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Biomedical subjects

B R Duling

Publications and source records attributed to B R Duling.

At least 55 records · Page 3Linked to original sources
PubMed1982

Adenosine and free-flow functional hyperemia in striated muscle.

Striated muscle arteriolar responses to 1.5 min of 1-Hz contraction and/or increased tissue O2 partial pressure (PO2) were observed during exposure of the tissue interstitial space to adenosine deaminase (ADA) to evaluate the role of adenosine (ADO) as a regulator for blood flow. The microvasculature of the hamster cremaster muscle was continuously superfused with a bicarbonate buffer containing 11 micrograms ADA/ml and equilibrated with 5% CO2 and various O2 concentrations. Arterioles (resting diameter less than 30 micrometers) constricted a maximum of 55% when the superfusate gas tension was increased from 0 to 95% O2, but ADA had no effect on this behavior. Arterioles dilated during exercise, but the diameter change was decreased 20-25% during exercise with ADA treatment at both normal and elevated tissue PO2. As ADA had no effect on either the vasodilation to 2-chloroadenosine or resting arteriolar diameter, it was probably specific in its action. Assuming that all extracellular ADO was accessible to ADA and that ADA neutralized most newly formed ADO, we conclude that ADO is one component of a multifactor system mediating short periods of free-flow exercise hyperemia and that the release of ADO is not necessarily dependent on tissue hypoxia.

Adenosine↗
PubMed1982

A study of rat intracerebral arterioles: methods, morphology, and reactivity.

Penetrating, intracerebral arterioles from rat were isolated, cannulated, and studied in vitro. Vessel wall elements were found to consist of an endothelial cell layer, one smooth muscle cell layer, and a thin adventitial layer or leptomeningeal sheath. Smooth muscle cell nuclei were oriented perpendicular to the vessel's longitudinal axis; endothelial cell nuclei were parallel to the axis. Mean vessel diameter with the smooth muscle inactivated (passive diameter) was 36.7 +/- 1.6 (SE) micrometer. Spontaneous smooth muscle tone developed at 37 degrees C and reduced vessel diameter to 70 +/- 4% of passive diameter. Vessels were activated by the extraluminal application of 140 mM KCl solution at pH 8.00, which produced a transient contraction that decayed within 30 s to a steady contraction of somewhat less intensity. Changes in intravascular pressure were used to alter wall tension of the vessels. Tension in the vessel wall was computed, and length-tension curves for the arteriolar smooth muscle were approximated. Length-tension relationships similar to those seen in other smooth-muscle preparations were found with maximal estimated force development of 1.29 x 10(-5) N . m-2. Alterations of bath pH caused changes in vessel diameter that were inversely related to extraluminal pH and varied by approximately 77% in the range from pH 6.85 to 8.00. Adenosine dilated vessels to 140 +/- 6% of control diameter at a concentration of 10(-5) M. The mechanical characteristics and the reactivity to H+, K+, and adenosine of these vessels were quantitatively consistent with in vitro data from larger cerebral vessels and in vivo data from pial arteries.

Adenosine↗
PubMed1982

Direct measurement of microvessel hematocrit, red cell flux, velocity, and transit time.

A method is presented for the in vivo study of red cell flow dynamics. The method permits direct measurement of the red cell volume fraction in microvessel blood without resort to in vitro calibration curves. Furthermore, the method does not require extensive mathematical manipulation and can be applied to any microvascular network in any tissue. The method also enables direct measurement of red cell velocity, flux, and capillary transit time. Fluorescently labeled erythrocytes in tracer quantities, but known concentrations, are used as indicators of the behavior of the total cell population. Erythrocyte transit time across vascular networks and erythrocyte velocity are determined directly by following the behavior of the labeled cells. Hematocrit and red cell flux are measured by standard microcirculatory methods using labeled cells instead of the total cell population. Data are then converted to absolute values from the measured fraction of labeled cells. The method is thus absolutely dependent on the labeled cells being rheologically normal, and the conditions under which this requirement is satisfied are defined. Microvascular data obtained by the use of this method are presented for hamster cheek pouch and cremaster muscle.

