Biomedical subjects

P Gaehtgens

Publications and source records attributed to P Gaehtgens.

At least 19 recordsLinked to original sources

PubMed1995

Flow-dependent regulation of arteriolar diameter in rat skeletal muscle in situ: role of endothelium-derived relaxing factor and prostanoids.

1. Arteriolar diameter in the resting rat spinotrapezius muscle was studied by intravital video microscopy before and after blockade of the L-arginine-EDRF (NG-nitro-L-arginine, L-NNA) or the cyclo-oxygenase-prostacyclin (indomethacin) pathway. Blockade of either pathway leads to a decrease of arteriolar diameter of 25-40%, while the combined blockade of both results in vasoconstriction of 50-60%. 2. Alteration of blood flow velocity elicited by partial micropipette occlusion induces corresponding changes of vessel diameter. The flow-dependent diameter response is reduced by about 80% by L-NNA. By contrast, blockade of prostanoid production shows no significant influence on vessel response to blood flow alteration in the range tested. 3. Transient overshooting vasodilatation is seen for about 1 min following the sudden restoration of flow velocity subsequent to occlusion. In contrast to the initial phase of this response, the late phase is blocked by L-NNA. 4. The findings suggest that basal release of endothelium-derived relaxing factor (EDRF) and prostanoids leads to additive and independent dilator effects, and that flow-dependent diameter changes are primarily mediated by EDRF. 5. If present data are compared with literature reports, it appears that arterial flow sensitivity is most pronounced in the smallest vessels. In such vessels, flow-dependent dilatation will amplify even small changes of volume flow by more than four times.

Animals↗

PubMed1995

The leukocyte integrin Mac-1 (CD11b/CD18) contributes to binding of human granulocytes to collagen.

Adhesion of polymorphonuclear granulocytes (PMN) to extracellular matrix proteins has been shown to be important for their migration in vitro and is thought to participate in PMN recruitment to sites of inflammation. Isolated human PMN stimulated with PMA were found to adhere best to microtiter wells coated with the novel ECM glycoprotein undulin (27 +/- 3% of PMNs added), followed by fibrinogen (25 +/- 2%), collagen type VI (18 +/- 2%), fibronectin (16 +/- 2%), and laminin (15 +/- 3%). PMN adhesion to other collagens ranged between 3 and 11%. Monoclonal antibodies recognizing CD18 and CD11b subunits of Mac-1 inhibited adhesion of PMN to collagens by an order of magnitude more effectively than to all noncollagenous substrates. F(ab')2 fragments of the anti-CD18 antibody were also able to block adhesion to collagens. Anti-LFA-1 (CD11a) and anti-CD44 antibodies did not significantly reduce adhesion. PMN adhesion was also inhibited by soluble collagens type II and VI (ID50 approximately 75 micrograms/ml). Binding of soluble radiolabeled collagens type II and VI to PMNs was specific and saturable with apparent dissociation constants of 2.2 and 1.9 nM, respectively, and specific binding of collagens type II and VI was almost completely inhibited by anti-CD18, but not by control antibodies. These data indicate that Mac-1 function is required for binding of human PMN to collagens.

Antibodies, Monoclonal↗

PubMed1995

Effect of oxygen tension on regulation of arteriolar diameter in skeletal muscle in situ.

Skeletal muscle arterioles are known to constrict upon elevation of ambient PO2. While several studies have shown that the endothelium plays an important role in this response, it is not clear how this response is mediated. We examined the oxygen-induced constriction of arterioles in the rat spinotrapezius muscle. Elevation of superfusion solution PO2 from about 15 to 150 mm Hg caused arteriolar constriction by 25% (+/- 3%, n = 18). Inhibition of prostaglandin synthesis by superfusion of indomethacin (30 microM) produced vasoconstriction by 28% (+/- 9.5%, n = 5), but left the PO2 response unaffected. Blockade of the synthesis of endothelium-derived relaxing factor (EDRF) by NG-nitro-L-arginine (L-NNA, 35 mg/kg i.v.) caused arteriolar constriction by 31% (+/- 8%, n = 8). During application of L-NNA, the constrictor response to PO2 elevation was reduced to 3 +/- 2%. Administration of superoxide dismutase (SOD, 80,000 U/kg i.v.) did not affect the PO2 response. It is concluded that in small arterioles of skeletal muscle both EDRF and prostanoids sustain a significant basal dilatation. The dilatory effects of EDRF but not of prostaglandins are strongly dependent on PO2. The vasoconstriction in response to high ambient PO2 is not due to EDRF breakdown during its diffusion from endothelial to smooth muscle cells.

