Relationship between pathological findings and values of haematological and blood-chemistry variables in apparently healthy finishing pigs at slaughter.

The present study was performed to study possibilities of early decision making for appropriate conveyor-line at future slaughtering of normal, clinically healthy finishing pigs. Blood was collected at slaughter from barrows (n = 112). A meticulous examination for subclinical pathological lesions was performed, revealing 5 groups of subjects listed in order of increasing disease-activity: 1--no real disease-activity; 2--with mild subchronic lesions; 3--with subacute lesions; 4--with abscesses; and 5--with fibrinous-necrotic lesions. Significant differences for values of erythrocyte sedimentation rate (ESR), protein, albumin, globulins, and plasma viscosity appeared to occur in this series. It is suggested that measuring acute phase reactants in blood of slaughtered pigs in the near future may reveal appropriate modern tools for meat inspection and predicting slaughtered animal quality.

Blood platelets are concentrated near the wall and red blood cells, in the center in flowing blood.

Hematocrit and vessel wall shear rate are important factors in the transport and subsequent adherence of platelets to vessel wall subendothelium. When mass transport theory is applied to platelets in flowing blood, the blood is usually considered to be a fluid with platelet and red cell wall concentrations similar to the average tube concentration. With the laser-Doppler technique, we found how red blood cell ghosts and platelets were distributed radially for various hematocrits and wall shear rates. Red cell ghosts are crowded near the axis of the tube, with a local hematocrit higher than the average tube hematocrit, and they decrease steadily toward the wall. In the absence of ghosts, platelets exhibit the 'tubular pinch' effect (rigid particles crowding at 0.6 x tube radius). In the presence of ghosts, the platelets are expelled toward the wall region. This high concentration at the wall increases with higher average tube hematocrit and wall shear rates. Increasing the average tube platelet concentration 10 times causes the wall concentration to increase only three times. The increase in platelet adherence observed with increasing hematocrit and increasing wall shear rate can be partially ascribed to increased platelet concentration near the wall. The observation that the increased platelet concentration does not fully explain the platelet adherence data suggests that platelet transport may also be enhanced by a shear rate-dependent rotary motion.

An in vitro study of the adhesion of blood platelets onto vascular catheters. Part I.

The adhesion of human blood platelets onto vascular catheters was studied using a specially designed perfusion chamber. Polyurethane catheters were exposed to citrated human blood for different periods (up to 20 min) and at different wall shear rates (190, 260, 330 sec-1). The rate of platelet adhesion was determined using 111In-labeled platelets, while the morphology of adhering platelets was investigated using scanning electron microscopy. A linear increase in platelet adhesion was found within the first 10 min of perfusion, after which a plateau value was reached. The number of adhering platelets did not vary significantly with the shear rates applied, which may indicate that within the range of shear rates studied, the adhesion of platelets onto the catheter surface is mainly determined by the rate of the reaction between the platelets and the material surface. Catheters coated with a conjugate of heparin and albumin showed a four- to five-fold reduction in platelet adhesion as compared to uncoated catheters. This reduction in platelet adhesion was not only due to the presence of albumin moieties at the surface but also to the presence of heparin residues in the adsorbed albumin-heparin conjugate.

Increased red blood cell deformability due to isoxsuprine administration decreases platelet adherence in a perfusion chamber: a double-blind cross-over study in patients with intermittent claudication.

