thromboplastin and Kidney-Failure--Chronic

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

Topics: Abruptio Placentae; Adrenal Glands; Aminocaproates; Animals; Basement Membrane; Biopsy; Blood Coagulation Disorders; Blood Coagulation Factors; Blood Platelets; Brain; Female; Fibrin; Fibrinogen; Fibrinolysis; Fluorescent Antibody Technique; Hemorrhage; Hemorrhagic Disorders; Heparin; Humans; Hypertension, Malignant; Kidney Cortex Necrosis; Kidney Failure, Chronic; Kidney Glomerulus; Liver; Maternal Mortality; Microscopy, Electron; Myocardium; Placental Extracts; Pre-Eclampsia; Pregnancy; Rabbits; Shwartzman Phenomenon; Thromboplastin; Thrombosis

Heparin inhibits prothrombotic tissue factor (TF) and releases its inhibitor, tissue factor pathway inhibitor (TFPI), from the endothelium, but repeated administration of heparin depletes vascular stores of TFPI. We studied the anticoagulant effects of unfractionated heparin (UFH) vs low-molecular-weight enoxaparin-used for thrice-weekly maintenance haemodialysis (HD)-on plasma levels of total TF and TFPI and on those of an activated coagulation marker prothrombin fragment 1+2 (PF 1+2).. Twenty-five patients dialysed using a single injection of enoxaparin (at a mean dose of 0.68 mg/kg) were randomly assigned to either receive UFH administered as a mean bolus of 42.1 IU/kg and continuous infusion of 57.8 IU/kg (n=12) or to be maintained on enoxaparin (n=13), and were followed prospectively for 12 weeks. Plasma immunoreactive TF, TFPI and PF 1+2 were measured at the start and after 10 and 180 min of HD, and compared with values in 15 healthy controls.. Pre-dialysis TF, TFPI and PF 1+2 were higher than normal (all P<0.0001). TF and PF 1+2 did not change, while TFPI levels, compared with baseline, increased at each interval in enoxaparin-anticoagulated HD patients (all P<0.0001). TFPI increments correlated inversely with pre-dialysis TFPI (both P<0.0007). In patients switched to UFH, TF levels remained unchanged compared with pre-randomization values, TFPI increased at each interval of HD sessions (all P<0.035) and PF 1+2 increased pre-dialysis (P=0.015). The over-dialysis effects of UFH resembled those of enoxaparin. In contrast, baseline TFPI and its 10-min rise correlated inversely with the UFH loading dose (both P<0.040). Pre-dialysis PF 1+2 was inversely associated with TFPI increments (both P<0.034), and directly with pre-dialysis TFPI (P=0.018) and the UFH loading dose (P=0.045).. Depletion of heparin-releasable stores of TFPI is an untoward effect of repeated anticoagulation during maintenance HD therapy. The traditional UFH regimen is more prothrombotic than single enoxaparin injections, with high loading doses of UFH being involved in TFPI exhaustion and subsequent hypercoagulability.

Topics: Aged; Anticoagulants; Blood Coagulation; Enoxaparin; Female; Fibrinolytic Agents; Follow-Up Studies; Hemostatics; Heparin; Humans; Kidney Failure, Chronic; Lipoproteins; Male; Middle Aged; Peptide Fragments; Prospective Studies; Protein Precursors; Prothrombin; Renal Dialysis; Thromboplastin; Time Factors

Plasma von Willebrand factor antigen, soluble thrombomodulin, and tissue factor were increased in 31 patients with severe chronic renal failure (creatinine clearance <20 ml/min) under conservative treatment, whereas plasminogen activator inhibitor antigen did not differ significantly from healthy controls. No correlation among plasma levels of these proteins was found. Three patterns of relationship between endothelial cell markers and hemostatic defects were identified: 1) Plasma thrombomodulin, a marker of endothelium damage, was found an independent predictor of bleeding time and platelet aggregation, and secretion defects, and was also related to the severity of renal failure; 2) von Willebrand factor antigen, an index of endothelial cell activation and secretion, was significantly correlated with intravascular markers of thrombin and plasmin generation and with platelet adenosine triphosphate content, but not with plasma creatinine levels; and 3) tissue factor and plasminogen activator inhibitor antigen levels were not statistically correlated with the diverse hemostatic defects. Activation of coagulation and fibrinolysis, secondary to endothelial cell activation, appearing early during the evolution of chronic renal failure, is pathogenically related to the platelet dysfunction, and probably to development of atherosclerosis and thrombotic events in this disease. The progression of chronic renal failure, through endothelial cell damage, would lead to aggravation of the platelet functional defect potentiating the hemorrhagic risk.

