thromboplastin and Diabetes-Mellitus

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

Atherothrombosis--defined as atherosclerotic lesion disruption with superimposed thrombus formation--is a leading cause of death in patients with diabetes mellitus. Platelets play a pivotal role in atherothrombosis and platelets of diabetic patients are hyperreactive. Numerous studies have investigated the usefulness of antiplatelet therapy for primary and secondary prevention of atherothrombotic events in diabetic patients. However, there are limited evidences that aspirin may be effective in the reduction of atherothrombotic complication in this population. Additionally, dual antiplatelet therapy with aspirin and clopidogrel has been suggested to be harmful. In contrast, the role of antiplatelet therapy in secondary prevention after ischemic cardiac events is well established in diabetes. Glycoprotein IIb/IIIa receptor antagonists can reduce mortality in diabetic patients committed to undergo percutaneous coronary intervention (PCI). Upregulation of P2Y(12) signalling occurs in hyperglycemia, and the relevance of platelet P2Y(12) receptor inhibition with prasugrel in reducing adverse events following PCI has been recently suggested. Besides platelet activation, several other mechanisms may be involved in the pathophysiology of diabetic atherothrombosis. Tissue factor (TF)-bearing procoagulant microparticles (MPs) are a heterogeneous population of membrane-coated vesicles released by several cell lines upon activation or apoptosis. There is converging evidence that MPs and MP-associated TF activity are upregulated in patients with diabetes mellitus and can participate actively in promoting atherothrombotic complications. In this context, drugs that may reduce the release of microparticles and/or their thrombogenic capacity has the potential to improve upon current antiplatelet therapy, possibly resulting in lower adverse events rates in diabetic individuals.

Topics: Acute Coronary Syndrome; Apoptosis; Aspirin; Blood Platelets; Clopidogrel; Diabetes Complications; Diabetes Mellitus; Glycoproteins; Humans; Models, Biological; Piperazines; Platelet Activation; Prasugrel Hydrochloride; Receptors, Purinergic P2Y12; Thiophenes; Thromboplastin; Thrombosis; Ticlopidine

Atherothrombotic vascular disease is a complex disorder in which inflammation and coagulation play a pivotal role. Rupture of high-risk, vulnerable plaques with the subsequent tissue factor (TF) exposure is responsible for coronary thrombosis, the main cause of unstable angina, acute myocardial infarction, and sudden cardiac death. Tissue factor (TF), the key initiator of coagulation is an important modulator of inflammation. TF is widely expressed in atherosclerotic plaques and found in macrophages, smooth muscle cells, extracellular matrix and acellular lipid-rich core. TF expression can be induced by various stimulants such as C-reactive protein, oxLDL, hyperglycemia and adipocytokines. The blood-born TF encrypted on the circulating microparticles derived from vascular cells is a marker of vascular injury and a source of procoagulant activity. Another form of TF, called alternatively spliced has been recently identified in human and murine. It is soluble, circulates in plasma and initiates coagulation and thrombus propagation. Evidence indicates that elevated levels of blood-borne or circulating TF has been associated with metabolic syndrome, type 2 diabetes and cardiovascular risk factors and is a candidate biomarker for future cardiovascular events. Therapeutic strategies have been developed to specifically interfere with TF activity in the treatment of cardiovascular disease.

Topics: Coronary Artery Disease; Coronary Thrombosis; Diabetes Mellitus; Endothelium, Vascular; Humans; Hyperglycemia; Inflammation; Obesity; Thromboplastin

