Plasma Levels Of Tissue Factor Research Articles

Edoxaban, a Factor Xa-Specific Direct Oral Anticoagulant, Significantly Suppresses Tumor Growth in Colorectal Cancer Colon26-Inoculated BALB/c Mice.

Introduction Certain low-molecular-weight heparins have been reported to reduce tumor growth and metastasis in tumor cell-inoculated mouse models and cancer patients. Recently, direct oral anticoagulants (DOACs) have been widely used in patients with thromboembolism. This study was aimed at investigating the effect of DOACs, which target thrombin or factor Xa, on tumor growth in a syngeneic mouse model comprising BALB/c mice inoculated with colon cancer Colon26 cells. Materials and Methods DOACs targeting thrombin (dabigatran etexilate [DABE]) or factor Xa (rivaroxaban [RVX] and edoxaban [EDX]) were orally administered daily to male BALB/c mice inoculated with Colon26 cells, followed by analyses of tumor growth and plasma levels of coagulation- and tumor-related factors such as tissue factor (TF), plasminogen activator inhibitor-1 (PAI-1), interleukin-6 (IL-6), and matrix metalloproteinase-2 (MMP-2). Results Colon26 cells expressed significant amounts of functionally active TF. Tumor growth in Colon26-inoculated mice was significantly suppressed in DABE- or RVX-treated mice ( p <0.05) and was suppressed more significantly in EDX-treated mice ( p <0.01). Therefore, the antitumor mechanism of action of EDX was investigated next. Plasma levels of TF, PAI-1, IL-6, and MMP-2 were elevated in Colon26-inoculated mice but were significantly reduced in EDX-treated mice ( p <0.01). The expression of protease-activated receptor (PAR)1, PAR2, signal transducer and activator of transcription-3 (STAT3), cyclin D1, and Ki67 was increased in tumor tissue of Colon26-inoculated mice but (except for PAR1) was significantly decreased in tumor tissues of EDX-treated mice ( p <0.01). In addition, apoptotic cells and p53 protein levels were significantly increased in tumor tissues of EDX-treated mice. Conclusion The data suggest that among the tested DOACs, EDX significantly suppresses tumor cell proliferation via the factor Xa-PAR2 pathway, which is activated by coagulation and inflammation in Colon26-inoculated mice and induces tumor cell apoptosis.

COVID-19 coagulopathy - what should we treat?

New Findings What is the topic of this review? Overview of the coagulation abnormalities, including elevated D‐dimers widely reported with COVID‐19, often labelled as COVID coagulopathy. What advances does it highlight? The review highlights the changes in bronchoalveolar haemostasis due to apoptosis of alveolar cells, which contributes to acute lung injury and acute respiratory distress syndrome; the pathophysiological mechanisms, including endothelial dysfunction and damage responsible for thrombosis of pulmonary microcirculation and potential contribution to the hypoxaemia of COVID‐19 acute lung injury; and changes in coagulation proteins responsible for the hypercoagulability and increased risk of thrombosis in other venous and arterial beds. The rationale for anticoagulation and fibrinolytic therapies is detailed, and potential confounders that might have led to less than expected improvement in the various randomised controlled trials are considered. Coronavirus disease 19 (COVID‐19) causes acute lung injury with diffuse alveolar damage, alveolar–capillary barrier disruption, thrombin generation and alveolar fibrin deposition. Clinically, hypoxaemia is associated with preserved lung compliance early in the disease, suggesting the lack of excessive fluid accumulation typical of other lung injuries. Notably, autopsy studies demonstrate infection of the endothelium with extensive capillary thrombosis distinct from the embolic thrombi in pulmonary arteries. The inflammatory thrombosis in pulmonary vasculature secondary to endothelial infection and dysfunction appears to contribute to hypoxaemia. This is associated with elevated D‐dimers and acquired hypercoagulability with an increased risk of deep vein thrombosis. Hypercoagulability is secondary to elevated plasma tissue factor levels, von Willebrand factor, fibrinogen, reduced ADAMTS‐13 with platelet activation and inhibition of fibrinolysis. Multi‐platform randomised controlled studies of systemic therapeutic anticoagulation with unfractionated and low molecular mass heparins demonstrated a survival benefit over standard care with full‐dose anticoagulation in patients with non‐severe disease who require supplemental oxygen, but not in severe disease requiring ventilatory support. Late intervention and the heterogeneous nature of enrolled patients can potentially explain the apparent lack of benefit in severe disease. Improvement in oxygenation has been demonstrated with intravenous fibrinolytics in small studies. Inhaled anticoagulants, thrombolytic agents and non‐specific proteolytic drugs in clinical trials for decreasing alveolar fibrin deposition might benefit early disease. Essentially, COVID‐19 is a multi‐system disorder with pulmonary vascular inflammatory thrombosis that requires an interdisciplinary approach to combination therapies addressing both inflammation and intravascular thrombosis or alveolar fibrin deposits to improve outcomes.

