[Campylobacter jejuni meningitis]. / Méningite à Campylobacter jejuni.

Cases of Campylobacter jejuni meningitis are extremely rare. In the literature, less than ten cases have been described so far, although Campylobacter spp is one of the most common pathogens causing gastroenteritis in the world. Some common stigmata can be found across these cases such as rupture of the blood-brain barrier, immunosuppression, as well as the tropism of Camplylobacter jejuni for neurological parenchyma. Campylobacter jejuni bacteremia is certainly underestimated because Campylobacter is a thermophilic bacterium and special conditions are required to isolate this organism in blood cultures. PCR is thus an interesting alternative technique for diagnosis. In our case, a patient with a history of resected astrocytoma, had undergone treatment with chemotherapy and radiotherapy because of anaplastic transformation. The patient was admitted with gastroenteritis and Campylobacter jejuni meningitis. The diagnosis was obtained initially on stool cultures and then by PCR of cerebrospinal fluid. The evolution was favorable with meropenem.

Les cas de méningite à Campylobacter jejuni restent extrêmement rares. Dans la littérature, on décrit moins de 10 cas à ce jour, alors que l'infection à Campylobacter est cependant l'une des causes les plus répandues de gastro-entérite dans le monde. Le point commun de tous ces cas de méningite rapportés semble être la fragilité de la barrière hémato-encéphalique et l'immunodépression, ainsi que le tropisme du Campylobacter jejuni pour les tissus neuronaux. La bactériémie à Campylobacter jejuni est par ailleurs sous-estimée car le germe est thermophilique et des conditions particulières sont nécessaires pour isoler cet organisme dans les hémocultures. La PCR est une alternative intéressante pour le diagnostic microbiologique. Dans le cas décrit, le patient présentait des antécédents d'astrocytome pariéto-temporal droit opéré, avec une transformation anaplasique ayant bénéficié de chimio- et radiothérapie concomitantes. Le patient a été admis avec une gastro-entérite compliquée d'une méningite à Campylobacter jejuni. Le diagnostic a été posé initialement sur la coproculture et ensuite par la PCR du liquide céphalo-rachidien. L'évolution a été favorable sous méropénem.

Susceptibility to measles, mumps, and rubella in 5-year-old children in Flanders, Belgium.

UNLABELLED: The second dose of an MMR vaccine is a catch up for persons who did not receive the first dose or for primary vaccine failures. Catch up doses can be scheduled according to convenience into the program of the country. The second MMR dose is often administered at the age of 5 years, before school entry. Some countries chose to implement the second dose at the age of 10-13 years, as is the case for Belgium. The here presented long-term follow-up of a cohort of children, set up originally to analyze maternal antibodies against vaccine preventable diseases, offers a unique opportunity to evaluate ad interim the current long-interval MMR vaccination schedule in Belgium. After 1 MMR dose at 12 months of age, rubella immunity is almost intact at 5 years of age (94.5 % is seropositive), measles seropositivity scores 86.8 %, and mumps 32 %, measured with ELISA. A seroneutralization (SN) test for mumps antibodies reveals much higher seropositivity rates (88 %). Using a regression model on the log (IgG) titer for all antigens, no influence was found from any of the studied variables, except for girls who had a significantly higher rubella IgG titer (p=0.002) compared to boys. CONCLUSION: The data show considerable susceptibility to mumps and measles in 5-year-old children, confirming a previously conducted seroprevalence study (2006). Both advantages and disadvantages of shortening or enlarging the vaccine schedule are discussed.

[Advanced glycation end products: A risk factor for human health]. / Les produits de glycation avancée : un risque pour la santé humaine.

Advanced glycation end products (AGE) result from a chemical reaction between the carbonyl group of reducing sugar and the nucleophilic NH2 of a free amino acid or a protein; lysine and arginine being the main reactive amino acids on proteins. Following this first step, a molecular rearrangement occurs, rearrangement of Amadori resulting to the formation of Maillard products. Glycation can cause the clouding of the lens by inducing reactions crosslinking proteins. Specialized receptors (RAGE, Galectin 3…) bind AGE. The binding to the receptor causes the formation of free radicals, which have a deleterious effect because they are powerful oxidizing agents, but also play the role of intracellular messenger, altering the cell functions. This is especially true at the level of endothelial cells: the attachment of AGE to RAGE receptor causes an increase in vascular permeability. AGE binding to endothelium RAGE and to monocytes-macrophages, led to the production of cytokines, growth factors, to the expression of adhesion molecules, and the production of procoagulant activity. Diabetic retinopathy is related to excessive secretion of vascular growth factor (vascular endothelial growth factor [VEGF]). AGE-RAGE receptor binding causes the synthesis and secretion of VEGF. Increased permeability, facilitation of leukocyte migration, the production of reactive oxygen species, cytokines and VEGF suggest that the AGE could be an element of a cascade of reactions responsible for the diabetic angiopathy and vascular damages observed during aging and chronic renal failure. Balanced diet or some drugs can limit the deleterious effect of AGE.