Topics: Antigens; Biomarkers; Blood Coagulation; Chronic Disease; Disease Progression; Endothelium, Vascular; Fibrinolysis; Hemostasis; Humans; Kidney Failure, Chronic; Plasminogen Activator Inhibitor 1; Thrombomodulin; Thromboplastin; Uremia; von Willebrand Factor

The association of platelet and monocyte activity markers with long-term mortality was assessed in hemodialysis (HD) patients.. In 41 HD patients (25 male, 16 female), surface expression of CD40L and CD62P on platelets, tissue factor (TF) binding on monocytes, and platelet-monocyte aggregates were measured by flow cytometry. Plasma levels of MCP-1, IL-6, TNFα, and soluble CD40L were analyzed by enzyme linked immunosorbent assay. Cox proportional hazard regression analyses and Kaplan-Meier curve were calculated. The predefined endpoint was all-cause mortality.. The study follow-up was 11.54 years. Thirty-one patients (75.6%) died within the study period. Mean patient survival after study inclusion was 5.45 +/- 4.24 years. TF on monocytes above the median of the study population was significantly and independently associated with total mortality (HR (95% CI) 3.45 (1.32 - 9.07); p = 0.01). Cumulative mortality in patients with TF on monocytes above median was significantly higher compared to pa-tients with TF on monocytes below median value (log rank p < 0.01). Platelet-monocyte aggregates, the expression of CD40L and CD62P on platelets, and plasma levels of sCD40L, IL-6, MCP-1, and TNFα were not significantly correlated with mortality.. The present study confirms a high mortality in ESRD patients. TF binding on monocytes was significantly correlated with increased mortality and may identify a subgroup of patients at higher risk.

Topics: Aged; Atherosclerosis; Blood Platelets; Female; Humans; Kidney Failure, Chronic; Male; Middle Aged; Monocytes; Platelet Activation; Renal Dialysis; Thromboplastin

Plasma advanced oxidation protein products (AOPPs), a class of pro-inflammatory pathogenic mediators, accumulate in subjects with chronic kidney disease. Whether AOPPs contribute to coagulation abnormalities, which are frequently seen in uremic patients, is unknown. Here we report that AOPPs activate platelets via a CD36-mediated signaling pathway. Activation of signaling pathways by AOPP-platelet interaction resulted in the expression of several platelet activation markers and rapidly induced the expression of CD40 ligand, triggering platelet adhesion to endothelial cells and promoting endothelial tissue factor expression. AOPPs and serum tissue factor levels were considerably increased in end stage renal disease patients on hemodialysis and a significant correlation of AOPPs and serum tissue factor was found. Interestingly, serum levels of AOPPs and tissue factor were substantially lower in stable kidney transplant patients when compared with hemodialysis patients. Given that CD36 is known to transduce the effects of oxidized lipids into platelet hyperactivity, our findings reveal previously unknown pro-thrombotic activities of oxidized plasma albumin via a CD36 dependent pathway.

Topics: Adolescent; Adult; Advanced Oxidation Protein Products; Aged; Aged, 80 and over; Blood Platelets; CD36 Antigens; CD40 Ligand; Endothelial Cells; Female; Gene Expression Regulation; Humans; Kidney Failure, Chronic; Kidney Transplantation; Male; Middle Aged; Platelet Activation; Platelet Aggregation; Renal Dialysis; Serum Albumin; Signal Transduction; Thromboplastin

Disturbances in hemostasis are common complications of kidney diseases and correlate well with cardiovascular mortality. Little is known about the effects of fasudil on tissue factor (TF) and plasminogen activator inhibitor-1 (PAI-1) expression in peripheral blood mononuclear cells (PBMCs) in CAPD patients. PBMCs were isolated from 13 individuals with CAPD and 13 healthy subjects. After 4 h of incubation with or without LPS (10 ng/mL), TF and PAI-1 mRNA of PBMCs were detected by RT-PCR. The levels of TF and PAI-1 in culture supernatants of PBMCs were determined by ELISA. Compared with healthy controls, CAPD patients had increased TF, PAI-1 protein and mRNA expression by PBMCs at baseline and after stimulated by LPS (10 ng/mL) [p < 0.001]. The fasudil treatment resulted in a significant effect in decreasing TF and PAI-1 [p < 0.05] synthesis in PBMCs. TF and PAI-1 mRNA expression and activities in PBMCs were increased in CAPD patients. Fasudil reduced LPS-mediated TF and PAI-1 expression and activity in PBMCs. These effects may partially be relevant to the clinical benefits of fasudil in the treatment of CAPD patients.

Topics: 1-(5-Isoquinolinesulfonyl)-2-Methylpiperazine; Adult; Aged; Enzyme-Linked Immunosorbent Assay; Female; Follow-Up Studies; Gene Expression Regulation; Humans; Kidney Failure, Chronic; Leukocytes, Mononuclear; Male; Middle Aged; Peritoneal Dialysis, Continuous Ambulatory; Plasminogen Activator Inhibitor 1; Protein Kinase Inhibitors; Reverse Transcriptase Polymerase Chain Reaction; RNA, Messenger; Thromboplastin

