Walls Of Postcapillary Venules Research Articles

QUANTITATIVE MORPHOLOGICAL ANALYSIS OF THE FEATURES OF REMODELING OF HEMOMICROCIRCULATORY VESSELS IN THE TESTIS OF EXPERIMENTAL ANIMALS AT CONDITIONS OF ETHANOL INTOXICATION

Ethanol is classifi ed as a vascular poison, long-term intoxication of which negatively aff ects almost all organs and systems body of humans and experimental animals. Morphological changes in testes during ethanol poisoning have not been studied enough. The aim of the study was a quantitative morphological analysis of the features of the remodeling of blood vessels of the hemomicrocirculatory bed of the left and right testes at conditions of ethanol intoxication. The structures of the left and right testis of 30 white male rats, which were divided into 2 groups, were morphologically and morphometrically studied. The 1 group included 15 intact animals, the 2-15 rats, which were injected intragastrically with a 30 % ethanol solution at the rate of 2 ml per 100 g of body weight for 28 days once a day. A month after the start of the experiment, the experimental animals were euthanized by bloodletting under thiopental anesthesia. Histological and semithin sections were made from testes. The outer and inner diameters, wall thickness of arterioles, precapillary arterioles, hemocapillaries, postcapillary venules and venules were determined. Quantitative indicators were processed statistically.It was established that twenty- eight days of ethanol intoxication in laboratory sexually mature white male rats leads to pronounced remodeling of the blood vessels of the hemomicrocirculatory bed of the left and right testes, which is characterized by narrowing of the lumens and thickening of the walls of its arterial (arterioles, precapillary arterioles) and exchange (hemocapillaries) vessels. pronounced expansion and thinning of the walls of postcapillary venules and venules, venous hemoptysis, hypoxia, dystrophic- necrotic changes in cells, stromal structures, infi ltration and sclerosis. The revealed remodeling of blood vessels of the hemomicrocirculatory bed in conditions of ethanol poisoning dominates in the left testicle.

Progressive ascending telangiectasias

Progressive ascending telangiectasias

Oxidative stress activates red cell adhesion to laminin in sickle cell disease.

Vaso-occlusive crises are the hallmark of sickle cell disease (SCD). They are believed to occur in two steps, starting with adhesion of deformable low-dense red blood cells (RBC), or other blood cells such as neutrophils, to the wall of post-capillary venules, followed by trapping of denser RBC or leukocytes in the areas of adhesion because of reduced effective lumen-diameter. In SCD, RBC are heterogeneous in terms of density, shape, deformability and surface proteins, which accounts for the differences observed in their adhesion and resistance to shear stress. Sickle RBC exhibit abnormal adhesion to laminin mediated by Lu/BCAM protein at their surface. This adhesion is triggered by Lu/BCAM phosphorylation in reticulocytes but such phosphorylation does not occur in mature dense RBC despite firm adhesion to laminin. In this study, we investigated the adhesive properties of sickle RBC subpopulations and addressed the molecular mechanism responsible for the increased adhesion of dense RBC to laminin in the absence of Lu/BCAM phosphorylation. We provide evidence for the implication of oxidative stress in post-translational modifications of Lu/BCAM that impact its distribution and cis-interaction with glycophorin C at the cell surface activating its adhesive function in sickle dense RBC.

Surface Chemistry of Quantum Dots Determines Their Behavior in Postischemic Tissue

The behavior of quantum dots (QDs) in the microvasculature and their impact on inflammatory reactions under pathophysiological conditions are still largely unknown. Therefore, we designed this study to investigate the fate and effects of surface-modified QDs in postischemic skeletal and heart muscle. Under these pathophysiological conditions, amine-modified QDs, but not carboxyl-QDs, were strongly associated with the vessel wall of postcapillary venules and amplified ischemia-reperfusion-elicited leukocyte transmigration. Importantly, strong association of amine-QDs with microvessel walls was also present in the postischemic myocardium. As shown by electron microscopy and verified by FACS analyses, amine-modified QDs, but not carboxyl-QDs, were associated with endogenous microparticles. At microvessel walls, these aggregates were attached to endothelial cells. Taken together, we found that both the surface chemistry of QDs and the underlying tissue conditions (i.e., ischemia-reperfusion) strongly determine their uptake by endothelial cells in microvessels, their association to endogenous microparticles, as well as their potential to modify inflammatory processes. Thus, this study strongly corroborates the view that the surface chemistry of nanomaterials and the physiological state of the tissue are crucial for the behavior of nanomaterials in vivo.