Meglumine Cyclic Adenylate Improves Coagulation Abnormalities in Endotoxemic Rats

<p id="p00005"><strong>Background:</strong> Sepsis and endotoxemia are linked to the concurrent activation of inflammatory and hemostatic pathways, which contribute to organ dysfunction and death. Meglumine cyclic adenylate (MCA) has been found to inhibit NF-κB activity during endotoxemia in rats. This study aims to investigate the effect of MCA on coagulation abnormalities in endotoxemic rats. <p id="p00010"><strong>Methods:</strong> seventy-two male SD rats were randomly divided into three groups (n=24 per group): Normal saline group (NS), group LPS, and group MCA. Endotoxemia was induced by LPS 10 mg/kg injected via the femoral vein in the LPS and MCA groups. MCA 2mg/kg was injected 20 min before LPS injection in group MCA via the femoral vein. An equal volume of normal saline was given to NS and LPS groups. Prothrombin time (PT), partial activated thromboplastin time (APTT), platelet count, cAMP, plasminogen activator inhibitor-1(PAl-1), D-dimer, TNF-α, antithrombin III (ATIII), t-PA and Tissue factor (TF) were assessed before (baseline) and 2, 4, 6 h after LPS injection. <p id="p00015"><strong>Results:</strong> After LPS administration, PT and APTT were prolonged, ATIII concentrations, cAMP, and platelet count were significantly decreased, while plasma PAI-1, D-dimer, t-PA, TNF-α, and TF levels were significantly increased compared to group NS. However, these alterations induced by LPS were inhibited by MCA treatment. <p id="p00020"><strong>Conclusion:</strong> MCA amelio-rates coagulation abnormalities induced by LPS in endotoxemic rats.

Tissue factor and its procoagulant activity on cancer-associated thromboembolism in pancreatic cancer.

Pancreatic cancer frequently involves cancer‐associated thromboembolism, which is strongly associated with poor prognosis. Tissue factor, a blood coagulation factor largely produced in cancer patients as a component of extracellular vesicles, plays a key role in the incidence of cancer‐associated thromboembolism in patients with pancreatic cancer. However, no prospective studies have been published on the relationship between tissue factor and cancer‐associated thromboembolism or patient clinical characteristics, including recent chemotherapy regimens. Thus, we aimed to address this in a Japanese cohort of 197 patients and 41 healthy volunteers. Plasma tissue factor levels were measured by ELISAs preevaluated by tissue factor specificity. Multivariable analysis was used to identify independent predictors of cancer‐associated thromboembolism. We found that the cancer‐associated thromboembolism rate in the patient cohort was 6.6% (4.6%, venous thromboembolism; 2.0%, arterial thromboembolism). Tissue factor levels of 100 pg/mL or higher at patient registration were predictive of cancer‐associated thromboembolism, with positive and negative predictive values of 23.1% and 94.6%, respectively. Multivariable analysis showed that plasma tissue factor levels were an independent predictive factor for cancer‐associated thromboembolism, with a risk ratio of 5.54 (95% confidence interval, 1.02‐30.09). Unlike in healthy volunteers and patients without cancer‐associated thromboembolism, tissue factor levels were highly correlated with extracellular vesicles’ procoagulant activity in patients developing cancer‐associated thromboembolism. Taken together, our data show that the tissue factor levels at patient registration were a predictive factor for cancer‐associated thromboembolism in this cohort of patients with pancreatic cancer.

Biomarkers of Endothelial Function in Sepsis-Associated Disseminated Intravascular Coagulation

