr normothermic ischemia (i.e., 3 hr control: 0.24 mm/s +/- 0.01 compared to 0.07 mm s +/- 0.003 following 90 min reperfusion). In fact, VRBC was essentially zero 90 min following 4 hr normothermic ischemia (0.01 mm/s +/- 0.01). However, when the muscle was allowed to cool to 24 degrees C during 4 hr ischemia no significant change in either VRBC or CDper was measured compared to pre-ischemic controls. Evidence of edema was found after 3 and 4 hr normothermic ischemia. This study establishes a skeletal muscle model of I-R, which may be useful in testing hypotheses regarding mechanisms of I-R injury, and effectiveness of proposed treatments of I-R.

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Application of image analysis for evaluation of red blood cell dynamics in capillaries.

We have devised a method to display and directly evaluate red blood cell (rbc) dynamics in capillaries using the same dual camera intravital video microscopy system employed to determine rbc oxygen saturation (Ellis et al., 1990). Capillary images are recorded on videotape and an interactive graphics system is used for analysis. Data are sampled once a frame for 60 sec using a window (one pixel wide (0.93 micron) and 100 pixels high) positioned along the axis of a capillary. The resulting data are displayed as sequential space-time images 100 pixels high by 300 pixels wide (10 sec). The space-time images thus created represent the dynamics of the rbc's in a single comprehensive static image in which the rbc's appear as dark, diagonal bands separated by light bands representing plasma gaps. From these images one can obtain information on velocity of individual rbc's (micron/sec), lineal density of rbc's (rbc/mm), and rbc supply rate (rbc/sec). This information can be used to delineate the temporal and spatial heterogeneity of hemodynamics in capillary networks. These data can then be combined with coincident data on red blood cell oxygenation to provide a complete picture of oxygen transport in capillaries or it can be used alone as a tool for the evaluation of basic in vivo and in vitro rheological questions.

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Muscle capillary-to-fiber perimeter ratio: morphometry.

It is known that a substantial amount of capillary tortuosity is found in shortened muscles. However, the increased capillary length and surface area contributed by tortuosity and branching are seldom taken into account when capillarity is estimated and/or blood-tissue exchange is modeled in muscles. In this paper, we sought morphometric estimates of capillarity in transverse sections that incorporated data on capillary geometry. We derived equations to estimate capillary perimeter per fiber perimeter (i.e., capillary-to-fiber perimeter ratio) in transverse sections. We show how capillary-to-fiber perimeter ratio is related to capillary surface per fiber surface, i.e., to the amount of capillary surface available for exchange per muscle fiber surface area, and how it can be obtained by morphometry. Because capillary tortuosity and fiber perimeter are both a function of sarcomere length, the degree of extension or shortening of muscle samples obviously needs to be taken into account when capillary-to-fiber perimeter ratio is compared between muscles and/or samples. Using data currently available on capillary length and diameter with fiber shortening and extension, we show that it is a feature of capillary-to-fiber perimeter ratio to change relatively little with sarcomere length. As sarcomere length decreases from 2.80 to 1.58 microns in perfusion-fixed hindlimb muscles of rats, capillary and fiber perimeters in transverse sections increase substantially, whereas the ratio between the two variables, capillary-to-fiber perimeter ratio, changes only less than or equal to 10-15%.(ABSTRACT TRUNCATED AT 250 WORDS)

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Effect of sarcomere length on total capillary length in skeletal muscle: in vivo evidence for longitudinal stretching of capillaries.

It is generally assumed that when a muscle is shortened or extended the total length of capillaries does not change, implying that capillaries are nondistensible, longitudinally. On the basis of stereological estimates of capillary anisotropy versus sarcomere length, we propose that as long as capillaries are in a tortuous configuration muscle extension will merely decrease the tortuosity, leaving vessel length unaltered. Once capillaries have been pulled into a straight configuration, further extension of the muscle will cause the vessels to stretch. By means of intravital videomicroscopy we have demonstrated that stretching of individual capillaries does indeed occur over a sarcomere length range of 2.1 to 2.9 microns in rat extensor digitorum longus muscle. In vivo measurements of the lengths of six capillaries together with the sarcomere lengths of adjacent fibers were made in muscles positioned at various degrees of extension. Normalized data indicated that four capillaries stretched to the same degree as the muscle, one stretched more and another less. This may reflect differences in distensibility or tortuosity of capillaries in series with one another. The elastic stretching of capillaries during muscle activity may have important consequences in terms of shifts in permeability and increases in capillary surface area.

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Determination of red blood cell oxygenation in vivo by dual video densitometric image analysis.

