Nitric oxide mediates histamine induced down-regulation of H<Subscript>2</Subscript> receptor mRNA and internalization of the receptor protein (R1)

During agonist-dependent long-term stimulation of cells, histamine receptor subtypes are frequently down-regulated. However, the mechanisms underlying the modulation of receptor expression during long-term histamine stimulation have yet to be resolved. Based on our recently reported results showing an H₁-mediated down-regulation of histamine H₂ receptor mRNA in endothelial cells, our aim was to characterize the mechanism controlling rapid and long-term histamine-mediated modulation of H₂ receptor expression in more detail. We were able to show that the histamine-induced down-regulation of H₂ receptor mRNA and cell surface expression lasting for 24 h was accompanied by augmentation of the receptor protein level in the cytoplasmatic fraction of endothelial cells for this time period. Furthermore, changes in receptor protein levels in whole-cell lysate were negligible, indicating that the rapid and prolonged modulation of cell surface H₂ receptor levels by histamine was regulated solely via internalization. The role of nitric oxide (NO) as a key mediator in histamine-stimulated cell responses was underlined by subsequent studies showing the attenuation of histamine-induced H₂ receptor mRNA down-regulation and protein trafficking following NO synthase isozyme inhibition.Received 11 March 2003; received after revision 11 June 2003; accepted 17 June 2003     

Zum Wirkungsmechanismus des Histamins

Summary With the aid of the tensiometer, it can be proved that histamine reacts in a specific manner with primary phosphoric acid esters. This proof is brought about by strongly increasing the capillary activity of cetyl phosphate by histamine on the interface chloroform water. The mechanism of this reaction is discussed. Simple secondary phosphoric acid esters and bi-secondary phosphoric acid esters are not influenced under the same conditions. A pronounced antagonistic effect of calcium ions antihistamine compounds may be proved with the same assays.The point of attack of these compounds is the phosphate and not the histamine. The results of the tests confirm the assumption that the mode of action of the histamine and of the antihistamine compounds is connected with primary phosphoric acid esters.     

Calcium dobesilate (Doxium) as a prostaglandin synthetase inhibitor in pregnant human myometrium in vitro

This comparative study on the effect of calcium dobesilate and indomethacin on prostaglandin biosynthesis was performed on microsomal fractions of pregnant human myometrium. Both drugs inhibited prostaglandin synthesis, indomethacin being more potent. Calcium dobesilate inhibited, in a dose-dependent manner, the synthesis of 6-oxo-PGF1 alpha, PGF2 alpha, PGE2 and TXB2. Its inhibitory action is comparable to that of etamsylate.     

Shear-dependence of endothelial functions

Endothelial cells are subjected to shear forces which influence important cell functions. Shear stress induces cell elongation and formation of stress fibers, increases permeability, pinocytosis and lipoprotein internalization, is involved in the formation of atherosclerotic lesions, increases the production of tissue plasminogen activator, and enhances von Willebrand factor release and hence platelet aggregation. It decreases adherence of erythrocytes and leukocytes, and increases the release of prostacyclin, endothelium derived relaxing factor, histamine and other compounds, but decreases erythropoietin secretion. The mechanism of signal transduction to the endothelial cell is not known exactly; shear-sensitive ion channels seem to be involved. It is concluded that a better understanding of shear-dependent endothelial functions will influence pathophysiologic concepts and therapeutic interventions.     

Activation-induced cellular accumulation of histamine in immature but not mature murine mast cells

Mast cell activation involves the rapid release of inflammatory mediators, including histamine, from intracellular granules. The cells are capable of regranulation and multiple rounds of activation. The goal of this study was to determine if there are changes in the content of pre-formed mast cell mediators after a round of activation. After 24 h, the histamine content of bone marrow-derived mast cells (BMMC), but not that of peritoneal mast cells, exceeded the amount in resting cells. Accumulation of histamine in BMMC peaked at 72 h of activation, and returned toward preactivation levels by 96 h. The increase in histamine content was accompanied by an increase in the gene expression of histidine decarboxylase. No increases in beta hexosaminidase or murine mast cell protease-6 were observed. These findings indicate that BMMC respond to activation by increasing total cell-associated histamine content. This increase may be important to the response of these cells upon subsequent exposure to antigens.     