Platelet transport towards the vessel wall is influenced by the hematocrit, red blood cell (RBC) size, and shape. Recent in vitro studies have indicated that RBC deformability may also influence platelet transport. The observation that isoxsuprine, a known vasodilating drug, caused increased RBC deformability in vitro and decreased platelet transport in vitro prompted us to study the effects of this drug in vivo. The study was performed in a double-blind cross-over study of isoxsuprine v placebo in ten patients with peripheral arterial insufficiency. RBC deformability was estimated from viscosity measurements using the blood viscosity equation of Dintenfass and expressed as T value. Platelet transport was studied in an annular perfusion chamber according to Baumgartner. Human umbilical arteries were used as blood vessels. Perfusion studies were performed with whole blood or with RBCs of the patients mixed with normal platelets and plasma at a standardized hematocrit and platelet count. An increase in RBC deformability concomitant with a decrease in platelet adherence was observed in patients on isoxsuprine with a drop in T value of approximately 0.06 (from 0.91 toward 0.86), and a concomitant decrease in platelet adherence of 20% to 40%. These observations differed significantly from the results in the placebo group and showed a significant group-period interaction at the cross-over of medication (analysis of variance). The effects on platelet adherence were observed at high vessel wall shear rate (1,800 s-1) with perfusates consisting of patients' RBCs and donor plasma and platelets at standardized hematocrit and platelet count. No differences were observed under these conditions at a shear rate of 300 s-1. When whole blood of patients was used, nonsignificant effect was observed at shear rates of 300 s-1 and 1,800 s-1. This was probably caused by the added noise due to variations in hematocrit and platelet number. These data demonstrate that isoxsuprine increases RBC deformability, and they suggest the possibility of decreasing platelet-vessel wall interaction in vivo by manipulation of RBC deformability.

Fluid shear as a possible mechanism for platelet diffusivity in flowing blood.

Platelet transport theory is based on convection diffusion and describes adequately the influence of wall shear rate, platelet concentration and axial (down stream) position. Until now, the influence of the predominant factors affecting platelet adherence, the hematocrit and the red cell size, was not included in this theory. Their role remained hidden in the platelet diffusivity (Dw), which was assumed to be related to the shear rate (gamma) expressed in s-1 by a power law function Dw = m gamma n, in which m and n were thought to be constants. We have determined platelet diffusivity directly from platelet adherence to subendothelium as a function of axial distance in an in vitro perfusion system. Our results indicate that m is a constant with a value of (1.05 +/- 0.05) 10(-9) cm2 s-1 and that n is a function of the hematocrit (h) which is best approximated by a quadratic equation n = 0.297 + 1.29 h - 0.90 h2. The effect of red cell size was introduced by correcting the hematocrit containing factors in this quadratic equation for the square of the red cell diameter. This correction was made on the basis of theoretical considerations. The theoretically derived platelet adherence correlated closely with the previous experimental data regarding the effect of red cell size in which we found that the hemodynamic effect of red cells on platelet adherence decreases with decreasing red cell diameter.

Red blood cell deformability influences platelets--vessel wall interaction in flowing blood.

Hematocrit and red cell size are important factors for the transport of blood platelets toward subendothelium in flowing blood. We report that red cell deformability also influences platelet transport. Red cell deformability was estimated with Couette-flow viscosimetry at a shear rate of 130 s-1 and expressed as a 'T' factor--a dimensionless parameter relating the relative viscosity and the hematocrit derived from the relationship: T = (1 - mu -0.4 rel)/H, where mu rel is the relative viscosity and H is the hematocrit. The normal value of T was within a narrow range (0.91 +/- 0.02). Treatment of normal red cells with isoxsuprine and chlorpromazine caused decreased rigidity and decreased T. Cholesterol loading and treatment with diamide increased rigidity and increased T. In vitro perfusion experiments in an annular perfusion system with everted human umbilical arteries were performed with perfusates to which such treated red blood cells were added to investigate their influence on platelet adherence to artery subendothelium. Platelet adherence was well correlated with red cell rigidity, with increased adherence at increased rigidity and vice versa. A change in T of 0.10 corresponded to a change in platelet adherence of approximately 50%. These effects were more pronounced at a wall shear rate of 1,800 s-1 than at 300 s-1.

Velocity profiles in annular perfusion chamber measured by laser-Doppler velocimetry.

p6e fluid flow in the annular perfusion chamber of Baumgartner developed to study platelet vessel wall interaction was examined with laser-Doppler velocimetry. A laminar and stable flow with a Reynolds number of less than or equal to 50 was measured at flow rates up to 3 ml s-1. No turbulence was found. The wall shear rate directly determined from measured velocity profiles agreed well with theory. The experiments underlined the necessity to work with vessels of uniform thickness and a smooth surface.

A perfusion chamber developed to investigate platelet interaction in flowing blood with human vessel wall cells, their extracellular matrix, and purified components.

