Thrombomodulin Levels Research Articles

Hemostatic Abnormalities in Dementia: A Systematic Review and Meta-Analysis.

Alzheimer's disease (AD) is considered the most frequent cause of dementia. It is known that vascular risk factors play an important role in the development and progression of this condition. Alterations in vascular walls represent documented findings in patients with AD and other dementias affecting elderly people. The authors performed a systematic review and meta-analysis, aiming to synthesize observational studies that evaluated how the hemostatic system may contribute to cognitive decline in the elderly, using papers published until April 2018 and as indexed in Medline (PubMed), Scopus, Web of Science, ScienceDirect, Lilacs, Cinahl, PsycINFO, Cochrane Central Register of Controlled Trials, and Cochrane Database of Systematic Reviews. Among 5,278 studies identified, 32 were included in the final synthesis, and these included 485 patients with mild cognitive impairment, 568 with vascular dementia (VD), 1,781 with AD, and 2,855 participants without dementia. AD patients had increased plasma von Willebrand factor (VWF) (standardized mean difference [SMD]: 2.53; 95% confidence interval [CI]: 0.10-4.95), D-dimer (SMD: 0.50; 95% CI: 0.35-0.66), plasminogen activator inhibitor-1 (SMD: 3.34; 95% CI: 1.01-5.67), thrombomodulin (SMD: 1.08; 95% CI: 0.53-1.62), and homocysteine levels (SMD: 0.65; 95% CI: 0.15-1.15). In contrast, the VD group showed increased fibrinogen levels (SMD: 0.77; 95% CI: 0.13-1.41), activated factor VII (SMD: 0.36; 95% CI: 0.05-0.67), factor VIII (SMD: 0.57; 95% CI: 0.22-0.91), VWF (SMD: 2.34; 95% CI: 0.38-4.29), D-dimer (SMD: 1.14; 95% CI: 0.51-1.78), and homocysteine (SMD: 2.17; 95% CI: 1.67-2.68). AD showed an elevation in some markers of endothelial dysfunction, whereas VD presented mostly an involvement of coagulation cascade components.

Analytical and clinical performance evaluation and comparison of automated urinalysis analyzers

This study has investigated whether high levels of Reticulocytes-C4d (R-C4d) and Platelets-C4d (P-C4d) reflecting recent activity in SLE patients are correlated with changes in natural anticoagulation components, coagulation activation and endothelial injury markers.This study included three groups: 1) healthy women (control, n = 30); 2) women with low activity of the disease (SLEDAI 2 K ≤ 4, n = 30); 3) women with active disease (moderate or high activity) (SLEDAI 2 K > 4, n = 30). Median fluorescence intensity (MFI) of R-C4d and P-C4d were determined by flow cytometry using double labeling with specific monoclonal antibodies. Endothelial injury and hypercoagulability were evaluated by measuring Thrombomodulin and D-dimer levels.Higher MFI index of R-C4d were related to the recent activity of SLE, and higher expression of P-C4d indicated an elevated risk of thrombotic complications. Increased levels of soluble thrombomodulin and D-dimer were observed in patients with active SLE.R-C4d is helpful to monitor early disease activity and PC4-d may be an important tool to detect a prothrombotic phenotype in SLE. Elevated levels of D-dimer and thrombomodulin add value to P-C4d data and corroborate a hypercoagulable profile in women with SLE, contributing to an increased prothrombotic risk associated with inflammation.

Analysis of the role of thrombomodulin in all-trans retinoic acid treatment of coagulation disorders in cancer patients