Diabetes mellitus is characterized by chronic vascular disorder and presents a main risk factor for cardiovascular mortality. In particular, hyperglycaemia and inflammatory cytokines induce vascular circulating tissue factor (TF) that promotes pro-thrombotic conditions in diabetes. It has recently become evident that alterations of the post-transcriptional regulation of TF via specific microRNA(miR)s, such as miR-126, contribute to the pathogenesis of diabetes and its complications. The endothelial miR-19a is involved in vascular homeostasis and atheroprotection. However, its role in diabetes-related thrombogenicity is unknown. Understanding miR-networks regulating procoagulability in diabetes may help to develop new treatment options preventing vascular complications.. Plasma of 44 patients with known diabetes was assessed for the expression of miR-19a, TF protein, TF activity, and markers for vascular inflammation. High miR-19a expression was associated with reduced TF protein, TF-mediated procoagulability, and vascular inflammation based on expression of vascular adhesion molecule-1 and leukocyte count. We found plasma expression of miR-19a to strongly correlate with miR-126. miR-19a reduced the TF expression on mRNA and protein level in human microvascular endothelial cells (HMEC) as well as TF activity in human monocytes (THP-1), while anti-miR-19a increased the TF expression. Interestingly, miR-19a induced VCAM expression in HMEC. However, miR-19a and miR-126 co-transfection reduced total endothelial VCAM expression and exhibited additive inhibition of a luciferase reporter construct containing the F3 3'UTR.. While both miRs have differential functions on endothelial VCAM expression, miR-19a and miR-126 cooperate to exhibit anti-thrombotic properties via regulating vascular TF expression. Modulating the post-transcriptional control of TF in diabetes may provide a future anti-thrombotic and anti-inflammatory therapy.

Topics: 3' Untranslated Regions; Aged; Binding Sites; Blood Coagulation; Diabetes Mellitus; Endothelial Cells; Epigenesis, Genetic; Female; Gene Expression Regulation; Humans; Male; MicroRNAs; Middle Aged; THP-1 Cells; Thromboplastin; Thrombosis; Vascular Cell Adhesion Molecule-1

Diabetes mellitus involves vascular inflammatory processes and is a main contributor to cardiovascular mortality. Notably, heightened levels of circulating tissue factor (TF) account for the increased thrombogenicity and put those patients at risk for thromboembolic events. Here, we sought to investigate the role of micro-RNA (miR)-driven TF expression and thrombogenicity in diabetes mellitus.. Plasma samples of patients with diabetes mellitus were analyzed for TF protein and activity as well as miR-126 expression before and after optimization of the antidiabetic treatment. We found low miR-126 levels to be associated with markedly increased TF protein and TF-mediated thrombogenicity. Reduced miR-126 expression was accompanied by increased vascular inflammation as evident from the levels of vascular adhesion molecule-1 and fibrinogen, as well as leukocyte counts. With optimization of the antidiabetic treatment miR-126 levels increased and thrombogenicity was reduced. Using a luciferase reporter system, we demonstrated miR-126 to directly bind to the F3-3'-untranslated region, thereby reducing TF expression both on mRNA and on protein levels in human microvascular endothelial cells as well as TF mRNA and activity in monocytes.. Circulating miR-126 exhibits antithrombotic properties via regulating post-transcriptional TF expression, thereby impacting the hemostatic balance of the vasculature in diabetes mellitus.

Topics: 3' Untranslated Regions; Aged; Binding Sites; Cell Line; Diabetes Mellitus; Endothelial Cells; Female; Fibrinogen; Gene Expression Regulation; Genes, Reporter; Hemostasis; Humans; Hypoglycemic Agents; Male; MicroRNAs; Middle Aged; Monocytes; RNA Interference; Thromboplastin; Thrombosis; Time Factors; Transfection; Vascular Cell Adhesion Molecule-1

Circulating ('blood-borne') tissue factor (TF) is implicated in the pathogenesis of several chronic conditions, most notably cardiovascular disease, diabetes, and cancer. Full-length TF is an integral membrane protein, while alternatively spliced TF (asTF) can be secreted and, owing to its unique C-terminus, selectively detected in bio-specimens. The predictive and/or prognostic value of asTF in the circulation is unknown. In a retrospective study, we measured levels of circulating asTF in healthy subjects and individuals with acute coronary syndrome (ACS), diabetes mellitus (DM), ongoing ACS + DM, and pancreatic ductal adenocarcinoma (PDAC).. The prototype-tailored procedure (Diagnostica Stago) was used to measure asTF in plasma from 205 subjects.. There was no significant difference between the proportion of healthy subjects with asTF ≥200 pg/mL and those with ACS, DM, or ACS + DM. The proportion of pancreatic cancer patients (n = 43; PDAC: 42; pancreatic neuroendocrine tumor: 1) with asTF levels ≥200 pg/mL was significantly higher than in healthy subjects; asTF levels ≥200 pg/mL were detected more often in patients with unresectable disease irrespective of initial evaluation and/or preoperative carbohydrate antigen 19-9 (CA19-9) levels.. While asTF levels ≥200 pg/mL are not observed with increased frequency in patients with ACS and/or DM, they do occur more frequently in the plasma of patients with pancreatic cancer and are associated with lower likelihood of tumor resectability, irrespective of the preoperative diagnosis. asTF may thus have utility as a novel marker of aggressive pancreatic tumor phenotype.

