Biomedical subjects

Anatoliy A Gashev

Publications and source records attributed to Anatoliy A Gashev.

12 recordsLinked to original sources

PubMed2006

Length-tension relationships of small arteries, veins, and lymphatics from the rat mesenteric microcirculation.

The passive and active length-tension relationships of isolated rat mesenteric lymphatics ( approximately 150 microm ID), and adjacent small arteries ( approximately 240 microm) and veins ( approximately 275 microm) were compared under isometric conditions using a wire myograph. About 60% of the lymphatic vessels developed spontaneous contractions in physiological saline solution at nominal preload. To maximally activate smooth muscle, 145 mM K(+) + 5 x 10(-5) M norepinephrine was used for arteries, and 145 mM K(+) + 1 x 10(-6) M substance P was used for lymphatics and veins. In response, arteries exhibited monotonic force development to a plateau level, whereas lymphatics and veins showed biphasic force development, consisting of a transient force peak followed by partial relaxation to a plateau over approximately 5 min. The passive and the active length-tension curves were similar in shape among all three vessels. However, the maximal active tension of arteries (3.4 +/- 0.42 mN/mm) was significantly greater than peak active tension (0.59 +/- 0.04 mN/mm) or plateau tension (0.20 +/- 0.04 mN/mm) in small veins and greater than peak active tension (0.34 +/- 0.02 mN/mm) or plateau tension (0.21 +/- 0.02 mN/mm) in lymphatics. Maximal active medial wall stress was similar between lymphatics and veins but was approximately fivefold higher in small arteries. For lymphatics, the pressure calculated from the optimal preload was significantly higher than that found previously in isobaric studies of isolated lymphatics, suggesting the capacity to operate at higher than normal pressures for increased responsiveness. Our results represent the first mechanical comparisons of arterial, venous, and lymphatic vessels in the same vasculature.

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PubMed2006

Image correlation algorithm for measuring lymphocyte velocity and diameter changes in contracting microlymphatics.

Efforts have recently been made to estimate wall shear stress throughout the contractile cycle of mesenteric rat lymphatics with a high speed video microscopy system. This was prompted by reports in the literature that lymphatic pumping is related to wall shear stress. While one can estimate wall shear stress by tracking lymphocyte velocity, it is prohibitively tedious to manually track particles over a reasonable time frame for a good number of experiments. To overcome this, an image correlation method similar to digital particle imaging velocimetry was developed and tested on contracting lymphatics to measure both vessel diameter and fluid velocity. The program tracked temporal fluctuations in spatially averaged velocity with a standard error of prediction of 0.4 mm/s. From these studies we have measured velocities ranging from -2 to 4 mm/s. Diameter changes were also measured with a standard error of 7 microm. These algorithms and techniques could be beneficial for investigating various changes in contractile behavior as a function of changes in velocity and wall shear stress.

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PubMed2006

Intrinsic pump-conduit behavior of lymphangions.

Lymphangions, segments of lymphatic vessels bounded by valves, have characteristics of both ventricles and arteries. They can act primarily like pumps when actively transporting lymph against a pressure gradient. They also can act as conduit vessels when passively transporting lymph down a pressure gradient. This duality has implications for clinical treatment of several types of edema, since the strategy to optimize lymph flow may depend on whether it is most beneficial for lymphangions to act as pumps or conduits. To address this duality, we employed a simple computational model of a contracting lymphangion, predicted the flows at both positive and negative axial pressure gradients, and validated the results with in vitro experiments on bovine mesenteric vessels. This model illustrates that contraction increases flow for normal axial pressure gradients. With edema, limb elevation, or external compression, however, the pressure gradient might reverse, and lymph may flow passively down a pressure gradient. In such cases, the valves may be forced open during the entire contraction cycle. The vessel thus acts as a conduit, and contraction has the effect of increasing resistance to passive flow, thus inhibiting flow rather than promoting it. This analysis may explain a possible physiological benefit of the observed flow-mediated inhibition of the lymphatic pump at high flow rates.

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PubMed2006

Contraction-initiated NO-dependent lymphatic relaxation: a self-regulatory mechanism in rat thoracic duct.