Introduction: Sepsis-associated disseminated intravascular coagulation (DIC) is a complex clinical syndrome with high mortality. The pathogenesis of DIC involves significant interactions between multiple pathophysiologic processes including coagulation, inflammation, infection response, platelets and endothelial dysfunction. The role of the endothelium in DIC is multifaceted and may contribute significantly to its severity and outcome. In sepsis, increased inflammation can lead to activation of the endothelium and the development of a procoagulant phenotype, characterized in part by elevated levels of cell surface tissue factor (TF). The endothelial response to inflammation is also characterized by the production of other factors that may be descriptive of the status of the endothelium, including endocan. The endothelium is also largely responsible for the production of endogenous anticoagulants, such as Tissue Factor Pathway Inhibitor (TFPI) and Protein C. The derangement of the endogenous anticoagulant system is critical to the pathogenesis of DIC and may also be reflective of the state of the endothelium. Investigation of factors that may prove damaging to the endothelium, including extracellular nuclear material such as nucleosomes and the protein High Mobility Group Box 1 (HMGB-1) may also provide insight into endothelial status and associated patient prognosis. The purpose of this study was to quantify measures of endothelial function, including markers of activation (TF and endocan), endogenous anticoagulants (TFPI and Protein C), and damage-associated factors (nucleosomes and HMGB-1) in the plasma of patients with sepsis and DIC and to determine the relationship of these factors with severity of illness and outcome.Materials and Methods: Citrated plasma samples were prospectively collected from 103 patients with sepsis and suspected DIC at ICU admission (day 0) and again serially on ICU days 4 and 8 under an IRB approved protocol. DIC score was determined using the ISTH scoring algorithm (e.g. platelet count, PT/INR, fibrinogen, and D-Dimer). Plasma from healthy individuals was purchased from a commercial laboratory (George King Biomedical, Overland, KS). Plasma levels of endocan, TF, TFPI, nucleosomes, and HMGB-1 were evaluated using commercially available ELISA kits. Protein C levels were measured using a functional clot-based assay.Results: In the DIC patients, endocan, TF, TFPI, nucleosomes, and HMGB-1 were significantly elevated compared to normal on ICU days 0, 4, and 8. Protein C was significantly reduced compared to normal at all time points.Patients were stratified into groups of sepsis and DIC based on the ISTH DIC score, and levels of markers were compared between groups. Markers reflective of overall endothelial activation (endocan and TF) showed no variation based on DIC score. Protein C showed significant variation based on DIC score, with a marked drop in Protein C levels with increasing severity of illness; however, TFPI showed no variation. Increasing levels of factors associated with endothelial damage (nucleosomes and HMGB-1) were observed with increasing severity of illness. Results are shown in Table 1.Differences in biomarker levels between survivors and non-survivors were also evaluated. Endocan (p = 0.0252) and HMGB-1 (p = 0.0306) showed significant increases in non-survivors compared to survivors while Protein C showed a significant reduction (p = 0.0093) in non-survivors compared to survivors. TF, TFPI, and nucleosomes showed no variation based on survival.Conclusion: Overall, levels of markers of endothelial activation, endogenous anticoagulants, and factors contributing to endothelial damage were elevated in the plasma of patients with sepsis-associated DIC compared to the healthy population. In particular, factors associated with endothelial damage, namely the toxic nuclear factors nucleosomes and HMGB-1, showed elevated levels with increasing severity of coagulopathy, suggesting that the endothelial damage caused by these factors may contribute to the development of coagulation dysfunction. Levels of endocan, Protein C, and HMGB-1 also showed significant association with mortality, underscoring the relevance of all aspects of endothelial physiology to patient outcome in DIC. [Display omitted] DisclosuresNo relevant conflicts of interest to declare.

Hyperglycemia Potentiates Prothrombotic Effect of Aldosterone in a Rat Arterial Thrombosis Model

We investigated the role of aldosterone (ALDO) in the development of arterial thrombosis in streptozotocin-induced diabetic rats. To evaluate the effect of endogenous ALDO, the rats underwent adrenalectomy (ADX). ADX reduced the development of arterial thrombosis. A 1 h infusion of ALDO (30 μg/kg/h) enhanced thrombosis in adrenalectomized rats, while this effect was potentiated in diabetic rats. ALDO shortened bleeding time, increased plasma levels of tissue factor (TF) and plasminogen activator inhibitor, decreased plasma level of nitric oxide (NO) metabolites, and increased oxidative stress. Moreover, 2 h incubation of human umbilical vein endothelial cells (HUVECs) with ALDO (10−7 M) disrupted hemostatic balance in endothelial cells in normoglycemia (glucose 5.5 mM), and this effect was more pronounced in hyperglycemia (glucose 30 mM). We demonstrated that the acute ALDO infusion enhances arterial thrombosis in rats and hyperglycemia potentiates this prothrombotic effect. The mechanism of ALDO action was partially mediated by mineralocorticoid (MR) and glucocorticoid (GR) receptors and related to impact of the hormone on primary hemostasis, TF-dependent coagulation cascade, fibrinolysis, NO bioavailability, and oxidative stress balance. Our in vitro study confirmed that ALDO induces prothrombotic phenotype in the endothelium, particularly under hyperglycemic conditions.

