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Marcos Intaglietta

Publications and source records attributed to Marcos Intaglietta.

66 records · Page 4Linked to original sources
PubMed2003

Targeted O2 delivery by low-P50 hemoglobin: a new basis for O2 therapeutics.

To assess O2 delivery to tissue by a new surface-modified, polyethylene glycol-conjugated human hemoglobin [MP4; Po2 at 50% saturation of hemoglobin (P50); 5.4 mmHg], we studied microcirculatory hemodynamics and O2 release in golden Syrian hamsters hemodiluted with MP4 or polymerized bovine hemoglobin (PolyBvHb; P50 54.2 mmHg). Comparisons were made with the animals' hemodiluted blood with a non-O2 carrying plasma expander with similar solution properties (Dextran-70). Systemic hemodynamics (arterial blood pressure and heart rate) and acid-base parameters were not correlated with microhemodynamics (arteriolar and venular diameter, red blood cell velocity, and flow). Microscopic measurements of Po2 and the O2 equilibrium curves permitted analysis of O2 release in precapillary and capillary vessels by red blood cells and plasma hemoglobin separately. No significant differences between the groups of animals with respect to arteriolar diameter, flow, or flow velocity were observed, but the functional capillary density was significantly higher in the MP4-treated animals (67%) compared with PolyBvHb-treated animals (37%; P < 0.05) or dextran-treated animals (53%). In the PolyBvHb-treated animals, predominant O2 release (both red blood cells and plasma hemoglobin) occurred in precapillary vessels, whereas in MP4 animals most of the O2 was released from both red blood cells and plasma hemoglobin in capillaries. Base excess correlated directly with capillary O2 release but not systemic O2 content or total O2 release. Higher O2 extraction of both red blood cell and plasma hemoglobin in capillaries represents a new mechanism of action of cell-free hemoglobin. High O2 affinity appears to be an important property for cell-free hemoglobin solutions.

Acid-Base Equilibrium↗
PubMed2003

pO(2) measurements by phosphorescence quenching: characteristics and applications of an automated system.

An automated system for pO(2) analysis based upon phosphorescence quenching was tested. The system was calibrated in vitro with capillary samples of saline and blood. Results were compared to a conventional measuring procedure wherein pO(2) was calculated off-line by computer fitting of phosphorescence decay signals. PO(2) measurements obtained by the automated system were correlated (r(2) = 0.98) with readings simultaneously generated by a blood gas analyzer, accuracy being highest in the low (0-20 mm Hg) and medium pO(2) ranges (21-70 mm Hg). Measurements in in vivo studies in the hamster skin-fold preparation were similar to previously reported results. The automated system fits the phosphorescence decay data to a single exponential and allows repeated pO(2) measurements in rapid sequence.

Animals↗
PubMed2003

Measurement of the cardiac output in small animals by thermodilution.

Cardiac output (CO) measurements based on indicator dilution, microspheres, thermodilution and ultrasonic sensors are not suitable for small animals, because of limited blood volume, high heart rates and small caliber vessels that do not allow probe placement within the heart. We developed a modified thermodilution method to measure CO in awake animals weighing less than 100 g. Under anesthesia, the animal is instrumented with a jugular vein catheter placed proximal to the subclavian vein and a temperature probe in the carotid artery with the thermocouple positioned at the aortic arch. Two days after implantation, room temperature saline is injected (150 microl) into the jugular catheter and the temperature change recorded. This system uses the temperature probe as a digital feedback control: (1) to minimize recirculation error; (2) to adjust baseline temperature, thereby increasing sensitivity to small changes in temperature; and (3) to stabilize animal core temperature. The system was calibrated using a laboratory bench model with anatomically scaled components. CO was measured (n=29) in 16 hamsters (65-115 g), and was linearly related to body weight. Cardiac index (CI=CO/weight) was 197.0 +/- 18.8 (ml/min)/kg. Repeated measurements were made. This technique allows correlating systemic flow changes to be correlated to those measured in the microcirculation of window chamber preparations.

Animals↗
PubMed2003

Resuscitation with polyethylene glycol-modified human hemoglobin improves microcirculatory blood flow and tissue oxygenation after hemorrhagic shock in awake hamsters.