The objectives of this study were to evaluate the physiological importance of the flow and shear generated by phasic contractions of lymphatic vessels and the mechanisms responsible for the influences of such shear on lymphatic pumping. Lymphatic segments of the rat thoracic duct were isolated, cannulated and pressurized. The diastolic diameters were measured in phasically non-active segments. The diastolic and systolic diameters, half-relaxation time (HRT), contraction frequency, ejection fraction and fractional pump flow were determined in phasically active segments. Since imposed flow was excluded, flow and shear occurred only as a result of the intrinsic contractions in phasically active segments whereas in phasically non-active segments contraction-generated flow and shear were absent. The influences of incrementally increased transmural pressure (from 1 to 5 cmH(2)O) were examined in control conditions and after NO synthase blockade (l-NAME 10(-4) m) or cyclooxygenase blockade (indomethacin 10(-5) m). The spontaneous phasic contractions produced a flow-dependent diastolic relaxation. This reduction of the lymphatic tone is a regulatory mechanism that maintains pumping in thoracic duct in an energy-saving/efficient mode: it improves diastolic filling (enhanced lusitropy - lowering HRT), makes lymphatic contractions stronger (enhanced inotropy - higher contraction amplitude) and propels more fluid forward during each contraction (elevated ejection fraction) while decreasing contraction frequency (reduced chronotropy). The findings also demonstrated that the NO pathway, not the cyclooxygenase pathway is responsible for this reduction of lymphatic tone and is the prevailing pathway responsible for the self-regulatory adjustment of thoracic duct pumping to changes in lymph flow pattern.

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PubMed2006

Inhibition of active lymph pump by simulated microgravity in rats.

During spaceflight the normal head-to-foot hydrostatic pressure gradients are eliminated and body fluids shift toward the head, resulting in a diminished fluid volume in the legs and an increased fluid volume in the head, neck, and upper extremities. Lymphatic function is important in the maintenance of normal tissue fluid volume, but it is not clear how microgravity influences lymphatic pumping. We performed a detailed evaluation of the influence of simulated microgravity on lymphatic diameter, wall thickness, elastance, tone, and other measures of phasic contractility in isolated lymphatics. Head-down tail suspension (HDT) rats were used to simulate the effects of microgravity. Animals were exposed to HDT for 2 wk, after which data were collected and compared with the control non-HDT group. Lymphatics from four regional lymphatic beds (thoracic duct, cervical, mesenteric, and femoral lymphatics) were isolated, cannulated, and pressurized. Input and output pressures were adjusted to apply a range of transmural pressures and flows to the lymphatics. Simulated microgravity caused a potent inhibition of pressure/stretch-stimulated pumping in all four groups of lymphatics. The greatest inhibition was found in cervical lymphatics. These findings presumably are correlated to the cephalic fluid shifts that occur in HDT rats as well as those observed during spaceflight. Flow-dependent pump inhibition was increased after HDT, especially in the thoracic duct. Mesenteric lymphatics were less strongly influenced by HDT, which may support the idea that lymph hydrodynamic conditions in the mesenteric lymphatic during HDT are not dramatically altered.

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PubMed2006

Automated measurement of diameter and contraction waves of cannulated lymphatic microvessels.

BACKGROUND: Studies of lymphatic function often employ collecting lymphatic vessels that exhibit large-amplitude, spontaneous contractions. Data from such preparations have been analyzed using cardiac pump analogies that require accurate determination of vascular dimensions, including external (OD) and internal (ID) diameters. These measurements would be facilitated by an accurate automated measurement system. METHODS AND RESULTS: A computer-based diameter tracking system was developed specifically for lymphatic vessels, with advantages over previous automated systems. The system also permits continuous diameter tracking at two axial locations, enabling the measurement and analysis of contraction wave conduction. The method was validated using spontaneously contracting segments of rat thoracic duct which sometimes exhibited conducted contraction waves. In such preparations, conduction wave velocity was modulated by the axial flow rate and could be easily measured by the tracking system. CONCLUSIONS: The method offers improvement and increased convenience over manual diameter measurements in lymphatic vessels, with little or no sacrifice in accuracy. It should be a useful tool for general studies of collecting lymphatic function as well as for the analysis of contraction wave conduction and coordination.