Topics: Acute Coronary Syndrome; Alternative Splicing; Biomarkers, Tumor; Carcinoma, Pancreatic Ductal; Case-Control Studies; Diabetes Mellitus; Enzyme-Linked Immunosorbent Assay; Female; Follow-Up Studies; Humans; Lymphatic Metastasis; Male; Middle Aged; Neoplasm Staging; Pancreatic Neoplasms; Prognosis; Retrospective Studies; Thromboplastin

Diabetes predisposes to aortic stenosis (AS). We aimed to investigate if diabetes affects the expression of selected coagulation proteins and inflammatory markers in AS valves. Twenty patients with severe AS and concomitant type 2 diabetes mellitus (DM) and 40 well-matched patients without DM scheduled for valve replacement were recruited. Valvular tissue factor (TF), TF pathway inhibitor (TFPI), prothrombin, C-reactive protein (CRP) expression were evaluated by immunostaining and TF, prothrombin, and CRP transcripts were analyzed by real-time PCR. DM patients had elevated plasma CRP (9.2 [0.74-51.9] mg/l vs. 4.7 [0.59-23.14] mg/l, p = 0.009) and TF (293.06 [192.32-386.12] pg/ml vs. 140 [104.17-177.76] pg/ml, p = 0.003) compared to non-DM patients. In DM group, TF-, TFPI-, and prothrombin expression within valves was not related to demographics, body mass index, and concomitant diseases, whereas increased expression related to DM was found for CRP on both protein (2.87 [0.5-9]% vs. 0.94 [0-4]%, p = 0.01) and transcript levels (1.3 ± 0.61 vs. 0.22 ± 0.43, p = 0.009). CRP-positive areas were positively correlated with mRNA TF (r = 0.84, p = 0.036). Diabetes mellitus is associated with enhanced inflammation within AS valves, measured by CRP expression, which may contribute to faster AS progression.

Topics: Aged; Aortic Valve; Aortic Valve Stenosis; C-Reactive Protein; Diabetes Complications; Diabetes Mellitus; Disease Progression; Female; Humans; Inflammation; Lipoproteins; Male; Middle Aged; Prothrombin; RNA, Messenger; Thromboplastin

The abnormalities of tissue factor and its inhibitor system, increased oxidative stress and the presence of diabetes may be involved in the mechanism of thrombotic complication in peritoneal dialysis patients. We compared the plasma levels of tissue factor, its inhibitor and markers of oxidative stress (malondialdehyde and Cu/Zn superoxide dismutase) in 16 diabetic peritoneal dialysis patients, 40 nondiabetic peritoneal dialysis patients, and 20 healthy control individuals. We tried to establish whether there is an association between tissue factor, its inhibitor and oxidative stress in these patients. Compared with the control individuals, the patients both with and without diabetes showed a significant increase in plasma concentrations of Cu/Zn superoxide dismutase (P < 0.01 and P < 0.001, respectively), malondialdehyde (both P < 0.05), tissue factor (both P < 0.001) and tissue factor pathway inhibitor (P < 0.01 and P < 0.001, respectively). The differences in oxidative status and coagulation parameters between patients with and without diabetes were not statistically significant. In all peritoneal dialysis patients, both tissue factor and its inhibitor were positively related to Cu/Zn superoxide dismutase (r = 0.310, P < 0.05 and r = 0.460, P < 0.001, respectively) and malondialdehyde levels (r = 0.337, P < 0.05 and r = 0.361, P < 0.01, respectively). Our data suggest a relationship between increased oxidative stress and elevated tissue factor and its inhibitor levels in peritoneal dialysis patients, particularly those with diabetes.