Animals↗

PubMed1995

Does luminal release of EDRF contribute to downstream microvascular tone?

The contribution of luminally released endothelium-derived relaxing factor (EDRF) transported with the flowing blood to the control of smooth muscle tone of downstream arterioles was studied. The dilatory response of arterioles in the spinotrapezius muscle of the rat to superfusion of acetylcholine (ACH) was investigated by intravital microscopy before and after blockade of blood flow by micropipette occlusion upstream of the site of observation. Vessels were studied without treatment, after topical application of indomethacin (INDO, 3 10(-5) M) to inhibit prostanoid production and after local treatment with NG-nitro-L-arginine (LNNA, 2.9 10(-3) M) to suppress local production of EDRF. In untreated vessels and after INDO, responses to ACH were not significantly different in the presence or absence of blood flow. After LNNA, the responses to ACH were reduced to about 50%, but were again similar with or without flow. These results indicate that luminally transported EDRF does not play a significant role in controlling arteriolar tone in small skeletal muscle arterioles.

Acetylcholine↗

PubMed1995

Transient rheological behavior of blood in low-shear tube flow: velocity profiles and effective viscosity.

Velocity profiles of human blood flowing through vertical and horizontal glass tubes (25-100 microns ID) were measured as a function of time following a sudden reduction of wall shear stress (tau w) from a high value to values ranging from 2 to 100 mPa. Cell velocities at various radial positions were determined off-line from video recordings by digital image analysis. In vertical tubes, symmetric velocity profiles were obtained that developed increasing bluntness with time, particularly at lower tau w and in smaller tubes. In horizontal tubes, velocity profiles developed strong asymmetry as a function of time. Red blood cell (RBC) sedimentation was associated with uniform low flow velocities in the concentrating cell sediment, whereas faster flow and almost parabolic profiles were observed in the supernatant plasma region. Calculations of effective blood viscosity showed a decrease with time at low tau w in vertical tubes but an increase in horizontal tubes. The differences between profile shape and effective viscosity in vertical and horizontal tubes disappeared at tau w > 50 mPa. These findings are related to the cross-sectional distribution of RBC, which depends on RBC aggregation and sedimentation.

Blood Flow Velocity↗

PubMed1995

Postural vascular response vs. sympathetic vasoconstriction in human skin during orthostasis.

Sympathetic activation and local vascular smooth muscle reactions to vessel distension contribute to the increase in vascular resistance in the skin during orthostasis. The relative contribution of these two mechanisms to the changes of skin blood flow along the body axis on standing was investigated in healthy male subjects by laser-Doppler (LD) fluxmetry. Compared with recumbency, LD flux (LDF) in the standing subjects was reduced by -19.6 +/- 7.2% at the forehead and by -69.6 +/- 9.6% in the leg. In the absence of hydrostatic pressure changes, the LDF changes on standing averaged -29 +/- 13%, independent of skin region, reflecting the effect of vasoconstriction due to sympathetic activation. The postural vascular response, elicited by lowering the arm or the leg from heart level, was significantly attenuated in orthostasis compared with recumbency. The vessel reaction to local alteration of transmural pressure was studied in the skin of the forehead and lower leg by application of external pressure in supine subjects. No difference in vessel responsiveness to changes of transmural pressure was found between these skin sites. The findings suggest that the changes of skin perfusion in orthostasis result from a nonadditive interaction of height-dependent and -independent mechanisms.

Adult↗

PubMed1995

Structure and hemodynamics of microvascular networks: heterogeneity and correlations.