OBJECTIVE: To determine whether resuscitation with polyethylene glycol-modified human hemoglobin (MalPEG-Hb), an oxygen-carrying blood replacement fluid with 4 g/dL Hb, viscosity of 2.5 cP, colloid osmotic pressure of 49 mm Hg, and p50 of 5.5 mm Hg, improves systemic and microvascular variables after hemorrhage compared with shed blood (SB) and 5% hydroxyethyl starch (HES). SETTING: Laboratory. SUBJECTS: Golden Syrian hamsters. DESIGN: Prospective study. INTERVENTIONS: Hamsters implemented with a skin fold chamber were hemorrhaged 50% of blood volume and resuscitated with 50% shed blood volume (SB, HES, or MalPEG-Hb). MEASUREMENTS AND MAIN RESULTS: Shock and resuscitation were monitored for 1 hr each. Microvascular events were characterized in terms of vessel diameter, flow velocity, functional capillary density, and Po(2) in arterioles, venules, and extravascular tissue. Systemic variables include mean arterial pressure, heart rate, Po(2), Pco(2), pH, and base excess. MalPEG-Hb resuscitation increased functional capillary density to 64% vs. 44% for SB and 32% for HES relative to baseline before shock. Microvascular flow increased 16% for MalPEG-Hb relative to baseline and remained decreased by 44% for SB and 80% for HES. Hemoglobin concentration was 10.4 g/dL with SB, 7.5 (6.8 g/dL in red blood cells and 0.9 g/dL in plasma) with MalPEG-Hb, and 7.5 g/dL with HES, leading to tissue Po(2) of 19, 8, and 5 mm Hg respectively. Calculations of oxygen extraction show that 0.9 g/dL of MalPEG-Hb increased oxygen extraction per gram of red cell hemoglobin in the tissue analyzed compared with SB. These measurements correlate well with a systemic indicator of recovery, base excess, 5.4 +/- 4.7 (MalPEG-Hb), 1.7 +/- 3.8 (SB), and -0.3 +/- 5.7 (HES). CONCLUSION: The presence of 0.9 g/dL of high oxygen affinity MalPEG-Hb improves microvascular blood flow and oxygen transport during shock to a significantly greater extent than that attainable with blood or HES.

Animals↗
PubMed2003

Microcirculatory changes during chronic adaptation to hypoxia.

Microcirculatory changes in the window chamber preparation in Syrian golden hamsters, secondary to chronic hypoxia adaptation, are presented herein. Adaptation was attained by keeping animals in a 10% oxygen environment for 1 wk and 5% the following week. The following groups were studied: group 1, adapted to chronic hypoxia and kept in a 5% oxygen environment throughout the experiment; group 2, adapted to chronic hypoxia and kept in a 21% oxygen environment 24 h before and during the experiment; and group 3, control. Adaptation caused venule enlargement and hematocrit increase (68.6 +/- 2.44 in group 1, 70 +/- 2.66 in group 2, and 43.27 +/- 2.30 in group 3; P < 0.05). Whereas heart rate decreased in adapted animals, blood pressure remained constant. Group 1 presented alkalosis, hypocapnia, and hypoxemia. The adapted groups had decreased blood flow velocity in arterioles and veins. We found no difference in microvasculature oxygen tension between groups 2 and 3; however, the number of capillaries with flow was markedly reduced in group 1 but significantly increased in group 2. Our findings suggest that, as an adaptation to hypoxia, erythropoiesis may prove beneficial by increasing blood viscosity and shear stress, leading to vasodilatation, in addition to the increase in oxygen-carrying capacity. Calculations show that oxygen extraction in the tissue of the window chamber model was significantly lowered in adapted animals breathing 5% oxygen, but was unchanged from the control when breathing 21% oxygen, even though blood hemoglobin content was increased from 14.5 +/- 0.07 g/dl at control to 21.04 +/- 1.24 g/dl in the adapted animals (P < 0.05).

Adaptation, Physiological↗
PubMed2003

Oxygen gradients in the microcirculation.