Procoagulant Disorders in Patients with Newly Diagnosed Pancreatic Adenocarcinoma.

Background and objectives: Cancer coagulopathy is thought to be partially due to the up-regulation of tissue factor (TF), thrombin-antithrombin complex (TAT) and soluble P-selectin (sP-selectin). The purpose of this study was to evaluate the clinical significance of TF, TAT and sP-selectin in patients with pancreatic cancer. Materials and methods: The study included 93 subjects: 73 newly diagnosed patients with pancreatic adenocarcinoma (42 with stage I-III and 31 with metastatic cancer (stage IV)) and a control group of 20 healthy subjects. Analyzed patients were hospitalized in the Department of Digestive Tract Diseases, Medical University of Lodz or in the Department of Digestive Tract Surgery, Silesian University, Katowice, Poland. All laboratory parameters were measured using ELISA procedures. Results: TF plasma levels were detectable in all patients and were significantly higher in metastatic cancer compared to stage I-III patients and the control group (p < 0.05). In patients with pancreatic adenocarcinoma, the median levels of TAT were also elevated compared to the control group. Moreover, patients with metastases had significantly higher TAT concentration compared to the I-III cancer group. On the other hand, only the metastatic patients group showed significantly higher plasma sP-selectin levels compared to the controls (p = 0.009), whereas there was no difference between localized and metastatic cancer patients. Conclusions: The coagulation disorders are present in the majority of patients with pancreatic adenocarcinoma already at the diagnosis stage and reflect cancer progression and spread.

Changes of Tissue Factors and Tissue Factor Pathway Inhibitors in the Acute Phase of Cardiogenic Stroke in Patients With Atrial Fibrillation

Abstract Background To investigate changes of tissue factor (TF) and tissue factor pathway inhibitor (TFPI) in the acute phase of cardiogenic stroke in patients with atrial fibrillation (AF). Methods A total of 152 patients with nonvalvular persistent AF, 103 cardiogenic stroke patients with nonvalvular persistent AF, and 126 cases with noncardiogenic stroke were enrolled. Non-AF patients (n = 138) without stroke served as controls. The clinical data, biochemical indicators, and plasma levels of von Willebrand factor (vWF), TF, and TFPI were determined. Results Compared with controls, the plasma levels of vWF and TF were significantly higher, while TFPI and TFPI/TF ratio were lower in noncardiogenic stroke patients (all P &amp;lt; 0.05). Compared with cardiogenic stroke patients, noncardiogenic stroke subjects showed significantly higher plasma vWF and TF levels while TFPI levels and TFPI/TF ratio were significantly lower (all P &amp;lt; 0.05). Compared with AF patients, plasma levels of vWF, TF, and TFPI were higher while TFPI/TF ratios were decreased in the cardiogenic ischemic stroke subjects (all P &amp;lt; 0.05). Logistic regression analysis showed that a lower TFPI/TF ratio was an independent predictor of cardiogenic stroke in patients with nonvalvular AF (odds ratio = 3.78, 95% confidence interval: 1.86–7.31). Conclusions During the acute stage of stroke, TFPI/TF ratio is decreased in both cardiogenic and noncardiogenic stroke patients while plasma TFPI levels show divergent changes.

Interleukin-1β Mediates Arterial Thrombus Formation via NET-Associated Tissue Factor.

CANTOS reported reduced secondary atherothrombotic events in patients with residual inflammatory risk treated with the inhibitory anti-IL-1β antibody, Canakinumab. Yet, mechanisms that underlie this benefit remain elusive. Recent work has implicated formation of neutrophil extracellular traps (NETosis) in arterial thrombosis. Hence, the present study explored the potential link between IL-1β, NETs, and tissue factor (TF)—the key trigger of the coagulation cascade—in atherothrombosis. To this end, ST-elevation myocardial infarction (STEMI) patients from the Swiss multicenter trial SPUM-ACS were retrospectively and randomly selected based on their CRP levels. In particular, 33 patients with STEMI and high C-reactive protein (CRP) levels (≥ 10 mg/L) and, 33 with STEMI and low CRP levels (≤ 4 mg/L) were investigated. High CRP patients displayed elevated circulating IL-1β, NETosis, and NET-associated TF plasma levels compared with low CRP ones. Additionally, analysis of patients stratified by circulating IL-1β levels yielded similar results. Moreover, NETosis and NET-associated TF plasma levels correlated positively in the whole population. In addition to the above, translational research experiments provided mechanistic confirmation for the clinical data identifying IL-1β as the initial trigger for the release of the pro-coagulant, NET-associated TF. In conclusion, blunted TF presentation by activated neutrophils undergoing NETosis may provide a mechanistic explanation to reduced secondary atherothrombotic events as observed in canakinumab-treated patients in CANTOS.