BackgroundClinical studies have shown that all-trans retinoic acid (RA), which is often used in treatment of cancer patients, improves hemostatic parameters and bleeding complications such as disseminated intravascular coagulation (DIC). However, the mechanisms underlying this improvement have yet to be elucidated. In vitro studies have reported that RA upregulates thrombomodulin (TM) expression on the endothelial cell surface. The objective of this study was to investigate how and to what extent the TM concentration changes after RA treatment in cancer patients, and how this variation influences the blood coagulation cascade.ResultsIn this study, we introduced an ordinary differential equation (ODE) model of gene expression for the RA-induced upregulation of TM concentration. Coupling the gene expression model with a two-compartment pharmacokinetic model of RA, we obtained the time-dependent changes in TM and thrombomodulin-mRNA (TMR) concentrations following oral administration of RA. Our results indicated that the TM concentration reached its peak level almost 14 h after taking a single oral dose (110 frac{mg}{m^2} ) of RA. Continuous treatment with RA resulted in oscillatory expression of TM on the endothelial cell surface. We then coupled the gene expression model with a mechanistic model of the coagulation cascade, and showed that the elevated levels of TM over the course of RA therapy with a single daily oral dose (110 frac{mg}{m^2} ) of RA, reduced the peak thrombin levels and endogenous thrombin potential (ETP) up to 50 and 49%, respectively. We showed that progressive reductions in plasma levels of RA, observed in continuous RA therapy with a once-daily oral dose (110 frac{mg}{m^2} ) of RA, did not affect TM-mediated reduction of thrombin generation significantly. This finding prompts the hypothesis that continuous RA treatment has more consistent therapeutic effects on coagulation disorders than on cancer.ConclusionsOur results indicate that the oscillatory upregulation of TM expression on the endothelial cells over the course of RA therapy could potentially contribute to the treatment of coagulation abnormalities in cancer patients. Further studies on the impacts of RA therapy on the procoagulant activity of cancer cells are needed to better elucidate the mechanisms by which RA therapy improves hemostatic abnormalities in cancer.

Pediatric Patients Have Elevated Thrombomodulin Levels Prior to Onset of Transplant Associated Thrombotic Microangiopathy and Are Associated with Increased Treatment Related Mortality

Background The incidence of transplant-associated thrombotic microangiopathy (TMA) is 20-40%, likely triggered by endothelial injury, which may be due to conditioning regimen, neutrophil extracellular traps, drugs or infections. Several previous studies have associated thrombomodulin (THBD) single-nucleotide polymorphism (SNP) variants with TMA and TRM, but use of protein levels to identify endothelial injury and predict future risk of TMA has not been reported. Objectives Primary objective was to determine whether THBD levels 14 days post hematopoietic stem cell transplant (HSCT) predict increased risk of TMA. We hypothesized that elevated serum THBD levels are associated with higher incidence of TMA and TRM as a consequence of endothelial injury. Methods Serum and DNA of 106 pediatric patients, transplanted between 2014 and 2017 were collected prior to conditioning and at day 14. THBD levels were measured using human thrombomodulin/BDCA-3 quantikine elisa kits (R and D, Minneapolis). We compared HSCT outcomes to THBD levels above and below the median. TaqMan SNP genotyping assays determined THBD genotype. Results Results of our study are summarized in figure 1. We observed increased THBD levels at day 14 in children who later developed TMA at a median of 26 days (range 0-667 days) after HSCT (p=0.05, figure 2), compared with children who never developed TMA. Moreover, mortality (at a median of 248 days, range 60-1258 days) was increased in children with THBD levels above the median at day 14 (p=0.02 figure 3). Interestingly there was no statistical difference between THBD level in children with or without graft versus host disease (GVHD), although the point estimate was higher in children with GVHD. THBD levels prior to conditioning were not predictive of TMA or TRM (P=0.95 and 0.41). THBD levels were not increased in those with a high risk genotype compared with those with a low risk genotype at baseline or day +14 (p=0.86 and 0.16, Table 1). Of the 23 deaths, 3 succumbed to infection, 7 to endothelial injury (TMA or GVHD), and 13 to relapse. Conclusion Our data show elevation of THBD as early as day +14 was associated with increased TMA and TRM at later time points. Our data indicate the importance of events happening within 14 days after HSCT in “setting up” later events and offer a window of time for potential interventions to reduce later complications. We are currently studying the kinetics of THBD during transplant recovery and mechanisms of associations of THBD levels with later outcomes.