As arterialized blood transits from the central circulation to the periphery, oxygen exits through the vessel walls driven by radial oxygen gradients that extend from the red blood cell column, through the plasma, the vessel wall, and the parenchymal tissue. This exit determines a longitudinal gradient of blood oxygen saturation whose extent is inversely related to the level of metabolic activity of the tissue, being small for the brain and considerable for skeletal muscle at rest where hemoglobin is only half-saturated with oxygen when blood arrives to the capillaries. Data obtained by a variety of methods show that the oxygen loss is too great to be explained by diffusion alone, and oxygen gradients measured in the arteriolar wall provide evidence that this structure in vivo is a very large oxygen sink, and suggests a rate of oxygen consumption two orders of magnitude greater than seen in in vitro studies. Longitudinal gradients in the capillary network and radial gradients in surrounding tissue also show a dependence on the metabolic rate of the tissue, being more pronounced in brain than in resting skeletal muscle and mesentery. Mean PO2 values increase from the postcapillary venules to the distal vessels of this network while radial gradients indicate additional oxygen loss. This circumstance may be due to pathways with higher flow having higher oxygen content than low flow pathways as well as possible oxygen uptake from adjacent arterioles. Taken together, these newer findings on oxygen gradients in the microcirculation require a reexamination of existing concepts of oxygen delivery to tissue and the role of the capillaries in this process.

Animals↗
PubMed2002

Systemic and microvascular responses to hemorrhagic shock and resuscitation with Hb vesicles.

A phospholipid vesicle encapsulating hemoglobin (Hb vesicle, HbV) has been developed to provide O(2)-carrying capacity to plasma expanders. Its ability to restore systemic and microcirculatory conditions after hemorrhagic shock was evaluated in the dorsal skinfold window preparation of conscious hamsters. The HbV was suspended in 8% human serum albumin (HSA) at Hb concentrations of 3.8 g/dl [HbV(3.8)/HSA] and 7.6 g/dl [HbV(7.6)/HSA]. Shock was induced by 50% blood withdrawal, and mean arterial pressure (MAP) at 40 mmHg was maintained for 1 h by the additional blood withdrawal. The hamsters receiving either HbV(3.8)/HSA or HbV(7.6)/HSA suspensions restored MAP to 93 +/- 14 and 93 +/- 10 mmHg, respectively, similar with those receiving the shed blood (98 +/- 13 mmHg), which were significantly higher by comparison with resuscitation with HSA alone (62 +/- 12 mmHg). Only the HSA group tended to maintain hyperventilation and negative base excess after the resuscitation. Subcutaneous microvascular blood flow reduced to approximately 10-20% of baseline during shock, and reinfusion of shed blood restored blood flow to approximately 60-80% of baseline, an effect primarily due to the sustained constriction of small arteries A(0) (diameter 143 +/- 29 microm). The HbV(3.8)/HSA group had significantly better microvascular blood flow recovery and nonsignificantly better tissue oxygenation than of the HSA group. The recovery of base excess and improved tissue oxygenation appears to be primarily due to the increased oxygen-carrying capacity of HbV fluid resuscitation.

Animals↗
PubMed2002

A novel approach to blood plasma viscosity measurement using fluorescent molecular rotors.

Molecular rotors, a group of fluorescent molecules with viscosity-dependent quantum yield, were tested for their suitability to act as fluorescence-based plasma viscometers. The viscosity of samples of human plasma was modified by the addition of pentastarch (molecular mass 260 kDa, 10% solution in saline) and measured with a Brookfield viscometer. Plasma viscosity was 1.6 mPa x s, and the mixtures ranged up to 4.5 mPa x s (21 degrees C). The stimulated light emission of the molecular rotors mixed in the plasma samples yielded light intensity that was nonoverlapping and of significantly different intensity for viscosity steps down to 0.3 mPa x s (n = 5, P < 0.0001). The mathematical relationship between intensity (I) and viscosity (eta) was found to be eta = (kappaI)(nu). After calibration and scaling the fluorescence based measurement had an average deviation versus the conventional viscometric measurements that was <1.8%. These results show the suitability of molecular rotors for fast, low-volume biofluid viscosity measurements achieving accuracy and precision comparable to mechanical viscometers.

Blood Viscosity↗
PubMed2002

Effect of aggregation and shear rate on the dispersion of red blood cells flowing in venules.