Animals↗
PubMed1982

Augmented tissue oxygen supply during striated muscle contraction in the hamster. Relative contributions of capillary recruitment, functional dilation, and reduced tissue PO2.

To investigate the relative contributions of alterations in blood flow, capillary density, and tissue PO2 to elevated oxygen delivery in working muscle, we conducted experiments on the suffused hamster cremaster muscle, using in vivo microscopic techniques. Muscle PO2 was measured during striated muscle twitch contraction at 1 Hz. Tissue oxygenation was changed by using suffusion solutions equilibrated with 0%, 5%, 10%, 21%, or 50% oxygen. Contraction caused an increase in capillary density (capillary recruitment), whose magnitude was related to the equilibration gas and, thus, to the suffusate PO2. Capillary recruitment first increased as the oxygen content was raised, peaked with 10% oxygen, and then diminished with higher oxygen content. Arteriolar functional dilation was also observed; when oxygen was raised above 21%, dilation was decreased. The data suggest that oxygen supply is increased primarily by arteriolar conductance changes with low suffusion solution oxygen (0% to 5%), and by capillary recruitment and increased PO2 gradients above 10% oxygen. When vasomotor tone was increased by addition of norepinephrine to the suffusion medium, the changes observed were similar to those observed when oxygen was increased. Therefore, we propose that the altered microvascular responses during vasoconstriction are a function of vascular tone rather than the levels of tissue PO2. A model is proposed which may partially explain the relations among vascular tone, functional dilation, and capillary recruitment. Our data also suggest that tissue PO2 may not be precisely regulated about a narrowly defined set point in this striated muscle but that, instead, tissue PO2 is a dependent variable controlled by the integrated effects of capillary recruitment, functional vasodilation, and altered metabolism.

Animals↗
PubMed1982

A comparison of microvascular estimates of capillary blood flow with direct measurements of total striated muscle flow.

Relations between bulk flow into skeletal muscles and microscopically observed capillary flow are compared and disagreements between the two types of data are found. Mean capillary blood velocity was computed using data from a variety of literature sources and assuming uniform distribution of flow through a parallel array of capillaries. The average ratio of measured red cell velocity to computed mean blood velocity was 4.4 +/- 1.0. This is statistically different from the measured value reported in the literature of 1.3. In the cremaster muscle, bulk flow of red cells into the tissue was not statistically different from the measured flux of single red cells through capillaries observed microscopically. The factors which contribute to this apparent agreement of mass balance are not understood, however. Capillary hematocrit is very low and the low capillary hematocrit has been explained by others by the presence of nonuniform flow distribution among the capillaries (shunts or flow heterogeneity). However, for the cremaster data sample, red cells were accounted for in observed capillary flow and red cells were distributed rather homogeneously through the capillaries. This suggests that low capillary hematocrit is the result neither of shunting of red cells around the capillaries nor of nonuniform capillary red cell flow alone. There does not appear to be any well-accepted phenomenon which will explain the findings of: 1) low apparent mean blood velocity; 2) low and variable hematocrit; and 3) apparent conservation of red cell mass. The findings can be reconciled, however, if the capillary rheology is more complex than heretofore anticipated, with a stabilized layer of plasma on the inner surface of the capillary in the order of a 1-micrometer thickness. While there is little direct evidence for such a layer at this time, data which are consistent with the possible existence of such a layer are presented.

Animals↗
PubMed1981

Methods for isolation, cannulation, and in vitro study of single microvessels.

