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Biomedical subjects

L Drouet

Publications and source records attributed to L Drouet.

At least 37 records · Page 2Linked to original sources

Human mesothelial cells express tissue factor when switched to proliferating state. Pharmacological modulation in vitro.

Covering the inner surface of small-diameter arterial prostheses with endothelial cells (ECs) has been proposed as a means of improving biocompatibility and thrombosis resistance. Because the availability of autologous ECs is limited, autologous human mesothelial cells (HMCs) have been suggested as a substitute for ECs. However, HMCs express tissue factor (TF) in vitro, a deleterious characteristic in vivo. We investigated the distribution of TF antigen and of its inhibitor, tissue factor pathway inhibitor, on HMCs and the effect of pharmacological agents on TF expression. TF antigen was measured by enzyme-linked immunosorbent assay and localized by confocal microscopy. Three distinct pools of TF antigen were demonstrated: within the cells, at the cell surface, and in the extracellular matrix. The effects of ilomedin (10 microg/ml) and heparin (500 U/ml), known to affect procoagulant activity, were evaluated by incubating HMCs for 24 h with or without these agents. Ilomedin, but not heparin, decreased TF antigen expression by 30% (P < 0.05). Despite the theoretical potential of HMCs as a vascular prosthesis lining, TF expression by HMCs remains a major drawback. A technique capable of blocking TF expression until the HMCs return to their resting state is needed. Genetic manipulation of HMCs may hold promise for such a technique.

Cell Culture Techniques↗

Arterial stiffness and cardiovascular risk factors in a population-based study.

OBJECTIVE: To determine the relationships between pulse wave velocity (PWV), an estimate of arterial distensibility and cardiovascular risk factors. DESIGN: This cross-sectional population-based study was carried out from 1995 to 1997 to investigate these relationships. POPULATION AND METHODS: Some 993 subjects, aged 35-64 years (52.7% men), living in the south-west of France, were randomly selected from electoral rolls and participated in a cross-sectional study. Medical examinations were performed by specially trained medical staff. Carotid-femoral PWV was measured using a semiautomatic device (Complior, Garges les Gonesse, France). The relationships between PWV and risk factors were assessed, first in subjects not treated with hypolipidaemic, antidiabetic and antihypertensive drugs and then in treated subjects. In subjects not treated for cardiovascular risk factors, age, gender, systolic blood pressure (SBP) and heart rate (P< 0.001) were the variables significantly associated with PWV. In treated patients, age (P < 0.01), SBP (P < 0.001), heart rate (P < 0.001), apolipoprotein B (P< 0.05) and the number of treated cardiovascular risk factors (P< 0.05) were positively correlated with PWV. CONCLUSION: This study shows that, in a sample of subjects at high risk, the cumulative influence of risk factors, even treated, is an independent determinant of arterial stiffness. These results suggest that PWV may be used as a relevant tool to assess the influence of cardiovascular risk factors on aortic stiffness in high-risk patients.

Adult↗

[Venous thromboembolic pathology. New acquired risk factors or new data on acquired risk factors].