[Molecular basis of red blood cell adhesion to endothelium]. / Bases moléculaires de l'adhérence des globules rouges à l'endothélium.

The extent of red blood cell adhesion is correlated with the incidence of vascular complications and the severity of the disease. Patients with sickle cell anemia (HbSS) experience vasoocclusive episodes. The adhesion of RBCs from HbSS patients is increased and related to VLA-4 exposure, which binds to vascular cell adhesion molecule (VCAM-1). Inter Cellular Adhesion Molecule (ICAM-1), CD31, CD36 and glycans are potential receptors for PfEMP1 of RBCs parasited by plasmodium falciparum. The incidence of vascular complications is very high in patients with diabetes mellitus. RBC adhesion is increased and statistically correlated with the severity of the angiopathy. Glycation of RBC membrane proteins is responsible for binding to the receptor for advanced glycation end products (RAGE). Polycythemia Vera (PV) is the most frequent myeloproliferative disorder and characterized by a high occurrence of thrombosis of mesenteric and cerebral vessels. PV is due to a mutation of the Janus kinase 2 (JAK2 V617F). This mutation stimulates erythropoiesis and is the cause of Lu/BCAM (CD239) phosphorylation, which potentiated the interaction with laminin alpha 5. The couple laminin alpha 5 endothelial and phosphorylated Lu/BCAM explained the increased adhesion of RBCs from patients PV to endothelium.

Advanced glycation end products regulate extracellular matrix protein and protease expression by human glomerular mesangial cells.

Advanced glycation end products (AGEs) may play a role in the pathogenesis of diabetic nephropathy, by modulating extracellular matrix turnover. AGEs are known to activate specific membrane receptors, including the receptor for AGE (RAGE). In the present study, we analyzed the various receptors for AGEs expressed by human mesangial cells and we studied the effects of glycated albumin and of carboxymethyl lysine on matrix protein and remodelling enzyme synthesis. Membrane RAGE expression was confirmed by FACS analysis. Microarray methods, RT-PCR, and Northern blot analysis were used to detect and confirm specific gene induction. Zymographic analysis and ELISA were used to measure the induction of tPA and PAI-1. We show herein that cultured human mesangial cells express AGE receptor type 1, type 2 and type 3 and RAGE. AGEs (200 microg/ml) induced at least a 2-fold increase in mRNA for 10 genes involved in ECM remodelling, including tPA, PAI-1 and TIMP-3. The increase in tPA synthesis was confirmed by fibrin zymography. The stimulation of PAI-1 synthesis was confirmed by ELISA. AGEs increased PAI-1 mRNA through a signalling pathway involving reactive oxygen species, the MAP kinases ERK-1/ERK-2 and the nuclear transcription factor NF-kappaB, but not AP-1. Carboxymethyl lysine (CML, 5 microM), which is a RAGE ligand, also stimulated PAI-1 synthesis by mesangial cells. In addition, a blocking anti-RAGE antibody partially inhibited the AGE-stimulated gene expression and decreased the PAI-1 accumulation induced by AGEs and by CML. Inhibition of AGE receptors or neutralization of the protease inhibitors TIMP-3 and PAI-1 could represent an important new therapeutic strategy for diabetic nephropathy.

Enteropathogens in paediatric gastroenteritis: comparison of routine diagnostic and molecular methods.

