thromboplastin and Cardiovascular-Diseases

Microparticles (MPs) are small membrane vesicles that are released from many different cell types by exocytotic budding of the plasma membrane in response to cellular activation or apoptosis. MPs may be involved in both physiological processes and clinical treatments because they express phospholipids, which function as procoagulants. Elevated levels of platelet-derived MPs, endothelial cell-derived MPs, and monocyte-derived MPs are observed in almost all thrombotic diseases occurring in venous and arterial beds. Several studies have shown that the quantity, cellular origin, and composition of circulating MPs depend on the type of disease, the disease state, and medical treatment. Although MPs were initially thought to be small particles with only procoagulant activity, they are now known to have many different functions. An increasing number of studies have identified new implications of elevated MPs in clinical disorders. On the basis of evidence available till date, the present review suggests that MPs may be a useful biomarker in identifying atherothrombosis.

Topics: Animals; Apoptosis; Biomarkers; Blood Platelets; Cardiovascular Diseases; Cell Membrane; Cell-Derived Microparticles; Coagulants; Diabetes Mellitus, Type 2; Endothelial Cells; Enzyme-Linked Immunosorbent Assay; Flow Cytometry; Humans; Oxidative Stress; Phospholipids; Shear Strength; Stress, Mechanical; Thromboplastin; Thrombosis

The term microparticle (MP) identifies a heterogeneous population of vesicles playing a relevant role in the pathogenesis of vascular diseases, cancer and metabolic diseases such as diabetes mellitus. MPs are released by virtually all cell types by shedding during cell growth, proliferation, activation, apoptosis or senescence processes. MPs, in particular platelet- and endothelial-derived MPs (PMPs and EMPs), are increased in a wide range of thrombotic disorders, with an interesting relationship between their levels and disease pathophysiology, activity or progression. EMP plasma levels have been associated with several cardiovascular diseases and risk factors. PMPs are also shown to be involved in the progressive formation of atherosclerotic plaque and development of arterial thrombosis, especially in diabetic patients. Indeed, diabetes is characterised by an increased procoagulant state and by a hyperreactive platelet phenotype, with enhanced adhesion, aggregation, and activation. Elevated MP levels, such as TF+ MPs, have been shown to be one of the procoagulant determinants in patients with type 2 diabetes mellitus. Atherosclerotic plaque constitutes an opulent source of sequestered MPs, called "plaque" MPs. Otherwise, circulating MPs represent a TF reservoir, named "blood-borne" TF, challenging the dogma that TF is a constitutive protein expressed in minute amounts. "Blood-borne" TF is mainly harboured by PMPs, and it can be trapped within the developing thrombus. MP detection and enumeration by polychromatic flow cytometry (PFC) have opened interesting perspectives in clinical settings, particularly for the evaluation of MP numbers and phenotypes as independent marker of cardiovascular risk, disease and outcome in diabetic patients.

Topics: Acute Coronary Syndrome; Atherosclerosis; Biomarkers; Blood Platelets; Cardiovascular Diseases; Cell-Derived Microparticles; Diabetes Mellitus; Diabetes Mellitus, Type 2; Diabetic Cardiomyopathies; Endothelial Cells; Humans; Plaque, Atherosclerotic; Risk Factors; Stroke; Thromboplastin

Clinical and epidemiological studies support a connection between obesity and thrombosis, involving elevated expression of the prothrombotic molecules plasminogen activator inhibitor-1 and tissue factor (TF) and increased platelet activation. Cardiovascular diseases and metabolic syndrome-associated disorders, including obesity, insulin resistance, type 2 diabetes, and hepatic steatosis, involve inflammation elicited by infiltration and activation of immune cells, particularly macrophages, into adipose tissue. Although TF has been clearly linked to a procoagulant state in obesity, emerging genetic and pharmacologic evidence indicate that TF signaling via G protein-coupled protease-activated receptors (PAR2, PAR1) additionally drives multiple aspects of the metabolic syndrome. TF-PAR2 signaling in adipocytes contributes to diet-induced obesity by decreasing metabolism and energy expenditure, whereas TF-PAR2 signaling in hematopoietic and myeloid cells drives adipose tissue inflammation, hepatic steatosis, and insulin resistance. TF-initiated coagulation leading to thrombin-PAR1 signaling also contributes to diet-induced hepatic steatosis and inflammation in certain models. Thus, in obese patients, clinical markers of a prothrombotic state may indicate a risk for the development of complications of the metabolic syndrome. Furthermore, TF-induced signaling could provide new therapeutic targets for drug development at the intersection between obesity, inflammation, and thrombosis.

Topics: Adipocytes; Adipose Tissue; Animals; Cardiovascular Diseases; Gene Expression Regulation; Humans; Inflammation; Insulin Resistance; Macrophages; Metabolic Syndrome; Mice; Mice, Obese; Models, Biological; Obesity; Plasminogen Activator Inhibitor 1; Receptor, PAR-1; Receptor, PAR-2; Risk Factors; Signal Transduction; T-Lymphocytes, Regulatory; Thrombophilia; Thromboplastin; Thrombosis

Following the molecular cloning of human Tissue Factor (TF) in mid-1980's, great strides have been made in the understanding of TF biology, TF's crucial roles in the initiation of blood coagulation and embryonic development, and TF's contribution to the pathobiology of various disease states. The 21st century brought about a rather unexpected turn in the "TF journey"--a few years back it was reported that the TF gene produces not one, but two proteins with distinct structural and functional characteristics. The so-called "full-length TF" (flTF) - a much-studied integral membrane glycoprotein long presumed to be, and experimentally handled as "the TF" in hundreds of laboratories around the world - is now known to be one of the two TF forms naturally occurring in humans as well as mice. The other, recently discovered form is termed alternatively spliced TF (asTF) which, unlike flTF, lacks a transmembrane domain and can thus be secreted. In this review, we summarize the literature on asTF by discussing asTF's biologic roles as they are currently understood, tackling a number of questions pertaining to asTF's evident and proposed biologic properties, and briefly covering the emerging field of regulated TF pre-mRNA processing.

Topics: Alternative Splicing; Animals; Cardiovascular Diseases; Endothelium, Vascular; Genetic Engineering; Humans; Mice; Mice, Knockout; Mice, Transgenic; Monocytes; Neoplasms; Neovascularization, Physiologic; RNA, Messenger; Thromboplastin

1. Obesity is a major risk factor for cardiovascular disease. An increased body mass index (BMI) is associated with venous thromboembolism, myocardial infarction, stroke and stent thrombosis after percutaneous interventions. Studies in mouse models of obesity and induced arterial or venous thrombosis have provided insights into the mechanisms involved. 2. In addition to elevated circulating levels of fibrinogen, factor VII and plasminogen activator inhibitor (PAI)-1, changes in platelet biology and function may underlie the increased (athero) thrombotic risk in obesity. These include elevated platelet counts, an increase in mean platelet volume, an increased platelet aggregatory response to agonists and a reversible resistance to the anti-aggregatory effects of nitric oxide and prostacyclin I(2) . 3. Specific adipokines mediate the prothrombotic state in obesity. Of these, leptin enhances both arterial and venous thrombosis by promoting platelet adhesion, activation and aggregation. Leptin also induces tissue factor expression by human neutrophils and other cells. C-Reactive protein enhances the formation of monocyte-platelet aggregates and also promotes P-selectin expression and platelet adhesion to endothelial cells. Further, the adipose tissue is a significant source of tissue factor and PAI-1. Conversely, the circulating levels of adiponectin, a hormone that exerts vasculoprotective, anti-atherosclerotic and antithrombotic effects, are reduced in obese individuals. 4. A better understanding of the interactions of the adipose tissue with circulating and vascular cells and the dissection of the mechanisms linking adipokines to arterial and venous thrombosis may identify obese individuals at particularly high cardiovascular risk and indicate promising vasculoprotective and therapeutic targets.

Topics: Adipokines; Adipose Tissue; Animals; Blood Platelets; Body Mass Index; C-Reactive Protein; Cardiovascular Diseases; Epoprostenol; Female; Humans; Male; Mice; Neutrophils; Nitric Oxide; Obesity; P-Selectin; Platelet Adhesiveness; Platelet Aggregation Inhibitors; Thromboplastin; Thrombosis

Heightened activity of circulating Tissue Factor (TF) has been linked to a variety of macro- and microvascular cardiovascular complications commonly observed in diabetes mellitus. Systemic and localized vascular abnormalities comprise the most debilitating feature of diabetic pathophysiology. Blood monocytes are chronically activated in diabetes, and serve as the major source of bioactive intravascular TF. This review examines recent literature on this subject, with a special emphasis on the abnormal monocyte physiology in diabetes and the structural and functional diversity of circulating TF.

Topics: Cardiovascular Diseases; Diabetes Complications; Diabetes Mellitus, Type 2; Humans; Thromboplastin

The coagulation cascade represents a system of proteases responsible to maintain vascular integrity and to induce rapid clot formation after vessel injury. Tissue factor (TF), the key initiator of the coagulation cascade, binds to factor VIIa and thereby activates factor IX and factor X, resulting in thrombus formation. Different stimuli enhance TF gene expression in endothelial and vascular smooth muscle cells. In addition to these vascular cells, TF has recently been detected in the bloodstream in circulating cells such as leukocytes and platelets, as a component of microparticles, and as a soluble, alternatively spliced form of TF. Various cardiovascular risk factors like hypertension, diabetes, and dyslipidemia, increase levels of TF. In line with this observation, enhanced vascular TF expression occurs during atherogenesis, particularly in patients with acute coronary syndromes. (Circ J 2010; 74: 3 - 12).

Topics: Atherosclerosis; Blood Coagulation; Cardiovascular Diseases; Endothelium, Vascular; Hemostasis; Humans; Risk Factors; Thromboplastin

Full-length tissue factor (flTF) initiates coagulation, but also exerts non-hemostatic functions such as inflammation and angiogenesis through protease activated receptors (PARs). In 2003 a soluble variant of flTF was described which results from alternative splicing. Since its discovery the role of alternatively spliced tissue factor (asTF) in coagulation has been debated. asTF may have pro-coagulant properties but due to structural differences when compared to flTF, asTF coagulant function may be relatively low. Nevertheless, similar to flTF, asTF appears to have non-hemostatic properties; asTF expression in tumors correlates with increased tumor size, vessel number and poor survival in some cancer types, and drives tumor growth in animal models. Interestingly, unlike flTF, asTF does not promote angiogenesis through activating PARs but rather via integrin ligation. flTF is a critical determinant in cardiovascular disease but little is known about asTF in cardiovascular disease. asTF is produced by monocytes and macrophages, thus macrophage-derived asTF may contribute to atherosclerotic disease. In conclusion, unraveling asTF's non-hemostatic properties may generate new insights in the pathophysiology and diagnostics of cancer and cardiovascular disease.

Topics: Alternative Splicing; Blood Coagulation; Cardiovascular Diseases; Hemostasis; Humans; Neoplasms; RNA Precursors; Thromboplastin

Tissue factor (TF) is the major trigger of the coagulation cascade and thereby crucially involved in the maintenance of vascular hemostasis. By binding factor VIIa, the resulting TF:VIIa complex activates the coagulation factors IX and X ultimately leading to fibrin and clot formation. In the vessel wall, TF expression and activity is detectable in vascular smooth muscle cells and fibroblasts and, at a much lower level, in endothelial cells and can be induced by various stimuli including cytokines. In addition, TF is found in the bloodstream in circulating cells such as monocytes, in TF containing microparticles, and as a soluble splicing isoform. Besides its well-known extracellular role as a trigger of coagulation, TF also functions as a transmembrane receptor, and TF-dependent intracellular signaling events regulate the expression of genes involved in cellular responses such as proliferation and migration. TF indeed appears to be involved in the pathogenesis of neointima formation and tumor growth, and increased levels of TF have been detected in patients with cardiovascular risk factors or coronary artery disease as well as in those with cancer. Therefore, pharmacological or genetic inhibition of TF may be an attractive target for the treatment of cardiovascular disease and cancer. Different strategies for inhibition of TF have been developed such as inhibition of TF synthesis and blockade of TF action. Clinical applications of such strategies need to be tested in appropriate trials, in particular for evaluating the advantages of targeted versus systemic delivery of the inhibitors.

Topics: Animals; Blood Coagulation; Blood Coagulation Factors; Cardiovascular Diseases; Fibrinolytic Agents; Humans; Thromboplastin

Alternative pre-mRNA splicing is an essential mechanism regulating protein diversity and functional plasticity of the proteome in response to environmental changes. Several factors are involved in this regulatory mechanism, such as serine/arginine-rich (SR) proteins, the Cdc2-like kinase (Clk) family, the dual-specificity tyrosine phosphorylation regulated kinases, the SR protein kinases (SRPK) 1 and 2, the protein kinase B (PKB,Akt), and the DNA topoisomerase I (DNA topo I). Dynamic changes of the phosphorylation state of SR proteins, mediated by the kinases mentioned previously, play an important role in alternative splicing regulation. Through alternative splicing of the tissue factor (TF) pre-mRNA, two naturally occurring forms of TF, the primary initiator of blood coagulation, are expressed in humans-soluble alternatively spliced (as)TF and membrane-bound "full length" (fl)TF. Both isoforms are known to circulate in blood. flTF, rather than asTF, appears to be the major contributor to the thrombogenicity of vascular wall and blood. asTF has been linked more closely to increased cell survival and angiogenesis. We found the expression of asTF and flTF to be reduced in the myocardium of patients with dilated cardiomyopathy, indicating a role of TF in maintaining myocardial structure. Moreover, we demonstrated proinflammatory cytokines to immediately upregulate the expression of both TF isoforms, which was differentially regulated by SR proteins as well as Clks and DNA topo I. We and others have shown that asTF induces cell proliferation, survival, and angiogenesis via signaling pathways different from flTF-induced effects. These data indicate that both TF isoforms influence diverse processes in cardiovascular (patho)biology and are potential targets for antithrombotic, pro-survival, and proangiogenic therapeutic strategies.

Topics: Alternative Splicing; Blood Coagulation; Cardiovascular Diseases; Cardiovascular Physiological Phenomena; Humans; Protein Isoforms; Proteomics; RNA Precursors; Thromboplastin

Microparticles are submicron vesicles shed from plasma membranes in response to cell activation, injury, and/or apoptosis. The measurement of the phospholipid content (mainly phosphatidylserine; PSer) of microparticles and the detection of proteins specific for the cells from which they are derived has allowed their quantification and characterization. Microparticles of various cellular origin (platelets, leukocytes, endothelial cells) are found in the plasma of healthy subjects, and their amount increases under pathological conditions. Endothelial microparticles (EMP) not only constitute an emerging marker of endothelial dysfunction, but are also considered to play a major biological role in inflammation, vascular injury, angiogenesis, and thrombosis. Although the mechanisms leading to their in vivo formation remain obscure, the release of EMP from cultured cells can be caused in vitro by a number of cytokines and apoptotic stimuli. Recent studies indicate that EMP are able to decrease nitric-oxide-dependent vasodilation, increase arterial stiffness, promote inflammation, and initiate thrombosis at their PSer-rich membrane, which highly co-expresses tissue factor. EMP are known to be elevated in acute coronary syndromes, in severe hypertension with end organ damage, and in thrombotic thrombocytopenic purpura, all conditions associated with endothelial injury and pro-thrombotic state. The release of EMP has also been associated with endothelial dysfunction of patients with multiple sclerosis and lupus anticoagulant. More recent studies have focused on the role of low shear stress leading to endothelial cell apoptosis and subsequent EMP release in end-stage renal disease. Improved knowledge of EMP composition, their biological effects, and the mechanisms leading to their clearance will probably open new therapeutic approaches in the treatment of atherothrombosis.

Topics: Animals; Apoptosis; Biomarkers; Blood Vessels; Cardiovascular Diseases; Cell-Derived Microparticles; Cytokines; Endothelial Cells; Humans; Inflammation; Kidney Failure, Chronic; Lupus Coagulation Inhibitor; Multiple Sclerosis; Neovascularization, Pathologic; Nitric Oxide; Phosphatidylserines; Shear Strength; Thromboplastin

TF (tissue factor) is the main trigger of the coagulation cascade; by binding Factor VIIa it activates Factor IX and Factor X, thereby resulting in fibrin formation. Various stimuli, such as cytokines, growth factors and biogenic amines, induce TF expression and activity in vascular cells. Downstream targets of these mediators include diverse signalling molecules such as MAPKs (mitogen-activated protein kinases), PI3K (phosphoinositide 3-kinase) and PKC (protein kinase C). In addition, TF can be detected in the bloodstream, known as circulating or blood-borne TF. Many cardiovascular risk factors, such as hypertension, diabetes, dyslipidaemia and smoking, are associated with increased expression of TF. Furthermore, in patients presenting with acute coronary syndromes, elevated levels of circulating TF are found. Apart from its role in thrombosis, TF has pro-atherogenic properties, as it is involved in neointima formation by inducing vascular smooth muscle cell migration. As inhibition of TF action appears to be an attractive target for the treatment of cardiovascular disease, therapeutic strategies are under investigation to specifically interfere with the action of TF or, alternatively, promote the effects of TFPI (TF pathway inhibitor).