Many acquired risk factors may be identified to avoid the scholarly nature of these interminable lists, they may be reclassified with respect to their originality or their mechanism of action and those of current interest, whose data is still often hypothetical but recent, can be underlined. The following order may be proposed: risk factors which cannot be changed: age (which remains the principal factor) and gender (women being at higher risk than men); true acquired risk factors such as cancer, dysimmune conditions (more specifically, the antiphospholipid syndrome) and hormone replacement therapy (oestroprogestative contraception which has been updated by the debate about "third generation pills" and the risk related to progesterone-like substances themselves; hormone replacement therapy of the menopause which still has no clinical trials to assess "our" forms with natural hormones administered transdermally or transmucosally). Smoking has also been accused of being a risk factor for venous thrombosis in the latest clinical trials. Metabolic factors increase the risk of thrombosis: this is established for obesity, still suspected for hyperhomocystonaemia, the abnormalities being the result of complex gene-environment interactions. Other dysmetabolic conditions (diabetes, hypercholesterolaemia, hypertriglyceridaemia), responsible for arterial complications, are not clearly related to increased venous thromboembolic risk although a preventive effect of statins (yet another I) has just been reported. Similarly to these metabolic factors, the origin of which, genetic or environmental, is difficult to establish, interest has recently been shown in quantitative and functional changes in blood clotting factors. This has been established for arterial disease for fibrinogen but, in addition to this factor which slightly increases the risk of venous thrombosis, increases of factor VIII independent of inflammatory conditions, of blood group and Von Willebrand factor, which all influence the level of factor VIII, an increase by 150% of the normal increases the risk of venous thromboembolic disease by 3 or 4 times. As for factor VIII, increases in factor IX, factor XI, and resistance to activated C protein (independently of the Leiden mutation on the gene for factor V), are also associated in increased venous thromboembolic risk. Without knowing into which category to classify them, previous personal and family history of thromboembolic disease, in the absence of the already mentioned hereditary risk factors, must be noted. Finally, amongst the acquired risk factors, the authors also list conditions of blood stasis and vascular lesions with or without hypercoagulability (surgery, prolonged hospital stays, cardiac failure, paralysis, pregnancy...). Of these acquired conditions which increase the risk of thrombotic complications, particular attention has been given over the last few years to forced immobilisation in uncomfortable positions as in certain forms of transport. Although clinical reports have discordant results, it would seem that the risk is increased and the benefits of supportive elastic stockings have been confirmed. If the acquired risk is identified and quantified for a patient, it allows evaluation of global risk and the installation of appropriate therapeutic measures.

Animals↗

High plasma serotonin levels in primary pulmonary hypertension. Effect of long-term epoprostenol (prostacyclin) therapy.

Elevated plasma serotonin is associated with primary pulmonary hypertension (PPH). To test whether this elevation could be related to platelet activation, the 2 pools of blood serotonin (platelets and plasma) and plasma 5-hydroxyindoleacetic acid (5-HIAA) as well as markers of platelet activation (alpha(IIb)beta(3), CD36, P-selectin, and CD63 membrane epitopes) were measured in 16 patients with severe PPH (group 1) before and at days 10 and 40 of treatment with a continuous infusion of epoprostenol (prostacyclin). The same biological parameters were also measured in 19 healthy subjects (group 2) and in 10 patients after cardiovascular surgery with extracorporeal circulation (group 3), a condition known to profoundly activate the platelets. Twelve PPH patients showed hemodynamic and clinical improvement, 3 remained stable, and 1 had the treatment stopped because of clinical aggravation. At day 0, mean plasma serotonin (5-hydroxytryptamine [5-HT]) concentration was much higher in PPH patients than in normal subjects (34.4+/-21.2 versus 9.1+/-6.0 nmol/L, respectively; P:<0.001) and positively correlated with total pulmonary resistance. The mean platelet 5-HT content was not significantly different in PPH compared with normal individuals. Mean plasma 5-HIAA concentrations were much higher in PPH than in normal patients (162+/-57 versus 61+/-7 nmol/L, respectively; P<0.001). These parameters did not significantly change during epoprostenol treatment. There was no correlation between the changes in plasma 5-HT during treatment and clinical or hemodynamic improvement. In PPH patients, the mean platelet volume significantly decreased (ANOVA, P<0.01) during treatment. Positive correlations were evidenced between the size of platelets and the number of alpha(IIb)beta(3) and CD36 epitopes. When compared with control platelets, the number of alpha(IIb)beta(3) epitopes detected on PPH platelets at day 0 tended to be higher, but this difference did not reach a statistical significance (41 300+/-7140 for PPH patients versus 36 010+/-3930 for control subjects, P=0.069). The number of CD36 epitopes, in the range of controls at day 0 (11 590+/-5080 for PPH patients versus 11 900+/-1790 for control subjects), decreased during treatment (ANOVA, P=0.038) and became significantly low at day 40 (8660+/-3520, P<0.001). The number of CD63 epitopes was not elevated, and P-selectin was never detected at any time point on PPH platelets. This glycoprotein profile indicates that the platelets of PPH patients were not highly activated but had an accelerated turnover and returned to normal under epoprostenol treatment without change of the elevated plasma serotonin, characteristic of PPH. In conclusion, neither platelet activation nor a significant alteration of the 5-HT endothelial metabolism explains the high level of plasma 5-HT in PPH patients. The 5-HT plasma concentration is not a predictive marker of the severity of PPH, and its evolution is independent of the clinical and hemodynamic status. Treatment by a potent antiaggregating agent, epoprostenol, does not affect the increase of plasma 5-HT, despite a therapeutic benefit.