707P - Cancer-associated thrombosis in patients with pancreatic cancer and its correlation with plasma tissue factor level: A Japanese prospective cohort study

BackgroundCancer-associated thrombosis (CAT) is a well–known complication of cancer, especially pancreatic cancer. One possible cause of CAT is secretion of tissue factor (TF)-positive extracellular vesicles from cancer cells. The previous cohort of Western countries showed that patients (pts) with a high level of TF tended to develop CAT with 20–40% incidence in pancreatic cancer. This prospective cohort study aimed to assess the efficacy of TF level for predicting the incidence of CAT in Japanese pts with pancreatic cancer. MethodsWe registered consented 197 pts from June 2013 to November 2016, who had pathologically confirmed pancreatic cancer and no history of treatment for the current disease. Plasma samples were collected from pts at the time of registration, 3, 6, and 12 months after. The TF level was evaluated using the Quantikine® ELISA kit (R&D systems) as we found that TF levels in multiple cancer cell lines evaluated using this kit correlated well with those analyzed by western blotting experiments. We estimated the positive (PPV) and negative predictive values (NPV) for the incidence of CAT by using obtained plasma TF levels (cutoff value: 100pg/ml) and D-dimer levels (cutoff value: 1.2μg/ml). ResultsOf all the patients, 189 patients were with unresectable disease. The median of the initial level of TF for these pts was 47.8pg/ml (range: 0.0—318.7). The incidence of CAT was observed in 13 pts (6.6%) at the median follow-up time of 325 days. The median TF level at the time of registration was 148.5pg/ml in CAT+ group and 53.5pg/ml in CAT- group with a p–value of<0.0001. PPV and NPV estimated by TF levels at the time of registration were 23.1% and 94.6%, and by the D-dimer levels were 7.2% and 92.4%, respectively. In pts with a D-dimer level >1.2 µg/ml, NPV by TF<100pg/ml was 95%. Similar results were obtained when TF levels of 3 months before the onset of CAT were used. ConclusionsPresent study revealed that the incidence of CAT in Japanese pts was lower than that of Western countries. It was also found that TF levels alone could not predict CAT observed in Japanese pts with pancreatic cancer: however, TF levels were found to contribute to high NPV even in pts with a high level of D–dimer. Legal entity responsible for the studyThe authors. FundingHas not received any funding. DisclosureAll authors have declared no conflicts of interest.

The effects of citrate dialysate in hemodialysis on polymorphonuclear elastase interaction with tissue factor and its inhibitor.

This study aimed to investigate whether hemodialysis (HD) affects tissue factor (TF), tissue factor pathway inhibitor (TFPI), and polymorphonuclear elastase (PMNE) in endstage renal disease (ESRD) patients when eliminating the effects of heparin. Also, to explore the interaction of TF, TFPI, and PMNE throughout a single HD session. We enrolled 57 ESRD patients who had undergone hemodialysis for >3 months as an experimental group. Plasma levels of TF, TFPI and PMNE were measured by ELISA in 24 ERSD patients on intermittent HD using low-molecular-weight heparin (LMWH) as anticoagulation (LMWH group) and 33 ESRD patients using citrate as anticoagulation (citrate group) at the start and at 1, 2 and 5 h of the HD session. Meanwhile,28 ESRD patients not on dialysis were enrolled as a control group and fasting venous blood samples were taken in the morning. Compared with the control group, the plasma TFPI levels of the LMWH group and the citrate group were significantly higher (P=0.000, P=0.002, respectively) under baseline conditions as well as the plasma PMNE levels (P=0.001, P=0.02, respectively), whereas TF showed no difference (P=0.186). During HD with citrate, plasma TFPI decreased slightly (P=0.012) and PMNE increased significantly (P=0.008) at 1 h. The plasma TFPI levels of the citrate group correlate with PMNE at 2 and 5 h (P=0.001, P=0.008, respectively). ESRD patients on HD have significantly higher TFPI and PMNE levels compared to patients not on HD under baseline conditions, while TF levels were similar between the three groups. TFPI and PMNE are differently regulated, but the plasma levels correlated during HD in the citrate group. It might be possible that PMNE plays a role in anticoagulative activity through TFPI.

