The effect of inhibition of endothelial cell cyclooxygenase on arterial thrombosis

Arterial platelet thrombosis was induced in vivo in a branch of the mesenteric artery of the white Wistar rat by topical superfusion by adenosine diphosphate following local de-endothelialization of the dissected segment by means of a small electrical current. Detection of the thrombotic phenomena was performed by projection of the arterial segment onto a set of light sensitive elements, wich allows the registration of several discriminating parameters. The addition of arachidonic acid to the superfusing mixture results in an increase in thromboformation; this increasing effect can be blocked completely by prior inhibition od the vessel wall cyclooxygenase activity by inhibitors such as flurbiprofen or indomethacin. We therefore concluded that the arachidonate cascade, triggered within the endothelial cells, is involved in platelet-vessel wall interaction leading ultimately to local thrombosis.     

Viral IL-10 gene transfer decreases inflammation and cell adhesion molecule expression in a rat model of venous thrombosis

Post-thrombotic inflammation probably contributes to chronic venous insufficiency, and little effective treatment exists. IL-10 is an anti-inflammatory cytokine that previously has been shown to decrease perithrombotic inflammation and thrombosis. We investigated in a rat model whether local expression of viral IL-10 (vIL-10) in a segment of vein that undergoes thrombosis would confer an anti-inflammatory effect and how this effect might be mediated. Rats underwent inferior vena cava isolation, cannulation, and instillation of saline or adenovirus encoding either beta-galactosidase or vIL-10. Two days after transfection, thrombosis was induced, 2 days after this the rats underwent gadolinium (Gd)-enhanced magnetic resonance venography exam, and the vein segments were harvested. Tissue transfection was confirmed by either RT-PCR of vIL-10 or positive 5-bromo-4-chloro-3-indolyl beta-d-galactopyranoside (X-Gal) staining. vIL-10 significantly decreased both leukocyte vein wall extravasation and area of Gd enhancement compared with those in controls, suggesting decreased inflammation. Immunohistochemistry demonstrated decreased endothelial border staining of P- and E-selectin, while ELISA of vein tissue homogenates revealed significantly decreased P- and E-selectin and ICAM-1 levels in the vIL-10 group compared with those in controls. Importantly, native cellular IL-10 was not significantly different between the groups. However, neither clot weight nor coagulation indexes, including tissue factor activity, tissue factor Ag, or von Willebrand factor levels, were significantly affected by local vIL-10 expression. These data suggest that local transfection of vIL-10 decreases venous thrombosis-associated inflammation and cell adhesion molecule expression, but does not directly affect local procoagulant activity.     

Differences in distensibility between the anterior and posterior walls of the left ventricle in dogs

Nonuniformity of myocardial systolic and diastolic performance in the normal left ventricle has been recognized by a number of investigators. Lack of homogeneity in diastolic properties might be caused by or related to differences in the distensibility of different regions of the left ventricular (LV) wall. Thus, we compared the end-diastolic transmural pressure-strain relations in both the anterior and posterior LV walls in seven anesthetized dogs during two interventions (pulmonary artery constriction and aortic constriction). Transmural pressure was defined as the difference between LV intracavitary pressure and local pericardial pressure. LV pressure was measured using a micromanometer; pericardial pressures over the LV anterior and posterior walls were measured with balloon transducers. Circumferentially oriented pairs of sonomicrometer crystals were implanted in the midwall of the anterior and posterior walls of the LV to measure segment lengths. Strains were calculated as (L-L0)/L0, where L was the instantaneous segment length and L0 was the segment length when transmural pressure was zero. The pattern of end-diastolic transmural pressure--strain relations was similar in all dogs. The change in strain in the posterior wall was always greater than that in the anterior wall. Opening the pericardium did not affect the difference in distensibility of the anterior and posterior walls. The results suggest that the posterior wall is more compliant than the anterior wall (that is, for a given difference in transmural pressure, the local segment length change of the posterior wall was greater). This seems consistent with other observations, which suggest that the posterior wall might make a greater contribution to diastolic filling.     