Adult↗

Feasibility and reliability of ankle/arm blood pressure index in preventive medicine.

Despite its potential usefulness for assessing preclinical atherosclerosis and cardiovascular risk, the ankle/arm blood pressure index (AAI) has not yet been the matter of study evaluating its feasibility and reliability by nonspecialist doctors in a general population. This study was planned for two steps. In step 1, the measurement of AAI, (ratio between Doppler systolic pressure at the ankle for each lower limb and the highest value of Doppler systolic pressure of the two upper limbs), should be performed by 50 general practitioners (GPs), 50 social security center physicians, and 50 occupational health physicians in 3,000 male smokers, 40 to 59 years, without clinical cardiovascular disease. In step 2, AAI measurement, coupled with echography-Doppler of iliofemoral arteries, should be repeated by a specialist in all subjects with decreased AAI (<0.90) and the first two subjects with normal AAI recruited in step 1 by each nonspecialist. The number of physicians and subjects participating in step 1 was lower than planned (80 physicians and 962 subjects) with the greatest defect for GPs (six physicians and 35 subjects) and the prevalence of decreased AAI was low (28 subjects). AAI measurement was repeated in step 2 in only 12 subjects with decreased AAI in step 1 and in 124 subjects with normal AAI in step 1. Five of the six subjects with decreased AAI in step 2 also had decreased AAI in step 1 and 123 of the 130 subjects with normal AAI in step 2 also had normal AAI in step 1. As regards echographic stenosis, decreased AAI had a sensitivity of 44% and a specificity of 98%. AAI seems more feasible for occupational health physicians and social security center physicians and AAI is also reliable for nonspecialists previously trained, but its predictive value as regards echographic stenosis is poor in asymptomatic subjects, which may limit its usefulness for detecting preclinical atherosclerosis.

Adult↗

[Thrombosis and its therapeutic targets].

Pathophysiological factors of thrombosis (flow, exposed surface and blood reactivity) explain when, how and why a thrombosis does develop. The same factors associated to the fibrinolytic capacity of the vessel wall explain the spontaneous evolution of the thrombosis. Among these factors, blood flow determines the relative participation of platelets and that of coagulation cascade. In arterial thrombosis, atherosclerotic plaque rupture is the triggering factor and platelets are the main actors in the thrombotic reaction: in consequence platelets are the main target and antiplatelet agents the main drugs to use for preventing atherothrombotic ischaemic events. In venous thrombosis, flow (stasis) is the main factor (with an additional role of blood coagulability and of venous wall lesion), and blood coagulation is the main contributor: anticoagulants are the logical option to prevent venous thromboembolic events. Thrombolytic agents can be used in arterial thrombosis only when a fibrin network completes and stabilizes the occluding platelet thrombosis (in myocardial infarction and not in unstable angina) and this has to be applied very early so that the physiological dethrombosis can still be possible.

Blood Coagulation Disorders↗

Risk factors and outcomes for atherothrombotic disease in French patients: the RIVAGE study. RIsque VAsculaire Group d'Etude.