Role of calcium in the histamine release induced by D-galactosamine from rat mast cells

Rat peritoneal mast cells were isolated and purified by differential centrifugation in Ficoll. Cells pooled from three to four rats were suspended at approximately 10(6) cells/ml in a buffered salt solution and incubated for 1 h at 37 degrees C in 300 microliter volumes in the absence or presence (9 X 10(-4) M) of calcium chloride. Addition of D-galactosamine hydrochloride (DGM; 2.8 X 10(-4)M) caused (in addition to basal release) a mean +/- SEM percent histamine release of 15.7 +/- 5.2 in the presence of Ca++ and 19 +/- 4.9 in the absence of Ca++ (p greater than 0.05). It is suggested that D-galactosamine does not require extracellular Ca++ for the release of histamine from the rat mast cell.     

Effect of vasoactive amines on Weibel-Palade bodies in capillary endothelial cells

The presence and distribution of Weibel-Palade bodies in stomach and colonic mucosal microvessels after the administration of vasoactive amines (serotonin and histamine), the serotonin depletor reserpine, and the von Willebrand factor secretagogue thrombin, was studied by transmission electron microscopy. These agents elevated the number of Weibel-Palade bodies in all microvascular endothelial cells and especially in capillaries. It is concluded that vasoactive amines enhance the synthesis and secretion of large von Willebrand protein multimers by endothelial cells.     

Effect of vasoactive amines on Weibel-Palade bodies in capillary endothelial cells

The presence and distribution of Weibel-Palade bodies in stomach and colonic mucosal microvessels after the administration of vasoactive amines (serotonin and histamine), the serotonin depletor reserpine, and the von Willebrand factor secretagogue thrombin, was studied by transmission electron microscopy. These agents elevated the number of Weibel-Palade bodies in all microvascular endothelial cells and especially in capillaries. It is concluded that vasoactive amines enhance the synthesis and secretion of large von Willebrand protein multimers by endothelial cells.     

Calcium dobesilate (Doxium) as a prostaglandin synthetase inhibitor in pregnant human myometrium in vitro

Summary This comparative study on the effect of calcium dobesilate and indomethacin on prostaglandin biosynthesis was performed on microsomal fractions of pregnant human myometrium. Both drugs inhibited prostaglandin synthesis, indomethacin being more potent. Calcium dobesilate inhibited, in a dose-dependent manner, the synthesis of 6-oxo-PGF₁, PGF₂, PGE₂ and TXB₂. Its inhibitory action is comparable to that of etamsylate.     

Choroidal and iris angioarchitecture of the newt: A scanning electron-microscopic study of vascular corrosion casts

The corrosion cast technique provided for the first time an excellent three-dimensional visualization of the vascular pattern of the choroid and iris in the newt eye. The results show the presence of a single arterial afference to the choroidal and iris capillaries: the ophthalmic artery is the origin fo both ciliary arteries and the long posterior ciliary artery. Slightly behind the equatorial circumference of the eyeball the venous drainage consists of a single vessel on the dorsal side and two distinct vessels on the ventral one. It receives blood from both iris and choroid. The surface of the plastic endocasts shows some details of fine luminal structures of the endothelial cells. Shallow depressions may be regarded as imprints of endothelial cell nuclei, and they are distinctly different for arteries and capillaries. The angioarchitecture of the newt eye differs from that of brain in that hairpin-shaped capillary loops are not observed at all.     