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PubMed2004

Regional variations of contractile activity in isolated rat lymphatics.

OBJECTIVE: To evaluate lymphatic contractile activity in different regions of the lymphatic system in a single animal model (the rat thoracic duct, mesenteric, cervical, and femoral lymphatics) in response to changes in lymph pressure and flow. METHODS: The systolic and diastolic diameters of isolated, cannulated, and pressurized lymphatic vessels were measured. Contraction frequency, ejection fraction, and fractional pump flow were determined. The influences of incrementally increased transmural pressure (from 1 to 9 cm H2O) and imposed flow (from 1 to 5 cm H2O transaxial pressure gradient) were investigated. RESULTS: The authors determined regional differences in lymphatic contractility in response to pressure and imposed flow. They found the highest pumping (at the optimal pressure levels) in mesenteric lymphatics and lowest pumping in thoracic duct. All lymphatics had their optimal pumping conditions at low levels of transmural pressure. Different degrees of the flow-induced inhibition of the pump were observed in the different types of lymphatics. During high flow, the active lymph pumps in thoracic duct and cervical lymphatics were almost completely abolished, whereas mesenteric and femoral lymphatics still exhibited significant active pumping. CONCLUSIONS: The active lymph pumps in different regions of the rat body express variable relative strengths and sensitivities that are predetermined by different hydrodynamic factors and regional outflow resistances in their respective locations.

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PubMed2003

Lymphatic muscle: a review of contractile function.

A recent surge in lymphangiogenesis research has led to a greater understanding of lymphatic endothelial cell biology. However, a general understanding of lymphatic muscle cell biology lags far behind its endothelial counterpart. Lymphatics at the level of the collecting vessels and higher contain muscular walls capable of both tonic and phasic contractions, which both generate and regulate lymph flow. Because lymphatic contraction is crucial to lymphatic function, a solid understanding of lymphatic muscle development and function is necessary to understand lymphatic biology. This review summarizes the current body of lymphatic muscle research and addresses important questions that are currently unanswered.

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PubMed2002

Inhibition of the active lymph pump by flow in rat mesenteric lymphatics and thoracic duct.

There are only a few reports of the influence of imposed flow on an active lymph pump under conditions of controlled intraluminal pressure. Thus, the mechanisms are not clearly defined. Rat mesenteric lymphatics and thoracic ducts were isolated, cannulated and pressurized. Input and output pressures were adjusted to impose various flows. Lymphatic systolic and diastolic diameters were measured and used to determine contraction frequency and pump flow indices. Imposed flow inhibited the active lymph pump in both mesenteric lymphatics and in the thoracic duct. The active pump of the thoracic duct appeared more sensitive to flow than did the active pump of the mesenteric lymphatics. Imposed flow reduced the frequency and amplitude of the contractions and accordingly the active pump flow. Flow-induced inhibition of the active lymph pump followed two temporal patterns. The first pattern was a rapidly developing inhibition of contraction frequency. Upon imposition of flow, the contraction frequency immediately fell and then partially recovered over time during continued flow. This effect was dependent on the magnitude of imposed flow, but did not depend on the direction of flow. The effect also depended upon the rate of change in the direction of flow. The second pattern was a slowly developing reduction of the amplitude of the lymphatic contractions, which increased over time during continued flow. The inhibition of contraction amplitude was dependent on the direction of the imposed flow, but independent of the magnitude of flow. Nitric oxide was partly but not completely responsible for the influence of flow on the mesenteric lymph pump. Exposure to NO mimicked the effects of flow, and inhibition of the NO synthase by N (G)-monomethyl-L-arginine attenuated but did not completely abolish the effects of flow.

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PubMed2002

Physiologic aspects of lymphatic contractile function: current perspectives.