OBJECTIVES: Studies of acute gastroenteritis (AGE) are hampered by the lack of routine diagnostic methods with good sensitivity and specificity. Molecular methods are increasingly used for clinical purposes, but the clinical significance of a positive result remains a challenge. In this study we aimed to compare results of routine diagnostic methods and molecular methods in symptomatic children and asymptomatic controls. METHODS: Patients presenting to the pediatric emergency departments of two university hospitals in Brussels with AGE were recruited prospectively from May 2015 to October 2016; asymptomatic controls were recruited from the same hospitals. Stool analyses were performed for all participants for common pathogenic bacteria (culture), virus (immunochromatography) and parasites (microscopy). Stools were also analysed with the Luminex Gastrointestinal Pathogen Panel, a multiplex-PCR for common enteropathogens. RESULTS: Stools from 178 patients and 165 controls were analysed. An enteropathogen was detected in 62.4% (111/178) of cases when combining the two methods (56.2% (100/178) by Luminex, 42.7% (76/178) with routine methods) and 29.1% (48/165) of controls (24.2% (40/165) by Luminex and 10.3% (17/165) by routine methods). Some pathogens were detected more often with Luminex than with routine methods, such as Salmonella (16.3% (29/178) with Luminex and 3.9% (7/178) with routine method, p < 0.05), whereas others identified by culture methods, such as Campylobacter, Shigella, Yersinia, were missed by Luminex. CONCLUSIONS: Molecular tools seem attractive methods, providing high positivity and a rapid turn-around time for the diagnosis of AGE. However, high rates of positivity in both cases and controls highlight the difficulty in interpreting results. Pathogens missed by Luminex but detected by culture methods raise more questions about the true clinical interest of the technique for our patients.

Prospective evaluation of diagnostic tools for respiratory viruses in children and adults.

AIM: To compare the performances of molecular and non-molecular tests to diagnose respiratory viral infections and to evaluate the pros and contras of each technique. METHODS: Two hundred ninety-nine respiratory samples were prospectively explored using multiplex molecular techniques (FilmArray Respiratory Panel, Clart Pneumovir), immunological techniques (direct fluorescent assay, lateral flow chromatography) and cell cultures. FINDINGS: Molecular techniques permitted the recovery of up to 50% more respiratory pathogens in comparison to non-molecular methods. FilmArray detected at least 30% more pathogens than Clart Pneumovir which could be explained by the differences in their technical designs. The turnaround time under 2 hours for the FilmArray permitted delivery of results when patients were still in the emergency room.

Contribution of the FilmArray Respiratory Panel in the management of adult and pediatric patients attending the emergency room during 2015-2016 influenza epidemics: An interventional study.

AIM: To evaluate the contribution of a multiplex PCR for respiratory viruses on antibiotic and antiviral prescription, ancillary test prescription, admission and length of stay of patients. METHODS: Two hundred ninety-one adult and pediatric patients visiting the emergency department during the 2015-2016 influenza epidemic were prospectively included and immediately tested 24/7 using the FilmArray Respiratory Panel. The results were communicated to the practitioner in charge as soon as they became available. Clinical and biological data were gathered and analyzed. FINDINGS: Results from the FilmArray Respiratory Panel do not appear to impact admission or antibiotic prescription, with the exception of a lower admission rate for children who tested positive for influenza B. Parameters that account for the clinical decisions evaluated are CRP level, white blood cell count, suspected or proven bacterial infection and, for adult patients only, signs of respiratory distress. Length of stay is also not significantly different between patients with a positive and a negative result. A rapid influenza test result permits a more appropriate prescription of oseltamivir.

Severity of diabetic microvascular complications is associated with a low soluble RAGE level.

AIMS: The receptor for advanced glycation end-products (RAGE) has been implicated in diabetic microvascular complications, but several lines of evidence suggest that the soluble isoform of RAGE (sRAGE) may protect against AGE-mediated vessel damage. The characterized AGE Nepsilon-carboxymethyllysine (CML) is associated with diabetic microvascular complications. In the present study, we measured blood levels of sRAGE and CML-protein in diabetic patients with and without microvascular complications. METHODS: Thirty patients with type-2 diabetes were recruited into the study, comprising 20 who had no microvascular complications, and 10 who had both retinal and renal complications. sRAGE was measured in serum by ELISA, and CML by competitive ELISA. RESULTS: sRAGE blood levels were similar in both the controls and diabetic patients without microvascular complications. In patients with complications, the mean sRAGE blood level was significantly decreased (1068+/-231pg/mL) compared with diabetic patients without complications (P=0.028). CML-protein was increased in all diabetic patients, but to a higher extent in those who had microvascular complications. CONCLUSION: The association of low sRAGE with high CML-protein levels in diabetic patients who developed severe diabetic complications supports the hypothesis that sRAGE protects vessels against AGE-mediated diabetic microvascular damage.