The objective of this study was to quantify the heterogeneity of topological, morphological, and hemodynamic parameters in microvascular networks and to identify functionally relevant correlations among these parameters. Seven networks in the rat mesentery (383-913 vessel segments per network) were examined, and measurements were made of segment generation, diameter, length, and hematocrit in all segments (n = 3,129) and of flow velocity (only in 3 networks, 1,321 segments). In addition, hematocrit, flow rate, and pressure were derived for all segments from a mathematical simulation. All parameters obtained exhibit heterogeneous distributions with coefficients of variation ranging from 0.28 (capillary diameter) to > 1.5 (volume flow and pressure gradient). Several strong correlations exist between parameters, e.g., discharge hematocrit increases with vessel diameter, and shear rate increases with intravascular pressure. Because of such correlations, the extrapolation from average values for "typical vessels" to network properties can lead to substantial errors. For example, the mean network transit time estimated based on averaged quantities is 6.5 s, which is about 60% higher than the true value (4.08 s). Simplified models of the vascular bed may therefore be inadequate to describe functional properties of the microcirculation.

Animals↗

PubMed1995

Design principles of vascular beds.

Hemodynamic parameters were determined in each vessel segment of six complete microvascular networks in the rat mesentery by using a combination of experimental measurements and theoretical stimulations. For a total number of 2592 segments, a strong unified dependence of wall shear stress on intravascular pressure for arterioles, capillaries, and venules was obtained. All three types of segments exhibit an essentially identical variation of shear stress from high to low values (from approximately 100 to 10 dyne/cm2) as intravascular pressure falls from 70 to 15 mm Hg. On the basis of these observations, it is proposed that vascular beds grow and adapt so as to maintain the shear stress in each vessel at a level that depends on local transmural pressure. In contrast to Murray's classic 'minimum-cost' hypothesis, which implies uniformity of wall shear rate throughout the vasculature, the proposed design principle provides an explanation for the functionally important arteriovenous asymmetry of wall shear rates and flow resistance in the circulation.

Animals↗

PubMed1995

Fluid, electrolyte, and packed cell volume shifts in racing greyhounds.

Arterial blood samples were obtained at rest, just before, and 5 minutes after a 704-m race, to quantify changes in hematologic variables, plasma electrolyte and protein concentrations, osmolality, and acid/base variables. Changes in plasma volume were estimated from the change in plasma protein concentration. Immediately prior to the race, plasma volume decreased by 10% from rest and total circulating RBC volume increased by 60%, attributable to increased RBC number rather than size. Increases in blood volume (VB) by 24% and PCV by 29% also were detected before the race. Five minutes after the race, plasma volume was 21% below the resting value and total circulating RBC volume had increased 73% above the resting value, resulting in a 40% increase in PCV. Contraction of the spleen appeared responsible for increased PCV and VB before the race and maintenance of VB after the race. Plasma chloride concentration was the same before and after the race; the chloride content of the plasma decreased by the same fraction (22%) as did the plasma volume, indicating Cl- loss from the plasma. Plasma Na+ content decreased by a smaller fraction (13%), causing Na+ concentration to increase from 151 mEq/L at rest to 167 mEq/L after the race. Assuming that Na+ concentration was the same throughout the extracellular fluid, H2O likely moved into the intracellular compartment. As a consequence of these changes, the inorganic strong ion difference in plasma increased by about 16 mEq/L, tending to minimize the acid/base disturbance induced by the 33 mEq/L increase in lactate concentration.(ABSTRACT TRUNCATED AT 250 WORDS)

Acid-Base Equilibrium↗

PubMed1994

Cross-linking of CD18 in human neutrophils induces an increase of intracellular free Ca2+, exocytosis of azurophilic granules, quantitative up-regulation of CD18, shedding of L-selectin, and actin polymerization.