We have developed a new video microspectrophotometric system for the in vivo determination of oxygen saturation in red blood cells in striated muscle capillaries. This method allows one to quantify changes in the oxygenation of small groups of red blood cells as they traverse the capillary. Simultaneous images of a single microscopic field are recorded using two silicon-intensified target cameras and high-resolution video recorders. One image is recorded at an oxygen-dependent wave-length (431 nm) and the other at an isosbestic wavelength (420 nm). Light intensities from 10 adjacent pixels aligned along the axis of the capillary from identical 10-s segments of the video-tapes are digitized once per frame. Both sets of data are redisplayed simultaneously as two-dimensional images (10 pixels high x 300 frames wide) using a graphics system. These images show alternating bright and dark bands corresponding to plasma gaps and red blood cells. Light intensities in the presence and absence of red blood cells are determined by positioning a window over the appropriate region of the graphics image. Optical densities of single red blood cells at the two wavelengths, OD431 and OD420, are computed as is their ratio (OD431/OD420), which is linearly related to oxygen saturation. In vivo calibration studies in capillaries of the hamster retractor muscle indicate that the error in measuring oxygen saturation with this technique is approximately 2.7% saturation for a group of 10 cells.

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Temporal distributions of red cell supply rate to individual capillaries of resting skeletal muscle, in frog and rat.

Microvascular red cell (RBC) supply in resting skeletal muscle fluctuates with time and varies from capillary to capillary. Our aim was to quantitate this heterogeneity in two species (frog, rat) with very different oxygen requirements. Video recordings were made of RBC flow in capillaries associated with superficial fibers of frog sartorius and rat gracilis muscles, for periods of 2-10 min. RBC supply rate (cell/s) to individual capillaries was computed as the product of RBC 'content' (measured as lineal density, cells/mm, by a video-densitometric method) and RBC velocity (mm/s, measured by a spatial correlation technique). The videotapes were analyzed at rates of 6 to 15 samples per second. Regression analysis of the mean data showed little or no correlation between RBC content and RBC velocity (r2: frog 0.095, rat 0.002), both parameters contributing equally to variations in RBC supply rate. Temporal distributions of the three supply parameters demonstrate (1) the wide range of values present within each capillary, and (2) the symmetry of the distributions at high mean values versus a pronounced skewness as mean values approached zero. The wide range of mean values of each parameter, found in both species, suggests considerable 'spatial' heterogeneity of RBC supply among capillaries in resting muscle. Temporal heterogeneity of RBC supply rate to individual capillaries increased markedly as either velocity or content (or both) approached zero (overall mean coefficient of variation: frog, 62%; rat, 100%). Both species show remarkably similar rates of convective transport of RBCs per capillary in resting muscle (e.g. mean supply rates 3.2 and 6.2 RBCs/s in frog and rat, respectively).

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Variation in axial velocity profile of red cells passing through a single capillary.

We have used an analysis of the velocity of individual red cells as the cells pass through a capillary in order to estimate the variability in cross-sectional area of the capillary lumen available for flow along the length of the vessel. The purpose of the study was to determine if there were irregularities of sufficient magnitude and frequency to support Secomb's hypothesis that local constrictions in the capillary lumen could hinder blood flow at low driving pressure, due to the energy required to deform red cells as they pass through the constriction. All capillary segments analyzed to date, in both rat and frog, have shown regions where the velocity of individual cells is consistently faster or slower than that of the mean velocity of all other cells in the same segment. There are approximately two constrictions per 100 microns in the rat and one per 100 microns in the frog. On average these constrictions appear to reduce the cross-sectional area by 30% in the rat and 16% in the frog. These results provide evidence in support of Secomb's hypothesis. In addition, our results from one bifurcation indicate that the capillary lumen increases in cross-sectional area as one moves from the parent vessel to the region of the bifurcation. Downstream of the bifurcation the lumen rapidly decreases in area by 45 to 54%. Thus a red cell must undergo even greater deformation as it passes through a capillary bifurcation than it will in most other sections of the capillary network.(ABSTRACT TRUNCATED AT 250 WORDS)

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Localized heterogeneity of red cell velocity in skeletal muscle at rest and after contraction.