Role of Lu/BCAM in abnormal adhesion of sickle red blood cells to vascular endothelium.

Lutheran (Lu) blood group and Basal Cell Adhesion Molecule (BCAM) antigens are both carried by two glycoprotein (gp) isoforms of the immunoglobulin superfamily representing receptors for laminin alpha5 chain. They are expressed in red blood cells, in endothelial cells of vascular capillaries and in epithelial cells of several tissues. Lu/BCAM gps are overexpressed in sickle red blood cells (SS RBCs). Stimulation of SS RBCs by epinephrine activates the PKA depending signaling pathway and induces reinforced Lu/BCAM-mediated adhesion to laminin10/11. We have analyzed the phosphorylation state of Lu/BCAM long isoform cytoplasmic tail and showed that it is phosphorylated by CKII, GSK3b and PKA. Phosphorylation of this isoform in transfected K562 cells is stimulated by effectors of the PKA pathway and induces cell adhesion to laminin10/11. Lu/BCAM gps are highly expressed in endothelial cells and exhibit potential integrin binding motifs. We showed that they interact with integrin alpha4beta1, the unique integrin expressed on the surface of young reticulocytes. Adhesion assays under flow conditions showed that SS RBCs adhere to primary human endothelial cells (HUVEC) after selective activation of intergin alpha4beta1 and that this adhesion is mediated by endothelial Lu/BCAM gps. Our studies show that Lu/BCAM gps expressed either on erythroid or on endothelial cells are involved in SS RBC-endothelium interactions and could play a role in the abnormal adhesion of SS RBCs to vascular endothelium contributing to the vaso-occlusive crises reported for sickle cell disease patients.

Red cell and endothelial Lu/BCAM beyond sickle cell disease.

Recent studies shed new lights on the biological function of blood group antigens, such as the adhesion properties of the Lutheran (Lu) blood group antigens carried by the Lu/BCAM glycoproteins. The Lu/BCAM adhesion glycoproteins were first identified as laminin-10/11 erythroid receptors involved in RBC adhesion to endothelium in sickle cell anemia. Lu/BCAM mediated cell adhesion to laminin is stimulated by epinephrine, a physiological stress mediator, and is dependent of phosphorylation by protein kinase A. More recently, we demonstrated that constitutive phosphorylation of Lu/BCAM is also involved in abnormal RBC adhesion to endothelium in patients with polycythemia vera (PV), a frequent myeloproliferative disorders associated with the V617F mutation of the tyrosine kinase JAK2 leading to continuous stimulation of erythropoiesis. This observation suggests that Lu/BCAM could participate to the high incidence of vascular thrombosis that also characterizes PV disease. In mice, which do not express Lu/BCAM in erytroid tissues, invalidation of the Lu/BCAM gene provided evidence that Lu/BCAM gps, as laminin-alpha5 receptors, are involved in vivo in the maintenance of normal basement membrane organization in different non erythroid tissues since up to 90% of the mutant kidney glomeruli exhibited a reduced number of visible capillary lumens and irregular thickening of the glomerular basement membrane, while intestine exhibited smooth muscle coat thickening and disorganization. All these results further illustrate that minor blood group antigens might have important role under physiological and physiopathological conditions in erythroid and non erythroid tissues as well.

Receptor-mediated endothelial cell dysfunction in diabetic vasculopathy. Soluble receptor for advanced glycation end products blocks hyperpermeability in diabetic rats.

Dysfunctional endothelium is associated with and, likely, predates clinical complications of diabetes mellitus, by promoting increased vascular permeability and thrombogenicity. Irreversible advanced glycation end products (AGEs), resulting from nonenzymatic glycation and oxidation of proteins or lipids, are found in plasma, vessel wall, and tissues and have been linked to the development of diabetic complications. The principal means through which AGEs exert their cellular effects is via specific cellular receptors, one of which, receptor for AGE (RAGE), is expressed by endothelium. We report that blockade of RAGE inhibits AGE-induced impairment of endothelial barrier function, and reverse, in large part, the early vascular hyperpermeability observed in diabetic rats. Inhibition of AGE- and diabetes-mediated hyperpermeability by antioxidants, both in vitro and in vivo, suggested the central role of AGE-RAGE-induced oxidant stress in the development of hyperpermeability. Taken together, these data support the concept that ligation of AGEs by endothelial RAGE induces cellular dysfunction, at least in part by an oxidant-sensitive mechanism, contributing to vascular hyperpermeability in diabetes, and that RAGE is central to this pathologic process.