Cerebral cavernous malformations form an anticoagulant vascular domain in humans and mice

AbstractCerebral cavernous malformations (CCMs) are common brain vascular dysplasias that are prone to acute and chronic hemorrhage with significant clinical sequelae. The pathogenesis of recurrent bleeding in CCM is incompletely understood. Here, we show that central nervous system hemorrhage in CCMs is associated with locally elevated expression of the anticoagulant endothelial receptors thrombomodulin (TM) and endothelial protein C receptor (EPCR). TM levels are increased in human CCM lesions, as well as in the plasma of patients with CCMs. In mice, endothelial-specific genetic inactivation of Krit1 (Krit1ECKO) or Pdcd10 (Pdcd10ECKO), which cause CCM formation, results in increased levels of vascular TM and EPCR, as well as in enhanced generation of activated protein C (APC) on endothelial cells. Increased TM expression is due to upregulation of transcription factors KLF2 and KLF4 consequent to the loss of KRIT1 or PDCD10. Increased TM expression contributes to CCM hemorrhage, because genetic inactivation of 1 or 2 copies of the Thbd gene decreases brain hemorrhage in Pdcd10ECKO mice. Moreover, administration of blocking antibodies against TM and EPCR significantly reduced CCM hemorrhage in Pdcd10ECKO mice. Thus, a local increase in the endothelial cofactors that generate anticoagulant APC can contribute to bleeding in CCMs, and plasma soluble TM may represent a biomarker for hemorrhagic risk in CCMs.

Cooling therapy upregulates protein C activation in heat stressed rats: An experimental study.

Protein C (PC) pathway homeostasis is implicated in heat stress (HS). This study determines whether cooling could improve the PC pathway in HS. Fifty-six anesthetized rats were warmed to achieve HS (rectal temperature [Tr] 42°C). These rats were divided into seven groups: (a) control group:sacrifice immediately 15 min after HS; (b) HS+I:sacrifice immediately after 15 min ice-water treatment or (c) 3 hr after HS; (d) HS+C:sacrifice immediately after 15-min cold-water treatment or (e) 3 hr after HS; (f) HS: sacrifice immediately 15 min after HS or (g) 3 hr after HS. Plasma PC, activated protein C (APC), and soluble thrombomodulin (sTM) levels were tested at both time points. After cooling, Tr in the HS+I and HS+C groups significantly decreased, when compared with the HS group, and Tr was significantly lower in the HS+I group than in the HS+C group ( p < 0.05). Furthermore, sTM levels were highest in the HS group among the groups at both time points. Plasma PC and APC levels increased after HS. In the HS+I and HS+C groups, plasma APC levels and the APC/PC ratio significantly increased at both time points. The proportions were significantly higher in the HS+I group than in the HS+C group, and there was no significant increase in APC/PC ratio in the HS group. Cooling exerts an anticoagulant effect following HS by increasing APC levels. Ice-water blanket therapy is more effective than cold-water blanket therapy in increasing APC levels.

Plasma Thrombomodulin Levels are Associated with Endothelial Injury in Patients with Bacterial Infections.

Endothelial injury is the key to the occurrence and development of bacterial infections. The aim of this study was to discuss the relationship between the molecular marker of endothelial damage, thrombomodulin (TM), and infectious disease severity, and prognosis. From January 2017 to April 2018, 296 patients with bacterial infections and 163 controls were recruited from our hospital. The concentrations of thrombomodulin and other routine coagulation and inflammatory factors were quantified. Plasma levels of thrombomodulin were obviously increased in infection group compared with control group (8.30 (7.23 - 9.68) vs. 15.83 (10.60 - 22.20) TU/mL, p < 0.001) and logistic regression analysis showed that the thrombomodulin was an independent risk factor for bacterial infection (OR, 1.189 (1.079 - 1.311)). In the infection group, patients with elevated thrombomodulin levels (> 75th percentile of its distribution, n = 71) experienced a higher level of coagulation factors (p < 0.05) and inflammatory factors (p < 0.05) than patients with levels below this cutoff. Multiple linear regression analysis showed that there was a positive correlation among the plasma thrombomodulin and D-dimer, white blood cells, and procalcitonin (β coefficient = 0.590, 0.220, and 0.208, p = 0.004, 0.027, and 0.025, respectively). With the increase of severity of disease, thrombomodulin levels gradually rose (13.58 ± 0.47 TU/mL vs. 25.07 ± 2.01 TU/mL vs. 31.34 ± 2.56 TU/mL, respectively, p < 0.001). Furthermore, there was an abnormal increase of plasma thrombomodulin in patients with bacterial infections and poor prognosis (p < 0.05). The area under curve of thrombomodulin as diagnosis for organ failure and non-survivor was 0.867 and 0.778, respectively. Plasmatic level of thrombomodulin could be considered as a diagnostic tool for bacterial infections. An increase in thrombomodulin plasmatic level was associated with poor outcome.