Evaluation of Tissue Factor -603 A/G Promoter Gene Polymorphism in Thrombotic Processes in Patients with Antiphospholipid Syndrome

<strong>Objective</strong>: Major clinical manifestations of Antiphospholipid syndrome (APS) are trombosis and thrombosis related complications. Tissue factor (TF) serves primary initiatory of the in vivo coagulation. TF gene expression may be induced in endothelial cells to initiate thrombus formation. In this study, we investigated the frequency of TF 603A/G promoter gene polymorphism in patients with antiphospholipid syndrome (APS) and its effects on TF expression and thrombotic events. <br> <strong>Materials and Methods</strong>: This study included 64 patients with APS (APS patient with thrombosis arterial and venous (n=38), antiphospholipid antibody-positive patients without thrombosis (n=26) and 44 healthy controls (HCs). Plasma TF levels and antiphospholipid antibodies were measured by ELISA method. Lupus anticoagulant was measured by activated partial thromboplastin time and kaolin clotting time. TF-603 A/G polymorphism was investigated by PCR method. <br> <strong>Results</strong>: Our results have showed that patients with APS, TF levels were found in patients significantly higher than the healthy control group (p: 0.006, p&lt;0.001, respectively). Compared between the two patient groups, TF level was significantly higher in the patient with thrombosis group compared to the patient without thrombosis group (p: 0.027). <br> <strong>Conclusion</strong>: In this study, we suggest that thrombotic complications observed in patients with APS are associated with increased TF expression.&nbsp;

Elevated plasma levels of tissue factor as a valuable diagnostic biomarker with relevant efficacy for prediction of breast cancer morbidity.

The aim of this study was to evaluate the concentrations of tissue factor (TF) and its inhibitor (TFPI), vascular endothelial growth factor A (VEGF-A), soluble forms of VEGF receptors type 1 and 2 (sVEGFR1 and aVEGFR2) in patients diagnosed with luminal A breast cancer (BrC) and in healthy individuals and to find associations of analyzed factors with demographic, clinical and pathological characteristics in a homogeneous breast cancer group. Study group consisted of 60 women aged 40 - 69 years, diagnosed with luminal-A subtype of BrC, without distant metastases (M0). Control group comprised 40 healthy women aged 45 - 63 years. Blood samples were collected from each patient in order to determine plasma levels of TF, TFPI, VEGF-A and sVEGFR1 and sVEGFR2. The examined parameters were measured by enzyme-linked immunosorbent assay (ELISA). The capacity of angiogenic and hemostatic parameters in predicting neoplasm disease was analyzed using receiver operating characteristic (ROC) curve analysis. According to ROC curve analysis, the optimum cut-off point for TF was 304.58 pg/ml, with 100% sensitivity and 100% specificity, which was calculated to discriminate between controls and malignancy patients. In luminal A BrC patients there were significantly higher concentrations of VEGF-A and TF than in controls. On the contrast the levels of sVEGF receptors type 1 and 2 as well as TFPI in luminal-A BrC cases were significantly lower in respect to healthy volunteers. Levels of examined factors in the study group varied depending on age, menopausal status, lymph node involvement and histological type. We concluded that altered levels of examined factors in patients diagnosed with luminal-A breast cancer indicate increased activation of angiogenesis and hemostasis. The results obtained may be indicative of a mutual connection between angiogenesis and hemostasis processes in tumor development and progression. Clinical and pathological parameters may possibly affect levels of angiogenic and coagulation factors.

Evaluation of plasma levels of tissue factor and tissue factor pathway inhibitor in patients with psoriasis.

IntroductionPsoriasis is a chronic, recurrent, inflammatory skin disorder with systemic involvement. It has recently been established that psoriasis is associated with an increased cardiovascular risk. Chronic skin-specific inflammation may promote atherosclerosis. Myocardial infarction or stroke can also be a result of underlying haemostasis disorders. Disorders in fibrinolysis and thrombosis in patients with psoriasis have been observed by many authors.AimThis study points to the key role played by the tissue factor (TF) and tissue factor pathway inhibitor (TFPI) in the extrinsic pathway of blood coagulation and the potential influence of microvascular disorders in inflamed psoriatic skin on TF and TFPI activity.Material and methodsThe study included 47 patients with active psoriasis vulgaris, hospitalized in the Dermatological Ward of the Regional Specialist Hospital, Research and Development Centre in Wroclaw, as well as 18 people from the control group.ResultsThere were significant differences in the blood concentrations of TF and TFPI in patients with psoriasis when compared to the control group. A low TFPI concentration in psoriatic patients may indicate an increased risk of atherosclerosis. Interpretation of a decreased level of TF in patients with psoriasis is difficult because it seems to be at odds with observations among patients with other atherosclerosis risk factors such as hypertension, hyperlipidaemia, diabetes or smoking.ConclusionsIt appears that further studies are necessary to explain this problem, perhaps to include an evaluation of TF levels in psoriatic skin.