Topics: Adult; Biomarkers; Diabetes Mellitus; Female; Humans; Lipoproteins; Male; Malondialdehyde; Middle Aged; Oxidative Stress; Peritoneal Dialysis; Superoxide Dismutase; Thromboplastin

Advanced atherogenesis is characterized by the presence of markers of enhanced prothrombotic capacity, attenuated fibrinolysis, and by clinical conditions associated with defective coagulation. Diabetes may be associated with enhanced lesion instability and atherosclerotic plaque rupture. Plaques obtained from 206 patients undergoing carotid endarterectomy were divided into diabetic (type 2) and nondiabetic and analyzed by Western blotting and immunohistochemistry to detect tissue factor (TF), metalloproteinases (MMP)-2, -8, -9, and fibrin/fibrinogen related antigens, and in situ zymography to detect MMP activity. Plasma samples were quantified for TF procoagulant activity, C-reactive protein, fibrinogen and D-dimer. Diabetic and symptomatic patients with hypoechogenic plaques had increased plasma TF activity and D-dimer, compared with those with hyperechogenic plaques (p = 0.03, p = 0.007, respectively). Diabetic, symptomatic patients had higher plasma D-dimer levels than asymptomatic patients (p = 0.03). There was a significant correlation between intramural TF levels and D-dimer in diabetic patients with symptomatic disease (p = 0.001, r2 = 0.4). In diabetic patients, plasma fibrinogen levels were higher in patients with hypoechogenic plaques (p = 0.007). Diabetic patients with ulcerated plaques had higher plasma D-dimer and MMP-8 levels than those with fibrous plaques (p = 0.02, p = 0.01, respectively). This data suggests that currently available circulating markers may be clinically useful to select diabetic patients at higher risk of atherothrombosis. Increased procoagulant activity in diabetic patients may be linked to increased mural remodeling.

Topics: Aged; Atherosclerosis; Biomarkers; Carotid Artery Diseases; Diabetes Complications; Diabetes Mellitus; Endarterectomy, Carotid; Female; Fibrin Fibrinogen Degradation Products; Humans; Male; Matrix Metalloproteinase 2; Matrix Metalloproteinase 8; Matrix Metalloproteinase 9; Thromboplastin

Coagulative function of saliva derives from the thromboplastin found in saliva. It may establish hemostasis in the mouth. Salivary disfunction and changes in salivary composition and are frequent complications of diabetes. This study investigated the influence of some local etiologic and systemic factors on salivary thromboplastic activity (STA) in diabetics. In this study, cytological smears and biochemical tests were used. STA was measured by Quick's one stage method, serum glucose by the glucose oxidase method, and salivary protein by the method of Lowry. STA was almost the same in the diabetic and control groups. The only statistically significant difference within the diabetic group was found to be due to antibiotic usage. STA, i.e. clotting time, was 30% longer (114 s) ( p<0.05) and salivary protein (4.07 mg ml(-1)) ( p<0.1) was lower in diabetics not taking antibiotics than in those taking them. No such differences were observed in the healthy controls. Significant linear correlations ( p<0.05) with respect to STA were with salivary protein in the control group (r=0.61) and in the diabetic group (r=0.51) and with antibiotic usage (r=0.29), with leukocyte cell count (r=0.27) in the diabetic group. It can be concluded that salivary cells, proteins and antibiotic usage are important for STA.

Topics: Adolescent; Adult; Aged; Analysis of Variance; Anti-Bacterial Agents; Blood Coagulation; Blood Glucose; Case-Control Studies; Chi-Square Distribution; Diabetes Mellitus; Female; Humans; Hydrogen-Ion Concentration; Male; Middle Aged; Saliva; Salivary Proteins and Peptides; Secretory Rate; Statistics, Nonparametric; Thromboplastin