Using intravital video microscopy, the present study focussed on a detailed analysis of Vrbc heterogeneity in a 2.4 x 1.8 x 0.15 surface volume of a frog sartorius muscle, before and after supramaximal contraction. Heterogeneity of Vrbc was evaluated (1) for an entire population of capillaries seen in this volume, (2) for a series of optical cross-sections, (3) along a series of longitudinal muscle strips, and (4) in terms of an asymmetry ratio for pairs of concurrent capillaries surrounding a muscle fibre. All four types of analysis showed an increased Vrbc homogeneity after contraction. Velocities became more homogeneous along rather than across muscle fibres. The mean asymmetry ratio became significantly larger during post-contraction hyperemia suggesting that each fibre receives a more uniform blood supply that will contribute to an improved exchange of materials across the capillary wall. The analysis of localized Vrbc heterogeneity provides new means of pinpointing the sources of perfusion heterogeneity. It enables, therefore, a specific experimental intervention that is aimed at an improved perfusion under both normal and abnormal conditions.

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Capillary configuration and fiber shortening in muscles of the rat hindlimb: correlation between corrosion casts and stereological measurements.

It has been a matter of discussion whether the dramatic increase in capillary tortuosity visualized in shortened muscles by vascular cast represents in vivo situations. In this study, we combined vascular cast and stereological methods in the same samples, in order to obtain (1) measurements of sarcomere length in the same muscles from which corrosion casts were prepared, and (2) scanning electron micrographs of the three-dimensional arrangement of capillaries in the same muscles where capillary anisotrophy was estimated by morphometry. Various rat skeletal muscles (soleus, gastrocnemius, and gracilis) were examined at lengths ranging from full shortening to full extension. We found a very good correlation between capillary geometry in material prepared for vascular casts and in muscles perfusion-fixed in situ. All muscles, cast and noncast, showed the same progressive curvilinear decrease in capillary anisotropy with decreasing sarcomere length. Capillary tortuosity visualized by corrosion casts in shortened muscles is a consequence of fiber shortening, within physiological sarcomere lengths; it does not represent an artifact related to the casting procedure.

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Respiratory enthalpy changes in mine rescue workers exercising under heat stress.

Respiratory enthalpy change, rectal temperature, and heart rate of mine rescue workers exercising at a metabolic energy production rate of 4 met (1 met-58.15 W.m-2) in a 40 degrees C saturated environment, wearing closed-circuit breathing apparatus, were continuously measured in 10 volunteer subjects. The effects of using liquid O2 and compressed O2 apparatus were compared in each subject. Evaporative heat exchange was much greater with the liquid O2 type of apparatus, causing a significantly lower rate of rise of rectal temperature and heart rate. Convective heat exchange was negligible. Mean values for evaporative heat loss (maximum) were 61 +/- 16 (SD) W with liquid O2 and 20 +/- 18 W with compressed O2 (P less than 0.0001, 2-sided t test). Mean values for rectal temperature (rate of increase) were 0.022 +/- 0.009 (SD) degrees C.min-1 for liquid O2 and 0.036 +/- 0.015 degrees C.min-1 for compressed O2 (P less than 0.005, 2-sided paired t test). Mean values for heart rate (rate of increase) were 2.64 +/- 0.74 (SD) min-2 for liquid O2 and 3.27 +/- 0.89 min-2 for compressed O2 (P less than 0.02, 2-sided paired t test). This study quantifies, for the first time, the respiratory enthalpy change in exercising heat-stressed mine rescue workers and shows, from a physiological point of view, that the liquid O2 apparatus is clearly superior to the compressed O2 apparatus.

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Measurement of hemoglobin oxygen saturation in capillaries.

We present a computer-aided videodensitometric method for the determination of oxygen saturation in red blood cells flowing through capillaries of the hamster cheek pouch retractor muscle. The optical density (OD) of red blood cells is determined at two wavelengths. At the first, 431 nm, there is a maximum difference between absorption by oxygen deoxyhemoglobin. At the second, 420 nm, absorption is equal for the two absorbing species (isosbestic wavelength). In capillaries of the retractor muscle a relationship between oxygen saturation (S) and the following OD ratio was obtained as S = -1.71 (OD431/OD420) + 2.20. The error (95% confidence interval) in oxygen saturation associated with a determination of the OD ratio is estimated to be +/- 4.8%. The computerization of the method employs a frame-by-frame analysis of the light intensity over a selected capillary segment. The light intensity waveform along the segment is digitized and the minimum (I) and maximum (I0) light intensities are used to compute an optical density (OD = log10 [I0/I]). These minimum and maximum intensities correspond to the presence and absence of a red blood cell, respectively. The method permits the off-line analysis of videotaped scenes and provides a means of assessing the extent of temporal and spatial heterogeneity of oxygen saturation in selected capillary networks. The method has been developed for use in capillaries in transilluminated striated muscle but should be generally applicable to the measurement of capillary oxygen saturation in other tissues.

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