Topics: Blood Coagulation; Cardiovascular Agents; Cardiovascular Diseases; Humans; Metabolic Diseases; Thromboplastin

Topics: Blood Coagulation Factors; Cardiovascular Diseases; Disease Susceptibility; Fibrinogen; Fibrinolysis; Humans; Polymorphism, Single Nucleotide; Prothrombin; Risk Factors; Thromboembolism; Thrombophilia; Thromboplastin; Thrombosis

Topics: Animals; Biomarkers; C-Reactive Protein; Cardiovascular Diseases; Humans; Lipoproteins; Mice; Muscle, Smooth, Vascular; Myocytes, Smooth Muscle; Thromboplastin

Tissue factor is the principal activator of blood coagulation in vivo. The existence of extravascular tissue factor has been recognized for over a century, but a rational role as a cell-based enzymatic cofactor in blood coagulation was first proposed by Paul Morawitz in 1905. By the close of the last century, very low levels of circulating tissue factor had been identified in the intravascular compartment, but its role in health and in hemostatic and thrombotic disorders continues to be debated. Nonetheless, ongoing research suggests that tissue factor may be a rational therapeutic target in a number of human disease states.

Topics: Animals; Blood Coagulation; Cardiovascular Diseases; Humans; Thromboplastin

Angiogenesis, the growth of new blood vessels from preexisting ones, is a necessary component of embryogenesis, wound healing, and the proliferative phase of the female reproductive cycle. Angiogenesis also plays a critical role in important pathologic processes such as cancer and cardiovascular complications. In addition, clinical, laboratory, and pharmacologic evidence has shown a link between angiogenesis, coagulation, hemostasis, and thrombosis in the settings of these pathologies. Recent studies in our laboratory revealed that thrombin has a significant stimulatory effect on angiogenesis. This effect of thrombin is independent of fibrin formation and can be attributed mainly to the activation of protease-activated receptor-1 (PAR-1). PAR-1 is widely expressed in vascular cells and is involved in cardiovascular complications such as atherosclerosis, restenosis, and neointimal formation. It is also expressed in many cancer cells contributing to induction of tumor growth and metastasis. In this review, we will summarize our present-day understanding of the role of thrombin and PAR-1 in angiogenesis and the potential therapeutic utility of targeting PAR-1 in angiogenesis-related disease, such as atherosclerosis, restenosis, and cancer.

Topics: Animals; Apoptosis; Blood Coagulation; Blood Vessels; Cardiovascular Diseases; Endothelium, Vascular; Humans; Neoplasm Metastasis; Neoplasms; Neovascularization, Pathologic; Neovascularization, Physiologic; Receptor, PAR-1; Thrombin; Thromboplastin

Tissue factor (TF) plays a role in thrombogenesis. TF initiates blood coagulation resulting in the generation of protease coagulant mediators (FVIIa, FXa, and FIIa) and fibrin production. TF hypercoagulablility directly contributes to thrombus formation resulting from the major events of fibrin deposition and FIIa-induced platelet activation/aggregation. In addition, blood coagulation indirectly promotes thrombogenicity via the coagulation-inflammation cycle in which TF plays a diverging and converging role. As the consequence of coagulation-dependent inflammation in which protease-activated receptor (PAR) mediates the coagulant signaling to elicit cytokines, selectins, and growth factors, such inflammation facilitates thrombosis by platelet aggregation and leukocyte recruitment. As TF hypercoagulability concerned, anti-thrombotic strategies involve the prevention by anticoagulation and PAR antagonism. Anticoagulants block the direct and indirect thrombotic contributions, while PAR antagonists arrest coagulation-dependent inflammation. With respect to both thrombosis and inflammation being cardiovascular risk factors, such strategies offer diverse benefits to cardioprotection.

Topics: Animals; Blood Coagulation; Blood Vessels; Cardiovascular Diseases; Factor VIIa; Factor Xa; Fibrin; Humans; Inflammation; Models, Biological; Myocardium; Peptides; Platelet Aggregation; Prothrombin; Receptors, Proteinase-Activated; Thromboplastin; Thrombosis

Tissue factor (TF), formerly known as thromboplastin, is the key initiator of the coagulation cascade; it binds factor VIIa resulting in activation of factor IX and factor X, ultimately leading to fibrin formation. TF expression and activity can be induced in endothelial cells, vascular smooth muscle cells, and monocytes by various stimuli such as cytokines, growth factors, and biogenic amines. These mediators act through diverse signal transduction mechanisms including MAP kinases, PI3-kinase, and protein kinase C. Cellular TF is present in three pools as surface, encrypted, and intracellular protein. TF can also be detected in the bloodstream, referred to as circulating or blood-borne TF. Elevated levels of TF are observed in patients with cardiovascular risk factors such as hypertension, diabetes, dyslipidemia, and smoking as well as in those with acute coronary syndromes. TF may indeed be involved in the pathogenesis of atherosclerosis by promoting thrombus formation; in addition, it can induce migration and proliferation of vascular smooth muscle cells. As a consequence, therapeutic strategies have been developed to specifically interfere with the action of TF such as antibodies against TF, site-inactivated factor VIIa, or recombinant TF pathway inhibitor. Inhibition of TF action appears to be an attractive target for the treatment of cardiovascular diseases.

Topics: Animals; Blood Coagulation; Cardiovascular Diseases; Endothelium, Vascular; Gene Expression Regulation; Humans; Thromboplastin

Overexpression and exposition of tissue factor (TF) in atherosclerotic plaques and/or arterial thrombi are critical events in atherothrombosis. TF, the receptor for factor VII (FVII) and activated factor VII (FVIIa), is the principal initiator of blood coagulation and induces thrombin generation leading to fibrin formation and platelet activation. TF also plays a major role in cell migration and angiogenesis. TF activity is downregulated by Tissue Factor Pathway Inhibitor (TFPI), a Kunitz-type inhibitor, which forms a neutralizing complex with TF, FVIIa and activated factor X. In physiological conditions, TF is absent from vascular cells which come into contact with flowing blood and is present as an inactive pool in fibroblasts and smooth muscle cells (SMC). In contrast, TF is widely expressed in atherosclerotic plaques and is found in macrophages, SMCs, and foam-cells and also in extracellular matrix and acellular lipid-rich core. TF expression is up-regulated by inflammatory cytokines and oxidized lipids. Plaque thrombogenicity is directly correlated to their TF content. After fibrous cap disruption, TF is exposed on plaque surface and triggers thrombus formation leading to arterial lumen occlusion and/or downstream embolization. In coronary and carotid plaques, TF content was found to be higher in plaques from symptomatic than asymptomatic patients. Soluble forms of TF and microparticles of monocyte and platelet origin, and bearing TF, constitute "blood-born TF". The contribution of this TF pool to arterial thrombosis is still under discussion. TF pathway is a target for new therapeutic agents that can decrease TF activity, such as active site-inactivated factor VIIa, recombinant TFPI and antibodies against TF or peptides interfering with TF-FVIIa complex activity.

Topics: Arteriosclerosis; Cardiovascular Diseases; Cell Movement; Fibroblasts; Hemostatics; Humans; Inflammation; Muscle, Smooth; Neovascularization, Pathologic; Thromboplastin

The adhesion receptor P-selectin has long been known to support leukocyte rolling and emigration at sites of inflammation. Recently, P-selectin was also revealed to be a key molecule in hemostasis and thrombosis, mediating platelet rolling, generating procoagulant microparticles containing active tissue factor and enhancing fibrin deposition. Elevated levels of plasma P-selectin are indicative of thrombotic disorders and predictive of future cardiovascular events. Because the interaction between P-selectin and its receptor P-selectin glycoprotein ligand-1 (PSGL-1) represents an important mechanism by which P-selectin induces the formation of procoagulant microparticles and recruits the microparticles to thrombi, anti-thrombotic strategies are currently aimed at inhibiting this interaction. Recent developments also suggest that the procoagulant potential of P-selectin could be used to treat coagulation disorders such as hemophilia A.

Topics: Animals; Blood Coagulation Disorders; Cardiovascular Diseases; Cell Adhesion; Fibrin; Humans; Inflammation; Leukocytes; Membrane Glycoproteins; Mice; Models, Biological; P-Selectin; Phenotype; Protein Binding; Protein Structure, Tertiary; Thromboplastin; Thrombosis

Topics: Adrenomedullin; Biomarkers; Cardiovascular Diseases; Cardiovascular System; Humans; Lipoproteins; Mucous Membrane; Neurosecretory Systems; Peptides; Thromboplastin; Tissue Distribution

The development of anticoagulants for treating patients with atherothrombotic disorders of the arterial circulatory system has focused, either directly or indirectly, on thrombin - a pleuripotential effector enzyme with prothrombotic and proinflammatory properties. The pivotal role of factor (f) Xa in thrombin generation, coupled with its direct cellular effects and widely recognized limitations of currently available anticoagulants, has led to the development of pharmacologic inhibitors of this important protease. The following review focuses on DX-9065a - first in a class of direct, selective and reversible fXa antagonists - and its potential applications in the management of patients with cardiovascular disease.

Topics: Animals; Anticoagulants; Area Under Curve; Blood Coagulation; Cardiovascular Diseases; Clinical Trials as Topic; Coronary Artery Disease; Disease Models, Animal; Dose-Response Relationship, Drug; Factor Xa; Factor Xa Inhibitors; Humans; International Normalized Ratio; Mice; Models, Biological; Models, Chemical; Naphthalenes; Pilot Projects; Platelet Aggregation Inhibitors; Propionates; Rats; Thrombin; Thromboplastin; Thrombosis; Time Factors

Tissue factor (TF) is a transmembrane glycoprotein, currently considered as being the major regulator of the coagulation cascade and the initiator of thrombogenesis in vivo. When TF comes in contact with blood, it forms a high-affinity complex with factors VII/VIIa, activating factors IX and X and thus leading to the formation of an insoluble fibrin clot. The regulation of TF-VIIa activity plays a key role in blood-vessel wall interactions. Selective patterns of cellular expression of TF are observed in tissues. TF is constitutively localized only on the surface of cells anatomically separated from the blood, where it plays an essential role in hemostasis by limiting hemorrhage after vessel wall injury. A number of pathophysiologic stimuli are capable of inducing TF transcription and activity in endothelial cells and monocytes. An aberrant TF expression in contact with blood is implicated in thrombotic complications of atherosclerosis, including acute myocardial infarction. Recent findings have demonstrated cell-derived microparticles containing TF in the circulating blood of patients with acute coronary syndromes, capable of triggering and propagating thrombus growth. This observation suggests a new view of thrombosis that does not necessarily require the exposure of vessel wall-derived TF at the site of vascular injury to initiate and propagate thrombosis.

Topics: Angina, Unstable; Animals; Arteriosclerosis; Blood Coagulation; Cardiovascular Diseases; Disease Models, Animal; Factor IX; Factor VII; Factor VIIa; Factor X; Fibrinolytic Agents; Helminth Proteins; Hemostasis; Humans; Myocardial Infarction; Syndrome; Thromboplastin; Thrombosis

Topics: Animals; Binding Sites; Blood Coagulation; Cardiovascular Diseases; Catalysis; Clinical Trials as Topic; Dogs; Endothelial Growth Factors; Enzyme Activation; Factor VIIa; Gene Expression Regulation; Granulocytes; Humans; Kinetics; Lymphokines; Macromolecular Substances; MAP Kinase Signaling System; Models, Molecular; Neoplasm Metastasis; Neoplasm Proteins; Protein Binding; Protein Conformation; Protein Structure, Tertiary; Signal Transduction; Structure-Activity Relationship; Substrate Specificity; Thermodynamics; Thromboplastin; Thrombosis; Vascular Endothelial Growth Factor A; Vascular Endothelial Growth Factors

Human tissue factor pathway inhibitor (TFPI) is a modular protein comprised of three Kunitz type domains flanked by peptide segments that are less structured. The sequential order of the elements are: an N-terminal acidic region followed by the first Kunitz domain (K1), a linker region, a second Kunitz domain (K2), a second linker region, the third Kunitz domain (K3), and the C-terminal basic region. The K1 domain inhibits factor VIIa complexed to tissue factor (TF) while the K2 domain inhibits factor Xa. No direct protease inhibiting functions have been demonstrated for the K3 domain. Importantly, the Xa-TFPI complex is a much more potent inhibitor of the VIIa-TF than TFPI by itself. Furthermore, the C-terminal basic region of TFPI is required for rapid physiologic inhibition of coagulation and is needed for the inhibition of smooth muscle cell proliferation. Although a number of additional targets for attachment have been reported, the C-terminal basic region appears to play an important role in binding of TFPI to cell surfaces. A primary site of TFPI synthesis is endothelium and the endothelium-bound TFPI contributes to the antithrombotic potential of the vascular endothelium. Further, increased levels of plasma TFPI under septic conditions may represent endothelial dysfunction. We have proposed that the extravascular cells that synthesize TF also synthesize TFPI providing dual components necessary for the regulation of clotting in their microenvironment. Like the TF synthesis in these cells is augmented by serum, so is the case with the TFPI gene expression. TFPI gene knock out mice reveal embryonic lethality suggesting a possible role of this protein in early development. Since TF-induced coagulation is thought to play a significant role in many disease states, including disseminated intravascular clotting, sepsis, acute lung injury and cancer, recombinant TFPI may be a beneficial therapeutic agent in these disease states to attenuate pathologic clotting. The purpose of this review is to outline recent developments in the field related to the structural specificity and biology of TFPI.

Topics: Acute Disease; Amino Acid Sequence; Amino Acids; Antiphospholipid Syndrome; Blood Coagulation; Cardiovascular Diseases; Endothelium, Vascular; Humans; Lipoproteins; Lung Diseases; Models, Biological; Models, Molecular; Molecular Sequence Data; Neoplasm Metastasis; Neoplasms; Protein Conformation; Protein Structure, Tertiary; Sepsis; Sequence Alignment; Sequence Homology, Amino Acid; Structure-Activity Relationship; Thrombophilia; Thromboplastin

An occlusive thrombus in the coronary arteries is the critical pathological event that immediately precedes most cases of myocardial infarction. Often the thrombus originates with a bleed from a fissured atheroma. Atheroma formation, therefore, creates risk of thrombosis; asymptomatic episodes of thrombosis and healing contribute to the pathogenesis of atherosclerosis and the development of atherosclerotic plaques. Based largely on in vitro and animal model evidence, infectious agents and their products can activate the coagulation cascade enzymatically or by up-regulating tissue factor. By initiating a procoagulant response, infectious agents can indirectly trigger a prothrombotic response. Alternatively, some microbes can directly trigger platelet aggregation in vitro and in animal models, suggesting direct prothrombotic potential in human cardiovascular disease. Activation of coagulation and thrombosis characterizes the pathological response to infectious agents in human disseminated intravascular coagulation and infective endocarditis. Given the underlying biological plausibility, the cumulative lifetime burden of chronic pathogens may be expected to create risk of atherosclerosis and thrombosis, and, indirectly, signs of cardiovascular disease.

Topics: Animals; Antigens, Bacterial; Bacteremia; Bacterial Proteins; Blood Coagulation; Cardiovascular Diseases; Collagen; Coronary Artery Disease; Coronary Thrombosis; Disease Models, Animal; Disseminated Intravascular Coagulation; Endocarditis, Bacterial; Humans; Platelet Aggregation; Risk Factors; Thromboplastin; Thrombosis

Topics: Adipocytes; Animals; Cardiovascular Diseases; Fatty Acids, Nonesterified; Gene Expression Regulation; Humans; Insulin; Obesity; Plasminogen Activator Inhibitor 1; RNA, Messenger; Serine Proteinase Inhibitors; Thromboplastin; Thrombosis; Transforming Growth Factor beta; Triglycerides; Tumor Necrosis Factor-alpha

Clinical trials of 3-hydroxy-3-methylglutaryl coenzyme A (HMG-CoA) reductase inhibitors or statin therapy have demonstrated that baseline or treated low-density lipoprotein (LDL) cholesterol levels are only weakly associated with net coronary angiographic change or cardiovascular events. The beneficial effects of statins on clinical events may involve nonlipid mechanisms that modify endothelial function, inflammatory responses, plaque stability, and thrombus formation. Experimental animal models suggest that statins may foster stability through a reduction in macrophages and cholesterol ester content and an increase in volume of collagen and smooth muscle cells. The thrombotic sequelae caused by plaque disruption is mitigated by statins through inhibition of platelet aggregation and maintenance of a favorable balance between prothrombotic and fibrinolytic mechanisms. These nonlipid properties of statins may help to explain the early and significant cardiovascular event reduction reported in several clinical trials of statin therapy.