Neutrophil Adhesion and Activation under Flow

Neutrophil recruitment into inflamed tissue in response to injury or infection is tightly regulated. Reduced neutrophil recruitment can result in a reduced ability to fight invading microorganisms. During inflammation, neutrophils roll along the endothelial wall of postcapillary venules and integrate inflammatory signals. Neutrophil activation by selectins and chemokines regulates integrin adhesiveness. Binding of activated integrins to their counter-receptors on endothelial cells induces neutrophil arrest and firm adhesion. Adherent neutrophils can be further activated to undergo cytoskeletal rearrangement, crawling, transmigration, superoxide production, and respiratory burst. Signaling through G-protein-coupled receptors, selectin ligands, Fc receptors and outside-in signaling through integrins are all involved in neutrophil activation, but their interplay in the multistep process of recruitment is only beginning to emerge. This review provides an overview of signaling in rolling and adherent neutrophils.

Cisplatin up‐regulates ICAM‐1 expression in endothelial cell via a NF‐κB dependent pathway

The anticancer drug cis-diammindichloroplatin (cisplatin) can cause severe side-effects, but to date, the mechanisms of action of these dangerous side-effects have not been completely elucidated. Since cellular adhesion molecules (CAMs), by mediating the recruitment of circulating leukocytes to the blood vessel wall and their subsequent migration into the subendothelial spaces, play a crucial role in several pathophysiologic processes, we sought potential proof for CAMs in the pathophysiology of cisplatin-induced vascular damage. In vitro, human umbilical vein endothelial cells (HUVECs) were subjected to various concentrations of cisplatin, considerable up-regulation of intercellular adhesion molecule-1 (ICAM-1) but not P-selectin, E-selectin, and vascular cell adhesion molecule 1 at both messenger mRNA and protein expression levels were observed. Electrophoretic mobility shift assays and Western blotting analysis revealed that cisplatin up-regulates ICAM-1 expression in HUVECs via an NF-kappaB-dependent pathway. Further intravital microscopy study demonstrated that significantly higher (P < 0.01) numbers of rolling and sticking leukocytes on the wall of postcapillary venules in male Golden Syrian hamster's cheek pouch bearing a human cervical carcinoma were observed, while inhibition of ICAM-1 by using specific anti-ICAM-1 antibody can attenuate cisplatin-stimulated leukocyte/endothelium interactions. These data suggest that ICAM-1 involves in the pathophisiologic process of cisplatin-induced vascular toxicity and may be exploited for therapeutic advantage.

Mechanisms and Consequences of Neutrophil Interaction with the Endothelium

Leukocyte recruitment into inflamed tissue proceeds in a cascade-like fashion. The first contact of neutrophils with the endothelium is mediated by selectins and their counterreceptors, followed by rolling of neutrophils along the endothelial wall of postcapillary venules and integrin-mediated arrest. While rolling, neutrophils collect different inflammatory signals that can activate several pathways. In addition to activation of neutrophils by ligation of G-protein-coupled receptors with chemokines and other chemoattractants, integrins and selectin ligands are also able to signal into the cell, where they initiate neutrophil extravasation, promote cytoskeletal rearrangement, and ultimately induce superoxide production and degranulation. These signaling pathways may be targeted by therapeutic interventions to inhibit specific functions of neutrophils without affecting others. This Review is focused on the signaling events during the interaction of neutrophils with the endothelium.