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

The tissue factor (TF) plays a key role in triggering the coagulation system in vivo. Our study was designed to determine whether or not the plasma levels of TF and its pathway inhibitor (TF pathway inhibitor; TFPI) in patients with chronic renal failure (CRF) treated by peritoneal dialysis (PD) (1) are pathologically altered; (2) differ between diabetics and nondiabetics, and (3) depend on the metabolic disorders associated with CRF and/or diabetes.. Using ELISA, plasma TF and TFPI levels were measured once in 21 PD patients (10 with diabetes, 11 without diabetes) and in 21 healthy subjects.. As compared with healthy subjects (TF 282 pg/ml; TFPI 73 ng/ml), both TF and TFPI levels were significantly higher in PD patients with diabetes (485 pg/ml, p < 0.001, and 106 ng/ml, p < 0.01, respectively) and in PD patients without diabetes (480 pg/ml, p < 0,001, and 121 ng/ml, p < 0.001, respectively). The difference between diabetics and nondiabetics was not significant. In stepwise regression analysis, the TF levels depended on serum creatinine (partial correlation 0.39, p < 0.05), glycemia (0.43, p < 0.01), and insulin (-0.43, p < 0.05), and the TFPI levels depended on creatinine (partial correlation 0.67, p < 0.001), apolipoprotein B (0.46, p < 0.01), and plasma fibrinogen (0.43, p < 0.01).. CRF patients on PD show increased plasma TF and TFPI levels. There is no difference between diabetics and nondiabetics. The TF and TFPI levels depend significantly on the renal function, as assessed by serum creatinine, and on some metabolic disorders. Elevated TF and TFPI levels may be related to thrombosis and atherosclerosis in CRF patients on PD.

Topics: Adult; Aged; Case-Control Studies; Diabetes Mellitus; Female; Hemostasis; Humans; Kidney Failure, Chronic; Lipoproteins; Male; Metabolic Diseases; Middle Aged; Peritoneal Dialysis; Thromboplastin; Time Factors

Changes of the tissue factor (TF) pathway of blood coagulation have been described in diabetes and could be involved in its vascular complications. In order to evaluate the influence of the type of diabetes and of the obesity index and age on these changes, factor VII coagulant activity, factor VII antigen, activated factor VII, monocyte TF expression, and plasma Tissue Factor Pathway Inhibitor (TFPI) were examined in 18 Type 1 and 16 Type 2 diabetic patients compared to non-diabetic control subjects matched for age, sex, and obesity index (Types 1 and 2 controls, respectively). Multicomplicated patients were excluded. FVIIc, FVIIAg, and FVIIa were higher in Type 2 diabetic patients and controls than in Type 1 diabetic patients and controls (P< .03). However, FVIIc and FVIIAg were lower in diabetic patients than in their matched controls (P< .03). Monocyte expression of TF was not different between Types 1 and 2 diabetic patients and their matched controls except for LPS-stimulated monocyte TF activity which was lower in Type 2 diabetic patients than in Type 2 controls (P< .05). Plasma TFPI was slightly but significantly higher in Type 1 diabetic patients than in Type 1 controls (P= .01) and was correlated to glycemia. However, both in Type 2 diabetic patients and controls, TFPI was higher than in Type 1 controls and was correlated with BMI (P< .0003). These results indicate that in not multicomplicated patients, the increase of FVII and TFPI was highly dependent on obesity index and age rather than on diabetes by itself.

Topics: Adult; Age Factors; Antigens; Apolipoproteins; Body Mass Index; Cholesterol; Cholesterol, HDL; Cholesterol, LDL; Diabetes Mellitus; Diabetes Mellitus, Type 1; Diabetes Mellitus, Type 2; Factor VII; Factor Xa Inhibitors; Female; Glycated Hemoglobin; Humans; Lipoproteins; Male; Middle Aged; Monocytes; Obesity; Reference Values; Thromboplastin

Obesity and insulin resistance are associated with accelerated macrovascular and microvascular coronary disease, cardiomyopathic phenomena, and increased concentrations and activity in blood of plasminogen activator inhibitor type 1 (PAI-1), the primary physiological inhibitor of fibrinolysis.. To determine whether hypofibrinolysis in blood and tissues and its potential sequelae could be attenuated pharmacologically, we studied genetically modified obese mice. By 10 weeks of age, obese mice exhibited increases in left ventricular weight and glucose and immunoreactive insulin in blood. PAI-1 activity in blood measured spectrophotometrically was significantly elevated as well. The difference compared with values in lean controls widened by 20 weeks of age. Perivascular fibrosis in coronary arterioles and small coronary arteries was evident in obese mice 10 and 20 weeks of age, paralleling increases in PAI-1 and tissue factor expression evident by immunohistochemical image analysis, in situ hybridization, and reverse transcription-polymerase chain reaction. Inhibition of ACE activity initiated in obese mice 10 weeks of age and continued for 20 weeks arrested the increase in PAI-1 activity in blood and in cardiac PAI-1 and tissue factor mRNA as well as coronary perivascular fibrosis.. Thus, inhibition of proteo(fibrino)lysis and augmented tissue factor expression in the heart precede and may contribute to the coronary perivascular fibrosis seen with obesity and insulin resistance. Furthermore, inhibition of ACE activity can attenuate all 3 phenomena.