Topics: Animals; Antithrombins; Cardiovascular Diseases; Cholesterol, LDL; Clinical Trials as Topic; Coronary Disease; Endothelium, Vascular; Fibrinogen; Humans; Hydroxymethylglutaryl-CoA Reductase Inhibitors; Hypolipidemic Agents; Inflammation; Lovastatin; Muscle, Smooth, Vascular; Platelet Aggregation; Pravastatin; Simvastatin; Thromboplastin; Thrombosis

VEGF has been proposed to participate in normal and pathological vessel formation. Surprisingly, lack of only a single VEGF allele resulted in embryonic lethality due to abnormal formation of intra- and extra-embryonic vessels. Homozygous VEGF-deficient embryos, generated by tetraploid aggregation, revealed an even more severe defect in vessel formation. These results (1) suggest a tight regulation of early vessel development by VEGF and, indirectly, the presence of other VEGF-like molecules; (2) reveal an unprecedented lethal phenotype associated with heterozygous deficiency of an autosomal gene, and (3) demonstrate that tetraploid aggregation was a valid and the only method to study the phenotype of the homozyogous VEGF-deficient embryos. The dominant and strict dose-dependent role of VEGF in vivo renders this molecule a desirable therapeutic target for promoting or preventing angiogenesis. Tissue factor (TF) is the principal cellular initiator of coagulation and its deregulated expression has been related to thrombogenesis in sepsis, cancer, and inflammation. However, TF appears to be also involved in a variety of non-hemostatic functions including inflammation, cancer, brain function, immune response, and tumor-associated angiogenesis. Surprisingly, TF deficiency resulted in embryonic lethality due to abnormal extra-embryonic vessel development and defective vitelloembryonic circulation. The abnormal yolk sac vasculature is reminiscent of that observed in embryos lacking VEGF, possibly suggesting that both gene functions are interconnected. These targeting studies extend the recently documented role of TF in tumor-associated angiogenesis and warrant further study of its role in angiogenesis during other pathological disorders. The plasminogen system, via its triggers, tissue-type plasminogen activator (t-PA) and urokinase-type plasminogen activator (u-PA) and its inhibitor, plasminogen activator inhibitor-1 (PAI-1), has been implicated in thrombosis, arterial neointima formation, and atherosclerosis. Studies in mice with targeted gene inactivation of t-PA, u-PA, PAI-1, the urokinase receptor (u-PAR), and plasminogen (Plg) revealed (1) that deficiency of t-PA or u-PA increase the susceptibility to thrombosis associated with inflammation and that combined deficiency of t-PA:u-PA or deficiency of Plg induces severe spontaneous thrombosis; (2) that vascular injury-induced neointima formation is reduced in mice lacking u-PA-mediated plasmin proteolysis, un

Topics: Animals; Cardiovascular Diseases; Cardiovascular Physiological Phenomena; Cardiovascular System; Endothelial Growth Factors; Gene Targeting; Gene Transfer Techniques; Humans; Lymphokines; Mice; Plasminogen; Receptors, Cell Surface; Thromboplastin; Vascular Endothelial Growth Factor A; Vascular Endothelial Growth Factors

Although biotechnology has been useful in the development of new diagnostic methods and drugs for the management of cardiovascular disorders, there are several issues which raise certain questions on the global use of biotechnology based drugs and diagnostic methods (Piascik, 1991; Fareed, 1993a; Fareed, 1994a). The cost is rather prohibitive in the development of this type of technology. Most diagnostic methods and drugs developed utilizing biotechnology based methods are relatively expensive. The second important consideration is the equivalence of the newer biotechnology derived drugs to the natural products. Many of the biotechnology derived drugs are obtained in prokaryotic systems (E. coli). Post-transcriptional modifications such as glycosylation are often important in determining the function of various proteins. On the other hand, biotechnology based diagnostic methods exhibit somewhat different specificity in comparison to conventional methods. Thus, it is rather important to assess the developments in this area in a careful manner. Furthermore, validation of the clinical, diagnostic and therapeutic efficacy of biotechnology derived diagnostic devices and drugs is a prerequisite for their use in cardiovascular medicine.

Topics: Amino Acid Sequence; Antibodies, Monoclonal; Anticoagulants; Biomarkers; Biotechnology; Cardiovascular Diseases; Hirudin Therapy; Hirudins; Humans; Lipoproteins; Molecular Sequence Data; Recombinant Proteins; Risk Factors; Thromboplastin

Topics: Cardiovascular Diseases; Factor VII; Factor VIIa; Follow-Up Studies; Hemostasis; Humans; Mutagenesis, Site-Directed; Sensitivity and Specificity; Thromboplastin; Thrombosis

Topics: Adult; Aged; Animals; Cardiovascular Diseases; Cerebrovascular Disorders; Cohort Studies; Cross-Sectional Studies; Epidemiologic Methods; Europe; Factor VII; Female; Fibrinogen; Hemostasis; Humans; Hyperlipidemias; Lipoproteins; Male; Middle Aged; Myocardial Infarction; Prospective Studies; Risk Factors; Thromboplastin; Triglycerides; United States

Microparticles (MP) are shed extracellular vesicles that express the prothrombotic tissue factor (TF). Aerobic exercise may reduce MP count and TF expression. This study investigated the impact of acute running or rest followed by standardized meal consumption on MP phenotypes and TF expression.. Fifteen males (age, 22.9 ± 3.3 yr; body mass, 81.9 ± 11.4 kg; V˙O2max, 54.9 ± 6.5 mL·kg·min; mean ± SD) completed 1 h of running (70% V˙O2max) or rest at 9:00 AM and consumed a standardized meal (1170 kcal, 43% CHO, 17% PRO, 40% fat) at 10:45 AM. Venous blood samples were taken at 9:00 AM, 10:00 AM, and 11:30 AM. The MP concentration, diameter, phenotypes, and TF expression were assessed using nanoparticle tracking analysis and flow cytometry.. Nanoparticle tracking analysis identified no changes in MP concentration or diameter in response to time or trial. Flow cytometry revealed total MP count increased from 9:00 AM to 10:00 AM (1.62 ± 2.28 to 1.74 ± 2.61 × 10 L, P = 0.016, effect size (η) = 0.105), but was unaffected by trial. TF platelet-derived MP % reduced from 9:00 AM to 10:00 AM (44.0% ± 21.2% to 21.5% ± 9.3%, P = 0.001, η = 0.582) after exercise only (control, 36.8% ± 18.2% to 34.9% ± 11.9%; P = 0.972). TF neutrophil-derived MP percentage reduced from 9:00 AM to 11:30 AM (42.3% ± 17.2% to 25.1% ± 14.9%; P = 0.048, η = 0.801) in the exercise trial only (control, 28.5% ± 15.7% to 32.2% ± 9.6%; P = 0.508).. Running induced a significant reduction in %TF platelet and neutrophil MP, suggesting a transient reduction in cardiovascular risk via reduced TF-stimulated thrombosis. This requires further investigation over longer periods in cardiovascular disease populations.

Topics: Adult; Cardiovascular Diseases; Cell-Derived Microparticles; Cross-Over Studies; Exercise; Flow Cytometry; Humans; Male; Meals; Risk Factors; Thromboplastin; Young Adult

Aging is associated with a vasoconstrictive, pro-coagulant, and pro-inflammatory profile of arteries and a decline in the bioavailability of the endothelium-derived molecule nitric oxide. Dietary nitrate elicits vasodilatory, anti-coagulant and anti-inflammatory effects in younger individuals, but little is known about whether these benefits are evident in older adults. We investigated the effects of 140 mL of nitrate-rich (HI-NI; containing 12.9 mmol nitrate) versus nitrate-depleted beetroot juice (LO-NI; containing ≤0.04 mmol nitrate) on blood pressure, blood coagulation, vascular inflammation markers, plasma nitrate and nitrite before, and 3 h and 6 h after ingestion in healthy older adults (five males, seven females, mean age: 64 years, age range: 57-71 years) in a randomized, placebo-controlled, crossover study. Plasma nitrate and nitrite increased 3 and 6 h after HI-NI ingestion (

Topics: Aged; Aging; Beta vulgaris; Biomarkers; Blood Platelets; Blood Pressure; Cardiovascular Diseases; CD11b Antigen; Cross-Over Studies; Diet; Double-Blind Method; Female; Fruit and Vegetable Juices; Granulocytes; Hemostasis; Humans; Inflammation; Male; Middle Aged; Monocytes; Nitrates; Nitrites; P-Selectin; Plant Roots; Prothrombin; Thromboplastin; Waist Circumference

Circulating microparticles (cMPs) are small phospholipid-rich microvesicles shed by activated cells that play a pivotal role in cell signalling related to the pathogenesis of atherothrombosis. We aimed to investigate the prognostic value of cMPs released from different vascular cells for cardiovascular event (CVE) presentation in asymptomatic patients at high cardiovascular risk factors under nutritional and pharmacologic treatment. This is a nested case-control study of 50 patients from the five-year follow-up prospective PREDIMED trial enrolled in the nuts arm of the Mediterranean diet (MedDiet-nuts). We randomly selected 25 patients who had suffered a CVE during follow-up and pair-matched them for sex, age, and classical CV risk factors to 25 patients who remained asymptomatic (no-CVE). Total Annexin V-(AV)+ cMPs and cMPs from cells of the vascular compartment were quantified by flow cytometry at baseline and after one year follow-up. MedDiet-nuts and pharmacological treatment neither modified levels nor source of MP shedding in CVE patients. However, no-CVE patients showed 40-86 % decreased total AV+, PAC-1+/AV+, CD61+/AV+, CD142+/CD61+/AV+, CD62P+/AV+, CD146+/AV+, CD63+/AV+ and CD11a+/AV+ cMPs at one year follow-up (p≤0.046, all). CD142+/CD61+/AV+, CD146+/AV+ and CD45+/AV+ cMPs were decreased in no-CVE patients compared to CVE patients. A ROC-curve clustered model for CD142+/CD61+/AV+, CD45+/AV+ and CD146+/AV+ cMPs predicted a future CVE [p<0.0001, AUC=0.805 (0.672 to 0.938)]. In patients at high CV risk profile treated with a controlled MedDiet supplemented with nuts and receiving up-to-date CV drug treatment, reduced cMPs derived from activated platelets, leukocytes and endothelial cells are predictive of protection against CVE within the next four years.

Topics: Aged; Aged, 80 and over; Cardiovascular Diseases; Case-Control Studies; CD146 Antigen; Cell-Derived Microparticles; Diet, Mediterranean; Female; Humans; Integrin beta3; Leukocyte Common Antigens; Male; Middle Aged; Nuts; Prognosis; Prospective Studies; Risk Factors; Thromboplastin

Increased coagulation activation may contribute to the high incidence of cardiovascular complications observed in obese and type 2 diabetes (T2D) subjects. Although tissue factor (TF), the primary initiator of coagulation is increased in obesity, its expression in adipose tissues and its association with metabolic parameters are unclear. We sought to compare TF expression in plasma and adipose tissues of obese subjects with and without T2D, its correlation with metabolic parameters, and regulation in response to antidiabetic drugs.. Subjects were recruited from diabetes clinics and adipose tissue was obtained by needle biopsy of lower subcutaneous abdominal depot. For the intervention study, subjects were randomized into treatment groups with rosiglitazone or metformin for 4 months.. Plasma TF antigen, activity, and adipose TF mRNA were greater in obese T2D subjects compared with obese nondiabetics. Plasma TF activity correlated with fasting insulin, glucose, and free fatty acids, (FFAs), and adipose TF mRNA correlated with plasma FFA. Plasma TF activity was reduced by metformin and increased with rosiglitazone treatment.. Specific diabetes-related metabolic parameters, but not obesity per se, are correlated with TF expression. Regulation of TF activity by different classes of antidiabetic drugs may relate to protective or adverse cardiovascular outcomes.

Topics: Adult; Blood Glucose; Cardiovascular Diseases; Diabetes Mellitus, Type 2; Diabetic Angiopathies; Female; Gene Expression Regulation; Humans; Hypoglycemic Agents; Male; Metformin; Middle Aged; Obesity; Rosiglitazone; Thiazolidinediones; Thromboplastin

Abnormal levels of inflammation are associated with cardiovascular disease and mortality in human immunodeficiency virus (HIV)-infected patients. Microbial translocation, which may cause inflammation, is decreased by sevelamer in patients undergoing hemodialysis. In this single-arm study, we evaluated the effects of 8 weeks of sevelamer therapy on 36 HIV-infected subjects who were not receiving antiretroviral therapy. Sevelamer did not significantly change markers of microbial translocation, inflammation, or T-cell activation. During sevelamer treatment, however, levels of soluble tissue factor, low-density lipoprotein (LDL) cholesterol, and oxidized LDL cholesterol decreased significantly, whereas D-dimer levels increased. Thus, in this study population, sevelamer did not reduce microbial translocation but may have yielded cardiovascular benefits.. NCT 01543958.

Topics: Adult; Bacterial Translocation; Cardiovascular Agents; Cardiovascular Diseases; Cholesterol, LDL; HIV Infections; Humans; Lipopolysaccharide Receptors; Lipopolysaccharides; Lipoproteins, LDL; Male; Middle Aged; Polyamines; Sevelamer; Thromboplastin; Treatment Outcome; Young Adult

Metabolic syndrome (MS) represents a cluster of cardiovascular risk factors and an increased risk of cardiovascular events. The carotid intima-media thickness (CIMT) is correlated with coronary and carotid atherosclerosis, and is a significant predictor of cardiovascular events. Tissue factor (TF) is an initiator of the extrinsic coagulation cascade and is expressed on peripheral blood monocytes and macrophages in atherosclerotic plaques. TF plays important roles in both thrombosis and atherosclerosis. No study has investigated the relationship between monocyte TF activity and CIMT in MS patients.. Peripheral blood mononuclear cells (PBMCs) were collected from 39 normal subjects and 110 patients with MS. The procoagulant activity (PCA) in monocytes was measured using a one-stage clotting assay and is expressed as the mean±SD (mU TF/10(6) PBMCs).. The PCA in monocytes in MS patients was significantly higher than in normal subjects (86.2 ±69.5 vs. 52.4±9.9 mU TF/10(6) PBMCs, p < 0.001). In multivariate analysis, patient age (β coefficient= 0.373, p < 0.001), high-density lipoprotein cholesterol (β coefficient=-0.307, p = 0.001) and PCA (β coefficient= 0.422, p =0.002) were each significantly and independently associated with CIMT.. These data indicate that the upregulation of monocyte TF activity is significantly associated with CIMT in MS patients.

Topics: Aged; Cardiovascular Diseases; Carotid Arteries; Case-Control Studies; Female; Humans; Male; Metabolic Syndrome; Monocytes; Thromboplastin; Tunica Intima; Up-Regulation

Men have a higher incidence of cardiovascular disease (CVD) than women of similar age, and it has been suggested that testosterone may influence the development of CVD. Recently, we demonstrated that elderly men with low testosterone levels had lower plasma levels of free tissue factor pathway inhibitor (TFPI) Ag associated with shortened tissue factor (TF)-induced coagulation initiation in a population based case-control study. Our hypothesis was that one year of testosterone treatment to physiological levels in elderly men would increase the levels of free TFPI Ag in plasma and have a favorable effect on TF-induced coagulation. Twenty-six men with low testosterone levels (< or =11.0 nM) were randomly assigned to treatment with intramuscular testosterone depot injections (testosterone undecanoate 1,000 mg) or placebo in a double-blinded study. Each participant received a total of five injections, at baseline, 6, 16, 28 and 40 weeks, and TF-induced thrombin generation ex vivo and plasma free TFPI Ag were measured after one year. At the end of the study total and free testosterone levels were significantly higher in the testosterone treated group (14.9 +/- 4.5 nM vs. 8.1 +/- 2.4 nM; p < 0.001, and 363.3 +/- 106.6 pM vs. 187.3 +/- 63.2 pM; p < 0.001, respectively). Testosterone treatment for one year did neither cause significant changes in TF-induced thrombin generation ex vivo nor changes in plasma levels of free TFPI Ag. In conclusion, normalising testosterone levels by testosterone treatment for 12 months in elderly men did not affect TF-induced coagulation or plasma TFPI levels. The potential antithrombotic role of testosterone therapy remains to be elucidated.