Stent thrombosis (ST), a postinterventional complication with a mortality rate of 50%, has an incidence that rises precipitously in patients at risk. Chronic renal failure and end-stage renal disease have emerged as particularly strong ST risk factors, yet the mechanism remains elusive. Tissue factor (TF) is a crucial mediator of injury-related thrombosis and has been implicated for ST. We posit that uremia modulates TF in the local vessel wall to induce postinterventional thrombosis in patients with end-stage renal disease.. As a model of the de-endothelialized, postinterventional state, we exposed primary human vascular smooth muscle cells (vSMCs) pretreated with uremic serum (obtained from ESRD patients on hemodialysis) to coronary-like blood flow. vSMC TF expression, activity, stability, and posttranslational modification were examined after vSMCs were treated with uremic serum or solutes. We found significantly greater clot formation after uremic serum exposure, which was substantially reduced with the prior treatment with anti-TF neutralizing antibody. Uremic sera induced 2- to 3-fold higher TF expression and activity in vSMCs independent of diabetes mellitus. Relevant concentrations of isolated uremic solutes such as indole-3-acetic acid (3.5 μg/mL), indoxyl sulfate (25 μg/mL), and uric acid (80 μg/mL) recapitulated these effects in cell culture and the flow loop model. We show further that TF undergoes ubiquitination at baseline and that uremic serum, indole-3-acetic acid, and indoxyl sulfate significantly prolong TF half-life by inhibiting its ubiquitination.. The uremic milieu is profoundly thrombogenic and upregulates vSMC TF levels by increasing TF stability and decreasing its ubiquitination. Together, these data demonstrate for the first time that the posttranslational regulation of TF in uremia may have a causative role in the increased ST risk observed in uremic patients. These data suggest that interventions that reduce vSMC TF may help to prevent ST and that uremic solutes should be considered as novel risk factors for ST in patients with chronic renal failure.

Topics: Adult; Aged; Cell Line; Coronary Vessels; Endothelium, Vascular; Female; Humans; In Vitro Techniques; Indican; Indoleacetic Acids; Kidney Failure, Chronic; Male; Middle Aged; Myocytes, Smooth Muscle; Renal Dialysis; Risk Factors; Serum; Stents; Thromboplastin; Thrombosis; Ubiquitination; Uremia

Topics: Female; Humans; Kidney Failure, Chronic; Male; Myocytes, Smooth Muscle; Serum; Stents; Thromboplastin; Thrombosis; Uremia

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

Nutritional compounds that potentially limit inflammation and tissue factor expression may decrease the progression of chronic kidney disease (CKD) and associated cardiovascular disease. This project aimed to determine the effect of curcumin, bovine colostrum, and fish oil on inflammatory cytokine and tissue factor procoagulant activity of peripheral blood mononuclear cells (PBMCs) from patients with CKD before dialysis.. Peripheral blood mononuclear cells from patients with CKD before dialysis (n = 13) and age- and sex-matched healthy controls (n = 12) were cultured alone and with low and high doses of the nutritional compounds for 24 h. Cells were cultured with and without lipopolysaccharide. Supernatants were analyzed for tumor necrosis factor-α, interleukin (IL)-6, IL-8, monocyte chemoattractant protein-1, IL-1β, C-reactive protein, and tissue factor procoagulant activity.. The production of C-reactive protein, monocyte chemoattractant protein-1, IL-6, and IL-1β by PBMCs was inhibited by low- and high-dose fish oil in the CKD group (P < 0.05). Curcumin decreased secretion of IL-6 (P = 0.015) and IL-1 β (P = 0.016). Curcumin was more effective than colostrum at decreasing the procoagulant activity of PBMCs in the CKD and control groups (P < 0.019).. Fish oil decreased inflammatory cytokine secretion from CKD PBMCs. In addition, the beneficial effects of curcumin were demonstrated in decreasing inflammation in vitro, often to a similar magnitude as fish oil.

Topics: Adult; Animals; Anti-Inflammatory Agents; C-Reactive Protein; Case-Control Studies; Cattle; Chemokine CCL2; Colostrum; Curcuma; Curcumin; Female; Fish Oils; Humans; Inflammation; Inflammation Mediators; Interleukin-1beta; Interleukin-6; Kidney Failure, Chronic; Leukocytes, Mononuclear; Lipopolysaccharides; Male; Middle Aged; Phytotherapy; Thromboplastin

Thrombosis is a common complication of end-stage renal disease, particularly in patients on hemodialysis. Although substantial progress has been made in preventing thrombotic complications in various other groups of patients, the mechanisms of thrombosis during hemodialysis require clarification. In this report, we demonstrate that complement activation triggered by hemodialysis biomaterials, and the subsequent generation of the complement anaphylatoxin C5a, results in the expression of functionally active tissue factor (TF) in peripheral blood neutrophils. Because TF is a key initiator of coagulation in vivo, we postulate that the recurring complement activation that occurs during long-term hemodialysis contributes to thrombosis in dialyzed end-stage renal disease patients. Furthermore, we found that complement contributed to the induction of granulocyte colony-stimulating factor, which has been implicated in the pathogenesis of thrombosis in patients treated with the recombinant form of this molecule. Importantly, the inhibition of complement activation attenuated the TF expression and granulocyte colony-stimulating factor induction in blood passing through a hemodialysis circuit, suggesting that the complement system could become a new therapeutic target for preventing thrombosis in patients with chronic renal failure who are maintained on hemodialysis.