Topics: Angiotensin-Converting Enzyme Inhibitors; Animals; Blood Glucose; Body Weight; Coronary Vessels; Diabetes Mellitus; Fibrinolysis; Fibrosis; Heart Ventricles; Immunohistochemistry; In Situ Hybridization; Male; Mice; Mice, Inbred C57BL; Mice, Obese; Myocardium; Obesity; Organ Size; Peptidyl-Dipeptidase A; Plasminogen Activator Inhibitor 1; RNA, Messenger; Thiazepines; Thromboplastin

Platelet activation is known to participate to the pathogenesis of acute coronary syndromes. Aminophospholipid exposure and microparticles shedding are hallmarks of full platelet activation and may account for the dissemination of prothrombotic seats. Using flow cytometry analysis of annexin V binding to externalized aminophospholipids, we followed platelet procoagulant activity (PPA) and platelet microparticles (PMP) shedding in venous and coronary whole blood samples from 30 patients with unstable angina before and after percutaneous coronary angioplasty (PTCA) and stent implantation. Baseline values of PPA and PMP were significantly more elevated in patients than in control subjects (p < 0.005). PMP percentage was significantly higher in coronary than in venous blood, and in coronary blood of patients with proximal instead of mid/distal lesions of coronary arteries. No enhancement of platelet reactivity to TRAP and collagen was induced by procedure. Whereas activated GpIIb-IIIa and P-selectin expression decreased 24 h and 48 h after procedure, PPA and PMP remained as elevated as before. Thus, flow cytometry is a reliable method for detection of fully activated platelets in whole blood samples. Annexin V binding analysis demonstrates the persistance of in vivo platelet activation, despite the use of antiaggregating agents.

Topics: Aged; Angina, Unstable; Angioplasty, Balloon, Coronary; Annexin A5; Biomarkers; Collagen; Combined Modality Therapy; Comorbidity; Coronary Angiography; Coronary Stenosis; Coronary Vessels; Diabetes Mellitus; Female; Flow Cytometry; Humans; Hypercholesterolemia; Male; Middle Aged; Obesity; P-Selectin; Platelet Activation; Platelet Count; Platelet Glycoprotein GPIIb-IIIa Complex; Proteins; Receptors, Thrombin; Risk Factors; Stents; Thromboplastin; Veins

Tissue factor (TF) plays a central role in the initial activation of the extrinsic coagulation pathway and is thought to be involved in the development of atherosclerosis and thrombosis. The effect of advanced glycosylation end products (AGEs) on TF expression and its mechanism were assessed by flow cytometric analysis. Human macrophage-like U937 cells, which were shown to contain mRNA encoding the receptors of advanced glycosylation end products (RAGE), expressed TF in a dose-dependent manner on incubation with AGE-albumin. AGE-albumin-induced TF expression was completely inhibited by the anti-oxidant agents, catalase and probucol. TF expression in peripheral monocytes from normal volunteers was also increased by AGE-albumin. Finally, TF expression in monocytes from individuals with diabetes mellitus, in whom the concentration of circulating AGEs is reported to be increased, was higher than that in monocytes from normal controls. These results suggest that AGE-induced TF expression in macrophages/monocytes is mediated by oxidant stress. AGEs may promote thrombosis and the development of atherosclerosis by inducing TF expression in monocytes in patients with diabetes mellitus.