Topics: Aged; Antigens; Cardiovascular Diseases; Case-Control Studies; Delayed-Action Preparations; Double-Blind Method; Factor VII; Follow-Up Studies; Humans; Hypogonadism; Injections, Intramuscular; Lipids; Lipoproteins; Male; Middle Aged; Risk Factors; Testosterone; Thrombin; Thrombophilia; Thromboplastin; Time Factors

To determine the relationship between abnormalities in blood coagulation and prevalent or incident cardiovascular complications in Type 2 diabetes.. Prospective cohort study of 128 patients with Type 2 diabetes in whom blood samples were collected at baseline and after 1 year of follow-up. All cardiovascular complications at baseline and follow-up were recorded. Forty-three healthy, age-matched subjects served as a control group.. Logistic analysis revealed an independent relationship between soluble tissue factor (TF) and microvascular disease [per pg mL(-1) TF: Exp(B) = 1.008; CI(95%)1.002-1.014], or neurogenic disease [Exp(B) = 1.006; CI(95%)1.001-1.011]. The highest levels of soluble TF were observed in patients with microvascular and neurogenic disease (P < 0.001). Patients with Type 2 diabetes having a soluble TF concentration >300 pg mL(-1) are at a 15-fold higher risk for the presence of microvascular disease and at a 10-fold higher risk for the presence of neurogenic disease compared with the patients with concentrations below 100 pg mL(-1). Soluble TF was correlated with tissue type plasminogen activator, von Willebrand factor antigen, systolic blood pressure and age. Levels of F1' + 2, D-dimer, FVIII activity, t-PA and vWFag were not different among patients with micro-, macro- or neurogenic complications compared with patients without those complications. Forty-eight new micro-, macro- and/or neurogenic complications were diagnosed after 1 year follow-up. With the exception of higher F1 + 2 levels after 1 year all other markers remained unchanged. A trend toward higher soluble TF levels was observed in patients with new microvascular events (P = 0.056).. Soluble TF is associated with existing microvascular and neurogenic complications in patients with Type 2 diabetes and is a candidate marker for progression of microvascular disease.

Topics: Adult; Aged; Aged, 80 and over; Biomarkers; Blood Coagulation; Cardiovascular Diseases; Diabetes Mellitus, Type 2; Diabetic Angiopathies; Disease Progression; Female; Humans; Male; Middle Aged; Thromboplastin

The nature of dietary fats affects the postprandial activation of the hemostatic system.. We investigated whether the ratio of oleic to palmitic acid [and that of monounsaturated to saturated fatty acids (MUFA:SFA)] in the diet affects postprandial concentrations of triacylglycerols, tissue factor (TF), fibrinogen, tissue-type plasminogen activator (t-PA), and plasminogen activator inhibitor 1 (PAI-1).. We studied the effects of diets enriched in olive oil (ROO), high-palmitic sunflower oil (HPSO), butter, or a mixture of vegetable and fish oils (VEFO) on circulating concentrations of the aforementioned factors in 14 healthy men. The fats had ratios of oleic to palmitic acid (MUFA:SFA) of 6.83 (5.43), 2.36 (2.42), 0.82 (0.48), and 13.81 (7.08).. The largest and longest-lasting postprandial changes in plasma triacylglycerol concentrations were found with the butter-based diet (all P < 0.05). No correlation was observed between the net incremental area under the curve (netAUC) for triacylglycerol and the ratio of oleic to palmitic acid (or MUFA:SFA) in the dietary fats. The netAUCs for TF and PAI-1, however, were inversely related to the ratio of oleic to palmitic acid (and MUFA:SFA) in ROO, HPSO, butter, and VEFO. Similar results were found for the fibrinogen netAUC when VEFO was omitted from the analysis. The netAUC for t-PA was inversely correlated with postprandial concentrations of triacylglycerol.. Postprandial concentrations of TF, fibrinogen, and PAI-1 are associated with the ratio of oleic to palmitic acid (MUFA:SFA) in dietary fats. The postprandial t-PA response is related to postprandial concentrations of triacylglycerol.

Topics: Adult; Area Under Curve; Cardiovascular Diseases; Cross-Over Studies; Dietary Fats; Dietary Fats, Unsaturated; Enzyme Inhibitors; Fibrinogen; Humans; Male; Oleic Acid; Palmitic Acid; Plasminogen Activator Inhibitor 1; Plasminogen Activators; Postprandial Period; Risk Factors; Thromboplastin; Thrombosis; Triglycerides

Elevated homocysteine levels are associated with an increased cardiovascular disease (CVD) risk, but the underlying mechanism is still unclear. High homocysteine might affect the endothelium, and consequently lead to impaired haemostasis. In a randomized placebo controlled trial among 276 older adults with plasma total homocysteine concentrations above 13 mM at screening, we investigated the effect of homocysteine lowering by folic acid supplementation (0.8 mg/day) for 1 year on markers of endothelial function (von Willebrand factor), coagulation (tissue factor, factor VIIa, fragments 1+2), and fibrinolysis (fibrin degradation products, tissue-type plasminogen activator), and inflammation (C-reactive protein). Despite a 24% reduction in plasma homocysteine concentration and four-fold increase in serum folate concentration in the folic acid group compared to the placebo group, there was no clear change in any of the haemostasis markers, nor CRP. Although homocysteine is associated with vascular disease risk in the general population, marked lowering of slightly elevated homocysteine concentrations by one-year folic acid supplementation does not influence haemostasis markers.

Topics: Aged; C-Reactive Protein; Cardiovascular Diseases; Dietary Supplements; Factor VIIa; Female; Fibrin; Folic Acid; Hemostasis; Homocysteine; Humans; Inflammation; Male; Middle Aged; Placebos; Risk; Thromboplastin; Time Factors; von Willebrand Factor

We assessed the effects of estrogen on vascular dilatory and other homeostatic functions potentially affected by nitric oxide (NO)-potentiating properties in type II diabetic postmenopausal women.. There is a higher cardiovascular risk in diabetic women than in nondiabetic women. This would suggest that women with diabetes do not have the cardioprotection associated with estrogen.. We administered placebo or conjugated equine estrogen, 0.625 mg/day for 8 weeks, to 20 type II diabetic postmenopausal women in a randomized, double-blinded, placebo-controlled, cross-over design.. Compared with placebo, estrogen tended to lower low-density lipoprotein (LDL) cholesterol levels by 15 +/- 23% (p = 0.007) and increase high-density lipoprotein (HDL) cholesterol levels by 8 +/- 16% (p = 0.034). Thus, the ratio of LDL to HDL cholesterol levels significantly decreased with estrogen, by 20 +/- 24%, as compared with placebo (p = 0.001). Compared with placebo, estrogen tended to increase triglyceride levels by 16 +/- 48% and lower glycosylated hemoglobin levels by 3 +/- 13% (p = 0.295 and p = 0.199, respectively). However, estrogen did not significantly improve the percent flow-mediated dilatory response to hyperemia (17 +/- 75% vs. placebo; p = 0.501). The statistical power to accept our observation was 81.5%. Compared with placebo, estrogen did not significantly change E-selectin, intercellular adhesion molecule-1, vascular cell adhesion molecule-1, monocyte chemoattractant protein-1 or matrix metalloproteinase-9 levels. Compared with placebo, estrogen tended to decrease tissue factor antigen and increase tissue factor activity levels by 7 +/- 46% and 5 +/- 34%, respectively (p = 0.321 and p = 0.117, respectively) and lower plasminogen activator inhibitor-1 levels by 16 +/- 31% (p = 0.043).. The effects of estrogen on endothelial, vascular dilatory and other homeostatic functions were less apparent in type II diabetic postmenopausal women, despite the beneficial effects of estrogen on lipoprotein levels.

Topics: Brachial Artery; Cardiovascular Diseases; Chemokine CCL2; Cholesterol, HDL; Cholesterol, LDL; Cross-Over Studies; Diabetes Mellitus, Type 2; Double-Blind Method; Drug Synergism; E-Selectin; Estrogens, Conjugated (USP); Female; Fibrinolysis; Glycated Hemoglobin; Hemostasis; Homeostasis; Humans; Intercellular Adhesion Molecule-1; Matrix Metalloproteinase 9; Middle Aged; Nitric Oxide; Plasminogen Activator Inhibitor 1; Postmenopause; Thromboplastin; Triglycerides; Vascular Cell Adhesion Molecule-1; Vasodilation

Various methods have been described to evaluate efficacy of anticoagulant therapy using the international normalized ratio (INR). We compared the following approaches: (1) total INR's or the most recent measurement; (2) percent time within therapeutic range, with INR changing directly or halfway between visits; and (3) total observation time assuming INR changing linearly. The study population comprised 1700 post myocardial infarction patients. Treatment comprised 3725 patient-years. There were 61,471 INR assessments with target therapeutic level of 2.8-4.8. Acenocoumarol as well as phenprocoumon were employed. Therapeutic achievement in the first months of treatment was low: less than 60% of INR's were in range. Treatment stabilized after 6 months. Patients on acenocoumarol were within range 70% of the time compared to 80% for phenprocoumon. Method 3 is preferred because it incorporates time and is capable of calculating incidence rates at different INR levels. Our findings call for an urgent improvement of standard of anticoagulant control in the first months following commencement of treatment.

Topics: Acenocoumarol; Aged; Anticoagulants; Cardiovascular Diseases; Convalescence; Double-Blind Method; Female; Follow-Up Studies; Hemorrhage; Humans; Male; Middle Aged; Myocardial Infarction; Phenprocoumon; Prothrombin Time; Quality Control; Reference Standards; Thromboplastin; Treatment Outcome

 Several evidence show that elevated plasma levels of uric acid (UA) are associated with the increased risk of developing atherothrombotic cardiovascular events. Hyperuricemia is a risk factor for endothelial dysfunction (ED). ED is involved in the pathophysiology of atherothrombosis since dysfunctional cells lose their physiological, antithrombotic properties. We have investigated whether UA might promote ED by modulating the tissue factor (TF)/TF pathway inhibitor (TFPI) balance by finally changing the antithrombotic characteristics of endothelial cells..  Human umbilical vein endothelial cells were incubated with increasing doses of UA (up to 9 mg/dL). TF gene and protein expressions were evaluated by real-time polymerase chain reaction (PCR) and Western blot. Surface expression and procoagulant activity were assessed by FACS (fluorescence activated cell sorting) analysis and coagulation assay. The mRNA and protein levels of TFPI were measured by real-time PCR and Western blot. The roles of inflammasome and nuclear factor-κB (NF-κB) as possible mechanism(s) of action of the UA on TF/TFPI balance were also investigated..  UA significantly increased TF gene and protein levels, surface expression, and procoagulant activity. In parallel, TFPI levels were significantly reduced. The NF-κB pathways appeared to be involved in modulating these phenomena. Additionally, inflammasome might also play a role..  The present in vitro study shows that one of the mechanisms by which high levels of UA contribute to ED might be the imbalance between TF/TFPI levels in endothelial cells, shifting them to a nonphysiological, prothrombotic phenotype. These UA effects might hypothetically explain, at least in part, the relationship observed between elevated plasma levels of UA and cardiovascular events.

Topics: Cardiovascular Diseases; Fibrinolytic Agents; Human Umbilical Vein Endothelial Cells; Humans; Inflammasomes; NF-kappa B; Thromboplastin; Uric Acid

Proprotein convertase subtilisin kexin 9 (PCSK9) is a serin protease synthesized mainly in the liver that binds the receptor of low-density lipoprotein and promotes its degradation in lysosomes. PCSK9 is considered a promising target for the development of new therapies for the treatment of hypercholesterolemia and related cardiovascular diseases. Extracellular vesicles represent a heterogeneous population of vesicles, ranging in size between 0.05 and 1 μm involved in numerous pathophysiological processes, including blood coagulation. We investigated whether PCSK9 stimulation induces the release of procoagulant extracellular vesicles from human mononuclear cells (PBMCs) and THP-1 cells.. PBMCs and THP-1 cells were stimulated whit PCSK9, the generation of EV was assessed by the prothrombinase assay and by cytofluorimetric analysis. EV-associated tissue factor activity was assessed by a one-stage clotting assay. PCSK9 induced an increase in extracellular generation by PBMCs and THP-1 cells as well as an increase in extracellular vesicle-associated tissue factor. Pre-treatment with inhibitors of the toll like receptor, TLR4 (C34), and of NF-κB signaling (BAY 11-7082), downregulated PCSK9-induced extracellular vesicle generation and of extracellular- bound tissue factor. Similar effect was obtained by an anti-PCSK9 human-monoclonal antibody.. PCSK9-mediated generation of procoagulant EV could contribute to increase the prothrombotic status in patients with cardiovascular diseases.

Topics: Cardiovascular Diseases; Extracellular Vesicles; Humans; Proprotein Convertase 9; Receptors, LDL; Subtilisins; Thromboplastin

Membrane microparticles (MP) are released by activated or damaged cells and are able to accelerate blood clotting (coagulation). MP possess coagulation activity since all of them contain on their surface phosphatidylserine (PS), a substrate for the assembly of coagulation complexes, and some of them tissue factor (TF), the primary initiator of coagulation cascade reactions. We compared the coagulation activity and amount of MP in the blood of healthy donors (n=34) and patients with myocardial infarction (MI) (n=32), advanced atherosclerosis (AA) (n=32) and idiopathic pulmonary arterial hypertension (IPAH) (n=19). Total MP fraction was obtained from blood plasma by sedimentation at 20000 g, 30 min. The coagulation activity of PM isolated from 100 μl of donor and patient plasma was determined using a modified recalcification test. MP were added to substrate plasma devoid of endogenous MF, plasma was recalcified, and clotting was recorded by changes in optical density (A450), determining lag phase (min) and maximum rate (Vmax, %A450/min). MP were counted by flow cytometry as PS+ particles (lactadgerin-FITC staining) smaller than 1 μm and their concentration was expressed as 105 MP/μl plasma. MP in all patient groups accelerated plasma clotting more effectively than donor MP. Lag phase compared with donors (11.8 [11.0-13.1] median and interquartile range) was shorter in patients with AA (8.8 [7.0-10.3], p.. Membrannye mikrochastitsy (MCh) vysvobozhdaiutsia aktivirovannymi ili povrezhdennymi kletkami i obladaiut sposobnost'iu uskoriat' svertyvanie (koaguliatsiiu) krovi. Koaguliatsionnaia aktivnost' MCh opredeliaetsia tem, chto vse oni soderzhat na svoeĭ poverkhnosti fosfatidilserin (FS), kotoryĭ sluzhit substratom dlia sborki koaguliatsionnykh kompleksov, a nekotorye iz nikh tkanevoĭ faktor (TF) — pervichnyĭ initsiator reaktsiĭ koaguliatsionnogo kaskada. My sravnivali koaguliatsionnuiu aktivnost' i kolichestvo MCh v krovi zdorovykh donorov (n=34) i patsientov s infarktom miokarda (IM) (n=32), rasprostranennym aterosklerozom (RA) (n=32) i idiopaticheskoĭ legochnoĭ arterial'noĭ gipertenzieĭ (ILAG) (n=19). Obshchuiu fraktsiiu MCh iz plazmy krovi poluchali osazhdeniem pri 20000 g, 30 min. Kontsentratsiiu MCh i stepen' ékspressii na ikh poverkhnosti FS opredeliali metodom protochnoĭ tsitometrii. Koaguliatsionnuiu aktivnost' MCh otsenivali s pomoshch'iu modifitsirovannogo testa rekal'tsifikatsii plazmy. Kontsentratsiia MCh po sravneniiu so zdorovymi donorami byla vyshe u patsientov s RA, nizhe u patsientov s IM i dostoverno ne otlichalas' u patsientov s ILAG. MCh vo vsekh gruppakh patsientov uskoriali svertyvanie plazmy éffektivnee, chem MCh zdorovykh donorov. Lag-faza u patsientov s RA, IM i ILAG byla koroche po sravneniiu so zdorovymi donorami, a Vmaks v sravnenii s zdorovymi donorami byla vyshe u patsientov s RA i ILAG, no ne IM. Vo vsekh gruppakh patsientov otmechena dostovernaia korreliatsiia mezhdu kontsentratsieĭ MCh i parametrami 1/lag-faza i Vmaks svertyvaniia plazmy. Poluchennye dannye pokazyvaiut, chto uvelichenie obshcheĭ koaguliatsionnoĭ aktivnosti MCh mozhet byt' ob"iasneno povysheniem ikh kontsentratsii tol'ko u patsientov s IM. Poskol'ku anti-TF antitela ne izmeniali parametry svertyvaniia v prisutstvii MCh kak zdorovykh donorov, tak i patsientov, TF, ochevidno, ne uchastvuet v realizatsii koaguliatsionnykh svoĭstv testiruemykh MCh. V to zhe vremia vo vsekh gruppakh patsientov nabliudali dostovernoe uvelichenie ékspressii FS na poverkhnosti MCh po sravneniiu so zdorovymi donorami, chto mozhet byt' odnoĭ iz prichin ikh povyshennoĭ koaguliatsionnoĭ aktivnosti.