Continuous intra-arterial application of substance P induces signs and symptoms of experimental complex regional pain syndrome (CRPS) such as edema, inflammation and mechanical pain but no thermal pain

Substance P is involved in nociception in both the peripheral nervous system and the CNS and has been documented to play a crucial role in the complex regional pain syndrome (CRPS). So far, however, most experimental animal models are restricted to the effect of neurokinin-1 receptor blockers to inhibit substance P and do not directly evaluate its action. Thus, this study was conducted to test the hypothesis that local application of substance P causes signs and symptoms of CRPS. For this purpose rats received a continuous infusion of either substance P or saline over 24 h delivered by a mini-osmotic pump connected to an intrafemoral catheter. Animals were analyzed at either day 1 ( n=6, each group) or day 4 ( n=5, each group) after start of infusion. Substance P application caused a significant and long-lasting decrease in paw withdrawal thresholds upon mechanical stimulation, while animals did not present with thermal allodynia at days 1 and 4 after onset of infusion. In addition, severe s.c. edema was observed in all animals receiving substance P. In vivo fluorescence microscopy of the extensor digitorum longus muscle of the affected hind paw revealed enhanced leukocyte–endothelial cell interaction with a significant rise in the number of leukocytes both rolling along and firmly adhering to the wall of postcapillary venules, while saline-exposed animals were free of this local inflammatory response. Muscle cell apoptosis, as assessed by in vivo bisbenzimide staining, terminal deoxynucleotidyl transferase nick end labeling analysis and caspase 3-cleavage, could not be observed in either of the animals. In summary, the present study indicates that substance P is responsible for neurogenic inflammation, including local cell response, edema formation and mechanical pain, while it seems not to contribute to the generation of thermal allodynia.

56-Year-Old Man With Rash, Abdominal Pain, and Arthralgias

56-Year-Old Man With Rash, Abdominal Pain, and Arthralgias

Noninvasive characterization of light‐induced acute inflammation in the Pallid bat

It has been observed that ultraviolet (UV) and visible light induce vasodilation (i.e. photodilation) and increases local blood flow in skin. It is, however, not known whether the resulting vasodilation is a primary, NO-mediated effect, or a secondary effect of ultraviolet light phototoxicity. To address this question, we utilized the Pallid bat wing model at TAMU. The wing is thin and transparent, allowing for intravital microscopy without invasive procedures (leading to confounding effects of anesthesia and trauma). TAMU, however, has the only extant colony of bats dedicated to vascular research. To access unique animal and laboratory resources, we employed a novel use of Internet technology we refer to as ‘e-Research’, consisting of logging into a remote computer capable of manipulating positioning of an intravital microscope. To characterize an acute inflammatory response, we exposed the Pallid bat wing to brightfield and ultraviolet light and counted interactive leukocytes along the walls of post-capillary venules. We observed a measurable, but not significant, increase in leukocyte activity. In this study, we showed that light-induced vasodilation is not a consequence of an acute inflammatory response (i.e. tissue damage). Our unique approach allows us to explore basic microvascular function and acute inflammatory responses with well-established intravital microscopic methods.

Leukocyte phosphoinositide-3 kinase γ is required for chemokine-induced, sustained adhesion under flow in vivo

During inflammation, leukocytes roll along the wall of postcapillary venules scanning the surface for immobilized CXCL1, a chemokine that triggers firm adhesion by activating CXCR2 on the neutrophil. PI-3K are signaling molecules important in cellular processes, ranging from cellular differentiation to leukocyte migration. PI-3Kgamma can be activated directly by the betagamma dimer of heterotrimeric G proteins coupled to CXCR2. Here, we used in vivo and ex vivo intravital microscopy models to test the role of PI-3Kgamma in leukocyte arrest. PI-3Kgamma null mice showed an 80% decrease in CXCL1-induced leukocyte adhesion in venules of the exteriorized mouse cremaster muscle. In wild-type mice, rolling leukocytes showed rapid and sustained adhesion, but in PI-3Kgamma(-/-) mice, adhesion was not triggered at all or was transient, suggesting that absence of PI-3Kgamma interferes with integrin bond strengthening. Wild-type mice reconstituted with PI-3Kgamma null bone marrow showed a 50% decrease in CXCL1-induced leukocyte adhesion. In a blood-perfused micro-flow chamber, leukocytes from PI-3Kgamma(-/-) mice showed a defect in adhesion on a P-selectin/ICAM-1/CXCL1 substrate, indicating that leukocyte PI-3Kgamma was required for adhesion. The adhesion defect in PI-3Kgamma(-/-) mice was as severe as that in mice lacking LFA-1, the major integrin responsible for neutrophil adhesion. We conclude that the gamma isoform of PI-3K must be functional in leukocytes to allow efficient adhesion from rolling in response to chemokine stimulation.