The effects of wall thickness, axial strain and end proximity on the pressure-area relation of collapsible tubes

Segments of silicone rubber tube were suspended between rigid pipes and subjected to slowly varying transmural pressure covering a range from slight distension to collapse with osculation. The local inside cross-sectional area at a chosen axial site was simultaneously measured via catheter by an electrical impedance method. Pressure-area relations were recorded thus at various axial sites, under varying conditions of axial tube wall tension, in tubes of two different wall thickness (0.3 and 0.4 of mean radius). Unsupported tube segment length was also varied by means of an insert device. The relations were used to calculate the variation of wave velocity with area according to Young's equation. First opposite wall contact during collapse was shown to occur at a smaller fraction of undistended circular cross-sectional area than in the thin-walled tubes investigated previously by others.     

Characteristics of the microcirculation and blood rheology in arterial and venous forms of mesenteric vascular occlusion

The functional properties of microcirculation and rheology of blood were studied in dogs subjected to arterial and venous occlusion of mesenteric vessels (cranial mesenteric artery and cranial mesenteric vein). It was found that a local alterations of microvascular bed of intestinal wall are quite different in case of arterial or venous occlusion. The degree of hemorheological and microvascular deviations is higher in case of acute venous thrombosis than during the acute occlusion of cranial mesenteric artery.     

Towards a concept of thrombosis in accelerated flow: rheology, fluid dynamics, and biochemistry

The predilection sites of arterial thrombosis are characterized by local increase in wall shear stress, flow separation with eddy formation and stagnation point flow. The defenders of high shear, as well as those of low shear theory of thrombogenesis, point to correlations of predilection sites and the respective flow abnormalities. Experimental evidence is provided, that high shear rates can damage both red cells and platelets, that lysed red cells constitute a potent platelet stimulant, due to their content of adenine nucleotides, and that platelets do not adhere to surfaces unless transported onto them by convective motion, the effectiveness of the platelet-wall interaction being enhanced by platelet activation. Based on these facts, a resolution of the contrast between high and low shear theory of thrombosis is attempted in a way, that the different flow regimens, with blood cells sequentially passing them, are each considered important and interdependent steps on the way to thrombosis.     

Numerical 3D-simulation of pulsatile wall shear stress in an arterial T-bifurcation model

The structure of pulsatile blood flow and wall shear stress in a 90° T-bifurcation model is analysed numerically. The nonlinear Navier-Stokes equations for time-dependent incompressible Newtonian fluid flow are approximated using a newly developed pressure correction, finite element method. The wall shear stress is calculated from the finite element velocity field. The investigation shows viscous flow phenomena such as flow separation and stagnation and the distribution of high and low wall shear stress during the pulse cycle. Furthermore, the effect of a sharp corner the bifurcation edge on the wall shear stress is analysed. Detailed local flow investigation is required to examine fluid dynamic contribution to the development of arterial diseases such as atherosclerosis and thrombosis.     

Simulation of particle-hemodynamics in a partially occluded artery segment with implications to the initiation of microemboli and secondary stenoses.

Computational results of laminar incompressible blood-particle flow analyses in an axisymmetric artery segment with a smooth local area constriction of 75 percent have been presented. The flow input waveform was sinusoidal with a nonzero average. The non-Newtonian behavior of blood was simulated with a modified Quemada model, platelet concentrations were calculated with a drift-flux model, and monocyte trajectories were described and compared for both Newtonian and Quemada rheologies. Indicators of "disturbed flow" included the time-averaged wall shear stress (WSS), the oscillatory shear index (OSI), and the wall shear stress gradient (WSSG). Implications of the vortical flow patterns behind the primary stenosis to the formation of microemboli and downstream stenoses are as follows. Elevated platelet concentrations due to accumulation in recirculation zones mixed with thrombin and ADP complexes assumed to be released upstream in high wall shear stress regions, could form microemboli, which are convected downstream. Distinct near-wall vortices causing a local increase in the WSSG and OSI as well as blood-particle entrainment with possible wall deposition, indicate sites susceptible to the onset of an additional stenosis proximal to the initial geometric disturbance.     