Topics: Blood Coagulation; Cardiovascular Diseases; Cell-Derived Microparticles; Humans; Phosphatidylserines; Thromboplastin

Thrombin generation may be a potential tool to improve risk stratification for cardiovascular diseases. This study aims to explore the relation between thrombin generation and cardiovascular risk factors, cardiovascular diseases, and total mortality. For this study, N=5000 subjects from the population-based Gutenberg Health Study were analysed in a highly standardized setting. Thrombin generation was assessed by the Calibrated Automated Thrombogram method at 1 and 5 pM tissue factors trigger in platelet poor plasma. Lag time, endogenous thrombin potential, and peak height were derived from the thrombin generation curve. Sex-specific multivariable linear regression analysis adjusted for age, cardiovascular risk factors, cardiovascular diseases and therapy, was used to assess clinical determinants of thrombin generation. Cox regression models adjusted for age, sex, cardiovascular risk factors and vitamin K antagonists investigated the association between thrombin generation parameters and total mortality. Lag time was positively associated with obesity and dyslipidaemia for both sexes (p<0.0001). Obesity was also positively associated with endogenous thrombin potential in both sexes (p<0.0001) and peak height in males (1 pM tissue factor, p=0.0048) and females (p<0.0001). Cox regression models showed an increased mortality in individuals with lag time (1 pM tissue factor, hazard ratio=1.46, [95% CI: 1.07; 2.00], p=0.018) and endogenous thrombin potential (5 pM tissue factor, hazard ratio = 1.50, [1.06; 2.13], p=0.023) above the 95th percentile of the reference group, independent of the cardiovascular risk profile. This large-scale study demonstrates traditional cardiovascular risk factors, particularly obesity, as relevant determinants of thrombin generation. Lag time and endogenous thrombin potential were found as potentially relevant predictors of increased total mortality, which deserves further investigation.

Topics: Blood Coagulation Tests; Cardiovascular Diseases; Female; Humans; Male; Plasma; Thrombin; Thromboplastin

Topics: Adult; Anti-HIV Agents; Antigens, CD; Antigens, Differentiation, Myelomonocytic; Biomarkers; Blood Coagulation; Cardiovascular Diseases; Case-Control Studies; Cell Proliferation; Cross-Sectional Studies; Female; Forkhead Transcription Factors; HIV Infections; Humans; Lipids; Lymphocyte Activation; Macrophage Activation; Male; Matrix Attachment Region Binding Proteins; Membrane Proteins; Middle Aged; Monocytes; Receptors, Cell Surface; Risk Factors; T-Lymphocyte Subsets; Thromboplastin

Topics: Adult; Aged; Cardiovascular Diseases; Chronic Disease; Female; Humans; Male; Middle Aged; Migraine Disorders; Premenopause; Risk; Risk Factors; Thromboplastin

Plasma levels of the fibrinogen degradation product D-dimer are higher among African Americans (AAs) compared with those of European ancestry and higher among women compared with men. Among AAs, little is known of the genetic architecture of D-dimer or the relationship of D-dimer to incident cardiovascular disease.. These results highlight D-dimer as an important predictor of cardiovascular disease risk in AAs and suggest that sex-specific and African ancestral genetic effects of the

Topics: Adult; Aged; Aged, 80 and over; Biomarkers; Black or African American; Cardiovascular Diseases; Female; Fibrin Fibrinogen Degradation Products; Genetic Predisposition to Disease; Genome-Wide Association Study; Hemoglobins, Abnormal; Humans; Incidence; Male; Middle Aged; Molecular Epidemiology; Phenotype; Prognosis; Prospective Studies; Risk Assessment; Risk Factors; Sex Factors; Sickle Cell Trait; Thromboplastin; Time Factors; United States; Young Adult

During chronic kidney disease progression, kidney-specific risk factors for cardiovascular disease come into play. The present study investigated the impact of indoxyl sulfate, dietary tryptophan-derived uremic toxin, accumulated in the blood of patients with chronic kidney disease on hemostatic parameters, markers of inflammation, oxidative stress and monocyte to macrophage transition.. Fifty-one CKD patients not undergoing hemodialysis were enrolled in the study. Coagulation factors, fibrinolytic parameters, adhesion molecules, endothelial dysfunction markers, oxidative stress as well as inflammation markers were examined using immune-enzymatic method. Indoxyl sulfate levels were assessed using high-performance liquid chromatography. Biochemical parameters were determined by routine laboratory techniques using an automated analyzers. All assessed parameters were compared with controls and subjected to cross-sectional statistical analysis.. Elevated concentrations of indoxyl sulfate, the vast majority of parameters affecting hemostasis, and markers of renal insufficiency conditions were observed. Part of hemostatic factors, namely tissue factor, von Willebrand factor, thrombomodulin, soluble urokinase-type plasminogen activator receptor, soluble intercellular adhesion molecule-1, soluble vascular cell adhesion protein were correlated with the fraction of indoxyl sulfate. A significant quantity of assessed parameters showed strong correlations with superoxide-dismutase, renal insufficiency rate, C-reactive protein, and neopterin. Levels of indoxyl sulfate were independently associated with markers of impaired endothelial function (thrombomodulin, adhesion molecules), oxidative stress (superoxide-dismutase) and monocytes activation determinant (neopterin), which indicate unconventional links between these systems and the role of indoxyl sulfate. Furthermore, parameters that correlated with the levels of indoxyl sulfate (von Willebrand factor, soluble urokinase-type plasminogen activator receptor, soluble intercellular adhesion molecule-1) were positively associated with the prevalence of cardiovascular disease in a CKD patients.. The study demonstrated that in conditions of chronic exposure to uremic toxins, indoxyl sulfate seems to be one of the "missing links" between impaired renal function and prevalence of cardiovascular events, especially hemostatic disorders. The main functions of the action appear to be altered monocytes activation, intensified inflammatory process, and augmented oxidative stress by this uremic toxin.

Topics: Adult; Aged; C-Reactive Protein; Cardiovascular Diseases; Case-Control Studies; Female; Hemostasis; Humans; Hydrogen Peroxide; Indican; Inflammation; Intercellular Adhesion Molecule-1; Lipoproteins; Male; Middle Aged; Neopterin; Oxidative Stress; Peptide Fragments; Plasminogen Activator Inhibitor 1; Prevalence; Prothrombin; Renal Insufficiency, Chronic; Thrombin; Thromboplastin; Tissue Plasminogen Activator; Urokinase-Type Plasminogen Activator; Vascular Cell Adhesion Molecule-1; von Willebrand Factor

ESSENTIALS: Thrombosis is a major comorbidity in patients with chronic obstructive pulmonary disease (COPD). Roflumilast is a selective phosphodiesterase type-4 (PDE4) inhibitor approved for treatment of severe COPD. PDE4 blockade by roflumilast inhibits prothrombotic functions of neutrophils and monocytes. PDE4 inhibitors may reduce thrombotic risk in COPD as well as in other vascular diseases.. Roflumilast, an oral selective phosphodiesterase type 4 inhibitor, is approved for the treatment of severe chronic obstructive pulmonary disease (COPD). A recent meta-analysis of trials on COPD revealed that treatment with roflumilast was associated with a significant reduction in the rate of major cardiovascular events. The mechanisms of this effect remain unknown.. We tested the hypothesis that roflumilast N-oxide (RNO), the active metabolite of roflumilast, curbs the molecular mechanisms required for leukocyte-platelet (PLT) interactions and prevents the prothrombotic functions of polymorphonuclear leukocytes (PMNs) and monocytes (MNs).. Using well-characterized in vitro models, we analysed the effects of RNO on: (i) PMN adhesiveness; (ii) the release of neutrophil extracellular traps (NETs); and (iii) tissue factor expression in MNs. Key biochemical events underlying the inhibitory effects of RNO were defined.. In PMNs, RNO prevented phosphoinositide 3-kinase (PI3K)-dependent phosphorylation of Akt on Ser473, and Src family kinase (SFK)-mediated Pyk2 phosphorylation on Tyr579-580, while inducing protein kinase A-mediated phosphorylation of C-terminal Src kinase, the major negative regulator of SFKs. Modulation of these signaling pathways by RNO resulted in a significant impairment of PMN adhesion to activated PLTs or human umbilical vein endothelial cells, mainly mediated by inhibition of the adhesive function of Mac-1. Moreover RNO curbed SFK/PI3K-mediated NET release by PMNs adherent on fibrinogen-coated surfaces. In MNs interacting with activated PLTs, RNO curbed PI3K-mediated expression of tissue factor. The efficacy of RNO was significantly potentiated by formoterol, a long acting β-adrenergic receptor agonist. This study reveals novel antithrombotic activities by which roflumilast may exert protective effects against cardiovascular comorbodities in COPD.

Topics: Aminopyridines; Animals; Benzamides; Blood Platelets; Cardiovascular Diseases; Cyclic Nucleotide Phosphodiesterases, Type 4; Cyclopropanes; Extracellular Traps; Fibrinogen; Humans; Leukocytes; Macrophage-1 Antigen; Mice; Microscopy, Confocal; Monocytes; Neutrophils; P-Selectin; Phosphatidylinositol 3-Kinases; Phosphodiesterase 4 Inhibitors; Phosphorylation; Platelet Adhesiveness; Pulmonary Disease, Chronic Obstructive; Risk; Thromboplastin; Thrombosis

Tissue factor (TF) is the main in vivo initiator of the blood coagulation cascade. Active circulating TF was detected on small, negatively charged membrane vesicles, the so-called microvesicles (MVs), which are released upon cell activation and apoptosis from a variety of cells. Increased coagulation activation was found in morbidly obese patients, and elevated levels of TF-bearing MVs may contribute to the prothrombotic state in these patients.. To determine MV-associated TF activity levels in morbidly obese patients before and after weight loss due to bariatric surgery.. MV-TF activity was measured with a factor Xa generation assay in morbidly obese patients before and 2 years after bariatric surgery. In addition, clinical parameters were determined.. Seventy-four morbidly obese patients (mean age: 42 (±11) years; 61 females) were included in this study. After bariatric surgery, the body mass index decreased from (median, 25-75th percentile) 45.5 (42.3-50.2) to 30.5 (28.0-34.4 kg m(-2); P<0.001), and a significant improvement in metabolic parameters was observed. Preoperative MV-TF activity correlated with C-reactive protein levels (r=0.3; P=0.02). Postoperatively, the mean MV-TF activity decreased significantly from 0.20 pg ml(-1) (0.18-0.47) to 0.02 (0.00-0.28; P<0.01).. We could demonstrate a significant decrease in MV-TF activity after weight loss in morbidly obese patients. Decreased MV-TF activity might contribute to an improved coagulation profile in these patients after weight loss.

Topics: Adult; Austria; Bariatric Surgery; Biomarkers; Blood Coagulation; Body Mass Index; C-Reactive Protein; Cardiovascular Diseases; Down-Regulation; Female; Humans; Longitudinal Studies; Male; Middle Aged; Obesity, Morbid; Postoperative Period; Prospective Studies; Thromboplastin; Treatment Outcome; Weight Loss

Essentials Anti-activated factor X (Anti-Xa) monitoring is more precise than activated partial thromboplastin (aPTT). 20 804 hospitalized cardiovascular patients monitored with Anti-Xa or aPTT were analyzed. Adjusted transfusion rates were significantly lower for patients monitored with Anti-Xa. Adoption of Anti-Xa protocols could reduce transfusions among cardiovascular patients in the US.. Background Anticoagulant activated factor X protein (Anti-Xa) has been shown to be a more precise monitoring tool than activated partial thromboplastin time (aPTT) for patients receiving unfractionated heparin (UFH) anticoagulation therapy. Objectives To compare red blood cell (RBC) transfusions between patients receiving UFH who are monitored with Anti-Xa and those monitored with aPTT. Patients/Methods A retrospective cohort study was conducted on patients diagnosed with acute coronary syndrome (ACS) (N = 14 822), diagnosed with ischemic stroke (STK) (N = 1568) or with a principal diagnosis of venous thromboembolism (VTE) (N = 4414) in the MedAssets data from January 2009 to December 2013. Anti-Xa and aPTT groups were identified from hospital billing details, with both brand and generic name as search criteria. Propensity score techniques were used to match Anti-Xa cases to aPTT controls. RBC transfusions were identified from hospital billing data. Multivariable logistic regression was used to identify significant drivers of transfusions. Results Anti-Xa patients had fewer RBC transfusions than aPTT patients in the ACS population (difference 17.5%; 95% confidence interval [CI] 16.4-18.7%), the STK population (difference 8.2%; 95% CI 4.4-11.9%), and the VTE population (difference 4.7%; 95% CI 3.3-6.1%). After controlling for patient age and gender, diagnostic risks (e.g. anemia, renal insufficiency, and trauma), and invasive procedures (e.g. cardiac catheterization, hemodialysis, and coronary artery bypass graft), Anti-Xa patients were less likely to have a transfusion while hospitalized for ACS (odds ratio [OR] 0.16, 95% CI 0.14-0.18), STK (OR 0.41, 95% CI 0.29-0.57), and VTE (OR 0.35, 95% CI 0.26-0.48). Conclusion Anti-Xa monitoring was associated with a significant reduction in RBC transfusions as compared with aPTT monitoring alone.

Topics: Adolescent; Adult; Aged; Aged, 80 and over; Algorithms; Anticoagulants; Blood Transfusion; Cardiovascular Diseases; Comorbidity; Drug Monitoring; Erythrocyte Transfusion; Erythrocytes; Factor Xa; Factor Xa Inhibitors; Female; Heparin; Hospitalization; Humans; Infusions, Intravenous; Male; Middle Aged; Multivariate Analysis; Odds Ratio; Partial Thromboplastin Time; Propensity Score; Prospective Studies; Retrospective Studies; Thromboplastin; Young Adult

We previously reported that high levels of tissue factor (TF) can induce cellular apoptosis in endothelial cells. In this study, TF-mediated mechanisms of induction of apoptosis were explored. Endothelial cells were transfected to express wild-type TF. Additionally, cells were transfected to express Asp253-substituted, or Ala253-substitued TF to enhance or prevent TF release, respectively. Alternatively, cells were pre-incubated with TF-rich and TF-poor microvesicles. Cell proliferation, apoptosis and the expression of cyclin D1, p53, bax and p21 were measured following activation of cells with PAR2-agonist peptide. Greatest levels of cell proliferation and cyclin D1 expression were observed in cells expressing wild-type or Asp253-substituted TF. In contrast, increased cellular apoptosis was observed in cells expressing Ala253-substituted TF, or cells pre-incubated with TF-rich microvesicles. The level of p53 protein, p53-phosphorylation at ser33, p53 nuclear localisation and transcriptional activity, but not p53 mRNA, were increased in cells expressing wild-type and Ala253-substituted TF, or in cells pre-incubated with TF-rich microvesicles. However, the expression of bax and p21 mRNA, and Bax protein were only increased in cells pre-incubated with TF-rich microvesicle and in cells expressing Ala253-substituted TF. Inhibition of the transcriptional activity of p53 using pifithrin-α suppressed the expression of Bax. Finally, siRNA-mediated suppression of p38α, or inhibition using SB202190 significantly reduced the p53 protein levels, p53 nuclear localisation and transcriptional activity, suppressed Bax expression and prevented cellular apoptosis. In conclusion, accumulation of TF within endothelial cells, or sequestered from the surrounding can induce cellular apoptosis through mechanisms mediated by p38, and involves the stabilisation of p53.