Polymorphonuclear leukocytes (PMNs) exert most of their physiological functions while adherent to surfaces rather than in suspension. PMN adhesion is largely dependent on the function of the beta 2 integrins, CD11a,b,c/CD18. We mimicked engagement of beta 2 integrins by antibody cross-linking of CD18 on isolated human PMNs using both intact monoclonal antibody and F(ab')2 fragments. Within seconds of CD18 cross-linking, we observed a significant, transient rise of intracellular free Ca2+ concentration by 200-300 nM, which was largely due to Ca2+ mobilization from intracellular stores. The Ca2+ signal was blocked after pretreatment with phorbol myristate acetate, an activator of protein kinase C, but not with herbimycin A, a potent inhibitor of tyrosine kinases. In addition to the rise of intracellular free Ca2+ concentration, CD18 cross-linking induced exocytosis of azurophilic granules (release of 26% of total PMN elastase), which was significantly inhibited by herbimycin A. Moreover, 2.2-fold up-regulation of CD18 antigen and significant down-regulation of surface expression of the granulocyte adhesion molecule L-selectin were induced. Granulocyte F-actin content as measured by nitrobenzoxadiazole-phallacidin increased significantly 1 min after CD18 cross-linking. By contrast, CD18 cross-linking by soluble antibodies did not induce superoxide production, but PMNs bound to immobilized monoclonal antibodies against CD18 released significant amounts of superoxide. Initial signaling through beta 2 integrins does not appear to be mediated by a phospholipase C isoform activated through tyrosine phosphorylation, because the Ca2+ signal was not altered by herbimycin A. However, more complex cellular responses including exocytosis were found to require tyrosine phosphorylation. We show that engagement of beta 2 integrins provides an important stimulatory signal to PMNs inducing degranulation, modulation of L-selectin, and cytoskeletal changes.

Actins↗

PubMed1994

Resistance to blood flow in microvessels in vivo.

Resistance to blood flow through peripheral vascular beds strongly influences cardiovascular function and transport to tissue. For a given vascular architecture, flow resistance is determined by the rheological behavior of blood flowing through microvessels. A new approach for calculating the contribution of blood rheology to microvascular flow resistance is presented. Morphology (diameter and length), flow velocity, hematocrit, and topological position were determined for all vessel segments (up to 913) of terminal microcirculatory networks in the rat mesentery by intravital microscopy. Flow velocity and hematocrit were also predicted from mathematical flow simulations, in which the assumed dependence of flow resistance on diameter, hematocrit, and shear rate was optimized to minimize the deviation between measured and predicted values. For microvessels with diameters below approximately 40 microns, the resulting flow resistances are markedly higher and show a stronger dependence on hematocrit than previously estimated from measurements of blood flow in narrow glass tubes. For example, flow resistance in 10-microns microvessels at normal hematocrit is found to exceed that of a corresponding glass tube by a factor of approximately 4. In separate experiments, flow resistance of microvascular networks was estimated from direct measurements of total pressure drop and volume flow, at systemic hematocrits intentionally varied from 0.08 to 0.68. The results agree closely with predictions based on the above-optimized resistance but not with predictions based on glass-tube data. The unexpectedly high flow resistance in small microvessels may be related to interactions between blood components and the inner vessel surface that do not occur in smooth-walled tubes.

Animals↗

PubMed1994

Haemorrheology and long term exercise.

In general, the small number of haemorrheological studies concerning acute changes in long term exercise reveals remarkable small effects, given the substantial changes of pertinent cardiorespiratory parameters. To a large extent, this appears to result from careful maintenance of extra- and intravascular water balance and adequate control of electrolytes. Haemorrheological alterations during long term exercise depend on haematological parameters, especially haematocrit and plasma protein levels, which are acutely changed. During exercise, only small deviations of haematocrit from resting values are seen in the short term. This is explained by the usually constant plasma volume which depends on the amount of fluid intake during exercise. In contrast, the exercise-induced elevation of total intravascular protein content significantly increases the levels of plasma proteins, with the exception of fibrinogen. Although this leads to an increase of plasma viscosity, the absence of substantial haematocrit changes accounts for the remarkably small alterations, if any, of blood viscosity which are observed during long term exercise. Endurance training causes haemodilution by expansion of plasma volume, thus resulting in a reduction of blood and plasma viscosity. Red cell deformability is variable with training as well as during exercise. This appears to be related to methodological problems of measurement as well as to the type of endurance exercise performed. Physiological considerations suggest that the importance, for aerobic work capacity, of haemorrheological changes occurring in long term (submaximal) exercise may be limited. By contrast, this may be different during maximal exercise activity, when the entire cardiovascular reserve has been fully recruited.