An inelastic multi-mechanism constitutive equation for cerebral arterial tissue

Intracranial aneurysms (ICA) are abnormal saccular dilations of cerebral arteries, commonly found at apices of arterial bifurcations and outer walls of curved arterial segments. Histological evidence suggests the stages in ICA development include the deformation of a segment of arterial wall into a “bleb” with no identifiable neck region followed by the development of an aneurysm with a clear neck. Afterwards, the aneurysm may undergo further enlargement, possibly with significant biological response including calcification and thrombosis. Past studies of the biomechanics of cerebral aneurysm tissue have been directed at modeling elastic deformations of pre-existing aneurysms. Taking this approach, the aneurysm wall is treated as a different entity than the arterial tissue from which it developed. In the current work, a nonlinear, inelastic, dual-mechanism constitutive equation for cerebral arterial tissue is developed. It is the first to model the recruitment of collagen fibers and degradation of the internal elastic lamina, two important characteristics of early stage aneurysm formation.     

Finite element modelling of the common carotid artery in the elderly with physiological intimal thickening using layer-specific stress-released geometries and nonlinear elastic properties

To investigate the mechanical effects of tissue responses, such as remodelling, in the arteries of the elderly, it is important to evaluate stress in the intimal layer. In this study, we investigated a novel technique to evaluate the effect of layer-specific characteristics on stress in the arterial wall in an elderly subject. We used finite element analysis of a segment of carotid artery with intimal thickening, incorporating stress-released geometries and the stress–strain relationships for three separate wall layers. We correlated the stress–strain relationships and local curvatures of the layers with the stress on the arterial wall under physiological loading. The simulation results show that both the stress–strain relationship and the local curvature of the innermost stress-released layer influence the circumferential stress and its radial gradient. This indicates that intimal stress is influenced significantly by location-dependent intimal remodelling. However, further investigation is needed before conclusive inferences can be drawn.     

Analysis of particle trajectories in aortic artery bifurcations with stenosis

The fluid-particle dynamics in a two-dimensonal symmetric branching channel with local occlusions representing a diseased segment of an aortic artery bifurcation has been analyzed. The validated finite element model simulates the trajectories and landing or impact sites of spherical particles for laminar flow in bifurcation channels with generalized wall conditions. Two hypotheses relating critical wall shear stress levels and plaque formation, previously postulated by Kleinstreuer et al. (1988) and Nazemi et al. (1989), have been confirmed. Low shear stress may contribute to the onset of atherosclerotic lesions and areas of critically low and higher shear stresses are susceptible to accelerated growth of plaque.     

ACE inhibition and atherogenesis

Recent clinical studies such as HOPE, SECURE, and APRES show that angiotensin-converting enzyme (ACE) inhibitors like ramipril improve the prognosis of patients with a high risk of atherothrombotic cardiovascular events. Atherosclerosis, as a chronic inflammatory condition of the vascular system, can turn into an acute clinical event through the rupture of a vulnerable atherosclerotic plaque followed by thrombosis. ACE inhibition has a beneficial effect on the atherogenic setting and on fibrinolysis. Endothelial dysfunction is the end of a common process in which cardiovascular risk factors contribute to inflammation and atherogenesis. By inhibiting the formation of angiotensin II, ACE inhibitors prevent any damaging effects on endothelial function, vascular smooth muscle cells, and inflammatory vascular processes. An increase in the release of NO under ACE inhibition has a protective effect. Local renin-angiotensin systems in the tissue are involved in the inflammatory processes in the atherosclerotic plaque. Circulating ACE-containing monocytes, which adhere to endothelial cell lesions, differentiate within the vascular wall to ACE-containing macrophages or foam cells with increased local synthesis of ACE and angiotensin II. Within the vascular wall, angiotensin II decisively contributes to the instability of the plaque by stimulating growth factors, adhesion molecules, chemotactic proteins, cytokines, oxidized LDL, and matrix metalloproteinases. Suppression of the increased ACE activity within the plaque can lead to the stabilization and deactivation of the plaque by reducing inflammation in the vascular wall, thus lessening the risk of rupture and thrombosis and the resultant acute clinical cardiovascular events. The remarkable improvement in the long-term prognosis of atherosclerotic patients with increased cardiovascular risk might be the clinical result of the contribution made by ACE inhibition in the vascular wall.     