Topics: Amino Acid Substitution; Apoptosis; bcl-2-Associated X Protein; Breast Neoplasms; Cardiovascular Diseases; Cell Line, Tumor; Cell-Derived Microparticles; Cells, Cultured; Coronary Vessels; Cyclin D1; Endothelial Cells; Enzyme Activation; Female; Gene Expression Regulation; Genes, Reporter; Humans; Imidazoles; MAP Kinase Signaling System; Oligopeptides; p38 Mitogen-Activated Protein Kinases; Pyridines; Recombinant Fusion Proteins; RNA Interference; RNA, Messenger; RNA, Small Interfering; Thromboplastin; Transfection; Tumor Suppressor Protein p53

Circulating microparticles (cMPs) play important roles in cellular crosstalk and are messengers of cell activation. We have previously reported that platelet-released microparticles (pMPs) stimulate thrombosis and that lipid-lowering treatment as per guidelines in patients with familial hypercholesterolaemia (FH) is not sufficiently effective in reducing pro-inflammatory cell activation and, consequently, CD45+/CD3+-lymphocyte-derived cMP shedding. FH patients, due to life-long vascular exposure to high LDL-cholesterol levels, are at high cardiovascular risk (HCVR) and develop premature coronary artery disease. Our objectives were to investigate a) whether patients with HCVR have cMPs with a prothrombotic phenotype, and b) whether patients with magnetic resonance imaging (MRI) evidence of lipid-rich atherosclerotic lesions have a specific cMP profile regarding prothrombotic protein cargos. cMPs were isolated from HCVR-patients and from age/gender/treatment-matched control patients. cMP phenotype was characterised by triple-labelling flow cytometry. HCVR--patients have higher numbers of pMPs derived from activated platelets as well as of tissue factor-rich microparticles (TF+-cMPs) than controls (P< 0.0001). TF+-cMPs showed procoagulant activity, which associate with atherosclerotic plaque burden, indicating that TF in the cMPs is functional. In HCVR-patients, overall TF+-cMPs (monocyte-derived [CD142+/CD14+] and platelet-derived [CD142+/TSP1+]) and activated pMPs directly correlate with MRI-detected lipid-rich atherosclerotic plaques while inversely correlate with MRI-detected calcified plaques. C-statistics analysis showed that prothrombotic cMPs add significant prognostic value to a risk factor model for the prediction of lipid-rich plaques. In conclusion, the activation status of blood cells in HCVR-patients differed markedly from controls as shown by higher circulating levels of prothrombotic and TF+-cMPs. Prothrombotic cMP numbers identify subclinical atherosclerotic plaque burden.

Topics: Age Factors; Aorta, Thoracic; Aortic Diseases; Asymptomatic Diseases; Biomarkers; Blood Platelets; Calcinosis; Cardiovascular Diseases; Case-Control Studies; Cell-Derived Microparticles; Cohort Studies; Female; Heterozygote; Humans; Hypercholesterolemia; Hyperlipoproteinemia Type II; Lipids; Magnetic Resonance Imaging; Male; Middle Aged; Phenotype; Plaque, Atherosclerotic; Platelet Activation; Risk Factors; Thromboplastin; Thrombospondin 1

Systemic Lupus Erythematosus (SLE) and antiphospholipid syndrome (APS) are associated with a high prevalence of atherosclerosis. β2 glycoprotein I (β2GPI) represents a link between autoimmunity and endothelial dysfunction. Recently, β2GPI reactive T cells have been identified; however, their role in atherosclerosis is still under investigation. We evaluated early atherosclerosis in patients with SLE and APS and investigated T cell reactivity to β2GPI and its relationship with atherosclerotic process.. Fifty SLE, 18 patients with primary APS (PAPS), and 25 healthy controls were enrolled. Demographic and clinical data, including traditional cardiovascular risk factors, were recorded. Monocyte β2GPI and Tissue Factor (TF) expression and peripheral blood mononuclear cell response to β2GPI stimulation were evaluated. Doppler ultrasound was performed to investigate flow-mediated dilatation (FMD) and carotid intima-media thickness (IMT). We detected an increase in mean IMT and a decrease in FMD in patients with SLE versus controls (P<0.05 and P=0.0001, respectively) and a decrease in FMD in patients with PAPS versus controls (P<0.05). Monocyte β2GPI and TF expression was higher in patients with SLE and PAPS than in controls (P=0.006 and P=0.001, respectively); no correlation of monocyte β2GPI and TF with IMT or FMD was detected. β2GPI induced peripheral blood mononuclear cell proliferation in 32% of patients with SLE, 25% of patients with PAPS yet in none of the controls. Proliferative response to β2GPI correlated with a history of arterial thrombosis, thrombocytopenia, and IMT >0.9 mm.. A significant percentage of patients with SLE and PAPS show a β2GPI-specific T cell reactivity, which is associated with subclinical atherosclerosis.

Topics: Adult; Aged; Antibody Specificity; Antiphospholipid Syndrome; Atherosclerosis; Autoantibodies; Autoantigens; beta 2-Glycoprotein I; Blood Pressure; Cardiovascular Diseases; Carotid Intima-Media Thickness; Cell Division; Endothelium, Vascular; Female; Hemorheology; Humans; Interferon-gamma Release Tests; Lipoproteins, HDL; Lupus Erythematosus, Systemic; Male; Middle Aged; Monocytes; Risk Factors; T-Cell Antigen Receptor Specificity; Thromboplastin; Vasodilation; Young Adult

Obesity is a risk factor for both cardiovascular disease and cancer development. Leptin, a cytokine produced by adipose tissue, controls different processes in peripheral tissues, including cancer development and thrombotic disorders in patients with a variety of clinical disorders. Tissue factor (TF), the trigger of blood clotting, is abundant in the adipose tissue. Since TF, often expressed by cancer cells, is considered a hallmark of cancer progression, we investigated whether leptin could modulate TF in the human metastatic breast carcinoma cell line MCF-7.. MCF-7 cells were incubated with or without the different reagents at 37 °C. At the end of incubation, cells were tested for procoagulant activity by a one-stage clotting assay, TF and TNF-α antigen levels and mRNA by ELISA and real-time RT-PCR, respectively. Leptin receptor was studied by FACS.. Both TF activity and antigen constitutively expressed by MCF-7 were significantly increased by leptin in a dose-dependent fashion. TF mRNA levels were also enhanced indicating that leptin exerts its effect at the transcription level. The effect of leptin was specific and required binding to its receptor (Ob-R), which was found on the surface of the cells, since antibodies against leptin and Ob-R completely prevented TF expression upregulation. In addition, leptin enhanced both TNF-α mRNA synthesis and secretion from MCF7. An anti-TNF-α MoAb completely abolished the leptin-induced TF expression.. These data support the hypothesis that leptin, by its upregulation of TF, possibly mediated by TNF-α synthesis, may contribute to processes underlying both cancer and vascular cell disorders.

Topics: Breast Neoplasms; Cardiovascular Diseases; Cell Line, Tumor; Female; Humans; Leptin; Obesity; Receptors, Leptin; Thromboplastin; Up-Regulation

In patients with chronic renal failure (CRF), cardiovascular disease is the leading cause of increased morbidity and mortality. We hypothesized a role for endothelial activation and microparticle (MP) numbers and procoagulant activity in the pre-thrombotic state of these patients.. We analysed blood samples of 27 patients with CRF [8 chronic kidney disease Stage 4 (CKD4), 9 peritoneal dialysis (PD) and 10 haemodialysis (HD), samples taken before and after HD] and 10 controls. Degree and nature of endothelial activation were assessed by measuring mature von Willebrand factor (vWF) and vWF propeptide levels. Cellular MPs were characterized by flow cytometry and MP-specific thrombin generation (TG) measurements.. CRF was accompanied by chronic (CKD4 and PD) or acute (HD) endothelial activation. Patients with CRF had substantially higher MP numbers than controls (median 9400 versus 4350×10(6)/L, P=0.001), without significant differences between the treatment subgroups or between pre- and post-HD. The vast majority of MPs were platelet derived. Of the minor populations, endothelial MPs and tissue factor-bearing MPs were more abundant in CRF. MPs were procoagulant, but the increase in numbers was not reflected in a proportional increase in MP-specific TG.. Renal failure is accompanied by endothelial activation of a different nature in CKD4 and PD patients compared to HD patients, and results in all subgroups in an increase of mainly platelet-derived MPs that appear to be less procoagulant than in other disease states, possibly because of the uraemic functional defect of their cellular source.

Topics: Adult; Aged; Aged, 80 and over; Cardiovascular Diseases; Case-Control Studies; Cell-Derived Microparticles; Cross-Sectional Studies; Endothelium, Vascular; Female; Flow Cytometry; Humans; Kidney Failure, Chronic; Male; Middle Aged; Prognosis; Renal Dialysis; Risk Factors; Thrombin; Thromboplastin; von Willebrand Factor; Young Adult

To determine tissue factor procoagulant activity (TF-PCA) in patients with type 1 diabetes and to examine effects of hyperglycemia and hyperglycemia plus hyperinsulinemia on TF-PCA.. We have determined circulating TF-PCA and other coagulation factors under basal (hyperglycemic) conditions, after acute correction of hyperglycemia, in response to 24 h of selective hyperglycemia, and in response to 24 h of hyperglycemia plus hyperinsulinemia in nine type 1 diabetic patients and in seven nondiabetic control subjects.. As shown previously in patients with type 2 diabetes, basal TF-PCA and plasma coagulation factor VIIa (FVIIa) were higher in patients with type 1 diabetes than in nondiabetic control subjects. However, in contrast with type 2 diabetes, normalizing glucose did not decrease the elevated TF-PCA levels, and raising glucose or glucose plus insulin levels did not increase TF-PCA.. Patients with type 1 diabetes have elevated circulating TF-PCA and FVIIa levels and are in a procoagulant state that may predispose them to acute cardiovascular events. The mechanisms regulating TF-PCA in patients with type 1 and type 2 diabetes are different and should be further explored.

Topics: Adult; Analysis of Variance; Blood Coagulation; Cardiovascular Diseases; Diabetes Mellitus, Type 1; Diabetic Angiopathies; Factor VIIa; Female; Glucose Clamp Technique; Humans; Hyperglycemia; Hyperinsulinism; Male; Middle Aged; Thromboplastin; Young Adult

The metabolic syndrome (MetS) has been found to be closely related with thrombotic diseases. The mechanism, however, is far from elucidated.. This study was designed to investigate the relationship between endogenous resistin and thrombosis mediating factors, as well as its potential effects on the gene expression of cardiovascular disease biomarkers.. Ninety patients satisfied the MetS criteria, and 55 healthy subjects were recruited as part of a single-center clinical study. Plasma levels of resistin, tissue factor (TF), tissue factor pathway inhibitor (TFPI), tissue plasminogen activator (tPA), plasminogen activator inhibitor-1 (PAI-1) were measured by enzymelinked immunosorbent assays. The effect of resistin on the expression of cardiovascular disease biomarkers in human umbilical vein endothelial cells (HUVEC) was assayed by gene microarray.. 1) The average levels of resistin in MetS patients with or without acute myocardial or cerebral infarction were significantly higher than those of the controls. 2) The TF and TFPI increase was higher in MetS with infarction patients than in MetS patients. 3) In MetS with infarction patients, resistin was positively correlated with TF and PAI-1 (r=0.313, p=0.008; r=0.401, p=0.002, respectively). 4) In HUVEC, the microarray showed that apolipoprotein C-I, ACE, tumor necrosis factor receptor superfamily member 1A (TNFRSF1A) and member 5 (CD40) genes expression were dramatically increased by resistin.. In patients with MetS, resistin is strongly associated with hypercoagulative and hypofibrinolitic activities. Moreover, resistin may induce thrombotic complications via mediating the lipoprotein metabolism and stimulating inflammation.

Topics: Aged; Asian People; Biomarkers; Cardiovascular Diseases; Cells, Cultured; Endothelial Cells; Female; Humans; Lipoproteins; Male; Metabolic Syndrome; Microarray Analysis; Middle Aged; Plasminogen Activator Inhibitor 1; Resistin; Thromboplastin; Thrombosis; Tissue Plasminogen Activator

Topics: Cardiovascular Diseases; Factor VII; Factor VIIa; Humans; Plaque, Atherosclerotic; Thrombophilia; Thromboplastin; Thrombosis

The haemostatic and biochemical abnormalities participate in the progression of cardiovascular disease (CVD) in peritoneally dialysed (PD) patients. Recently, the role of kynurenine (KYN) pathway of tryptophan (TRP) degradation in the development of CVD has been postulated.. The present study was undertaken to investigate haemostatic parameters, biochemical profiles and kynurenines in PD patients both with and without CVD compared to age- and sex-matched healthy controls.. The multiple biochemical abnormalities were present in PD patients, particularly in those with CVD. Tissue factor (TF), its inhibitor (TFPI), prothrombin fragment 1+2 (F(1+2)), urokinase-type plasminogen activator (uPA), its soluble receptor (suPAR), plasmin/antiplasmin (PAP) complexes, KYN, kynurenic (KYNA) and quinolinic (QA) acids levels were significantly higher, whereas TRP was significantly lower in the PD patients than in the controls. Tissue-type plasminogen activator (tPA) and plasminogen activator inhibitor-1 (PAI-1) were higher in the patients with CVD than in the patients without CVD and controls. PD patients with CVD had higher F(1+2), and they had lower suPAR and KYNA levels compared with PD patients without CVD. KYNA was positively associated with TFPI, whereas its was inversely associated with F(1+2) both in the whole PD group and in CVD patients. Logistic regression analysis showed that low KYNA, high glucose, low HDL-cholesterol levels and the duration of dialysis treatment were independently associated with the presence of CVD in PD patients.. The present study suggests a relationship between kynurenine pathway of tryptophan degradation, haemostatic and biochemical disturbances and CVD prevalence in peritoneally dialyzed patients.

Topics: Adult; Antifibrinolytic Agents; Cardiovascular Diseases; Case-Control Studies; Cross-Sectional Studies; Fasting; Female; Fibrinolysin; Hemostasis; Humans; Kynurenic Acid; Kynurenine; Male; Middle Aged; Peptide Fragments; Peritoneal Dialysis, Continuous Ambulatory; Plasminogen Activator Inhibitor 1; Poland; Prevalence; Prothrombin; Quinolinic Acid; Receptors, Urokinase Plasminogen Activator; Thromboplastin; Tryptophan; Urokinase-Type Plasminogen Activator

Tissue factor (TF), key initiator of coagulation, has been ascribed roles not only in thrombosis but also in atherosclerosis. TF gene promoter haplotypes modulate TF expression, thereby potentially affecting atherosclerosis. The objective of this study was to evaluate functionally relevant TF promoter haplotypes as determinants of carotid intima-media thickness (C-IMT), a marker of atherosclerosis.. The haplotype-tagging TF A-603G polymorphism and C-IMT were determined in 611 subjects referred to cardiovascular risk prevention. Subjects were aged 59.7 (58.1-61.1) years (mean (95% C.I.)), 79% were male, and 74% in secondary prevention with a history of coronary, cerebrovascular, or peripheral atherosclerotic disease. TF plasma levels were measured in 120 subjects.. Significant dose-dependent associations were found between A-603G genotype and both IMT(max) and IMT(mean-max) after adjusting for potential confounders. IMT values were highest in A/A (n=173), intermediate in A/G (n=312) and lowest in G/G (n=126). IMT(max) values (average and 95% C.I., in mm) for A/A, A/G and G/G, were 2.22 (2.11-2.34), 2.09 (2.01-2.18) and 2.02 (1.90-2.15), respectively (p(trend)=0.019), and IMT(mean-max) values were 1.28 (1.23-1.32), 1.25 (1.22-1.28) and 1.21 (1.16-1.26), respectively (p(trend)=0.041). A significant interaction between A-603G genotype and prevention status was found (p=0.046 and p=0.042 for IMT(max) and IMT(mean-max), respectively). TF plasma levels did not differ between genotypes and were not determinants of C-IMT.. The haplotype-tagging TF A-603G polymorphism is associated with C-IMT, independently of TF levels in plasma. These findings provide clinical evidence of an involvement of TF in atherosclerosis, in addition to its well-known roles in hemostasis and thrombosis.