Blood Viscosity↗

PubMed1993

Fucoidin, but not yeast polyphosphomannan PPME, inhibits leukocyte rolling in venules of the rat mesentery.

Leukocyte rolling in venules is inhibited by several sulfated polysaccharides, by antibodies to the leukocyte adhesion receptor L-selectin (LECAM-1), and by recombinant soluble L-selectin. The sulfated fucose polymer fucoidin and the polyphosphomannan PPME bind to L-selectin and inhibit L-selectin-mediated lymphocyte adhesion to lymph node high endothelial venules (LN-HEV). We investigated whether fucoidin and PPME also inhibit leukocyte rolling. Rolling leukocyte flux was determined by intravital microscopy in 47 venules (diameter 21 to 50 microns) of the rat mesentery with and without micro-infusion of each reagent through 8-microns glass micropipettes. Micro-infusion (1 mg/mL) or intravenous (IV) injection (25 mg/kg) of fucoidin, but not vehicle, reduced leukocyte rolling by greater than 90%. The half-effective concentration was approximately 2.5 micrograms/mL. Stroboscopic fluorescence video microscopy showed that fucoidin decreased the fraction of rolling leukocytes from 44% of all leukocytes passing the venules in control to less than 1%. PPME micro-infusion (1 mg/mL) or IV injection (14 mg/kg) did not reduce leukocyte rolling. Hence, leukocyte rolling differs from lymphocyte homing with respect to the effect of PPME. This may be related to fucoidin binding to L-selectin with greater affinity than PPME. Alternatively, inflamed venular endothelium may express a ligand for L-selectin different from that constitutively expressed on LN-HEV.

Animals↗

PubMed1993

Time-dependent rheological behavior of blood at low shear in narrow vertical tubes.

The time-dependent flow behavior of normal human blood after a sudden reduction of wall shear stress from 5,000 mPa to a low level (2-100 mPa) was studied during perfusion of vertical tubes (internal diam 28-101 microns) at constant driving pressures. Immediately after the implementation of low-shear flow conditions the concentration of red blood cells (RBCs) near the tube wall started to decrease, and marginal plasma spaces developed as a result of the assembly of RBC aggregates. This was associated with a time-dependent increase of flow velocity by up to 200% within 300 s, reflecting a reduction of apparent viscosity. These time-dependent changes of flow behavior increased strongly with decreasing wall shear stress and with increasing tube diameter. A correlation between the width of the marginal plasma layer and relative apparent viscosity was obtained for every condition of tube diameter, wall shear stress, and time. Time-dependent changes of blood rheological properties could be relevant in the circulation, where the blood is exposed to rapid and repeated transitions from high-shear flow conditions in the arterial and capillary system to low-shear conditions in the venous system.

Blood Flow Velocity↗

PubMed1993

Postural vascular response in human skin: passive and active reactions to alteration of transmural pressure.

Laser-Doppler (LD) fluxmetry was performed in the palmar finger skin of healthy subjects to study the mechanisms contributing to the postural vascular response. Local transmural pressure in the skin blood vessels of the region studied was altered for 1 min in two experimental series either by passive movement of the arm to different vertical hand positions relative to heart level or by application of external pressure (-120-180 mmHg) to the finger. Heart and respiratory rate, arterial blood pressure, and LD flux in the contralateral finger (kept at heart level) were measured. The measurements suggest a compound reaction of local (myogenic) and systemic (neurogenic) mechanisms: the local regulatory component appears as a graded active vascular response elicited by passive vessel distension or compression. A systemic component, associated with a single deep inspiration, is frequently observed during the actual movement of the arm. In addition, prolonged holding of the test hand in a given vertical position also elicits a delayed vascular response in the control hand at heart level, which may be generated by volume receptors in the intrathoracic low-pressure system.

Adult↗

PubMed1992

Changes in haemorheology in the racing greyhound as related to oxygen delivery.