A method is described for the isolation and cannulation of microvessels (12-112 micrometers) that permits study, in vitro, of their physiology and pharmacology. Vessels from the hamster cheek pouch, testis, and mesentery and from rat brain have been isolated at 4 degrees C with specially prepared instruments and viewed with an inverted microscope. The vessels were cannulated at one end by equipment developed for renal tubular perfusion. The uncannulated end of the vessel is sealed, and experiments on reactivity and mechanics are carried out at fixed intravascular pressures. The isolated microvessels studied have a modulus of elasticity that is consistent with that observed in large vessels, and they display similar maximal active tension development (approximately 10(6) dyn/cm2). Reactivity to norepinephrine, acetylcholine, and adenosine are in the normal range for microvessels. Spontaneous tone is present, as evidenced by stable tonic contractions as well as phasic contractions in the frequency range of 3-30/min. The vessels display stress activation (myogenic response) consisting of contraction in response to increased intraluminal pressure. Our findings suggest that this preparation will be very useful in elucidating the physiology and pharmacology of the resistance vessels in the terminal vasculature.

Animals↗
PubMed1979

Measurements of the perivascular PO2 in the vicinity of the pial vessels of the cat.

PO2's in the environment of the pial micro-vessels of the cat were measured using recessed tip oxygen microelectrodes. Measurements were made on the surface of vessels with internal diameters ranging from 200 micrometers to 22 micrometers. Blood oxygen partial pressures were also measured inside these vessels by penetrating the vessels with sharpened electrodes. Both intravascular and extravascular PO2 values decreased progressively from the large arterial vessels down to the small arterioles. The observed values of intravascular PO2 showed a systematic longitudinal decrease from 98.5 +/- 10.7 (SEM) mm Hg in the largest vessels down to 72.6 +/- 3.6 mm Hg in the smallest vessels. In addition to the longitudinal gradient, a transmural gradient was observed across the walls of the microvessels. The difference between blood PO2 and vessel surface PO2 was 27.0 +/- 2.5 mm Hg in the largest vessels and 6.0 +/- 2.2 in the smallest. The mean wall thickness in these groups of vessels were 27.0 +/- 1.5 and 7.5 +/- 0.8 micrometers respectively. Measurements of the minimum tissue PO2 on the exposed surface of the cortex yielded a value of 25.4 +/- 6.6 mm Hg. Systemic arterial partial pressure of oxygen averaged 94.7 +/- 4.7 mm Hg. The data indicate that significant gradients for oxygen exist both longitudinally and radially in association with the pial vessels. The longitudinal gradients represent losses of oxygen from the precapillary vessels. The transmural gradients are apparently the result of both consumption by the microvessel wall and diffusional gradients due to oxygen flux into the extravascular space.

Animals↗
PubMed1979

Microvascular hematocrit and red cell flow in resting and contracting striated muscle.

Microvascular hematocrit and its possible relation to oxygen supply were systematically examined. We studied the red cell volume fraction (hematocrit) in arterial blood and in capillaries under a variety of circumstances. Control capillary hematocrit averaged 10.4 +/- 2.0% (SE) and arteriolar (14.2 micrometer ID) hematocrit averaged 13.9 +/- 1.2% in cremaster muscles of pentobarbital-anesthetized hamsters. Carotid artery hematocrit was 53.2 +/- 0.6%. The low microvessel hematocrit could not be entirely explained by a high red cell flux through arteriovenous channels other than capillaries (shunting). Hematocrit was not only low at rest, but varied with physiological stimuli. A 1-Hz muscle contraction increased capillary hematocrit to 18.5 +/- 2.4%, and maximal vasodilation induced a rise to 39.3 +/- 9.5%. The quantitative relations between capillary red cell flux, arterial hematocrit, and total blood flow could be explained by a two-element model of microvascular blood flow that incorporated a relatively slow-moving plasma layer (1.2 micrometer). Such a model would generate a low microvessel hematocrit and might reduce the diffusion capacity of individual capillaries, but would not reduce time-averaged red cell flux or alter steady-state vascular oxygen supply.

Animals↗
PubMed1978

Interrelations between contracting striated muscle and precapillary microvessels.