Topics: Aged; Anaphylatoxins; Blood Coagulation; Cells, Cultured; Complement Activation; Complement C5a; Female; Granulocyte Colony-Stimulating Factor; Humans; Kidney Failure, Chronic; Leukocytes; Male; Middle Aged; Receptor, Anaphylatoxin C5a; Renal Dialysis; Thromboplastin; Thrombosis; Time Factors

Patients with end-stage renal disease (ESRD) exhibit features of a hypercoagulable state, which may contribute to atherosclerosis. Kynurenines are the metabolites of tryptophan degradation in mammals. We examined the relationship between coagulation activation and kynurenines in 92 patients with ESRD on maintenance haemodialysis (HD) or continuous ambulatory peritoneal dialysis (CAPD) and 20 healthy controls. We measured the plasma levels of: tissue factor (TF), its pathway inhibitor (TFPI), the marker of coagulation activation - prothrombin fragments 1+2 (F(1+2)), kynurenine (KYN) and its metabolites: kynurenic (KYNA), anthranilic (AA) and quinolinic (QA) acids. The ratio of KYNA to KYN (kyna/kyn), AA to KYN (aa/kyn) and QA to KYN (qa/kyn), reflecting intensified activity of enzymes which converted KYN to its metabolites, were also determined. Measured coagulation parameters and kynurenines were significantly elevated in ESRD patients compared to controls. TF, TFPI and F(1+2) were significantly associated with AA, aa/kyn, QA and qa/kyn ratio. Multiple regression analysis showed that fibrinogen (p<0.01) and above mentioned KYN metabolites (all p<0.05) were the independent variables significantly associated with increased F(1+2) levels, reflecting hypercoagulability in ESRD patients. In conclusion, this study represents the first to investigate both the coagulation system and KYN pathway in ESRD patients. The coagulation was enhanced in dialysed uraemic patients compared with the healthy controls demonstrated by increased TF, TFPI and F(1+2) levels. These changes were correlated with activation of the KYN pathway. Finally, fibrinogen and KYN metabolites are independently and significantly associated with the hypercoagulable state in uraemic patients on CAPD and HD treatment.

Topics: Adult; Aged; Blood Coagulation; Female; Humans; Kidney Failure, Chronic; Kynurenine; Lipoproteins; Male; Middle Aged; Peptide Fragments; Peritoneal Dialysis, Continuous Ambulatory; Protein Precursors; Prothrombin; Renal Dialysis; Thrombophilia; Thromboplastin; Uremia

Patients with chronic kidney disease exhibit features of a hypercoagulable state and have endothelial dysfunction, which may contribute to their increased cardiovascular risk. We examined the relationship between coagulation activation and vascular function in patients with chronic kidney disease.. We measured parameters of the tissue factor pathway of blood coagulation (tissue factor, factor VIIc and factor X); natural inhibitors (tissue factor pathway inhibitor, protein C, free and total protein S, antithrombin III) and markers of coagulation activation (thrombin-antithrombin complexes, prothrombin fragment 1+2) in 66 stage 4&5 chronic kidney disease patients and 36 healthy controls. Their relationship with markers of vascular function (flow mediated dilatation, soluble E-selectin and thrombomodulin) and a mediator of inflammation (interleukin-6) was determined.. Up-regulation of the tissue factor pathway (increased tissue factor and factor VIIc), increased prothrombin fragment 1+2 and significant reductions in antithrombin III and the ratio of free protein S: total protein S were found in patients compared to healthy controls. Increased tissue factor antigen was significantly and independently correlated with creatinine and interleukin-6 (P<0.001). Factor X and antithrombin III were both reduced in chronic kidney disease and correlated (r=0.58; P<0.001). Changes in coagulation and anti-coagulation were independent of all measures of endothelial function.. Significant activation of the TF pathway of coagulation and depletion or reduction of some natural anticoagulants in chronic kidney disease was correlated with the degree of renal dysfunction, but not correlated with the abnormalities of vascular function. These data are consistent with a hypercoagulable state in chronic kidney disease that may be independent of endothelial based regulation but associated with an inflammatory state.

Topics: Aged; Antigens; Antithrombin III; Biological Phenomena; Biomarkers; Blood Coagulation; Creatinine; E-Selectin; Endothelium, Vascular; Factor VII; Factor X; Female; Humans; Interleukin-6; Kidney Failure, Chronic; Male; Middle Aged; Peptide Fragments; Protein S; Prothrombin; Renal Dialysis; Solubility; Thrombomodulin; Thrombophilia; 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

A moderate increase in plasma total homocysteine (t-hcy) is considered to be an independent risk factor for cardiovascular disease (CVD) in general population. One of the mechanisms by which hyperhomocysteinemia contributes to cardiovascular risk has been explained to be the increased thrombotic potential. Elevated t-hcy levels were also reported in chronic renal failure patients because the renal function is a major determinant of serum t-hcy levels.. We measured serum hcy and ADP-induced platelet aggregation and plasma tissue factor as a major activator of the coagulation cascade in hemodialysis (HD), peritoneal dialysis (PD) and early stage chronic renal failure (early stage CRF) patients who are not receiving dialysis and compared with those of control. In addition, we also determined serum vitamin B12 and folat levels which are the important factors regulating the metabolism of t-hcy.. Hcy levels in all patient groups were significantly higher (HD: 20.42 +/- 1.91 micromol/l, PD: 35.47 +/- 6.30, early stage CRF: 24.39 +/- 3.06) than the normal levels (10.74 +/- 0.74) in spite of standard multivitamin supplementation. The highest t-hcy values were found in peritoneal dialysis patients. Vitamin B12 levels in hemodialysis/peritoneal dialysis patients and folat levels in hemodialysis/early stage CRF patients were also significantly above those of control. On the other hand, the significant elevations in plasma tissue factor concentration were found in all patient groups (HD: 331.4 +/- 31.3 pg/ml, PD: 306.0 +/- 30.0, early stage CRF: 277.2 +/- 25.5 and. 69.5 +/- 13.5). t-hcy levels were positively correlated with creatinine (r: 0.791 p < 0.002) and tissue factor levels (r: 0.526 p < 0.05) in only early stage CRF group. The association between t-hcy and tissue factor persisted after these two parameters were adjusted for creatinine (r: 0.649 p < 0.05). On the other hand the same correlations were not observed in dialysis patient groups. In spite of the high tissue factor levels, ADP-induced platelet aggregations were found to be lower in all patient groups (HD: 102.6 +/- 6.7, PD: 98.6 +/- 7.6 and Early stage CRF: 84.9 +/- 7.6) than controls (154.9 +/- 13.7).. These results suggest that hyperhomocysteinemia and increased tissue factor level are present in patients with renal failure, despite supplementation with vitamin B6 and B12 and folat. However, elevated levels of these thrombogenic factors are not linked with platelet aggregation.