The lymphatic system plays an important role in fluid/macromolecular balance, lipid absorption, and immune functions, and is involved in many different pathologic conditions, like inflammation, spread of cancer cells, and lymphedema. There are several forces that drive lymph centripetally. Extrinsic driving forces, or the passive lymph pump, include lymph formation, arterial pulsations, skeletal muscles contractions, fluctuations of central venous pressure, gastrointestinal peristalsis, and respiration. Intrinsic forces, or the active lymph pump, are the result of coordinated contractions of lymphangions, the morpho-functional units of the lymphatic vessels, which include the valve and portion of the vessel extending to the next valve. The contractions of the lymphangions are initiated by the pacemaker activity of the smooth muscle cells of lymphangion wall. Transmural pressure is an important hydrodynamic factor that modulates pacemaking. Under conditions of low filling, lymphangions might produce negative intraluminal pressures and a suction effect. Because of the complicated hydrodynamic conditions in lymphatic beds, the passive and active lymph pumps sometimes work together to propel lymph centripetally. In other cases (i.e., under conditions of enhanced lymph flow), flow-mediated inhibition of the active lymph pump could serve to decrease lymphatic outflow resistance and save metabolic energy when the driving force of the passive lymph pump is enough to propel lymph. We have recently found that there are profound differences in the pressure and flow sensitivities of lymphatic vessels derived from different tissues, such as the thoracic duct and mesenteric lymphatics. Such results, when considered in light of the controversy surrounding some studies performed in different animals, lead to the idea that the active lymph pumps in humans may have greater regional differences in contractile function than has been seen in animals, because of the upright posture in bipedal humans. This posture creates an additional outflow resistance for lymphatics of the lower part of the body. Thus, despite the ongoing attempts to determine the mechanisms of lymphatic diseases and useful therapies to treat them, there are many disputable or unknown issues regarding the physiology of lymph transport in humans.

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PubMedDate not supplied

Lymph flow, shear stress, and lymphocyte velocity in rat mesenteric prenodal lymphatics.

OBJECTIVE: To measure lymphocyte velocity, lymphatic contraction, and shear stress in phasically contracting lymphatics in situ. METHODS: A high-speed video system was used to capture multiple contraction cycles in rat mesenteric lymphatic preparations. The images were analyzed to determine fluid velocity, volume flow rate, wall shear stress, and retrograde flow. RESULTS: Lymphocyte density and flux varied from 326 to 35,500 cells/microL and 206 to 2,030 cells/min, respectively. Lymphatics contracted phasically, with a mean diameter 91 +/- 9.0 microm and amplitudes of 39%. Lymph velocity varied with the phasic contractions in both direction and magnitude with an average of 0.87 +/- 0.18 and peaks of 2.2-9.0 mm/s. The velocity was approximately 180 degrees out of phase with the contraction cycle. The average lymph flow was 13.95 +/- 5.27 microL/h with transient periods of flow reversal. This resulted in an average shear of 0.64 +/- 0.14 with peaks of 4-12 dynes/cm(2). CONCLUSIONS: High-speed lymphocyte tracking provided the spatial and temporal resolution to measure lymphocyte flux throughout the phasic contraction. Poiseuille flow was a reasonable model for estimating wall shear stress through most of the phasic contraction cycle of the intervalvular lymphatic segments. Shear rate was low but had large variations in magnitude compared to that seen in blood vessels.

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PubMedDate not supplied

Measuring microlymphatic flow using fast video microscopy.

Despite advances in the measurement of lymphatic function, little is known about the actual velocities of flow in microlymphatic ( approximately 100 mum diam) vessels. In this work, video microscopy and particle tracking methods are adapted and integrated with an ultra-high-speed imaging camera to obtain measurements of lymph velocities throughout the entire lymphatic contraction cycle in the ratmesentery, something that previous systems were incapable of measuring. To determine the system's accuracy, calibration experiments are conducted across the hypothesized physiologically significant range of velocities for microlymphatic flow (up to 15 mmsec). The system shows high accuracy, less than 2% error, when comparing actual with measured velocities. Microspheres flowing through 140-mum-diam tubing are imaged to demonstrate the system's ability to determine flow rates in these small vessels by measuring particle velocities. To demonstrate biological applicability, mesenteric microlymphatics in loops of the small intestine of three male Sprague-Dawley rats are exteriorized and imaged with the high-speed system at a rate of 500 framessec for several contraction sequences. Lymph velocity fluctuates cyclically with the vessel wall contractions, ranging from -1 to 7 mmsec. These rates are higher than would be possible with standard video microscopy (3.75 mmsec maximum).

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