Sequential structural and fluid dynamic numerical simulations of a stented bifurcated coronary artery

Despite their success, stenting procedures are still associated to some clinical problems like sub-acute thrombosis and in-stent restenosis. Several clinical studies associate these phenomena to a combination of both structural and hemodynamic alterations caused by stent implantation. Recently, numerical models have been widely used in the literature to investigate stenting procedures but always from either a purely structural or fluid dynamic point of view. The aim of this work is the implementation of sequential structural and fluid dynamic numerical models to provide a better understanding of stenting procedures in coronary bifurcations. In particular, the realistic geometrical configurations obtained with structural simulations were used to create the fluid domains employed within transient fluid dynamic analyses. This sequential approach was applied to investigate the final kissing balloon (FKB) inflation during the provisional side branch technique. Mechanical stresses in the arterial wall and the stent as well as wall shear stresses along the arterial wall were examined before and after the FKB deployment. FKB provoked average mechanical stresses in the arterial wall almost 2.5 times higher with respect to those induced by inflation of the stent in the main branch only. Results also enlightened FKB benefits in terms of improved local blood flow pattern for the side branch access. As a drawback, the FKB generates a larger region of low wall shear stress. In particular, after FKB the percentage of area characterized by wall shear stresses lower than 0.5?Pa was 79.0%, while before the FKB it was 62.3%. For these reasons, a new tapered balloon dedicated to bifurcations was proposed. The inclusion of the modified balloon has reduced the mechanical stresses in the proximal arterial vessel to 40% and the low wall shear stress coverage area to 71.3%. In conclusion, these results show the relevance of the adopted sequential approach to study the wall mechanics and the hemodynamics created by stent deployment.     

Changes in composition of peptidoglycan during maturation of the cell wall in pneumococci.

An experimental system which allows the selective reisolation and structural analysis of a newly made (nascent) segment of pneumococcal peptidoglycan at various times after its incorporation into the preexisting old cell wall was developed. Age-related changes were observed in each one of the major nine wall peptide components resolvable by a high-performance liquid chromatography method. The nascent wall segment (made in 1.7% of a generation time) contained 60% of its peptides as the alanyl-isoglutamyl-lysine tripeptide monomer, 12% as the directly cross-linked peptide dimer (tri-tetra peptide), and a total of 2% as the two major peptide trimers. In the mature wall segment reisolated 1 h later (1 generation time), the proportion of the tripeptide monomer dropped to 40%, while the major dimer and trimers increased to 23% and 8%, respectively. The age-related structural changes were completely inhibited by cefotaxime. The observations indicate that covalent bonds in the structure of pneumococcal peptidoglycan undergo substantial secondary rearrangements after incorporation into the preexisting wall. These changes are likely to be related to the movement of the conserved cell wall segments within the cell surface during cell division.     

Structural Characteristics of Developing Nitella Internodal Cell Walls

The Nilella intermodal cell is formed by a division of the segment cell, the latter being a direct derivative of the shoot apical cell. The internodal cell is remarkable in that it elongates from an initial length of about 20 microns to a mature length of about 60 millimeters. The structures of the apical and segment cells, and the internodal cells in all stages of development were examined with the techniques of interference, polarization, and electron microscopy. The apical and segment cells were found to be isotropic. The upper part of the segment cell, destined to form a node, shows a curious pitted structure that was characteristic of certain node structures. The lower part of the segment cell, destined to become an internodal cell, shows a vague transverse arrangement of fibrils at the inner wall surface. The internodal cells, from the time they are first formed, show negative birefringence and a transverse arrangement of microfibrils at the inner wall surface. The elongation of the internodal cell is characterized by a rise, dip, and rise in both the optical thickness and retardation of the cell wall. The dip in both these variables coincides with the attainment of the maximum relative elongation rate. After the cessation of elongation, wall deposition continues, but the fibrils at .the inner surface of the wall are now seen to occur in fields of nearly parallel microfibrils. These fields, with varying fibrillar directions, may partly overlap each other or may merge with one another. Unlike the growing wall, this wall which is deposited after the end of elongation is isotropic.     