E-selectin, thymus- and activation-regulated chemokine/CCL17, and intercellular adhesion molecule-1 are constitutively coexpressed in dermal microvessels: a foundation for a cutaneous immunosurveillance system.

The success of the cutaneous immune system reflects its ability to rapidly and efficiently recruit leukocytes to areas of trauma and infection. Skin-homing memory T cells expressing cutaneous lymphocyte-associated Ag tether on the walls of postcapillary venules in inflamed skin via interaction with endothelial E-selectin and roll in response to the shear stress imparted by flowing blood. Rolling cells sample the vascular surface for chemoattractant compounds (e.g., thymus- and activation-regulated chemokine/CCL17 interacting with CCR4 on the leukocyte surface) and, if successfully stimulated, progress to firm arrest and transmigration mediated by LFA-1 and vascular ICAM-1. Although it is established that this sequence of events draws T cells into inflamed skin, the mechanisms directing trafficking of T cells to noninflamed skin are less well characterized. We hypothesized that basal expression and colocalization of E-selectin, chemokine (e.g., CCL17), and ICAM-1 in dermal vessels could serve to recruit T cells to noninflamed human skin. Immunohistochemical staining for E-selectin and CD31 demonstrated E-selectin expression in a restricted subset of dermal vessels in noninflamed human skin from three different sites. Confocal multicolor immunofluorescence imaging revealed a nonuniform distribution of E-selectin in dermal vessels as well as colocalization of E-selectin with CCL17 and ICAM-1. Coexpression of these molecules on blood vessels in noninflamed skin provides the basis for a model of cutaneous immunosurveillance system active in the absence of pathologic inflammation.

Hydrodynamic Interactions Between Rolling Leukocytes In Vivo

The aim of this study was to characterize the hydrodynamic interactions between rolling and free-stream leukocytes in an in vivo model of selectin-mediated rolling, and to identify those physical mechanisms that influence the dynamics of transient adhesion with the walls of postcapillary venules. Postcapillary venules of diameter 22-37 microm in the cheek pouch of anesthetized hamsters were visualized using intravital microscopy, with selectin-mediated rolling occurring in response to surgical preparation. Rolling velocity was found to be a strong function of the center-to-center separation with the nearest cell, and also was found to correlate strongly with the number of nearby cells. These effects are shown to be beyond that attributable to variations along the length of the vessel. Adherent leukocytes were observed to provide a nucleation site, precipitating further adhesion events of free-stream cells. The dynamics of the transient adhesion of leukocytes to the vessel wall in postcapillary venules is strongly dependent on the local concentration of adherent leukocytes, due to the complex hydrodynamics induced by their presence. The results are shown to agree well with theoretical considerations of the flow field induced by multiple nearby cells, suggesting a need for the future exploration of multicellular effects in the microcirculation.

Integration of inflammatory signals by rolling neutrophils.

In inflammation, neutrophils roll along the endothelial wall of postcapillary venules and sample inflammatory signals. Neutrophil activation is required to generate beta(2) integrin bonds with the endothelium that are strong enough to withstand the flow forces and thus achieve arrest from the rolling state. Unlike naïve T cells, neutrophils are not only activated by ligation of G-protein coupled receptors with chemokines and other chemoattractants but also receive signals from engagement of adhesion molecules including the selectins and beta(2) integrins. Rolling neutrophils integrate the sum total of inputs received while scanning the inflamed endothelium. In this process, the velocity of rolling neutrophils systematically decreases as a function of their contact time with the inflamed endothelium. If an activation threshold is reached, beta(2) integrins switch to the high-affinity conformation, redistribute on the cell surface, and trigger arrest and adhesion. Rolling cells that do not reach the activation threshold detach from the endothelium and are released back into the circulation. The role of chemokines, adhesion molecules, and other activating inputs involved in this response as well as signaling pathways are the subjects of ongoing investigations. This review provides a conceptual framework for neutrophil recruitment from the flowing blood.