Topics: Adult; Case-Control Studies; Chromatography, High Pressure Liquid; Enzyme-Linked Immunosorbent Assay; Female; Folic Acid; Homocysteine; Humans; Hyperhomocysteinemia; Kidney Failure, Chronic; Male; Platelet Aggregation; Renal Dialysis; Thromboplastin; Vitamin B 12; Vitamin B 6

Impaired haemostasis is common in patients with end-stage renal disease, and may cause either thrombotic or bleeding complications. The purpose of this study was to assess whether plasma markers of coagulation activation in patients undergoing chronic haemodialysis (HD) can identify high-risk individuals, and to test the relevancy of type of vascular access or dialysis filter. We measured plasma levels of prothrombin fragment 1+2, fibrin D-dimers and tissue factor in 82 HD patients before and after dialysis. Clinical endpoints during the year following blood sampling were thrombosis in blood access, changes in blood access, other thromboembolic events, bleeding complications, ischaemic vascular disease, or death. We found elevated baseline levels of all three parameters in HD patients, compared to normal reference ranges. Plasma levels of all parameters (particularly fibrin D-dimers) were significantly higher in patients with prosthetic grafts and central venous dialysis catheters than in patients with native vessels. Patients with AV-fistulas or grafts who had bleeding complications (n=7) had significantly higher plasma levels of fibrin D-dimer and prothrombin fragment 1+2. Bleeding complications also occurred more frequently among the patients with prosthetic grafts (3/18) or central venous dialysis catheters (3/11) compared with those with grafts from native vessels (1/53). Other than a bleeding tendency, our data do no show any correlation between coagulation parameters and other clinical complications during haemodialysis. In conclusion, we found elevated plasma levels of markers of coagulation activation among HD patients. High levels of D-dimers and prothrombin fragment 1+2 correlated to bleeding diathesis instead of thromboembolism, and this tendency was most pronounced in patients with prosthetic grafts.

Topics: Aged; Catheters, Indwelling; Female; Fibrin Fibrinogen Degradation Products; Hemorrhage; Humans; Kidney Failure, Chronic; Male; Middle Aged; Peptide Fragments; Prostheses and Implants; Prothrombin; Reference Values; Renal Dialysis; Thromboplastin; Thrombosis; Ultrafiltration

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

Enhanced oxidative stress (SOX), endothelial dysfunction and haemostatic abnormalities are common in end-stage renal failure patients undergoing maintenance haemodialysis (HD). We studied associations among circulating immunoreactive total lipid peroxides as a marker of short-time SOX, autoantibodies against oxidized LDL as a surrogate of prolonged SOX, copper/zinc superoxide dismutase (Cu/Zn SOD) as a major antioxidant enzyme, tissue factor (TF) as a principal initiator of extrinsic coagulation pathway counteracted by its inhibitor (TFPI), and prothrombin fragment 1+2 (F 1+2) as a surrogate of activated haemostasis.Pre-dialysis blood levels of all the markers studied were higher in 24 clinically stable HD patients compared to 11 healthy controls. Spearman's correlations among the three SOX markers were positive but nonsignificant in both HD patients and controls. In HD subjects, increased Cu/Zn SOD levels directly correlated with those of TF (rho=0.551, p=0.005) and TFPI (rho=0.501, p=0.001); the coagulation markers were also positively associated with each other (rho=0.663, p=0.0004). In healthy subjects, the relations between Cu/Zn SOD, TF and TFPI levels were inverse but not significant, and the direct association between TF and TFPI was nonsignificant either. In conclusion, increased plasma levels of Cu/Zn SOD, the antioxidant enzyme with emerging endothelial cell-protective and antithrombotic properties, may be a novel part of the system counteracting activated extrinsic coagulation system in maintenance HD patients.