Elevated blood plasma levels of tissue factor-bearing extracellular vesicles in patients with atrial fibrillation

BackgroundThe risk of thrombus formation in the left atrial appendage (LAA) in patients with atrial fibrillation (AF) may result from blood stasis, local endocardial changes, and/or changed blood composition. Extracellular vesicles (EVs), especially subtypes exposing tissue factor (TF), have procoagulant capacity. We hypothesized that blood concentrations of TF-bearing EVs and other procoagulant biomarkers are elevated in AF patients, particularly in the LAA lumen. MethodsFrom 13 AF patients and 12 controls a venous blood sample was drawn prior to cardiac surgery. Intraoperatively, venous blood and blood directly from the LAA was drawn. Plasma levels of EVs, including TF- and cell type specific antigen-bearing EVs, were measured using a protein microarray platform. Plasma levels of TF, von Willebrand factor (vWF), cell free deoxyribonucleic acid (cf-DNA), procoagulant phospholipids (PPLs), and total submicron particles were also evaluated. ResultsSignificantly higher EV levels, including a several-fold higher median level of TF-bearing EVs were measured in AF patients compared with controls. Median concentrations of TF and vWF were approximately 40% and 30% higher, respectively, in the AF group than in the control group, while no significant differences in levels of cf-DNA, PPLs, or total submicron particles were observed. No significant differences in levels of any of the measured analytes were observed between intraoperative venous and LAA samples. ConclusionsIncreased plasma concentrations of TF in AF patients are accompanied and probably at least partly explained by increased levels of TF-bearing EVs, which may be mechanistically involved in increased thrombogenicity in AF patients.

Clinical Significance of Detecting Tissue Factor and Tissue Factor Pathway Inhibitor in Thalassemia Patients

To investigate the changes of E-selectin, thrombin-antithrombin complex（TAT）, prothrombin fragment 1+2（F1+2）, tissue factor（TF）and tissue factor pathway inhibitor（TFPI）before and one year after splenectomy in thalassemia patients. A total of 30 thalassemia patients undergoing electric laparoscopic splenectomy and 30 normal controls（NC） were enrolled in the study．Plasma levels of E-selectin, TAT, F1+2, TF and TFPI were detected by enzyme-linked immuno sorbent assay（ELISA）. One year after splenectomy，the plasma concentrations of E-selectin, TAT, F1+2, TF, TFPI were significantly higher than those in both preoperative and NC groups．Levels of E-selectin, TAT, F1+2 before splenectomy were significantly higher than those in NC groups. In addition, there was a positive correlation between plasma TF and TFPI level before and after splenectomy, and the levels of TF and TFPI positively correlated with TAT and F1+2, respectively. After splenectomy, the platelet count increases, the activity of endothelial cells is injured, the procoagulant factor increases, the blood is in procoagulant state, the TF/TFPI shows an importent role in the thrombosis of thalassemia patieints after splenectomy and may be used to evaluate the prothrombotic state of this diasease.

Influence of tissue factor polymorphisms (603A>G and 5466A>G) on plasma tissue factor levels and their impact on deep vein thrombosis risk in young Indian population.

Deep vein thrombosis (DVT) is multifactorial disorder and well known to cause substantial morbidity and mortality. There is sparse data in the Asian population, particularly India regarding association of tissue factor (TF) gene single nucleotide polymorphisms (SNPs) with plasma TF levels in DVT. So, we analyzed the distribution of SNPs (603A>G and 5466A>G) in India, to evaluate their effect on TF levels in DVT patients. Plasma level and SNPs (603A>G and 5466A>G) of TF gene were screened in subjects (100 DVT patients and 100 controls). Patients had significantly higher TF levels than controls (patients: 84.95 ± 17.16pg/ml, controls: 70.55 ± 15.87pg/ml, p < 0.001). G allele of 603A>G polymorphism was significantly higher in patients than controls (patients: 40.5% controls: 27.5%, p = 0.004). Subjects with AG and GG genotype had significantly higher TF levels than AA genotype (p = 0.001). After multiple logistic regression analysis, risk of DVT was increased 1.398 fold (95% CI 0.738-2.651) and 4.41 fold (95% CI 1.404-13.884) with AG and GG genotype respectively. Allelic and genotypic frequencies of 5466A>G polymorphism was neither associated with TF levels nor with DVT. We found high TF level in patients with TF 603A>G polymorphism, which is an important predisposing factor in increasing risk of DVT in young Indians. Furthermore, GG genotype of 603A>G polymorphism augments the risk of thrombosis by 4.4 fold, thus highlighting the significance of this polymorphism in the development of DVT. So, we suggest that inclusion of 603A>G polymorphism in prothrombotic work-up may be helpful in making the treatment strategy in DVT patients.