Diabetes enhances leukocyte accumulation in the coronary microcirculation early in reperfusion following ischemia

Background: Diabetic hearts are particularly vulnerable to ischemia–reperfusion injury. For leukocytes to participate in ischemia–reperfusion injury, they must first sequester in the microcirculation. The aim of this study was to determine, by direct observation, if early leukocyte deposition was increased in the diabetic coronary microcirculation early in reperfusion following myocardial ischemia. Methods: Non-diabetic and streptozotocin (STZ)-induced diabetic rat hearts, subjected to 30 min of 37°C, no-flow ischemia, were initially reperfused with blood containing labeled leukocytes. The deposition of fluorescent leukocytes in coronary capillaries and venules was directly visualized and recorded using intravital fluorescence microscopy. In addition, flow cytometry was used to measure CD11b adhesion molecule expression on polymorphonuclear (PMN) leukocytes from non-diabetic and STZ-diabetic rats. Results: In the non-diabetic, control hearts, early in reperfusion, leukocytes trapped in coronary capillaries and adhered to the walls of post-capillary venules. In the diabetic hearts, leukocyte trapping in capillaries and adhesion to venules were both significantly increased ( P<0.05). PMN CD11b expression was also significantly increased in the diabetic blood compared to the non-diabetic blood ( P<0.05). Conclusions: Early in reperfusion following myocardial ischemia, leukocytes rapidly accumulate in greater numbers in the coronary microcirculation of the diabetic heart by both trapping in coronary capillaries and by adhering to venules. The enhanced retention of leukocytes in the diabetic coronary microcirculation increases the likelihood of inflammation-mediated reperfusion injury and may explain, in part, the poor recovery of diabetic hearts from an ischemic event.

Scanning electron microscopic studies on the route of neutrophil extravasation in the mouse after exposure to the chemotactic peptide N-formyl-methionyl-leucyl-phenylalanine (fMLP).

The present study was performed to demonstrate three-dimensionally the process of neutrophil extravasation induced by the bacterial chemotactic peptide N-formyl-methionyl-leucyl-phenylalanine (fMLP) in mice. Thirty to 40 min after the injection of fMLP to the mouse lip, the tissues were fixed with glutaraldehyde and examined by scanning electron microscopy (SEM) as well as by transmission electron microscopy (TEM). Observation of fMLP-injected tissues showed many neutrophils adhering to the inner wall of postcapillary venules. Some of these adherent neutrophils attached to each other to form groups of two to six. There were also many neutrophils migrating through the endothelium. Most of these neutrophils took a transcellular route, in that they penetrated the cytoplasm of the endothelial cells. The junction of two neighboring endothelial cells did not open, and endothelial pores free from migrating neutrophils were scarcely observed. There were bulging portions on the vascular wall which were probably produced by the presence of underlayed neutrophils. Thus, the present study gave direct evidence of neutrophil migration via a transcellular route in response to fMLP. Our findings also indicate that the pores of the endothelium close quickly after neutrophil extravasation.

Interactions of Lymphocytes from Patients with Psoriatic Arthritis or Healthy Controls and Cultured Endothelial Cells

Psoriatic arthritis (PA) is an inflammatory rheumatic disease that can concomitantly occur in patients with psoriasis vulgaris. Psoriatic synovitis shows alterations of the synovial microvasculature. Inflammatory cells adhere to endothelial cells (EC) and migrate through the vascular wall of postcapillary venules located in the subintimal layer of the synovial membrane. The aim of our study was to investigate, first, the phenotype of lymphocytes (LC) of PA patients using flow cytometry (FC) with regard to activation antigens and adhesion molecules; second, the adhesion of LC of PA patients on cultivated resting or activated (with thrombin, LPS, IFN-γ, or TNF-α) human umbilical vein endothelial cells (HUVEC) by counting the Feulgen-stained nuclei of both adherent LC and HUVEC using image analysis; and third, the synthesis of IL-6 and IL-8 in both LC and HUVEC 24 hr after cell contact. These cytokines were determined qualitatively by immunofluorescence and quantitatively at the single-cell level by FC as well as in the supernatants of the cultures using commercial cytokine ELISAs. Fourth, we investigated whether or not the LC adhesion on HUVEC as well as the cytokine production could be inhibited by monoclonal antibodies against LC- or EC-specific adhesion molecules. In contrast to controls PA patients showed an increased surface expression of CD11a, b, and c as well as of CD44 but a reduced surface expression of CD49d/CD29, CD49e/CD29, and cell-bound fibronectin on CD3 +LC. The activation markers CD25 and HLA-DR were found to be slightly enhanced in PA. The cell adhesion was generally enhanced in PA patients vs controls. It could be reduced with monoclonal antibodies (MoAbs) against CD11a and CD18 on IFN-γ- or TNF-α-activated HUVEC but was generally enhanced after treatment of HUVEC with MoAbs against CD54, CD62E, or CD106. Due to LC adhesion on HUVEC IL-6 and IL-8 were produced in significantly higher amounts in PA patients compared to controls. This effect occurred already in resting but was enhanced in activated HUVEC. While IL-6 is mainly produced by HUVEC but also in smaller quantities by LC, IL-8 is synthesized only by HUVEC and could be modified by preincubation with MoAbs against LC- or EC-specific adhesion molecules in parallel to the cell adhesion. The experiments show that the main adhesion pathway in LC homing of PA patients is the interaction of the LC adhesion molecule CD11a/CD18 with CD54 on EC followed by an enhanced synthesis of proinflammatory and chemotactic cytokines. These results favor the hypothesis that the pathological alterations of the microvasculature in PA patients are generated by altered homing processes.