Topics: Cardiovascular Diseases; Carotid Arteries; Carotid Artery Diseases; Female; Gene Frequency; Genetic Predisposition to Disease; Haplotypes; Heterozygote; Homozygote; Humans; Linear Models; Male; Middle Aged; Phenotype; Polymorphism, Single Nucleotide; Promoter Regions, Genetic; Risk Assessment; Risk Factors; Secondary Prevention; Thromboplastin; Tunica Intima; Tunica Media; Ultrasonography

Topics: Asian People; Cardiovascular Diseases; Gene Frequency; Genetic Predisposition to Disease; Haplotypes; Humans; Japan; Polymorphism, Genetic; Promoter Regions, Genetic; Risk Factors; Thromboplastin

Antiphospholipid (aPL) antibodies recognize receptor-bound beta(2) glycoprotein I (beta(2)GPI) on target cells, and induce an intracellular signaling and a procoagulant/proinflammatory phenotype that leads to thrombosis. Evidence indicates that annexin A2 (A2), a receptor for tissue plasminogen activator and plasminogen, binds beta(2)GPI on target cells. However, whether A2 mediates pathogenic effects of aPL antibodies in vivo is unknown. In this work, we studied the effects of human aPL antibodies in A2-deficient (A2(-/-)) mice. A2(-/-) and A2(+/+) mice were injected with immunoglobulin G (IgG) isolated from either a patient with antiphospholipid syndrome (IgG-APS), a healthy control subject (IgG-normal human serum), a monoclonal anti-beta(2)GPI antibody (4C5), an anti-A2 monoclonal antibody, or monoclonal antibody of irrelevant specificity as control. We found that, after IgG-APS or 4C5 injections and vascular injury, mean thrombus size was significantly smaller and tissue factor activity was significantly less in A2(-/-) mice compared with A2(+/+) mice. The expression of vascular cell adhesion molecule-1 induced by IgG-APS or 4C5 in explanted A2(-/-) aorta was also significantly reduced compared with A2(+/+) mice. Interestingly, anti-A2 monoclonal antibody significantly decreased aPL-induced expression of intercellular cell adhesion molecule-1, E-selectin, and tissue factor activity on cultured endothelial cells. Together, these data indicate for the first time that A2 mediates the pathogenic effects of aPL antibodies in vivo and in vitro APS.

Topics: Animals; Annexin A2; Antibodies, Antiphospholipid; Antiphospholipid Syndrome; Aorta; Blotting, Western; Cardiovascular Diseases; Carotid Arteries; Cells, Cultured; E-Selectin; Endothelium, Vascular; Enzyme-Linked Immunosorbent Assay; Humans; Immunoglobulin G; In Vitro Techniques; Intercellular Adhesion Molecule-1; Male; Mice; Mice, Inbred C57BL; Mice, Knockout; Middle Aged; Thromboplastin; Umbilical Veins; Vascular Cell Adhesion Molecule-1

Accumulating evidence suggests that cyclooxygenase-2 (COX-2) and prostaglandin E(2) (PGE(2)) may play an important role in colon carcinogenesis. Thus, blockage of this pathway may be a suitable strategy for colon cancer chemoprevention. Recent clinical studies suggest that COX-2 inhibitors cause adverse cardiovascular effects due to prostacyclin (PGI(2)) inhibition. To test our hypothesis that inhibition of PGE(2) signaling through E-prostanoid (EP) receptors may offer a safer cardiovascular profile than COX-2 inhibition, we analyzed expression of 6-keto PGF(1alpha), a hydrated form of PGI(2) and PGI(2) synthase, which was stimulated with cytokines in human umbilical vein endothelial cells (HUVECs) treated with the EP(1) receptor antagonist ONO-8711 or the COX-2 inhibitor celecoxib. ONO-8711 did not inhibit both 6-keto PGF(1alpha) production and PGIS expression, whereas celecoxib did in HUVECs. ONO-8711 also inhibited cytokine-induced tissue factor expression in HUVECs. These results suggest that ONO-8711 may be a safer chemopreventive agent with respect to cardiovascular events.

Topics: Blood Coagulation; Bridged Bicyclo Compounds; Caproates; Cardiovascular Diseases; Celecoxib; Cell Line; Cytokines; Dinoprostone; Endothelial Cells; Epoprostenol; Fibrinolytic Agents; Gene Expression Regulation; Humans; Peroxisome Proliferator-Activated Receptors; Pyrazoles; Receptors, Prostaglandin E; Response Elements; Signal Transduction; Sulfonamides; Thromboplastin

We examined the impact of C-reactive protein (CRP) on vascular smooth muscle cell (VSMC) expression of tissue factor (TF) and TF pathway inhibitor (TFPI).. TF mRNA, protein, and activity levels were significantly higher in VSMCs isolated from CRP-transgenic (Tg) mice than from wild-type (WT) mice. TFPI expression was significantly downregulated in CRP-Tg versus WT VSMCs. Transfection of human VSMCs with CRP expression plasmid significantly increased TF expression and decreased TFPI expression. Gene silencing of Fc gamma receptor IIIa (Fc gammaRIIIa) blocked the effect of CRP on VSMC TF expression. CRP activated p44/42, but not p38 or JNK MAP kinase (MAPK), and the effect of CRP on TF expression was blocked by pharmacological inhibitor of p44/42, but not p38 or JNK MAPK. Reactive oxygen species (ROS) scavengers blocked CRP-induced upregulation of VSMC TF expression. In vivo analyses revealed significant increases in TF expression and decreases in TFPI expression in carotid arteries of CRP-Tg mice versus WT mice.. CRP increases TF and decreases TFPI expression by VSMCs in vitro and in vivo. Induction of TF expression by CRP is mediated by Fc gammaRIIIa, p44/42 MAPK, and ROS generation. These data offer important insights into the role of CRP in the pathogenesis of arterial thrombosis.

Topics: Animals; Base Sequence; C-Reactive Protein; Cardiovascular Diseases; Carotid Arteries; DNA Primers; Humans; Lipoproteins; Male; MAP Kinase Signaling System; Mice; Mice, Congenic; Mice, Inbred C57BL; Mice, Transgenic; Muscle, Smooth, Vascular; Myocytes, Smooth Muscle; Oxidative Stress; Receptors, IgG; RNA, Small Interfering; Thromboplastin; Up-Regulation

Human urotensin II is an 11-aminoacid peptide with a controversial role in the human cardiovascular system. Indeed, urotensin effects on vascular reactivity and in heart failure are well documented, while its potential role in the pathophysiology of athero-thrombosis is still unknown. This study investigates the effects of urotensin on tissue factor (TF) and VCAM-1/ICAM-1 expression in human coronary endothelial cells (HCAECs).. Urotensin induced TF-mRNA transcription as demonstrated by real time PCR and expression of TF that was functionally active as demonstrated by procoagulant activity assay. In addition, urotensin induced expression of VCAM-1 and ICAM-1 as demonstrated by FACS analysis. VCAM-1 and ICAM-1 were functionally active because they increased leukocyte adhesivity to HCAECs. Urotensin-induced expression of TF and of VCAM-1/ICAM-1 was mediated through the Rho A-activation of the transcription factor, NF-kappaB, as demonstrated by EMSA. Indeed, lovastatin, an HMG-CoA reductase inhibitor, by modulating the Rho activation, and NF-kappaB inhibitors, suppressed the urotensin effects on TF and CAMs.. Data of the present study, although in vitro, describe the close relationship existing between urotensin II and athero-thrombosis, providing for the first time support for the view that this peptide might have not only vasoactive functions but it might be an effector molecule able to induce a pro-atherothrombotic phenotype in cells of the coronary circulation. Although future studies are required to clarify whether these mechanisms are also important in the clinical setting, this report supports an emerging new role for urotensin II in the pathogenesis and progression of cardiovascular disease.

Topics: Cardiovascular Diseases; Cell Adhesion; Cell Adhesion Molecules; Coronary Vessels; Endothelial Cells; Endothelium, Vascular; Gene Expression Regulation; Humans; Intercellular Adhesion Molecule-1; Leukocytes; RNA, Messenger; Thromboplastin; Urotensins; Vascular Cell Adhesion Molecule-1

An increased risk of death or severe injury due to late-morning thrombotic events is well established. Tissue factor (TF) is the initiator of the coagulation cascade, and endothelial stresses, coupled with production of pro-coagulant microparticles (MP) are also important factors in loss of haemostasis. TF and vascular cell adhesion molecule-1 (VCAM-1) -positive cell microparticles were assessed periodically over a 24-hour (h) period in healthy human subjects to ascertain if they followed a circadian rhythm. Eleven healthy male subjects were assessed in a temperature-controlled environment with dietary intake consistent between subjects. Blood samples were taken every 4 h by venipuncture, and TF and VCAM-1 positive microparticles were quantified by flow cytometry. A significant circadian rhythm was observed in VCAM-1 MP (p=or<0.0001), and a trend was shown, although not statistically significant (p=0.065) in TF microparticles. A peak was observed at 9 a.m. for VCAM-1 positive MP, followed by a decrease and subsequent peak at 9 p.m. and a minimum at 5 a.m. TF-positive MP followed a strikingly similar trend in both variation and absolute numbers with a delay. A circadian rhythm was observed in VCAM-1 and less so TF-positive MP. This has significant implications in terms of the well known increased risk of cardiovascular thrombotic events matching this data. To our knowledge this is the first such report of quantified measurements of these MP over a 24-h period and the only measurement of a 24-h variation of in-vivo blood-borne TF.

Topics: Adolescent; Adult; Biomarkers; Cardiovascular Diseases; Cell Membrane Structures; Circadian Rhythm; Enzyme-Linked Immunosorbent Assay; Flow Cytometry; Humans; Male; Reference Values; Thromboplastin; Vascular Cell Adhesion Molecule-1

Tissue factor (TF) and its specific inhibitor, tissue factor pathway inhibitor (TFPI), are important contributors to the initiation of the coagulation process.. To compare plasma levels of soluble TF (sTF) and free-TFPI (f-TFPI) between patients with stable angina pectoris (SAP) and acute coronary syndrome (ACS) and to assess the impact of the two variables on long-term prognosis.. Patients with SAPs (n = 1146) and acute coronary syndrome (n = 523) from the AtheroGene study were included and followed for 2.3 years. Because of the strong impact of unfractionated heparin (UFH) on f-TFPI levels, but not on sTF levels, patients having received UFH before blood drawing were excluded from the analyses on f-TFPI (n = 226).. On admission, no significant differences in sTF levels were observed between SAP and ACS patients. By comparison to patients with stable angina, f-TFPI levels significantly increased in patients with acute unstable angina and further increased in patients presenting with non-ST-elevation myocardial infarction and ST-elevation myocardial infarction (P < 10(-4)). Among the 1669 individuals with a coronary artery disease, 56 died from a cardiovascular cause. In prospective analyses, high sTF levels were independently associated with an increased risk of cardiovascular death in individuals with ACS (fully adjusted hazard ratio associated with one quartile increase = 2.06; 95% confidence interval 1.24-3.45; P = 0.006) but not in those with SAP (hazard ratio = 1.07; 95% confidence interval 0.78-1.46; P = 0.67). In SAP and ACS patients, high f-TFPI levels were not independently associated with an increased risk of cardiovascular death.. Plasma sTF levels were predictive of cardiovascular mortality in individuals with ACS, whereas f-TFPI levels were associated with the severity of myocardial damage on admission but were not independently related to outcome.

Topics: Aged; Angina Pectoris; Biomarkers; Cardiovascular Diseases; Cohort Studies; Coronary Stenosis; Female; Follow-Up Studies; Germany; Humans; Lipoproteins; Male; Middle Aged; Myocardial Infarction; Predictive Value of Tests; Prognosis; Proportional Hazards Models; Prospective Studies; Risk Assessment; Severity of Illness Index; Syndrome; Thromboplastin; Time Factors

Topics: Angina Pectoris; Biomarkers; Blood Coagulation; Cardiovascular Diseases; Coronary Artery Disease; Humans; Lipoproteins; Myocardial Ischemia; Predictive Value of Tests; Prognosis; Risk Assessment; Severity of Illness Index; Thromboplastin

The disturbances of haemostasis and enhanced oxidative stress (SOX) appear to contribute to the cardiovascular disease (CVD) in hemodialysis (HD) patients. The aim of the present study was to investigate if the disorders of coagulation/fibrinolysis system are associated with the presence of CVD in these patients.. We compared pre-dialysis levels of uPA, suPAR, tissue factor (TF) and its inhibitor (TFPI), prothrombin fragment F1+2 (F1+2); a marker of SOX-Cu/Zn superoxide dismutase (Cu/Zn SOD) and a surrogate of inflammation-high sensitivity C-reactive protein (hs CRP) in HD patients with and without CVD.. The uPA/suPAR system and hs CRP values were significantly greater in patients with CVD than in those without CVD; whereas TF, TFPI, F1+2 and Cu/Zn SOD levels were comparable in both patient groups. TF was positively correlated with both uPA (p<0.001) and suPAR levels (p<0.05). Logistic regression analysis showed that elevated levels of suPAR, TF and uPA were independently associated with the presence of CVD in HD patients.. The association between TF and uPA/suPAR system is significantly related to the presence of CVD in HD patients.

Topics: Adult; Aged; Biomarkers; C-Reactive Protein; Cardiovascular Diseases; Female; Fibrinolysis; Hemostasis; Humans; Kidney Failure, Chronic; Lipoproteins; Logistic Models; Male; Middle Aged; Oxidative Stress; Peptide Fragments; Prothrombin; Renal Dialysis; Superoxide Dismutase; Thromboplastin; Urokinase-Type Plasminogen Activator

Obesity is a major modifiable risk factor for cardiovascular disease. Leptin, the hormone synthesized and released primarily by adipose tissue and found increased in obese individuals, has been implicated in the regulation of inflammation and arterial and venous thrombosis.. To investigate the role of tissue factor (TF), the pivotal agonist of the clotting cascade, as a link between obesity and cardiovascular disease.. In 15 obese patients, plasma levels of leptin and TF as well as TF expression in resting and endotoxin-stimulated mononuclear leukocytes (MN) were increased when compared with healthy donors. In a selected sample of obese patients, loss of body weight led to decreased circulating leptin levels, accompanied by a reduction in plasma TF as well as in TF expression, both in resting and endotoxin-stimulated MN. In subsequent in vitro experiments, leptin was incubated with MN from healthy subjects. Leptin induced TF activity and antigen in a dose-dependent fashion, as assessed by clotting assay and ELISA, respectively. Increased migration of c-Rel/p65 into the nucleus, as determined by EMSA, and development of TF mRNA in monocytes, as assessed by RT-PCR, were observed. Experiments with mitogen-activated protein kinase (MAPK) inhibitors, indicated the involvement of p38 and ERK1/2 pathways.. The presence of TF-expressing MN in blood from obese subjects and the in vitro induction of TF by pharmacologic concentrations of leptin in MN from healthy subjects suggest that TF expression by leptin-stimulated monocytes may contribute to the cardiovascular risk associated with obesity.

Topics: Cardiovascular Diseases; Dimerization; Gene Expression Regulation; Humans; In Vitro Techniques; Leptin; Leukocytes, Mononuclear; Obesity; Proto-Oncogene Proteins c-rel; RNA, Messenger; Thromboplastin; Transcription Factor RelA

The oxidative modification of LDL may play an important role in the early events of atherogenesis. Thus the identification of antioxidative compounds may be of therapeutic and prophylactic importance regarding cardiovascular disease. Copper-chlorophyllin (Cu-CHL), a Cu(2+)-protoporphyrin IX complex, has been reported to inhibit lipid oxidation in biological membranes and liposomes. Hemin (Fe(3+)-protoporphyrin IX) has been shown to bind to LDL thereby inducing lipid peroxidation. As Cu-CHL has a similar structure as hemin, one may assume that Cu-CHL may compete with the hemin action on LDL. Therefore, in the present study Cu-CHL and the related compound magnesium-chlorophyllin (Mg-CHL) were examined in their ability to inhibit LDL oxidation initiated by hemin and other LDL oxidizing systems. LDL oxidation by hemin in presence of H(2)O(2) was strongly inhibited by both CHLs. Both chlorophyllins were also capable of effectively inhibiting LDL oxidation initiated by transition metal ions (Cu(2+)), human umbilical vein endothelial cells (HUVEC) and tyrosyl radicals generated by myeloperoxidase (MPO) in presence of H(2)O(2) and tyrosine. Cu- and Mg-CHL showed radical scavenging ability as demonstrated by the diphenylpicrylhydracylradical (DPPH)-radical assay and estimation of phenoxyl radical generated diphenyl (dityrosine) formation. As assessed by ultracentrifugation the chlorophyllins were found to bind to LDL (and HDL) in serum. The present study shows that copper chlorophyllin (Cu-CHL) and its magnesium analog could act as potent antagonists of atherogenic LDL modification induced by various oxidative stimuli. As inhibitory effects of the CHLs were found at concentrations as low as 1 mumol/l, which can be achieved in humans, the results may be physiologically/therapeutically relevant.