This prospective observational study was designed to delineate the course of atherosclerotic disease in a representative group of French patients receiving standard medical care and to look for clinical and laboratory factors predictive of recurrent cardiovascular events. The 2416 study patients (75.2% men and 24.8% women) had diagnoses of peripheral arterial disease (stage II or III), ischemic heart disease (stable angina or myocardial infarction), or cerebrovascular disease (transient ischemic attack or stroke); 2004 patients (82.9%) had only one of these diagnoses, and 412 (17.1%) had more than one. Among patients with a given stage of peripheral arterial disease, mean age was older in the women than in the men. Coronary disease and cerebrovascular disease were more severe in the men. During the 18-month follow-up, 408 cardiovascular events were recorded in 380 patients (15.7% of the overall study group). In patients who had a single clinical event at inclusion, subsequent clinical events usually occurred in the same vascular bed. The incidences of coronary and cerebral events were correlated with age and the incidence of peripheral events with smoking status. Fatal events were correlated with age but not with the baseline diagnosis, except for a weak relationship with peripheral arterial disease. In a subset of 411 patients who had laboratory tests, plasma fibrinogen level was the only independent predictor of recurrence for all cardiovascular events; this parameter was more closely correlated with fatal events than with all events.

Aged↗

Use of von Willebrand diseased kidney as donor in a pig-to-primate model of xenotransplantation.

BACKGROUND: The coagulation process in hyperacute and delayed xenograft rejection is essential and depends upon platelet adhesion and aggregation. The initial binding of platelets to the damaged endothelium is due to the interaction of the platelet receptor glycoprotein Ib with von Willebrand factor (vWF), which is present on activated endothelial cells and bound to the subendothelial matrix. We hypothesized that the use of organs from animals with homozygous von Willebrand disease (vWD), severely deficient in vWF, might prevent the thrombosis encountered in delayed xenograft rejection. METHODS: Ten baboons were treated by extracorporeal immunoadsorption of xenoreactive natural antibodies (XNA) through the donor pig liver to inhibit hyperacute rejection and received heterotopic vWD or control pig kidney xenografts. XNA levels, coagulation, and platelet activation markers were studied, and specimens of rejected kidneys were analyzed histologically. RESULTS: Although XNA depletion was comparable in both groups, neither kidney function nor survival times of control (n=5) or vWD (n=5) porcine kidneys showed any difference. Platelet and coagulation activation was evidenced in both groups after surgery and at rejection time. Immunohistochemical analysis revealed a weak endothelial vWF immunostaining in the rejected vWD kidneys, whereas it was undetectable in the nongrafted vWD kidneys, suggesting the deposition of baboon plasma vWF on the porcine vessels. CONCLUSIONS: The use of vWD organs did not improve the survival time of grafted kidneys in this xenotransplantation model. Further studies on the use of vWD organs, in association with other therapeutic approaches, such as complement inhibition, are nevertheless necessary to evaluate the usefulness of vWF deficiency as an adjunctive therapy to decrease the coagulation process during xenograft rejection.

Animals↗

Studies of platelet volume, chemistry and function in patients with essential thrombocythaemia treated with Anagrelide.

Anagrelide (imidazoquinazolin derivative) is a new compound proposed for the treatment of myeloproliferative disorders. In this study, Anagrelide was given to patients with essential thrombocythaemia (ET) in a compassionate-use protocol. The aim of this study was to test the effect of this drug not only on the platelet count but also on platelet volume, chemistry and function, which has not previously been reported. Thus, in ET, different functional or structural platelet abnormalities were reported: a shortening of the bleeding time, hypoaggregation to several agonists, and in particular a lack of response to adrenalin, an increase in the amount of total platelet glycoprotein IV (or CD36), and an abnormal migration of thrombospondin on electrophoresis. These different parameters were studied before and during therapy with Anagrelide. Although the platelet count was corrected, no functional or chemical abnormality was improved. Furthermore, platelet volume was shown to be constantly increased under Anagrelide. Thus, Anagrelide, in reducing the platelet count, may possibly decrease the risk of thrombosis and haemorrhage. Nevertheless, if the risk of thromboses and/or myelofibrosis is related not only to the platelet count but also to the platelet abnormalities, the persistence of a thrombocytopathy in patients treated with Anagrelide must be taken in consideration. Our data suggest that thromboses and myelofibrosis are clinical end-points which should be included in future large-scale use of Anagrelide.