Can we use biomarkers of coagulation to predict which patients with foot and ankle injury will develop deep vein thrombosis?

BackgroundOur aim was to determine whether plasma levels of Tissue Factor (TF), Vascular Cell Adhesion Molecule 1 (VCAM-1), Interleukin 6 (IL-6) or D-dimer after foot and ankle injury could predict which patients would develop deep vein thrombosis (DVT). MethodsPatients aged 18–60 years with acute foot and ankle injury had venous blood sample to measure TF, VCAM-1, IL-6 and D-dimer within 3 days of injury. Patients had bilateral lower limb venous ultrasound to assess for DVT on discharge from clinic. Results21 of 77 patients were found to have DVT (27%). There was no statistically significant association between levels of TF, VCAM-1, IL-6 or D-dimer and subsequent development of DVT. ConclusionTissue Factor (TF), Vascular Cell Adhesion Molecule-1 (VCAM-1), Interleukin-6 (IL-6) and D-dimer levels were not associated with development deep vein thrombosis in patients with acute foot and ankle injury.

Evaluation of venous thrombosis and tissue factor in epithelial ovarian cancer

ObjectiveOvarian clear cell carcinoma (OCCC) and high grade serous ovarian cancer (HGSOC) are associated with the highest risk of VTE among patients with epithelial ovarian cancer (EOC). Tissue factor (TF) is a transmembrane glycoprotein which can trigger thrombosis. We sought to evaluate if there is an association between VTE and tumor expression of tissue factor (TF), plasma TF, and microvesicle TF (MV TF) activity in this high-risk population. MethodsWe performed a case-control study of OCCC and HGSOC patients with and without VTE. 105 patients who underwent surgery at a tertiary care center between January 1995 and October 2013 were included. Plasma TF was measured with an enzyme-linked immunosorbent assay. A TF-dependent Factor Xa generation assay was used to measure MV TF activity. Immunohistochemical (IHC) analysis was performed to evaluate tumor expression of TF. Results35 women with OCCC or HGSOC diagnosed with VTE within 9months of surgery were included in the case group. Those with VTE had a worse OS, p<0.0001, with a greater than three-fold increase in risk of death, HR 3.33 (CI 1.75–6.35). There was no significant difference in median plasma TF level or MV TF activity level between patients with and without VTE. OCCC patients had greater expression of TF in their tumors than patients with HGSOC, p<0.0001. ConclusionsTFMV activity and plasma TF level were not predictive of VTE in this patient population. Given the extensive expression of TF in OCCC tumors, it is unlikely IHC expression will be useful in risk stratification for VTE in this population.

A Pilot Study of Tissue Factor-Tissue Factor Pathway Inhibitor Axis and Other Selected Coagulation Parameters in Broiler Chickens Administered in Ovo with Selected Prebiotics*.

The tissue factor (TF) - tissue factor pathway inhibitor (TFPI) axis plays a major role in hemostasis. Disorders of the coagulation system are commonly diagnosed with the help of screening tests such as prothrombin time (PT), activated partial thromboplastin time (aPTT), and plasma fibrinogen concentration (PFC). However, the effect of prebiotics on the hemostasis system has not been characterized in poultry yet. This study was designed to determine the effect of in ovo administration ofprebiotics on blood coagulation parameters of broiler chickens depending on their age. The study was conducted with 180 broiler chick embryos, the air cells of which were injected on day 12 of incubation with prebiotics (experimental groups: Bi2tos, DiNovoo and RFO) or physiological saline solution (control group). At 1, 21 and 42 days of rearing, blood was sampled from 15 broiler chickens from each group. An enzyme immunoassay was performed to determine plasma TF and TFPI levels, and PT, aPTT and PFC were determined in the chicken blood. We demonstrated that: 1) total TF levels increased with age in the experimental groups, 2) prebiotics had no significant effect on TF levels between the groups at a particular age, 3) total TFPI levels differed between both the type of in ovo injected substance and the broiler chicken age, 4) in the control group, PT and aPTT were found to increase with age whilst fibrinogen concentration decreased. The main conclusion from this pilot study is that total TF and TFPI levels change with age, however no clear patterns regarding TFPI were detected yet. The levels of PT, aPTT and PFC varied with the prebiotics administered in ovo as well as with the age of broiler chickens.