Previous in vitro studies of blood flow in small glass tubes have shown that red blood cells exhibit significant erratic deviations in the radial position in the laminar flow regime. The purpose of the present study was to assess the magnitude of this variability and that of velocity in vivo and the effect of red blood cell aggregation and shear rate upon them. With the use of a gated image intensifier and fluorescently labeled red blood cells in tracer quantities, we obtained multiple measurements of red blood cell radial and longitudinal positions at time intervals as short as 5 ms within single venous microvessels (diameter range 45-75 microm) of the rat spinotrapezius muscle. For nonaggregating red blood cells in the velocity range of 0.3-14 mm/s, the mean coefficient of variation of velocity was 16.9 +/- 10.5% and the SD of the radial position was 1.98 +/- 0.98 microm. Both quantities were inversely related to shear rate, and the former was significantly lowered on induction of red blood cell aggregation by the addition of Dextran 500 to the blood. The shear-induced random movements observed in this study may increase the radial transport of particles and solutes within the bloodstream by orders of magnitude.

Animals↗
PubMed2002

The unusual properties of effective blood substitutes.

Blood substitutes or oxygen carrying plasma expanders were originally formulated to simulate the transport properties of blood, particularly oxygen carrying capacity, viscosity, p50, and colloid osmotic pressure, under the hypothesis that blood is the most desirable fluid in volume restitution. However, changes introduced into the organism during hemorrhage adversely affect microvascular function due to reflex vasoconstriction which causes the fall of functional capillary density, and lowers tissue oxygenation, conditions that are not universally reversed with retransfusion of blood. The restoration of microvascular function is seldom complete upon retransfusion of blood. New formulations of hemoglobin molecules in solutions whose oncotic pressure is in the range of 60-100 mmHg, p50 is about 5 mmHg, viscosity 3-4 cP, and oxygen carrying capacity in the range of 4-7 g/dl equivalent hemoglobin deliver better microvascular function after resuscitation when compared to whole blood and oxygen carrying plasma expanders with transport properties similar to those of blood. The improved performance is in part due to the increased plasma viscosity which increases capillary transmural pressure which reverses capillary collapse induced during low perfusion pressures. High oncotic pressure reinforces this effect, since it brings more fluid into the circulation. Microvascular transport studies of the effects of resuscitation in shock show that functional capillary density is the primary determinant of survival, thus maintenance of an open and fully perfused microcirculation is more critical than insuring oxygen supply, since closed capillaries lead to the accumulation of slowly diffusing byproducts of metabolism which ultimately become toxic. The required combination of properties can be achieved by conjugating hemoglobin and polyethylene glycol. Resuscitation fluids based on hemoglobin containing vesicles may provide the next level of functional improvement in the formulation of volume restitution fluids since their biophysical properties can be specifically controlled through the inclusion of specialized compounds into the vesicles, and the formulation of the suspending medium.

Blood Circulation↗
PubMedDate not supplied

A computer-based method for determination of the cell-free layer width in microcirculation.

OBJECTIVES: The cell-free layer between the erythrocyte column and the vessel wall is an important determinant of hydrodynamic resistance in microcirculatory vessels. The authors report a method for continuous measurement of the width of this layer. METHODS: The light intensity of a linear array of pixels perpendicular to the vessel axis is continuously determined from a video image of a microcirculatory vessel. A threshold level based on Otsu's method is used to establish the interface between the cell-free layer and the erythrocyte column. To test the method, video images at 750-4500 frames/s were obtained from venules and arterioles in rat spinotrapezius muscle at normal and reduced arterial pressures before and after induction of erythrocyte aggregation with Dextran 500. The current measurements were compared to manual measurements of the same images. RESULTS: Values obtained by the manual and the new methods were in agreement within the 95% confidence limit by the Bland-Altman analysis and within 90-95% range by the correlation coefficient (R2). The more frequent measurements reveal substantial, rapid variations in cell-free layer width and changes in mean values with alteration of arterial pressure and red cell aggregability. CONCLUSIONS: A new, computer-based technique has been developed that provides measurements of rapid, time-dependent variations in the width of the cell-free layer in the microcirculation.

Animals↗