Topics: Aged; Biomarkers; Blood Coagulation; Endothelium, Vascular; Female; Humans; Kidney Failure, Chronic; Lipid Peroxidation; Lipoproteins; Lipoproteins, LDL; Male; Middle Aged; Oxidative Stress; Peptide Fragments; Prothrombin; Renal Dialysis; Superoxide Dismutase; 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

Topics: Adult; Aged; Angiotensin-Converting Enzyme Inhibitors; Biomarkers; Case-Control Studies; Endothelium, Vascular; Female; Humans; Kidney Failure, Chronic; Lipoproteins; Male; Middle Aged; Proteinuria; Thrombomodulin; Thromboplastin; von Willebrand Factor

Statins are effective in prevention of end-organ damage; however, the benefits cannot be fully explained on the basis of cholesterol reduction. We used an angiotensin II (Ang II)-dependent model to test the hypothesis that cerivastatin prevents leukocyte adhesion and infiltration, induction of inducible nitric oxide synthase (iNOS), and ameliorates end-organ damage.. We analyzed intracellular targets, such as mitogen-activated protein kinase and transcription factor (nuclear factor-kappaB and activator protein-1) activation. We used immunohistochemistry, immunocytochemistry, electrophoretic mobility shift assays, and enzyme-linked immunosorbent assay techniques. We treated rats transgenic for human renin and angiotensinogen (dTGR) chronically from week 4 to 7 with cerivastatin (0.5 mg/kg by gavage).. Untreated dTGR developed hypertension, cardiac hypertrophy, and renal damage, with a 100-fold increased albuminuria and focal cortical necrosis. dTGR mortality at the age of seven weeks was 45%. Immunohistochemistry showed increased iNOS expression in the endothelium and media of small vessels, infiltrating cells, afferent arterioles, and glomeruli of dTGR, which was greater in cortex than medulla. Phosphorylated extracellular signal regulated kinase (p-ERK) was increased in dTGR; nuclear factor-kappaB and activator protein-1 were both activated. Cerivastatin decreased systolic blood pressure compared with untreated dTGR (147 +/- 14 vs. 201 +/- 6 mm Hg, P < 0.001). Albuminuria was reduced by 60% (P = 0.001), and creatinine was lowered (0.45 +/- 0.01 vs. 0.68 +/- 0.05 mg/dL, P = 0. 003); however, cholesterol was not reduced. Intercellular adhesion molecule-1 and vascular cell adhesion molecule-1 expression was diminished, while neutrophil and monocyte infiltration in the kidney was markedly reduced. ERK phosphorylation and transcription factor activation were reduced. In addition, in vitro incubation of vascular smooth muscle cells with cerivastatin (0.5 micromol/L) almost completely prevented the Ang II-induced ERK phosphorylation.. Cerivastatin reduced inflammation, cell proliferation, and iNOS induction, which led to a reduction in cellular damage. Our findings suggest that 3-hydroxy-3-methylglutaryl coenzyme (HMG-CoA) reductase inhibition ameliorates Ang II-induced end-organ damage. We suggest that these effects were independent of cholesterol.

Topics: Albuminuria; Angiotensin II; Angiotensinogen; Animals; Animals, Genetically Modified; Blood Pressure; Cell Division; Cholesterol; Creatinine; Disease Models, Animal; Humans; Hydroxymethylglutaryl-CoA Reductase Inhibitors; Intercellular Adhesion Molecule-1; Kidney; Kidney Failure, Chronic; Leukocytes; Male; Mitogen-Activated Protein Kinases; NF-kappa B; Nitric Oxide Synthase; Nitric Oxide Synthase Type II; Organ Size; Phosphorylation; Plasminogen Activators; Pyridines; Rats; Rats, Sprague-Dawley; Renin; Thromboplastin; Transcription Factor AP-1; Urea; Vascular Cell Adhesion Molecule-1; Vasoconstrictor Agents

To investigate the expression of monocyte tissue factor (MTF) and adhesion molecules in patients with chronic renal failure (CRF) and to look for any correlation with thrombin generation and Lp(a) lipoprotein.. A study of MTF expression and adhesion molecules, prothrombin fragments 1+2 (PTf1+2), an index of thrombin generation, and lipoproteins in patients with CRF and in normal control subjects.. Patients with end stage renal failure have an increased risk of coronary artery disease despite advances in therapy. Stimulated monocytes are potent activators of blood coagulation through the generation of MTF, which was recently implicated in the aetiology of acute coronary ischaemic syndromes.. MTF expression and adhesion molecules were measured in whole blood using immunofluorescence of monocytes labelled with anti-tissue factor antibody and CD11b and c by flow cytometry. PTf1+2 and Lp(a) lipoprotein in plasma were measured by enzyme linked immunosorbent assay (ELISA).. 70 patients with CRF without documented coronary artery disease (30 patients with CRF undialysed, 20 patients undergoing chronic ambulatory peritoneal dialysis (CAPD), and 20 undergoing haemodialysis (HD)), together with 20 normal controls, were studied.. The (mean (SD)) increased MTF of CRF (48.0 (29) v 33.3 (7.2) mesf unit/100 monocytes in controls, p = 0.04) was more pronounced in patients undergoing dialysis (HD 73.1 (32.8) (p < 0.003) and CAPD 62.8 (28.9) mesf unit/100 monocytes, p < 0.04). MTF activity showed a positive correlation with both PTf1+2 and serum creatinine (p < 0.003) but not with Lp(a) lipoprotein. Lp(a) lipoprotein was significantly increased in both dialysis groups compared with controls (p < 0.005) and non-dialysis CRF groups (p < 0.02). Monocyte adhesion molecule (CD11b) was significantly higher in all three CRF groups than in the controls (p = 0.006).. This study has demonstrated a hypercoagulable state in patients with CRF. This was especially pronounced in the dialysis patients. These findings provide a possible explanation for the increased cardiovascular and cerebrovascular morbidity and mortality in these patients.