Topics: Albumins; Antioxidants; Catalase; Cells, Cultured; Diabetes Mellitus; Flow Cytometry; Glycation End Products, Advanced; Humans; Leukemia, Experimental; Macrophages; Monocytes; Probucol; Receptor for Advanced Glycation End Products; Receptors, Immunologic; Thromboplastin; Tumor Cells, Cultured

We measured plasma levels of tissue factor (TF), total tissue factor pathway inhibitor (TFPI) and free TFPI antigen in patients with diabetes mellitus (DM), hyperlipidemia and disseminated intravascular coagulation (DIC). The mean TF, total TFPI and free TFPI antigen concentrations were significantly higher in patients with DM than in controls and the plasma TF concentration was significantly higher in patients with retinopathy or nephropathy than in DM with no complications. The mean TF, total TFPI and free TFPI antigen concentrations were significantly higher in patients with hyperlipidemia than in controls. There was a significant positive correlation between levels of total TFPI and total cholesterol. In patients with hyperlipidemia, the level of total TFPI was significantly decreased compared to base line level by cholesterol lowering drug, however, free TFPI concentration did not change by cholesterol lowering drug. The TF and total TFPI concentrations were significantly higher in patients with DIC than in controls.

Topics: Anticoagulants; Antigens; Biomarkers; Cholesterol; Diabetes Mellitus; Disseminated Intravascular Coagulation; Humans; Hyperlipidemias; Lipoproteins; Thromboplastin

In the submitted pilot study we examined 37 patients suffering from diabetes mellitus without vascular complications and 15 healthy blood donors. The diabetic patients had not, as compared with the blood donors, significantly elevated values of the tissue factor (TF) and tissue factor pathway inhibitor (TFPI). The TFPI levels correlated with other markers of endothelial dysfunction, in particular thrombomodulin (r = 0.452, p < 0.01) and with triacylglycerol and cholesterol levels. They did not correlate with age, the C-peptide level and BMI.

Topics: Adult; Diabetes Mellitus; Factor Xa Inhibitors; Female; Hemostasis; Humans; Lipoproteins; Male; Middle Aged; Pilot Projects; Serine Proteinase Inhibitors; Thromboplastin; von Willebrand Factor

To investigate the clinical significance of determination of plasma tissue factor (TF) antigen, we have developed a highly sensitive enzyme-linked immunosorbent assay (ELISA) for plasma TF, using two different monoclonal antibodies against TF apoprotein, 6B4 (catching antibody) and 5G9 (detecting antibody), and tetramethyl benzidine/H2O2 as substrates. Titration curves of recombinant human TF in buffer containing Triton X-100 were linear within the range from 50 to 2000 pg/ml. The total assay time was 3 h. Ultracentrifugation and immunoblot analysis indicated that human plasma and urine contained 50,000 g sedimentable and non-sedimentable forms of TF, both of which were detected by our ELISA method. Plasma and urine concentrations of TF in healthy subjects and patients with various diseases were measured by the ELISA method. In healthy subjects, plasma and urinary TF levels were found to be 149 +/- 72 pg/ml (n = 30) and 175 +/- 60 pg TF/urine creatinine mg (n = 95), respectively. TF was increased in plasma of patients with disseminated intravascular coagulation (DIC), thrombotic thrombocytopenic purpura, vasculitis associated with collagen diseases, diabetic microangiopathy and chronic renal failure receiving haemodialysis, but not in the plasma of endotoxaemic patients without DIC. The plasma TF/serum creatinine ratio did not show a positive correlation. Measurement of TF antigen in plasma may be useful for evaluating the endothelial damage and cell destruction in TF-containing tissues.

Topics: Adult; Aged; Blood Coagulation Disorders; Diabetes Mellitus; Disseminated Intravascular Coagulation; Enzyme-Linked Immunosorbent Assay; Female; Hematologic Diseases; Humans; Immunoblotting; Male; Middle Aged; Purpura, Thrombotic Thrombocytopenic; Reference Standards; Thromboplastin; Ultracentrifugation; Vasculitis

Procoagulant albumin (P-Al) is present in normal human plasma and increases monocyte and endothelial cell expression of tissue factor activity. To develop a bioassay for P-Al, we partially purified plasma from healthy volunteers and several patient groups using BaCl2 and (NH4)2SO4 precipitation. The samples were assayed for tissue factor (TF) inducing activity, expressed as a percentage increase compared to a serum-free media control. Over six months, the assay was reproducible in stored samples and in serial samples from normal volunteers. The plasma P-Al activities of 35 volunteers averaged 141 +/- 8.2% (SEM). There was no diurnal variation. There was no difference in the P-Al activity after a 12 hour fast and 2 hours after a large meal in 4 healthy volunteers. There was no increase in activity (r = 0.16) with the subject's age. The average activity from 16 poorly-controlled diabetics was 131 +/- 11% (SEM). No alteration in activity was seen with samples from patients with uremia, liver dysfunction, hemophilia, thrombotic events, or adenocarcinoma. These results indicate that P-Al activity can be bioassayed in individual patient samples; however, pathologic states associated with abnormal P-Al-induced tissue factor activity presently remain unidentified.