Role of calcium in the histamine release induced by D-galactosamine from rat mast cells

Summary Rat peritoneal mast cells were isolated and purified by differential centrifugation in Ficoll. Cells pooled from three to four rats were suspended at approximately 10⁶ cells/ml in a buffered salt solution and incubated for 1 h at 37°C in 300 l volumes in the absence or presence (9×10^–4 M) of calcium chloride. Addition of D-galactosamine hydrochloride (DGM; 2.8×10^–4 M) caused (in addition to basal release) a mean ±SEM percent histamine release of 15.7±5.2 in the presence of Ca⁺⁺ and 19±4.9 in the absence of Ca⁺⁺ (p>0.05). It is suggested that D-galactosamine does not require extracellular Ca⁺⁺ for the release of histamine from the rat mast cell.A preliminary analysis of these results was presented at the International Symposium on calcium entry blockers and tissue protection, Rome, 15–16 March 1984.     

Allergic reactions, cyclic AMP and histamine release.

Both isobutylmethylxanthine and theophylline increased the level cyclic AMP in the mast cell. Theophylline reduced the allergic histamine release, whereas isobutylmethylxynthine caused a pronounced potentiation of the histamine release. This indicates that the hypothesis of an inverse relationship between the level of cyclic AMP in mast cells and histamine release is too simple.     

Electron microscopic study of secretion of chylomicrons in the rat by enterocytes in lymphatic capillary lumen]

Rats were fed on a 20% fat diet; 30 min. or 2 hrs later intestinal jejunal cells were fixed in situ; samples were taken and processed for subsequent study by electron microscopy. Chylomicron secretion was followed from the enterocyte to the lymphatic capillary lumen. Chylomicrons entered the lamina propria through breaks in the basal membrane at intercellular space level. Their penetration into the lymphatic capillary lumen occurred through the open junctions of the endothelial cells normally kept closed by desmosomal formations.     

Serglycin – Structure and biology

Serglycin is a proteoglycan found in hematopoietic cells and endothelial cells. It has important functions related to formation of several types of storage granules. In connective tissue mast cells the covalently attached glycosaminoglycan is heparin, whereas mucosal mast cells and activated macrophages contain oversulfated chondroitin sulfate (type E). In mast cells, serglycin interact with histamine, chymase, tryptase and carboxypeptidase, in neutrophils with elastase, in cytotoxic T cells with granzyme B, in endothelial cells with tissue-type plasminogen activator and in macrophages with tumor necrosis factor-α. Serglycin is important for the retention of key inflammatory mediators inside storage granules and secretory vesicles. Serglycin can further modulate the activities of partner molecules in different ways after secretion from activated immune cells, through protection, transport, activation and interactions with substrates or target cells. Serglycin is a proteoglycan with important roles in inflammatory reactions. Received 2 October 2007; received after revision 7 November 2007; accepted 12 November 2007     

Endothelial membrane reorganization during leukocyte extravasation

Leukocyte trafficking from the bloodstream to inflamed tissues across the endothelial barrier is an essential response in innate immunity. Leukocyte adhesion, locomotion, and diapedesis induce signaling in endothelial cells and this is accompanied by a profound reorganization of the endothelial cell surfaces that is only starting to be unveiled. Here we review the current knowledge on the leukocyte-mediated alterations of endothelial membrane dynamics and their role in promoting leukocyte extravasation. The formation of protein- and lipid-mediated cell adhesion nanodomains at the endothelial apical surface, the extension of micrometric apical membrane docking structures, which are derived from microvilli and embrace adhered leukocytes, as well as the vesicle-trafficking pathways that are required for efficient leukocyte diapedesis, are discussed. The coordination between these different endothelial membrane-remodeling events probably provides the road map for transmigrating leukocytes to find exit points in the vessel wall, in a context of severe mechanical and inflammatory stress. A better understanding of how vascular endothelial cells respond to immune cell adhesion should enable new therapeutic strategies to be developed that can abrogate uncontrolled leukocyte extravasation in inflammatory diseases.     