Model Studies of Leukocyte-Endothelium-Blood Interactions

Computational fluid dynamics (CFD) and large scale model experiments were used to analyze the hemodynamic impact of leukocytes adherent to the wall of post-capillary venules. Using a large scale model and, with the aid of a finite element package, so

Model studies of leukocyte-endothelium-blood interactions: II. Hemodynamic impact of leukocytes adherent to the wall of post-capillary vessels

Computational fluid dynamics (CFD) and large scale model experiments were used to analyze the hemodynamic impact of leukocytes adherent to the wall of post-capillary venules. Using a large scale model and, with the aid of a finite element package, solving the Navier Stokes equations for low Reynolds number flow in a cylinder past an adherent sphere, we have developed a dimensionless correlation which permits the estimation of the pressure drop across an adherent leukocyte in an in vivo vessel. This relationship is: f ⋅ Re ⁡ = exp ⁡ [ 2.877 + 4.630 ( d D ) 4 ] where f is the Fanning friction factor, Re is the Reynolds number and d/D is the leukocyte to vessel diameter ratio. The friction factor is proportional to the pressure drop across the leukocyte, and does not significantly increase until d/D is greater than 0.5, and then increases rapidly with increasing d/D. Computations indicate that the length of the disturbed flow region generated by an adherent leukocyte increases with decreasing vessel size. The average wall stress in the disturbed flow region remains constant, and equal to the wall stress in the undisturbed region for d/D less than approximately 0.5. For d/D greater than 0.5, the average wall stress in the disturbed flow region increases rapidly with increasing d/D. There is an even larger increase, up to five times greater than the average disturbed stress, in the peak wall stress in the disturbed flow region. This indicates that significant wall stress gradients can be generated by an adherent leukocyte in post-capillary size vessels. © 1997 Elsevier Science Ltd

Multiple ligands for cytoadherence can be present simultaneously on the surface of Plasmodium falciparum-infected erythrocytes.

A major virulence factor of Plasmodium falciparum is the adherence of parasitized erythrocytes to the wall of postcapillary venules via a specific interaction between parasite-derived erythrocyte surface ligands and receptors on endothelial cells. To study this phenomenon in vitro, we selected a parasite population that expressed at least two different ligands and demonstrated that parasitized cells may coexpress ligands with specificity for multiple receptors. This selected parasite line had several antigenic and cytoadherence characteristics that were different from those of the parent line. Single parasitized erythrocytes were able to adhere to three distinct receptors via at least two separate ligands; a trypsin-sensitive molecule mediated cytoadherence to CD36 and intercellular adhesion molecule 1 and a trypsin-insensitive molecule(s) was responsible for adherence to a third receptor on the surface of melanoma cells. We present evidence that this newly discovered receptor for cytoadherence is an N-linked glycosaminoglycan, as treatment of melanoma cells with endoglycosidase H abolished cytoadherence. These observations emphasize the adaptability of P. falciparum and the complexity of the cytoadherence phenomenon.