Dynamics of Soluble Thrombomodulin and Circulating miRNAs in Patients with Atrial Fibrillation Undergoing Radiofrequency Catheter Ablation.

Atrial fibrillation (AF) is the most common cardiac arrhythmia in the world and has a high risk of thromboembolism. The most effective approach, catheter ablation, requires evaluation by electrocardiography. The aim of our study was to investigate novel clinical markers that predict restoration of sinus rhythm (SR) after catheter ablation. Seventy-eight consecutive patients with AF underwent catheter ablation and were separated into 2 groups: restored SR and recurrent AF. The levels of 4 blood proteins (serum or plasma) and 3 mature microRNAs (miRNAs) and their primary miRNAs (pri-miRNAs) in serum were measured before and after ablation, and the associations between each parameter were analyzed statistically. Soluble thrombomodulin (s-TM) and plasminogen activator inhibitor-1 (PAI-1) levels increased above baseline after ablation in both the restored SR (s-TM 11.55 [2.92] vs 13.75 [3.38], P < .001; PAI-1 25.74 [15.25] vs 37.79 [19.56], P < .001) and recurrent AF (s-TM 10.28 [2.78] vs 11.67 [3.37], P < .001; PAI-1 26.16 [15.70] vs 40.74 [22.55], P < .001) groups. Levels of C-reactive protein and asymmetric dimethylarginine were not significantly changed. Pri-miR-126 levels significantly decreased after ablation in the recurrent AF group, but the other miRNAs and pri-miRNAs did not. The measurement of s-TM and pri-miR-126 in blood was a useful tool to reflect the condition of AF patients with catheter ablation.

THE ROLE OF ENDOTHELIAL DYSFUNCTION AS A PART OF SYSTEMIC INFLAMMATORY RESPONSE IN PATHOGENESIS OF AGGRESSIVE AND INDOLENT NON-HODGKIN’S LYMPHOMAS

Objective. To study of manifestation of angiogenic and hemostatic forms of endothelial dysfunction and systemic inflammatory response in patients with non-Hodgkin’s lymphomas at the onset of disease. Material and methods. A total of 99 patients were enrolled in this study, including 55 patients with aggressive variants and 44 patients with indolent variants. The average age was 60.71±10.63 years. The serum level of vascular endothelial growth factor (VEGF) was measured as a marker of angiogenic form of endothelial dysfunction, the serum levels of thrombomodulin, D-dimers, soluble fibrin-monomer complex (SFMC) and level of Willebrand factor activity were used for evaluation of hemostatic form of endothelial dysfunction. To assess the degree of systemic inflammation response the serum levels of proinflammatory cytokines – TNF-α, IL-1β, IL-6 were measured, as well as serum level of acute phase proteins – C-reactive protein (CRP), fibrinogen. The serum level of IL-4 was used to evaluate antinflammation resistance. Results and discussion. Proinflammatory cytokine (TNF-α, IL-1β and IL-6) content was higher in both studied groups compare to the control group. Serum concentration of anti-inflammatory cytokine IL-4 in studied groups did not differ from the level in the controls group. CRP and fibrinogen serum content was increased only in group of patients with aggressive variants. There was a significant correlation between the level of CRP and proinflammatory cytokines in aggressive lymphoma group. Serum content of VEGF, thrombomodulin, D-dimers, SFMC and Willebrand factor activity were increased in both studied group in comparison with the control group. Concentration of thrombomodulin, D-dimers, SFMC and Willebrand factor activity was higher in patients with aggressive than in patients with indolent lymphomas. Conclusion. The signs of angiogenic and hemostatic forms of endothelial dysfunction in patients with aggressive and indolent variants of non-Hodgkin’s lymphoma were detected, as well as manifestation of systemic inflammatory response and attenuation of anti-inflammatory resistance with development of cytokines imbalance with proinflammatory cytokines predominance. The manifestations of systemic inflammatory response and hemostatic type of endothelial dysfunction with activation of coagulation were displayed more prominently in the aggressive lymphoma group.