Arterioles and capillaries in the hamster cremaster muscle were observed during electrical stimulation of striated muscle fibers in order to characterize the microcirculatory basis of functional hyperemia. When contraction was restricted to single muscle fibers, responses were variable and frequently transient. Stimulation of either small bundles of muscle fibers or the entire cremaster muscle resulted in reproducible responses typified by: 1) a latency period, 2) an early, often transient phase of dilation, and 3) a second, slower phase of dilation. The latency varied inversely with contraction frequency, and the magnitude of the dilation varied directly with contraction frequency over the range 1--8/s. With stimulation of single fibers and small groups of fibers, arteriolar vasodilation was highly localized to regions of the arterioles that were in close apposition to the stimulated fibers. The number of capillaries with red blood cell flow increased during contraction, and the increase was graded with contraction frequency. The changes observed suggest that the vascular response during functional hyperemia is a two-part process and that the control processes are influenced by contraction frequency.

Animals↗
PubMed1978

Role of oxygen in arteriolar functional vasodilation in hamster striated muscle.

Small isolated groups of striated muscle cells were stimulated in the hamster cremaster muscle. During and after stimulation, oxygen microelectrodes were employed to determine the relationships among arteriolar vasodilation, tissue Po2, and periarteriolar Po2. Localized contraction produced a biphasic arteriolar vasodilation without associated alteration of Po2 on the surface of the arterioles (vascular smooth muscle Po2). In contrast, muscle contraction produced a decline in muscle tissue Po2 that was proportional to the contraction frequency over the range of 1--4 contractions per second. An increase in contraction frequency also produced a graded increase in arteriolar diameter, the magnitude of which was statistically correlated with the steady-state change in tissue Po2. However, arteriolar diameter changes preceded tissue Po2 changes, both with the initiation of functional dilation and during the recovery period. Tissue Po2 was manipulated at rest and during contraction by increasing the Po2 of the superfusion solution. Increasing the tissue Po2 caused a decrease in vascular diameter under both conditions and a reduction in the magnitude of the arteriolar vasodilation during contraction. Restoration of tissue Po2 to resting levels during muscle contraction produced only partial restoration of vascular diameters. The results are consistent with the hypothesis that at least three components are involved in the vascular control process during muscular activity: an early component independent of tissue oxygen levels, a late component independent of oxygen, and a late component associated with a decrease in muscle Po2, without an effect on vascular smooth muscle Po2. The evidence indicated that Po2 of the smooth muscle of the arterioles had no role in the dilation observed.

Animals↗
PubMed1975

A new method for the measurement of percent oxyhemoglobin.

A new method is presented for the spectrophotometric determination of percent oxyhemoglobin (percent saturation) in whole blood. The method is based on a theory of light absorption and scattering by particulate suspensions and requires the measurement of optical densities (D) of blood at three closely spaced wavelengths. The contribution of scattering to the optical density at each wavelength is determined from optical density values at two isosbestic wavelengths (546 and 520 nm) and the optical density at the third wavelength (555 nm) is related to the extent of oxygenation of the hemoglobin. The wavelength independence of the scattering contribution (B) induced by red cells was established from 510 and 575 nm. The optical density of red blood cell suspensions was measured with a spectrophotometer and a linear relationship was found between percent saturation and the corrected optical density ratio, (D555 B)/(D546 B). For a given saturation, this ratio was independent of optical path length (0.1-2 mm) and hematocrit (3-50%).

Erythrocytes↗
PubMed1975

Measurement of percent oxyhemoglobin in the microvasculature.

A new method is applied to the determination of percent oxyhemoglobin (percent saturation) in whole blood. The method is based on a theoretical treatment of light absorption and scattering by particulate suspensions and requires the measurement of optical densities (D) of blood at three closely spaced wavelengths. The optical density of red blood cell (RBC) suspensions was measured with a video microdensitometer and a linear relationship was found between percent saturation and the corrected optical density ratio, (D555 B)/(D546 B). For a given saturation, this ratio was independent of optical path length (12 mum-2 mm), hematocrit (3-50%), and RBC velocity (1.5-17 mm/s). The applicability to microvascular measurements has been assessed through the use of TV microdensitometry on micropipettes with flowing RBCs and on microvessels in the hamster cheek pouch.

Densitometry↗