Topics: Adult; Aged; Aged, 80 and over; Blood Coagulation; Blood Coagulation Factors; Cells, Cultured; Diabetes Mellitus; Endothelium, Vascular; Hemophilia A; Humans; Middle Aged; Pulmonary Edema; Serum Albumin; Thromboplastin; Thrombosis

We have carried out studies on cultured human fibroblasts in an attempt to trace the origins of age-dependent disorders to the cellular and molecular levels. Three interrelated areas are discussed. First, skin donors with diabetes mellitus (a disease complex that features inappropriate hyperglycemia) produce cultured fibroblasts with a moderate reduction in growth capacity, while two inherited disorders of inappropriate hyperglycemia and premature aging, progeria and Werner syndrome, yield fibroblast cultures with more severely impaired growth capacity. Second, there is a decreased response of progeria level and donor age; evidence is presented that this defective hormone responsiveness in aging cells may reside at the hormone receptor on the surface membrane, the cyclic AMP system, the intracellular enzymatic machinery, or all of these sites. Third, tissue factor, a procoagulant that activates the extrinsic clotting mechanism, is more abundant in cells from the premature aging syndromes of progeria and Werner syndrome. Fibroblast aging in vitro may help to explain various concomitants of normal aging and diabetes mellitus including cell dropout, impairment of hormone responsiveness, and increased atherothrombosis.

Topics: Aging; Blood Coagulation; Cell Division; Cell Survival; Cells, Cultured; Diabetes Mellitus; DNA; Glucose; Humans; Nonsuppressible Insulin-Like Activity; Progeria; Receptor, Insulin; Thromboplastin; Werner Syndrome

Topics: Blood Coagulation; Diabetes Mellitus; Female; Flurbiprofen; Humans; Platelet Adhesiveness; Propionates; Prothrombin Time; Thrombelastography; Thromboplastin

Topics: Adult; Animals; Arteriosclerosis Obliterans; Blood Coagulation; Diabetes Mellitus; Diabetic Angiopathies; Heparin; Humans; Methods; Middle Aged; Rabbits; Smoking; Thromboplastin

Topics: Adult; Aged; Angina Pectoris; Blood Coagulation; Blood Coagulation Tests; Blood Platelets; Coronary Disease; Diabetes Complications; Diabetes Mellitus; Female; Fibrinolysis; Humans; Lipids; Male; Middle Aged; Myocardial Infarction; Platelet Adhesiveness; Prothrombin Time; Thromboplastin

Topics: Adolescent; Adult; Blood Platelets; Diabetes Mellitus; Humans; Thromboplastin

Topics: Adolescent; Adult; Age Factors; Aged; Aminohippuric Acids; Biopsy; Blood Glucose; Blood Pressure; Cell Nucleus; Child; Child, Preschool; Cholesterol; Chronic Disease; Diabetes Complications; Diabetes Mellitus; Diabetic Nephropathies; Glomerulonephritis; Humans; Kidney Glomerulus; Lipids; Middle Aged; Nitrogen; Thromboplastin

Topics: Angina Pectoris; Blood Coagulation; Blood Coagulation Tests; Cholesterol; Diabetes Mellitus; Dietary Fats; Geriatrics; Humans; Linoleic Acid; Lipids; Myocardial Infarction; Palmitic Acid; Pharmacology; Stearic Acids; Thromboplastin; Triolein

Topics: Aged; Amputation, Surgical; Arteriosclerosis Obliterans; Blood Coagulation Disorders; Chronic Disease; Coumarins; Diabetes Mellitus; Follow-Up Studies; Humans; Hypercholesterolemia; Hypertension; Middle Aged; Myocardial Infarction; Thromboplastin