Topics: Case-Control Studies; Coronary Disease; Flow Cytometry; Humans; Kidney Failure, Chronic; Lipoprotein(a); Macrophage-1 Antigen; Monocytes; Peritoneal Dialysis, Continuous Ambulatory; Prothrombin; Regression Analysis; Renal Dialysis; Statistics, Nonparametric; Thrombin; Thromboplastin

Hemorrhagic complications are common among hemodialysis (HD) patients. The mechanisms by which HD perturbs the coagulation cascade are still being defined. This study evaluated the influence of HD serum on cellular expression of tissue factor (TF), a procoagulant membrane-associated protein that is a pivotal regulator of blood coagulation. Serum was collected immediately before dialysis and 15, 30, and 180 min into HD using polysulfone membranes. Serum was then assessed for its ability to influence basal and cytokine-stimulated TF activity in human umbilical vein endothelial cells and ECV304 cells. Predialysis serum did not influence basal levels of TF activity. HD was associated with the appearance of a serum factor that suppressed basal TF activity (TF units/microg protein: predialysis serum 8.2 +/- 0.9; 180-min dialysis serum 4.9 +/- 0.6; P < 0.05) and TF activity induced by the cytokine tumor necrosis factor-alpha (TNFalpha) (TF units/microg protein: TNFalpha alone 15.9 +/- 0.7; TNFalpha + 180-min dialysis serum 5.9 +/- 0.9; P < 0.01). This response was not mimicked by heparin, suggesting production of an endogenous inhibitor of TF activity during HD. Dialysis was associated with a striking increase in circulating levels of tissue factor pathway inhibitor (TFPI), a physiologic inhibitor of the TF/VIIa complex. The lack of temporal correlation between TFPI levels and suppression of TF activity, however, suggested the presence of additional TFPI independent pathway(s) for modulation of TF activity. Dialysis-related suppression of TF expression may contribute to hemorrhagic complications in HD patients.

Topics: Adult; Aged; Cells, Cultured; Endothelium, Vascular; Female; Humans; Kidney Failure, Chronic; Lipoproteins; Male; Middle Aged; Renal Dialysis; Thromboplastin; Tumor Necrosis Factor-alpha

The phenotypic alterations between blood monocytes from 11 patients with end-stage renal disease, who had been on peritoneal dialysis for less than one week, and blood monocytes from 10 healthy controls, were analyzed. In addition, peritoneal macrophages in the dialysate effluent were enclosed. Analysis of functional receptor density was performed using immunostaining and flow cytometry. The phenotypic characterization was selected to represent various biological functions such as adhesion, phagocytosis (CD11b/CD18, CD11c/CD18, CD16), antigen-presentation (HLA-DR, ICAM-1), differentiation (transferrin receptor, CD71), receptor for LPS (CD14) and initiation of the coagulation cascade (Tissue factor, CD142). The proportion of CD16-positive blood monocytes and the quantitative level of ICAM-1 were higher in the patient group, compared to healthy controls. A significant increase in the quantitative level of CD11b/CD18, CD11c/CD18, HLA-DR and ICAM-1, transferrin receptor, CD14 and CD16, was found on peritoneal macrophages, compared to monocytes, harvested both from the corresponding patients, as well as from healthy donors. In contrast, we did not find any significant differences in the expression of tissue factor between monocytes and peritoneal macrophages. In conclusion, phenotypic differences exist between monocyte populations in the blood circulation of CAPD patients, and healthy individuals. We also show that transmigration of monocytes into the peritoneal cavity implies a selective up-regulation of functional receptors, preferentially related to adhesion, and antigen-presentation in a steady-state situation in non-infected CAPD patients.

Topics: Adult; Aged; Aged, 80 and over; Antigens, CD; Cell Adhesion; Female; HLA-DR Antigens; Humans; Immunophenotyping; Kidney Failure, Chronic; Macrophages, Peritoneal; Male; Middle Aged; Monocytes; Peritoneal Cavity; Peritoneal Dialysis, Continuous Ambulatory; Phenotype; Receptors, IgG; Receptors, Transferrin; Thromboplastin

Tissue factor and tissue factor pathway inhibitor are important in extrinsic coagulation. We investigated their clinical significance in hemodialysis patients. We took blood samples, prior to initiation of routine hemodialysis, from 73 patients on hemodialysis (35 men and 38 women aged 56.1+/-11.7 years on dialysis for 82.1+/-61.0 months), and determined tissue factor and tissue factor pathway inhibitor levels by ELISA. In the patients the tissue factor level was 704.5+/-141.6 pg/ml and the tissue factor pathway inhibitor level was 44.5+/-23.3 ng/ml; both values were significantly higher than in normal controls (192.7+/-36.6 pg/ml and 18.6+/-5.7 ng/ml, respectively). In patients with shunt obstruction, tissue factor pathway inhibitor levels were significantly higher than in those without it. Therefore, the tissue factor pathway inhibitor level may be a marker of shunt obstruction.