Bleeding Time↗

In vivo regulation of von willebrand factor synthesis: von Willebrand factor production in endothelial cells after lung transplantation between normal pigs and von Willebrand factor-deficient pigs.

To evaluate the regulation of plasma von Willebrand factor (vWF) and its in situ production by endothelial cells (ECs), 12 swine leukocyte antigen (SLA)-compatible left lung transplantations were performed. Normal lungs were transplanted into 10 pigs homozygous for von Willebrand disease and into 2 normal pigs. Additionally, 1 normal pig underwent pneumonectomy, and 1 SLA-incompatible lung transplantation between normal pigs was performed. None of the transplanted animals received immunosuppressive therapy. Plasma vWF level was evaluated by ELISA and multimeric pattern. EC vWF content was assessed by immunohistochemistry. Global hemostasis was assessed by standardized ear bleeding time. Six of 12 SLA-compatible lung transplantations and the incompatible transplantation were successful and were used for the study. The functions and the viability of ECs, reflected by their ability to produce vWF and normal multimeric plasma vWF pattern, were preserved in SLA-compatible and -incompatible lung transplantations. vWF production was preserved in ECs that initially synthesized it. EC constitutive and storage pathways are modulated differently according to transplantation compatibility and severity of rejection. In SLA-compatible lung transplantations without histological evidence of rejection, the production of vWF was preserved, whereas constitutive vWF secretion appeared to be altered in cases with minor histological signs of rejection. In pigs with von Willebrand disease that were transplanted with normal lungs without sign of rejection, plasma vWF was significantly increased in an amount expected from the estimated production of a normal lung. In the transplanted normal lung, there was no vWF overexpression by the ECs and no recruitment of ECs that initially did not express vWF. In SLA-incompatible transplantation, ECs were morphologically normal with increased and blurred vWF labeling, whereas plasma vWF levels remained normal, reflecting that EC activation is associated with an increased vWF production with probable diversion to storage pathway. This model depicts the changes of EC regulation of vWF secretion in pig lung transplants. However, this model cannot be directly extrapolated to human organ transplantation because animals did not receive any immunosuppressive therapy, which may be toxic to ECs.

Acute Disease↗

Macro and microheterogeneity in normal endothelial cells: differential composition of luminal glycocalyx and functional implications.

Endothelial cells (EC) are involved in various physiological and pathological processes through the expression of their surface glycoproteins. They are covered by the glycocalyx, composed of glucidic residues from cell surface membrane glycoproteins, glycoplipids and proteoglycans. Glucidic sequences can be specifically characterized by their binding to lectins. Eight lectins were used to investigate the distribution and regulation of EC surface glucidic residues in various blood vessels of adult and newborn pigs. EC lectin binding was compared to von Willebrand factor (vWF) expression as EC reference marker. Six out of eight lectins (BSI-B4, DBA, EEA, HP, MAL I and PNA) were helpful for this determination. Considering only the intensity of labelings, vWF and DBA gave the best stainings of adult pig ECs. In newborn pigs, the best labelings were obtained with EEA and MAL I. Furthermore, the distribution of lectin binding to ECs and EC vWF expression was heterogeneous depending on the EC location along vascular tree and age. Beside this macroheterogeneity this study highlights a microheterogeneity of EC lectin binding and vWF expression in situ, defined as a staining of equal intensity by individual ECs, scattered among negative ones, in a given vascular segment. EC surface sugar residues were differently modulated in newborn and adult pig ECs and differently according to EC vWF expression. The functional involvement of EC glycocalyx was reflected by EC lectin binding in the spleen and liver. This study emphasizes the high level of EC heterogeneity for various markers. The EC macro- and microheterogeneity reflect the "plasticity" or "unstability" of EC phenotypes and suggests that ECs are subject to several levels of regulation and are probably grouped in functional clusters to best adjust their functions to microenvironmental requirements. This concept must be considered in further investigations notably in in vitro studies where EC phenotype can be altered.