Topics: Atherosclerosis; Biphenyl Compounds; Cardiovascular Diseases; Catalysis; Cells, Cultured; Chlorophyll; Chlorophyllides; Copper; Dose-Response Relationship, Drug; Endothelium, Vascular; Free Radical Scavengers; Free Radicals; Hemin; Humans; Hydrazines; Hydrogen Peroxide; Ions; Iron; Lipid Peroxidation; Lipids; Lipoproteins; Lipoproteins, LDL; Magnesium; Malondialdehyde; Models, Chemical; Octanols; Oxygen; Picrates; Protoporphyrins; Pyrazoles; Pyrimidines; Thiobarbituric Acid Reactive Substances; Thromboplastin; Time Factors; Tyrosine; Umbilical Veins; Water

High levels of the soluble fragment of CD40 ligand (sCD40L) have previously been associated with adverse cardiovascular outcomes. CD40L-CD40 interaction has been linked to the pathogenesis of atherothrombotic complications in cardiovascular disease (CVD). We sought to determine whether a "package of care" of intensified multifactorial cardiovascular risk intervention could reduce indices of platelet activation, inflammation, and coagulation in diabetes and whether patients with overt CVD would derive similar benefit compared with those without.. We measured plasma sCD40L, soluble P-selectin (sP-sel, an index of platelet activation), interleukin-6 (IL-6, a proinflammatory cytokine), and tissue factor (TF, an initiator of coagulation) in 97 patients with diabetes mellitus (41 with and 56 without overt CVD) and 39 comparable healthy control subjects. Thirty-six patients with and 32 without overt CVD then participated in a package of care of cardiovascular risk intervention over a period of 1 year. Plasma levels of sCD40L (P<0.001), sP-sel (P<0.001), IL-6 (P=0.001), and TF (P<0.001) were higher in patients with diabetes than in control subjects, with TF levels highest in patients with overt CVD. Multifactorial intervention was associated with significant reductions in sCD40L in both patient groups (both P<0.001), but reductions in sP-sel and TF were seen only in patients without overt CVD. There was no significant change in IL-6 levels in both patient groups.. Intensive multifactorial risk management can reduce high levels of sCD40L but can only partially correct abnormal platelet activation, inflammation, and coagulation in diabetes, particularly in patients with overt CVD.

Topics: Aged; Aspirin; Biomarkers; Blood Coagulation; Cardiovascular Diseases; CD40 Ligand; Cohort Studies; Diabetes Mellitus; Female; Follow-Up Studies; Humans; Hydroxymethylglutaryl-CoA Reductase Inhibitors; Hypercholesterolemia; Hypoglycemic Agents; Inflammation; Interleukin-6; Male; Middle Aged; P-Selectin; Platelet Activation; Platelet Aggregation Inhibitors; Risk Factors; Risk Management; Thromboplastin

Several studies suggest a role for an increased circulating pool of tissue factor (TF) in atherothrombotic diseases. Furthermore, certain cardiovascular risk factors, such as diabetes, hyperlipemia, and smoking, are associated with a higher incidence of thrombotic complications. We hypothesized that the observed increased blood thrombogenicity (BT) observed in patients with type 2 diabetes mellitus may be mediated via an increased circulating tissue factor activity. We have extended our study to smokers and hyperlipidemic subjects.. Poorly controlled patients with type 2 diabetes mellitus (n=36), smokers (n=10), and untreated hyperlipidemic subjects (n=10) were studied. Circulating TF was immunocaptured from plasma, relipidated, and quantified by factor Xa (FXa) generation in the presence of factor VIIa. BT was assessed as thrombus formation on the Badimon perfusion chamber. Patients with improvement in glycemic control showed a reduction in circulating TF (362+/-135 versus 243+/-74 pmol/L per min FXa, P=0.0001). A similar effect was observed in BT (15 445+/-1130 versus 12 072+/-596 microm/mm2, P=0.01). Two hours after smoking 2 cigarettes, TF was increased (217+/-72 versus 283+/-106 pmol/L per min FXa, P=0.003). Hyperlipidemic subjects showed higher TF (237+/-63 versus 195+/-44 pmol/L per min FXa, P=0.035) than healthy volunteers.. These findings suggest that high levels of circulating TF may be the mechanism of action responsible for the increased thrombotic complications associated with the presence of these cardiovascular risk factors. These observations strongly emphasize the usefulness of the management of the patients based on their global risk assessment.

Topics: Cardiovascular Diseases; Diabetes Mellitus, Type 2; Female; Humans; Hyperlipidemias; Male; Middle Aged; Risk Factors; Smoking; Thromboplastin; Thrombosis

Topics: Anemia; Biomarkers; Blood Proteins; Cardiovascular Diseases; Comorbidity; Endothelium, Vascular; Erythropoietin; Follow-Up Studies; Humans; Injections, Subcutaneous; Kidney Failure, Chronic; Lipoproteins; Recombinant Proteins; Renal Dialysis; Thromboplastin

This study was undertaken to test the hypothesis that transplant coronary vasculopathy (CV) is associated with increased myocardial protein expression of both tissue factor (TF) and integrin alphavbeta3.. The vitronectin receptor (integrin alphavbeta3) and TF have recently been found to play a key role in apoptotic cell death and vascular endothelial cell injury.. A total of 77 heart transplant recipients underwent simultaneous endomyocardial biopsy and intravascular ultrasound (IVUS) at one year of transplant. Patients with pre-existing donor coronary atherosclerosis (n = 35) or with acute rejection (grade >1A, n = 10) at the time of the IVUS were excluded from the analysis. The remaining 32 patients constitute the cohort of the present study. A computerized biopsy score was derived based on the duration and severity of cellular rejection. Both TF and alphavbeta3 expression in the heart biopsy specimens were evaluated by immunoperoxidase histochemistry and Western blot analysis.. Patients with CV (n = 24) had increased expression of alphavbeta3 (2.7-fold, p = 0.003) and TF (7.9-fold, p = 0.04) compared with patients without evidence of vasculopathy (n = 8). In the absence of myocardial fibrosis, alphavbeta3 expression correlated significantly with the cellular rejection score (r = 0.58, p = 0.02).. Transplant vasculopathy is associated with increased expression of both TF and alphavbeta3. The significant correlation of alphavbeta3 with cellular rejection suggests an important role for this integrin in serving as a mechanistic link between cellular rejection and vasculopathy.

Topics: Cardiovascular Diseases; Cohort Studies; Female; Follow-Up Studies; Graft Rejection; Heart; Heart Transplantation; Humans; Male; Middle Aged; Myocardium; Receptors, Vitronectin; Severity of Illness Index; Thromboplastin; Time Factors; Ultrasonography, Interventional; Up-Regulation

The aim of this study was to see if a short-term period of exposure to cold in young healthy subjects causes changes in hematological factors known to be associated with the promotion of thrombogenesis. Over a period of 48 hours, changes in the distribution of erythrocytes, granulocytes, and blood platelets, as well as several coagulation, inflammatory, and fibrinolytic parameters, were monitored in 11 young healthy male subjects following a short period (1 hour) of cold exposure (CE) (ambient temperature, 11 degrees C) or exposure to thermoneutral conditions (ambient temperature, 26 degrees C) in winter (November). The major findings were: (1) a CE-induced hemoconcentration as indicated by an increase in erythrocyte count (3.2% increase); (2) after appropriate adjustments for changes in hemoconcentration, a cold-induced mobilization of granulocytes (14.5% increase) and a cold-induced decrease in lymphocytes (7% decrease); (3) thromboxane B2 release following endotoxin stimulation of whole blood was increased by 27.4% in the CE experiments; (4) diurnal rhythms were observed in granulocytes, blood platelets, middle plate volume, tissue plasminogen activator, and plasma activator inhibitor; and (5) CE caused no significant changes in lipopolysaccharide-induced tissue factor, nor in the blood coagulation factor VII or cytokines, interleukin-6, and tumor necrosis factor. It is concluded that short-term cold exposure in young healthy subjects initiates a mild inflammatory reaction and a tendency for an increased state of hypercoagulability.

Topics: Adult; Blood Cell Count; Blood Coagulation; Cardiovascular Diseases; Cell Differentiation; Cold Temperature; Cytokines; Environmental Exposure; Fibrinogen; Fibrinolytic Agents; Hematocrit; Humans; Inflammation; Male; Norway; Pain Measurement; Plasminogen Activator Inhibitor 1; Risk Factors; Self-Assessment; Serine Proteinase Inhibitors; Thromboplastin; Thromboxane B2; Tissue Plasminogen Activator

The World Health Organization (WHO), the International Committee for Thrombosis and Hemostasis, and the International Committee for Standardization in Hematology, have strongly suggested that INR (International Normalized Ratio) values should be used to report a patient's level of coagulation. This paper discusses the use of the INR to monitor a patient's coagulation level, and reviews the recommendations on the INR levels at which dental extractions and similar oral surgical procedures may be performed safely. Studies are needed to determine whether new local hemostatic agents are sufficient for the safe management of patients at higher INR levels, thereby avoiding the need for hospitalization and heparin therapy.

Topics: Anticoagulants; Cardiovascular Diseases; Dental Care for Chronically Ill; Drug Monitoring; Humans; Oral Hemorrhage; Prothrombin Time; Reference Values; Sensitivity and Specificity; Thromboplastin; Warfarin

We investigated plasma activated factor VII (FVIIa) levels in uremic patients (nondialysis group: n = 38; dialysis group: n = 36) and healthy controls (n = 32). We also measured the plasma levels of thrombomodulin (an indicator of endothelial cell injury) and tissue factor. Plasma FVIIa showed a marked increase in the nondialysis group (mean [95% confidence interval]: 4.6 [4.1-5.1] ng/ml, p < 0.0001) with the progressive impairment of renal function, as indicated by the serum creatinine level, when compared with the 32 controls (2.8 [2.5-3.1] ng/ml), and was further increased in the dialysis group (6.1 [5.5-6.8] ng/ml, p < 0.001 vs. nondialysis group). Plasma levels of thrombomodulin and tissue factor were also higher in the nondialysis group than the control group, and were further increased in the dialysis group. Plasma tissue factor levels did not show any correlation with FVIIa or thrombomodulin in both the nondialysis and dialysis groups. Thus, circulating tissue factor appears to be released by a different mechanism from thrombomodulin and may not contribute to the direct activation of factor VII in uremic patients. On the other hand, the plasma level of thrombomodulin was positively correlated with that of FVIIa in the nondialysis group, and this correlation was independent of renal function. Thus, enhanced conversion of factor VII zymogen to FVIIa, probably related to endothelial cell injury, may be a risk factor for cardiovascular events in uremic patients.

Topics: Aged; Cardiovascular Diseases; Creatinine; Diabetic Nephropathies; Endothelium, Vascular; Factor VII; Factor VIIa; Female; Glomerulonephritis; Humans; Male; Middle Aged; Renal Dialysis; Risk Factors; Thrombomodulin; Thromboplastin; Uremia

We compared factor VII clotting activity (FVIIc) assays using different thromboplastins to determine which is the most sensitive for activated FVII (FVIIa) or for FVII antigen (FVIIag). FVIIc levels were measured using thromboplastins derived from bovine brain (FVIIc Bov), human placenta (FVIIc Hum), and rabbit brain (FVIIc Rab). FVIIa levels were measured by fluorogenic assays using human soluble tissue factor (rsTF) or bovine rsTF. We also measured FVII activity by an amidolytic assay (FVIIc:am Hum) using human thromboplastin and a chromogenic substrate for thrombin. FVIIag levels were determined by ELISA. In the FVIIa assay, the reaction time obtained from using bovine rsTF was shorter than that with human rsTF, suggesting that the interaction of plasma FVIIa with bovine rsTF was stronger than with human rsTF. The plasma FVIIa levels measured using human rsTF and bovine rsTF were almost the same (r = 0.947, p < 0.0001). Among the three FVIIc assays, FVIIc Bov had the strongest positive correlation with the plasma FVIIa level (r = 0.886, p < 0.0001), but had no correlation with FVIIag. An increase of 1 ng/ml in the plasma FVIIa level yielded a 27.9% increase of FVIIc Bov. Plasma FVIIc Hum and FVIIc:am Hum showed moderate correlations with both FVIIa (r = 0.520, p < 0.02 and r = 0.569, p < 0.01, respectively) and FVIIag (r = 0.438, p < 0.05 and r = 0.468, p < 0.05, respectively). FVIIc Rab had the lowest correlation with FVIIa (r = 0.367, p < 0.1), but had a moderate correlation with FVIIag (r = 0.436, p < 0.05).(ABSTRACT TRUNCATED AT 250 WORDS)

Topics: Adult; Aged; Animals; Antigens; Brain Chemistry; Cardiovascular Diseases; Cattle; Cold Temperature; Enzyme Activation; Enzyme-Linked Immunosorbent Assay; Factor VII; Factor VIIa; Female; Humans; Male; Middle Aged; Placenta; Rabbits; Risk Factors; Sensitivity and Specificity; Species Specificity; Thromboplastin; Time Factors

A simple chromogenic substrate assay for the quantitation of tissue factor pathway inhibitor (TFPI) activity in plasma or serum samples was developed. After immobilization on microtiter plates for 20 hours at 4 degrees C, a commercial thromboplastin was incubated for 1 hour at room temperature with 1 U/mL of a prothrombin complex concentrate (Protromplex). After washing, solid-phase Factor Xa activity was measured by a chromogenic substrate (S-2222). Factor Xa generation was progressively inhibited when increasing amounts (1-12 microL) of heated serum or plasma, and recombinant TFPI (1-5 ng/mL), were coincubated with Protromplex. Inhibition by serum or plasma was abolished by anti-TFPI polyclonal antibodies. Plasma levels of TFPI in 25 healthy volunteers were found to be 0.98 +/- 0.19 U/mL (range 0.71-1.52), with an intra- and inter-assay coefficient of variation of 10.7 and 11.1%, respectively. The use of a recombinant human thromboplastin improved the sensitivity and reproducibility of the assay. Plasma levels of TFPI were found to be normal in 10 women at the end of their pregnancies, in 10 patients receiving oral anticoagulant therapy, and in 10 diabetic patients. Significantly higher levels were detected in 10 patients with chronic liver disease and in 10 patients with unexplained juvenile thrombosis. In patients with cardiovascular disease, a 7-day treatment with subcutaneous standard heparin increased TFPI activity. The availability of a simple and rapid assay to measure TFPI that does not require purified coagulation proteins may facilitate studies of the pathophysiologic relevance of this inhibitor.

Topics: Adolescent; Adult; Cardiovascular Diseases; Chromogenic Compounds; Factor Xa Inhibitors; Female; Humans; Lipoproteins; Liver Diseases; Male; Middle Aged; Plasma; Recombinant Proteins; Reference Values; Sensitivity and Specificity; Serine Proteinase Inhibitors; Thromboplastin

The tissue factor coagulant activity of peripheral blood monocytes was measured in 20 patients on long-term anticoagulant treatment with warfarin and in 21 healthy individuals. Contrary to expectations, tissue factor activity was not reduced in the warfarin-treated group and was, in fact, elevated in several patients. No relationship existed between the daily warfarin dose or the degree of prolongation of the prothrombin time and the monocyte tissue factor activity. These results indicate that monocyte tissue factor activity is not necessarily reduced to below normal levels with warfarin treatment.

Topics: Cardiovascular Diseases; Female; Humans; Male; Monocytes; Phytohemagglutinins; Prothrombin Time; Thromboplastin; Warfarin

Topics: Acute Disease; Adaptation, Physiological; Animals; Blood Coagulation; Blood Coagulation Disorders; Blood Platelets; Cardiovascular Diseases; Chronic Disease; Convalescence; Coronary Disease; Fibrinolysin; Fibrinolysis; Humans; Myocardial Infarction; Rheumatic Diseases; Thrombelastography; Thromboplastin; Time Factors

Topics: Anticoagulants; Blood Coagulation Tests; Cardiovascular Diseases; Humans; In Vitro Techniques; Indicators and Reagents; Methods; Thrombin; Thromboplastin; Time Factors

Topics: Blood Cell Count; Blood Coagulation Disorders; Blood Coagulation Factors; Blood Coagulation Tests; Blood Platelets; Cardiovascular Diseases; Humans; Kaolin; Male; Platelet Adhesiveness; Prothrombin Time; Purpura; Radiation Injuries; Risk; Thromboplastin; Thrombosis; Thyroid Diseases

Topics: Anticoagulants; Blood Coagulation Tests; Cardiovascular Diseases; Embolism; Humans; Myocardial Infarction; Prothrombin; Thromboplastin; Thrombosis

Topics: Cardiovascular Diseases; Hemostatics; Humans; Prothrombin; Thromboplastin; Thrombosis; Vascular Diseases

Topics: Cardiovascular Diseases; Hematologic Diseases; Humans; Prothrombin Time; Research; Sclerosis; Thrombophilia; Thromboplastin