Endothelial cell injury is involved in atherosclerosis and lupus symptoms in gld.apoE- / - mice.

Cardiovascular complications related to atherosclerosis are major causes of morbidity and mortality in patients with systemic lupus erythematosus (SLE). However, the underlying mechanisms are not fully understood. Endothelial dysfunction has been identified as having involvement in pathogenesis of cardiovascular diseases and SLE. This study aims to evaluate endothelial cell injury in mice with the combination of lupus and atherosclerosis. The mouse model of accelerated atherosclerosis in lupus (gld.apoE- / - mouse) was generated from apolipoprotein E-deficient (apoE- / - ) and Faslgld C57BL/6 mice. The lupus-like autoimmunity and atherosclerotic lesions were evaluated. The endothelial cell injury was determined. The results showed that the double-mutant gld.apoE- / - mice were generated. Spleens from 5-month-old gld.apoE- / - mice were significantly enlarged compared with wild-type mice (WT mice). The gld.apoE- / - mice produced high levels of total immunoglobulin G (IgG) and IgM and showed marked increase of IgG and C3 deposits in the glomeruli. The gld.apoE- / - mice displayed a pattern of glomerulonephritis typically found in SLE. The gld.apoE- / - mice have high levels of serum creatinine. The total cholesterol, low-density lipoprotein cholesterol and triglycerides were significantly increased, while high-density lipoprotein cholesterol decreased in the double-mutant mice. The circulating endothelial progenitor cells were significantly decreased. The serum levels of thrombomodulin and vascular cell adhesion molecule-1 were significantly elevated in gld.apoE- / - mice. The gld.apoE- / - mice simultaneously exhibited SLE and atherosclerosis characteristics. Our findings indicated that endothelial cell injury might be a biomarker for evaluating risks of cardiovascular disease in SLE and targeting endothelial cell dysfunction might prevent and treat atherosclerosis in SLE.

New data on FII, FV, FIX and thrombomodulin defects: blood keeps clotting in normal and in peculiar ways

ABSTRACTObjective: To present the clinical and laboratory implications of defects or variants of some clotting factors and of thrombomodulin that were discovered during the past few years.Methods: Data concerning new aspects of FII, FV, FIX and thrombomodulin defects were investigated. This involved the dysprothrombinemias, the East Texas or short FV disorder, a FIX defect and a thrombomodulin abnormality.Results: the recently reported clotting defects or variants are: (1) the thrombophilic dysprothrombinemias due to Arg596 mutations (Prothrombin Yukuhashi, Belgrade and Padua 2) which are characterized by absence of bleeding and presence of venous thrombosis; (2) the short FV defects due to Ser356Gly (FV East Texas) or Ala863Gly (FV Amsterdam) mutations characterized by a mild bleeding tendency with normal FV and other clotting factors, increased TFPI and no thrombosis; (3) the abnormal FIX (FIX Padua) due to the Arg338Leu mutation which is associated with high levels of FIX activity, lack of bleeding and venous thrombosis; (4) the thrombomodulin Cys537Stop mutation associated with a mild bleeding tendency despite normal clotting factors but increased plasma levels of soluble thrombomodulin and no thrombosis.Conclusions: these new coagulation defects have great implications in the clinical and laboratory approach to the coagulation disorders. They have demonstrated that a prothrombin defect may be associated with thrombosis, that a mild bleeding tendency may occur despite normal Factor V levels and that high levels of plasmatic thrombomodulin may be associated with mild bleeding.