A flat perfusion chamber was developed to study the interaction of blood platelets in flowing blood with cultured human vessel wall cells, their connective tissue matrix, and isolated connective tissue components at defined shear rate conditions. A cover slip covered with endothelial cells or extracellular matrix components was introduced into the chamber. Laser-Doppler velocimetry showed a symmetrical flow profile at flow rates between 50 and 150 ml/min (wall shear rate 300 to 1100 sec-1). Platelet deposition was estimated by using blood platelets labeled with indium-111 or by a morphometric method. Blood platelets did not adhere to endothelial cells at wall shear rates of 765 sec-1 and the endothelial cells remained attached for at least 10 min of perfusion. In preconfluent cultures of endothelial cells, blood platelets adhered to extracellular material in areas between the cells. Removal of endothelial cells by treatment with 0.5% Triton X-100 induced increased platelet adherence with a preference for certain, as yet unidentified, fibrillar structures of the extracellular matrix. Platelet adherence to equine collagen was also studied after coating the cover slips by spraying of small collagen droplets followed by air drying. Platelet adherence and the subsequent platelet aggregate formation occurred predominantly along visible collagen fibers. These studies showed that this perfusion chamber has a laminar and symmetrical flow allowing qualitative and quantitative investigation of platelet interaction with endothelial cells, their extracellular matrix, and pure connective tissue components. A variety of wall shear rates and exposure times can be applied at controlled conditions without removing cells or extracellular material.

Red blood cell size is important for adherence of blood platelets to artery subendothelium.

The hematocrit is one of the main factors influencing platelet adherence to the vessel wall. Raising the hematocrit causes an increase of platelet accumulation of about an order of magnitude. Our studies concern the role of red cell size. We have studied this effect using an annular perfusion chamber, according to Baumgartner, with human umbilical arteries and a steady-flow system. Normal human red blood cells (MCV 95 cu mu) increased platelet adherence sevenfold, as the hematocrit increases from 0 to 0.6. Small erythrocytes from goats (MCV 25 cu mu) caused no increment in adherence in the same hematocrit range. Rabbit erythrocytes (MCV 70 cu mu) caused an intermediate increase in adherence. Red blood cells from newborns (MCV 110-130 cu mu) caused a larger increase in platelet adherence than normal red cells at hematocrit 0.4. These results were further confirmed with large red blood cells from two patients. Experiments with small red cells (MCV 70 cu mu) of patients with iron deficiency showed that platelet adherence was similar to normal red cells, provided the red cell diameter was normal. Small red blood cells of a patient with sideroblastic anemia caused decreased adherence. These data indicate that red cell size is of major importance for platelet adherence. Red cell diameter is more important than average volume. However, for size differences in the human range, the hematocrit remains the dominant parameter.

Phase transitions in phospholipid model membranes of different curvature.

1. Nuclear magnetic resonance, light scattering and freeze fracturing electron microscopic techniques were used to characterize the size of unilamellar phospholipid vesicles of 1,2-dimyristoyl-sn-glycero-3-phosphocholine. 2. Differential scanning calorimetric and light scattering analyses showed that very small unilamellar vesicles obtained by the sonication method exhibit a downward shifted, largely broadened phase transition with a slightly decreased enthalpy change when compared with multilayered liposomes. 3. The phase transition of vesicles with variable diameter as obtained by injection methods resembled the pattern of multilayered liposomes the more the diameter was increased. 4. Repeated cycling through the lipid phase transition was shown to have a progressive effect on a fusion process. This effect was strongly increased when the osmolarity of the medium was enhanced (e.g. by the addition of cryoprotectors). Furthermore it was shown that ice-water of the systems caused abrupt fusion of the lipid structures. 5. Controversial results in the literature on the thermotropic behavior of vesicles could be explained in terms of these fusion processes.

Dynamics of phospholipid aggregation in ethanol--water solutions.

The kinetics of liposome and vesicle formation of a synthetic lecithin has been studied by light scattering techniques. It is shown, that by evaporation of alcohol from a lipid--alcohol--water mixture, the aggregates formed undergo several changes in shape. A hypothesis is presented, visualising the formation of liposomes or vesicles from monomers, which is consistent with the experimental observations.