大鼠实验性血栓模型的建立及其评价

根据Ｖｉｒｃｈｏｗ血栓形成原理,采用机械性损伤内膜,同时降低血流速度方法制备大鼠实验性血栓模型。结果显示,利用此方法制备血栓模型成功率为７８．４％,血栓平均长度８．５３±１．４２ｍｍ,血栓形成后,腹主动脉的收缩功能明显下降。     

Flow patterns in an endovascular stent-graft for abdominal aortic aneurysm repair

Endovascular exclusion of the abdominal aortic aneurysm (AAA) has been carried out in selected patients during the past decade. The deployment of a complex multicomponent endovascular device in an aneurysmal aorta may alter the local haemodynamics and lead to thrombosis and intimal hyperplasia development. The aim of this in vitro study was to investigate the flow patterns using flow visualisation and laser Doppler anemometry in a commercial bifurcated stent-graft. Two configurations of the stent-graft, endo-stent and exo-stent, were investigated in an idealised planar AAA model. The flow structures in the main trunk in both configurations of the stent-graft are three-dimensional with complex secondary structures. However, these flow structures were not entirely caused by the stent-graft. The stent struts in the endo-stent configuration cause localised alteration in the flow pattern but the overall flow structures were not significantly affected. Low velocity regions in the main trunk and flow separation in the stump region and the curved segment of the iliac limbs were observed. These areas are associated with thrombosis in the clinical situation. Improvements in the design of endovascular devices may remove these areas of unfavourable flow patterns and lead to better clinical performance.     

A quantitative model of venous stasis thrombosis in rats.

A quantitative model of stasis-type of venous thrombosis in rats is described. The ligated bowel loop was used after provocation by an injection of kaolin. The mesenteric vessels of the loop were cut in a dish filled with distilled water and the extinction of escaped haemoglobin was measured photometrically. Heparin was highly effective in this model. Vessel wall lesion may be used or inducing thrombosis instead of kaolin.     

Certain aspects of acute arterial thrombosis]

An analysis included 228 patients with the acute arterial thrombosis in the extremities. Over 33% of patients reported to the treatment later than 24 hours after the onset. This delay markedly worsened the results of therapy. Sixty two percent of limbs was saved. Mortality rate was 15%. The author analyse the results of therapy in relation to the localization of thrombi and type of the treatment--surgical, use of vasodilators with heparin or streptokinase. In case of thrombosis localized in the end segment of aorta the results were less promising than in case of more peripheral arterial involvement. Excellent and favourable results in this group amounted to 32%, the limb was amputated in 13% of patients and 28% of patients died. Hundred eighty patients were operated. Arteriosclerotic lesions to the arterial wall were detected in 97%. Excellent and favourable results of surgery were achieved in 45%, amputations amounted to 22%, and mortality rate was 16%. Vasodilators combined with heparin produced an improvement in 13% of patients in whom surgery could be postponed. The best results were achieved in patients treated with fibrinolytic agents with subsequent surgery or without it. In this group excellent and favourable results amounted to 57%, amputations--24%, and mortality rate--8%.     

Resolution of smooth muscle and endothelial pathways for conduction along hamster cheek pouch arterioles

In the cheek pouch of anesthetized male hamsters, microiontophoresis of Ach (endothelium-dependent vasodilator) or phenylephrine (PE; smooth muscle-specific vasoconstrictor) onto an arteriole (resting diameter, 30-40 microm) evokes vasodilation or vasoconstriction (amplitude, 15-25 microm), respectively, that conducts along the arteriolar wall. In previous studies of conduction, endothelial and smooth muscle layers of the arteriolar wall have remained intact. We tested whether selective damage to endothelium or to smooth muscle would disrupt the initiation and conduction of vasodilation or vasoconstriction. Luminal (endothelial) or abluminal (smooth muscle) light-dye damage was produced within an arteriolar segment centered 500 microm upstream from the distal site of stimulation; conducted responses (amplitude, 10-15 microm) were observed at a proximal site located 1,000 microm upstream. Endothelial damage abolished local responses to ACh in the central segment without affecting those to PE. Nevertheless, ACh delivered at the distal site evoked vasodilation that conducted through the central segment and appeared unhindered at the proximal site. Smooth muscle damage inhibited responses to PE in the central segment and abolished the conduction of vasoconstriction but did not affect conducted vasodilation. We suggest that for cheek pouch arterioles in vivo, vasoconstriction to PE is initiated and conducted within the smooth muscle layer alone. In contrast, once vasodilation to ACh is initiated via intact endothelial cells, the signal is conducted along smooth muscle as well as endothelial cell layers.