Postprandial hyperglycemia alters inflammatory and hemostatic parameters.

Glucose or glucose derived products are increased in blood during the postprandial phase and are, to a certain extent, related to meal composition. Glucose and glucose derived products such as advanced glycation end products (AGEs) can be formed in the intracellular compartment but can also be absorbed as AGEs or AGE precursors present in food. Glucose, glucose metabolites and AGEs alter endothelial cell functions, induce adhesion molecule overexpression (ICAM-1, VCAM), cytokine release (IL-6, MCP-1) and tissue factor production. Tumor necrosis factor alpha systemic level is increased during the postprandial phase as are augmented C reactive protein and fibrinogen level. Hyperglycemia induced an increase in plasminogen activator inhibitor, and shortened fibrinogen half life. Hyperglycemia and AGEs provoked an oxidant stress. The formation of reactive oxygen intermediates perturbates NO (Nitric oxide) formation and are deleterious for cell functions. All the modifications observed in the postprandial phase are not too deleterious but their iterative characteristics may lead to vascular dysfunction.

Role of membrane sialic acid content in the adhesiveness of aged erythrocytes to human cultured endothelial cells.

Following our previous observation that the oldest normal red blood cells were the most adherent to human cultured endothelial cells, we attempted to simulate this age-related adherence. Among all the membrane modifications experienced by erythrocytes during their life-span, loss of sialic acids has attracted considerable attention. Using two different preparations of neuraminidase, we performed a sialic acid depletion on the youngest erythrocytes to reach a sialic acid content similar to that observed in physiologically aged erythrocytes. These pretreated youngest cells displayed limited increase in the adhesiveness to endothelial cells, lower than that found with intact oldest cells. To obtain an adhesiveness of pretreated cells similar to that of naturally aged cells, it was necessary to exceed 80% of sialic acid depletion. At this extent of desialation, modifications of the electrophoretic pattern of glycophorins were observed as well as the appearance of peanut agglutinin reactivity which were never found in physiologically aged erythrocytes. Therefore, the sialic acid loss cannot be considered as being a single determinant factor of the naturally aged red cell adhesiveness.

IgG binding to endothelial cells in systemic lupus erythematosus.

Endothelial-associated IgG were determined in 20 patients with systemic lupus erythematosis (SLE)-8 of whom had a lupus anticoagulant (LA) and 6 a history of thrombosis. The binding of IgG present in patient plasma to cultured human endothelial cells was detected using radiolabeled staphylococcal protein A. Thirteen samples gave positive results and a significant association between endothelial-associated IgG and lupus anticoagulant was found (p less than 0.05). No statistically significant relationship with a previous history of thrombosis was found. These results suggest that the lupus anticoagulant may be directly involved in immune vascular injury induced by either antibodies or immune complexes in SLE.

Modification of membrane properties of erythrocytes by PGI2.

The addition of prostaglandin I2 (PGI2) enhanced the adhesion of red cells from normals and patients with diabetes mellitus or sickle cell anemia (SCA) to human cultured endothelial cells (p less than 0.025). The maximal effect was reached with 10(-11) M PGI2 after 30 min incubation. Red cell adhesion was also increased by PGD2 but PGE1 and 6-keto-PGF1 alpha had no significant effect. Since enhanced adhesion of red cell to endothelium and increased red cell calcium content have been proposed to be related in SCA, we have investigated the calcium binding to human resealed normal erythrocyte membrane by using (45Ca) calcium in presence of the different PG which alter red cell adhesion or not. Calcium binding was time-dependent and potentiated in presence of PGI2 (p less than 0.01) but not of PGD2. The fact that erythrocyte adhesion is enhanced by both PGI2 and PGD2 while calcium binding is increased only by PGI2 suggests that the two phenomenon can be dissociated.

Plasma fibronectin-C1q complex formation and its effect on C1q inhibition of collagen-induced platelet aggregation.