Topics: Adult; Female; Humans; Kidney Failure, Chronic; Lipoproteins; Male; Middle Aged; Renal Dialysis; Thromboplastin

Activated partial thromboplastin time (aPTT) during heparin infusion in 10 patients on regular dialytic treatment was evaluated. At end-dialysis (3 hours) plasma heparinization ranged between 2000-4000 IU and from the clinical point of view dialytic performance was good. At end-dialysis aPTT was 53 +/- 14 seconds: this value is under the prescribed aPTT prolongation (1.5-2 times baseline values) to achieve a therapeutic heparin plasma level. Physiopathological implications of this aPTT "resistance" to heparin infusion are studied.

Topics: Adult; Aged; Antithrombin III; Fibrinogen; Heparin; Humans; Kidney Failure, Chronic; Middle Aged; Partial Thromboplastin Time; Prothrombin; Renal Dialysis; Thromboplastin

Over a period of 10 months hemodialyses in 20 patients with chronic renal failure were performed with a 40% lower heparin-dose. Blood coagulation was controlled from the activated partial thromboplastin time (APTT). There were no unfavourable side-effects, also the effectivity of the hemodialyses was not changed. However, we measured a significant increase in the hemoglobin-concentration and in the packed cell volume. Bleeding from the punctures was significantly shortened.

Topics: Adult; Aged; Blood Coagulation Tests; Female; Hematocrit; Hemoglobins; Heparin; Humans; Kidney Failure, Chronic; Male; Middle Aged; Renal Dialysis; Thromboplastin

A summarized account of our experiences over 1 1/2 years is given, in relation to a minimum heparinisation technique during haemodialysis. This novel technique was made possible by the introduction of an APTT bedside method. In a comparison between the techniques employed so far for preventing a heparin-caused risk of haemorrhage and our method, the clear advantage of our method was apparent. In addition to a discussion of our methods, we describe the cases treated so far. The amounts of heparin required for dialysis are so small that a necessary coagulation can occur even in a part of the organism where there is a danger of haemorrhage. Thereby it was possible to extend the range of indication for haemodialysis substantially. By using minimum heparinisation, it is possible to perform an immediate postoperative haemodialysis. The healing of wounds, which is impaired in cases of renal insufficiency, may be improved by early dialysis without the risk of haemorrhage. Our results show that the minimum heparinisation signifies a decisive achievement in acute dialysis therapy.

Topics: Acute Kidney Injury; Adult; Aged; Blood Coagulation Tests; Female; Hemorrhage; Heparin; Humans; Kidney Failure, Chronic; Male; Middle Aged; Monitoring, Physiologic; Renal Dialysis; Risk; Thromboplastin; Wounds and Injuries

The consistency of the anticoagulant effect of intravenously administered heparin was studied. The activated partial thromboplastin time (APTT) was measured for six patients hourly during three consecutive hemodialysis sessions each. Cephaloplastin was the plasma-activating agent. The time required to form a clot was measured by a light-sensitive electronic timer and confirmed within +/- 5% by the tilt tube method. Results are reported in second relative to units of heparin given to patients per kilogram body weight. The range of APTT's measured 55 minutes after each heparin dose greatly exceeded the range of technical variability of the assay method. The probably mechanisms and consequences for this variability after a constant heparin dose are discussed. The anticoagulation effect of heparin during hemodialysis in an otherwise stable clinical situation is not constant. The risks of having too much or too little anticoagulation are not eliminated by having determined a therapeutic heparin dose during one dialysis rung.

Topics: Blood Coagulation Tests; Body Weight; Female; Heparin; Humans; Kidney Failure, Chronic; Male; Renal Dialysis; Thromboplastin

Topics: Blood Cell Count; Blood Coagulation; Blood Coagulation Tests; Blood Platelets; Capillary Resistance; Child; Factor VIII; Fibrinogen; Hemolytic-Uremic Syndrome; Hemophilia A; Heparin; Hot Temperature; Humans; In Vitro Techniques; Infant; Kidney Failure, Chronic; Male; Nephrotic Syndrome; Plasminogen; Platelet Adhesiveness; Platelet Aggregation; Prothrombin Time; Serum Globulins; Thromboplastin

Topics: Adenine Nucleotides; Adenosine Diphosphate; Blood Platelets; Carbon Isotopes; Epinephrine; Humans; Kidney Failure, Chronic; Phenols; Platelet Adhesiveness; Serotonin; Thrombin; Thromboplastin

Topics: Analysis of Variance; Animals; Blood Coagulation Tests; Dogs; Female; Heparin; Humans; Injections, Intravenous; Kidney Failure, Chronic; Male; Methods; Monitoring, Physiologic; Renal Dialysis; Sodium; Thromboplastin; Time Factors

Topics: Adolescent; Adult; Blood Coagulation Disorders; Female; Fibrinolysis; Hemorrhagic Disorders; Humans; Kidney Failure, Chronic; Male; Middle Aged; Thromboplastin

Topics: Blood Coagulation; Blood Coagulation Tests; Blood Platelets; Hemorrhagic Disorders; Humans; Kidney Failure, Chronic; Kidney Transplantation; Thromboplastin; Transplantation, Homologous; Uremia