Syncytin is involved in breast cancer-endothelial cell fusions

Cancer cells can fuse spontaneously with normal host cells, including endothelial cells, and such fusions may strongly modulate the biological behaviour of tumors. However, the underlying mechanisms are unknown. We now show that human breast cancer cell lines and 63 out of 165 (38%) breast cancer specimens express syncytin, an endogenous retroviral envelope protein, previously implicated in fusions between placental trophoblast cells. Additionally, endothelial and cancer cells are shown to express ASCT-2, a receptor for syncytin. Syncytin antisense treatment decreases syncytin expression and inhibits fusions between breast cancer cells and endothelial cells. Moreover, a syncytin inhibitory peptide also inhibits fusions between cancer and endothelial cells. These results are the first to show that syncytin is expressed by human cancer cells and is involved in cancer-endothelial cell fusions. Received 2 May 2006; received after revision 7 June 2006; accepted 12 June 2006     

Proliferation status defines functional properties of endothelial cells

Homeostasis of solid tissue is characterized by a low proliferative activity of differentiated cells while special conditions like tissue damage induce regeneration and proliferation. For some cell types it has been shown that various tissue-specific functions are missing in the proliferating state, raising the possibility that their proliferation is not compatible with a fully differentiated state. While endothelial cells are important players in regenerating tissue as well as in the vascularization of tumors, the impact of proliferation on their features remains elusive. To examine cell features in dependence of proliferation, we established human endothelial cell lines in which proliferation is tightly controlled by a doxycycline-dependent, synthetic regulatory unit. We observed that uptake of macromolecules and establishment of cell–cell contacts was more pronounced in the growth-arrested state. Tube-like structures were formed in vitro in both proliferating and non-proliferating conditions. However, functional vessel formation upon transplantation into immune-compromised mice was restricted to the proliferative state. Kaposi’s sarcoma-associated herpes virus (KSHV) infection resulted in reduced expression of endothelial markers. Upon transplantation of infected cells, drastic differences were observed: proliferation arrested cells acquired a high migratory activity while the proliferating counterparts established a tumor-like phenotype, similar to Kaposi Sarcoma lesions. The study gives evidence that proliferation governs endothelial functions. This suggests that several endothelial functions are differentially expressed during angiogenesis. Moreover, since proliferation defines the functional properties of cells upon infection with KSHV, this process crucially affects the fate of virus-infected cells.     

Allergic reactions,cyclic AMP and histamine release

Summary Both isobutylmethylxantine and theophylline increased the level of cyclic AMP in the mast cell. Theophylline reduced the allergic histamine release, whereas isobutylmethylxynthine caused a pronounced potentiation of the histamine release. This indicates that the hypothesis of an inverse relationship between the level of cyclic AMP in mast cells and histamine release is too simple.This work was supported by grants from the Danish Medical Research Council (Grant No. 512-5270) and from Lundbeckfonden.     

Brain microvessel hexokinase: kinetic properties.

Kinetic differences between brain capillary and parenchymal hexokinase in the presence of glucose, ATP. fructose, potassium, sodium and different pH were established. Parenchymal hexokinase is more susceptible to glucose inhibition, can tolerate greater variations in the ATP concentration, is inhibited by increasing concentrations of fructose and potassium, and showed greater activity on the lower pH values. The data suggest that in brain parenchyma and endothelial cells of brain microvessels, there are 2 different enzymes with regard to the kinetics properties.     

Brain microvessel hexokinase: Kinetic properties

Summary Kinetic differences between brain capillary and parenchymal hexokinase in the presence of glucose, ATP, fructose, potassium, sodium and different pH were established. Parenchymal hexokinase is more susceptible to glucose inhibition, can tolerate greater variations in the ATP concentration, is inhibited by increasing concentrations of fructose and potassium, and showed greater activity on the lower pH values. The data suggest that in brain parenchyma and endothelial cells of brain microvessels, there are 2 different enzymes with regard to the kinetic properties.