Arterial blood samples were obtained from six greyhounds during rest, immediately before, and after a 704-m (7/16th mile) race. Measurements were made of various haematological (red cell count, haemoglobin, packed cell volume, white cell count, plasma proteins) and haemorheological variables. Blood and plasma viscosity were determined at high wall shear stresses (67-200 dynes.cm-2, 670-2000 microN.cm-2) in a 20-microns glass capillary device which was designed to take the diameter dependence of blood viscosity (Fahraeus-Lindqvist effect) into account. Compared to values at rest, substantial haemoconcentration occurred before the race, mainly due to splenic discharge of red cells. Additional haemoconcentration was found after the race. The increase of effective blood viscosity caused by elevation of packed cell volume was greater than the increase in O2 binding capacity resulting from the elevated haemoglobin concentration, suggesting that the haemoconcentration observed in the exercising greyhound does not enhance O2 delivery to skeletal muscle. The main physiological effect of red cell discharge from the contracting spleen appeared to be a consequence of the volume rather than the composition of the circulating blood.

Animals↗

PubMed1992

Differential adhesion of granulocytes to five distinct phenotypes of cultured microvascular endothelial cells.

Adhesion of isolated human polymorphonuclear granulocytes (PMNs) to five different phenotypes of cultured microvascular endothelial cells derived from bovine corpora lutea was investigated by measuring the myeloperoxidase content of cell lysates. Untreated and interleukin 1 (IL-1) -pretreated confluent monolayers were overlaid with unstimulated and phorbol ester (PMA)-stimulated PMNs in the absence and presence of the monoclonal antibody IB4 recognizing and functionally blocking beta 2 (CD18) of the leukocyte integrins. Unstimulated PMN adhesion was highest on type 4, followed by type 3 and 5 endothelial cells. This adhesion was not inhibited by treatment with IB4. IL-1 pretreatment of endothelial cells resulted in a significant increase of PMN adhesion on types 1, 2, and 4, most of which was also beta 2 integrin-independent. PMA-stimulation of PMNs increased adhesion to maximal values on cell types 1 and 5, which was largely blocked by IB4. Type 2 endothelial cells supported significantly less PMA-stimulated PMN adhesion than all other types. In the presence of IB4, adhesion of PMNs to untreated and IL-1-pretreated type 3 and 4 endothelial cells was significantly reduced by PMA. This reduction of beta 2 integrin-independent adhesion by PMA stimulation is compatible with possible shedding of the lectin-like leukocyte adhesion molecule, L-selectin, from PMNs. Differential PMN adhesion may reflect distinctive expression of endothelial adhesion molecules in different phenotypes of microvascular endothelial cells. Endothelial specialization within the microcirculation may have important functional consequences for the inflammatory response in vivo.

Animals↗

PubMed1992

Haemorheology and exercise: intrinsic flow properties of blood in marathon running.

Haemorheological and haematological parameters were studied in venous blood samples taken from 8 endurance-trained athletes before and after a marathon run. Viscometry was performed in a 20 microns glass capillary and in a Couette viscometer. Apparent blood viscosity was lower in the capillary than in the rotational viscometer, but significant differences between pre- and post-run samples were not observed in either system. This is presumably due to the absence of changes of haematocrit (pre-run [mean +/- SD]: 0.453 +/- 0.016, post-run: 0.456 +/- 0.018). Although protein concentration increased significantly from 70.8 +/- 2.1 g/l (pre-run) to 76.0 +/- 3.9 g/l (post-run), the difference of plasma viscosity was not statistically significant (pre-run: 1.26 +/- 0.03 cP, post-run: 1.30 +/- 0.07 cP). In all samples, plasma viscosity showed a significant positive correlation with total protein, globulin, albumin, but not with fibrinogen concentration. Cone-plate aggregometry and a filtration technique were used to evaluate red cell aggregation and deformability, two determinants of blood rheology which also were not different between pre- and post-run samples. The absence of viscosity changes in relation to the marathon run may be due to the relative stability of blood and plasma volume, which can be deduced from haematological parameters. Alterations of plasma composition (e.g. acidosis or hyperosmolality) seem to cause only minor, if any, changes of microrheological cell properties.

Adult↗