The first component of complement (C1q) contains a collagen-like structure which evidently accounts for its ability to bind to platelets and to inhibit collagen-induced platelet aggregation. This present investigation explored the interaction among C1q, plasma fibronectin (Pl Fn) and platelets. Gel filtration chromatography (Sepharose 2B and 4B) of preincubated mixtures of C1q and Pl Fn yielded evidence of complex formation. Preincubation of Pl Fn with platelets did not affect collagen-induced platelet aggregation nor did preincubation of Pl Fn with C1q significantly affect C1q-mediated inhibition of collagen-induced platelet aggregation. These observations may indicate that the site of interaction between Pl Fn and C1q is different from the site of interaction between C1q and platelets.

[Pathophysiologic aspects of diabetic angiopathy]. / Aspects physiopathologiques de l'angiopathie diabétique.

The high incidence of vascular complications in patients with diabetes mellitus prompted us to study the pathophysiology of diabetic angiopathy. Hyperglycaemia is a common feature resulting in several metabolic and endocrine alterations and the formation of advanced glycation end-products (AGE). AGE bind to different molecules and to a receptor (RAGE). RAGE interaction with AGE enhances receptor expression and initiates a feedback loop whereby RAGE occupancy triggers increased RAGE expression. In a model of accelerated atherosclerosis associated with diabetes in genetically-manipulated mice, the blockade of cell surface RAGE by infusion in a soluble truncated form completely suppressed enhanced formation of vascular lesions. Improvement of atherosclerosis in these diabetic-atherosclerotic animals through the use of soluble RAGE occurred in the absence of changes in plasma lipids or glycaemia, which emphasises the contribution of a lipid- and glycemia-independent mechanism to atherogenesis.

N(carboxymethyl)lysine as a biomarker for microvascular complications in type 2 diabetic patients.

AIMS: Hyperglycemia is linked to vascular dysfunction in patients with diabetes mellitus, either directly or through advanced glycation end product (AGE) formation. Experimental evidence has indicated the possible involvement of AGEs in the genesis of vascular complications. We investigated whether serum levels of AGEs and of the glycoxidation compound carboxymethyl-lysine (CML) were increased and correlated with vascular complications in type II diabetes mellitus. METHODS: Serum levels of AGEs and CML-human serum protein (CML-HSP) were measured by a specific immunoassay in 51 men and 26 women aged 58 +/- 6.1 years (mean +/- SD) who had been treated for type II diabetes mellitus for 11 +/- 8 years, and in a non-diabetic control group consisting of 39 men and 21 women aged 55.5 +/- 7.5 years. Patients with macroalbuminuria or abnormal creatinine clearance were excluded from the study. RESULTS: The serum levels of AGEs were significantly increased in patients with type II diabetes compared to controls (P<0.001). Blood levels of CML-HSP were significantly increased in diabetic patients compared to normal subjects [35.3 +/- 27.4 and 9.3 +/- 7.2 (mean +/- SD) pmol/mg of protein, respectively; P<0.0001]. In diabetic patients with retinopathy or microalbuminuria (urinary albumin excretion: UAE > 30 mg/24 h), CML-HSP levels were significantly higher (P<0.02), and even more elevated in patients with both complications. CONCLUSION: In patients with type II diabetes, CML-HSP levels that are at variance with the HbA(1c) index for blood glucose may be a biomarker of glycoxidation, and related to the development of microvascular complications.

Endothelial cells in culture: a model to study in vitro vascular toxicity.

This review discusses the importance of cultured endothelial cells in the evaluation of the potential toxicity of a drug and for understanding the toxic effects of some compounds on the vascular system. Vascular toxicity is observed when subjects are exposed to chemicals present in the air or after ingestion of xenobiotics or drugs. Furthermore, some drugs can lead to side-effects owing to an alteration of endothelial cell function. Endothelial cells of human and animal origin can be cultured and several of their properties can be studied using different experimental systems. Cyclosporin and penicillamine have been shown to reduce angiogenesis in vitro, as has also been reported for monocrotaline pyrrole. Other components, such as pyrrolizidine alkaloid, were found to be cytotoxic, as demonstrated by chromium-51 or lactate dehydrogenase release. More subtle changes can be detected in peroxidation, phospholipase activity and prostacyclin production. Endothelial cells cultured to confluency can be used to measure in vitro permeability to radiolabelled inulin or albumin. Tunicamycin, an inhibitor of glycosylation, increases permeability. Xenobiotics such as lead inhibit the production of plasminogen activator (t-PA) or by disrupting the thromboxane-A(2)/prostacyclin balance, which promotes a thrombotic process.