Animals↗

[Comparative study of the antithrombotic effect of aspirin and Bay U3405, antagonist of a thromboxane A2 receptor].

The antithrombotic activity of aspirin (acetylsalicylic acid), inhibitor of the synthesis of thromboxane A2 was compared to that of Bay U3405, a thromboxane A2 receptor antagonist. A model of experimental thrombosis induced by laser in guinea pig mesenteric arteries was used. Aspirin exhibited a dose-dependent antithrombotic activity with and inhibitory effect of 70% at the high dose of 100 mg/kg while Bay U 3405 was efficient at the dose of 10 mg/kg with a percent of inhibition that did not exceed 55%. These results showed that: first, the inhibition of the activity of thromboxane A2 by an antagonist was not more important than that by aspirin and second, the thromboxane A2 way was well implicated in the thrombosis in the guinea pig, suggesting that this species was a good model for the study of the mechanisms using the prostaglandins way in arterial thrombosis.

Animals↗

[Inhibition of thrombus formation by anti-Willebrand monoclonal antibodies in the guinea pig].

To experimentally evaluate the role of von Willebrand factor in thrombosis, a monoclonal antibody-712-, directed against the binding domain of von Willebrand factor to the platelet glycoprotein Ib and a control antibody D4H1, directed against the light chain of Factor VII C, were injected into guinea pigs. A model of experimental thrombosis induced by laser in guinea pig mesenteric arteries was used. The antibody 712 induced a dose-dependent reduction of the thrombus formation without affecting the bleeding time and the platelet number. Ex vivo, the ristocetin-induced platelet aggregation was inhibited by the antibody 712 in a dose-dependent manner while the adenosine diphosphate-induced platelet aggregation was not modified. In conclusion, this study showed that antibody-712- has a significant antithrombotic activity without inducing a hemorrhagic state, suggesting that the von Willebrand factor--glycoprotein Ib axis was a promising target for the strategy of a new antithrombotic therapy.

Animals↗

A new quantitative D-dimer assay appropriate in emergency: reliability of the assay for pulmonary embolism exclusion diagnosis.

Plasma D-dimers, degradation products of cross-linked fibrin, are elevated in several thrombotic diseases. In the last decade, their measurement has been performed with specific monoclonal antibody based ELISA assay, with a high negative predictive value of such pathologies. However these methods have a low clinical impact since they cannot be used in emergency because they are time consuming and require series. Recently, rapid tests have been proposed with similar accuracy and are feasible for use in emergency conditions. The aim of our study was to evaluate the potential value of a new quantitative rapid assay, based on agglutination of latex microparticles coated with two monoclonal antibodies specific for D-dimers (Liatest D-Di, Diagnostica Stago), in the exclusion diagnosis of pulmonary embolism (PE). Eighty-five consecutive suspected PE patients were included in the study. D-dimer determination was performed with both standard ELISA and Liatest D-Di just before pulmonary scintigraphy. Sixty-nine patients were free of PE and 16 had PE. Our results confirm the excellent sensitivity and negative predictive values of the conventional D-dimer ELISA. Using a cut-off value of 500 ng/ml, the sensitivity and the negative predictive values of Liatest D-Di were 94% and 96%, respectively. Only one case of PE had a D-dimer value at 480 ng/ml, close to the cut-off value. This study demonstrates that the Liatest D-Di assay is sensitive enough to be used as the first step in the assessment of PE. However, the best cut-off value has to be determined to get an exclusion diagnosis with certainty.

Adult↗