Correlation between active disease and hypercoagulability state in patients with systemic lupus erythematosus

ObjectiveThis study has investigated whether high levels of Reticulocytes-C4d (R-C4d) and Platelets-C4d (P-C4d) reflecting recent activity in SLE patients are correlated with changes in natural anticoagulation components, coagulation activation and endothelial injury markers. MethodsThis study included three groups: 1) healthy women (control, n = 30); 2) women with low activity of the disease (SLEDAI 2 K ≤ 4, n = 30); 3) women with active disease (moderate or high activity) (SLEDAI 2 K > 4, n = 30). Median fluorescence intensity (MFI) of R-C4d and P-C4d were determined by flow cytometry using double labeling with specific monoclonal antibodies. Endothelial injury and hypercoagulability were evaluated by measuring Thrombomodulin and D-dimer levels. ResultsHigher MFI index of R-C4d were related to the recent activity of SLE, and higher expression of P-C4d indicated an elevated risk of thrombotic complications. Increased levels of soluble thrombomodulin and D-dimer were observed in patients with active SLE. ConclusionR-C4d is helpful to monitor early disease activity and PC4-d may be an important tool to detect a prothrombotic phenotype in SLE. Elevated levels of D-dimer and thrombomodulin add value to P-C4d data and corroborate a hypercoagulable profile in women with SLE, contributing to an increased prothrombotic risk associated with inflammation.

Hemostatic Activation and Glycocalyx Shedding during Pregnancy

Background: Pregnancy has been described as a hypercoagulable state and is characterized by an increased risk of thrombotic events; yet the absolute incidence is low. The glycocalyx is a network of proteoglycans and glycoproteins projecting from the endothelial cell surface that plays a key role in maintaining endothelial integrity. Syndecan-1 is a component of the endothelial glycocalyx and is substituted with heparan sulfate chains which promote its interactions with key anticoagulants such as tissue factor pathway inhibitor and antithrombin. Proteolytic release of components of the glycocalyx (i.e. shedding) has been shown to occur in response to a number of physiologic challenges. Thrombomodulin, another endothelial cell transmembrane protein, is a key component of the primary dynamic anticoagulant mechanism limiting thrombus growth and is also subject to proteolytic release. In this study, we longitudinally evaluate hemostatic activation (D-dimer), glycocalyx shedding (syndecan-1) and thrombomodulin release in healthy women who became pregnant.Methods: Healthy nulliparous women (N=26) were prospectively enrolled in a longitudinal study at the University of Vermont Medical Center in a protocol approved by the Institutional Review Board. Study visits were conducted prior to pregnancy in the follicular phase (Visit 1), at the end of the first trimester (Visit 2), and the mid-third trimester (Visit 3). Blood was collected into citrate, and platelet poor plasma aliquots were prepared. Syndecan-1 and soluble thrombomodulin were measured on all samples by ELISA, and D-dimer was measured at Visits 1 and 3. For analyses, comparisons were made across Visits using a paired Student's t-test. Data are expressed as mean ± SD.Results: D-dimer levels were increased (3-13 fold across all women) significantly at Visit 3 (7.6 ± 3.9 μg/mL) compared to Visit 1 (1.2 ± 0.5 μg/mL), p < 0.001. Syndecan-1 levels were elevated at Visit 2 relative to Visit 1 in 25 of the 26 women; Visit 2 levels (43.7 ± 12.7 ng/mL) were significantly higher than Visit 1 (25.6 ± 18.0 ng/mL), p < 0.001. At Visit 3, an average 20-fold increase (range of 2.7 - 52.9-fold) in syndecan-1 levels (Visit 3, 518.5 ± 248.5 ng/mL) compared to Visit 1, p < 0.001 was observed. Soluble thrombomodulin did not change between Visit 1 (2.93 ± 0.5 μg/mL) and Visit 2 (2.88 ± 0.6 μg/mL); however there was a significant increase by Visit 3 (3.59 ± 0.7 μg/mL), p<0.001. Correlation analyses relating syndecan-1 and D-dimer levels and syndecan-1 and soluble thrombomodulin levels at each visit or relating fold changes in these markers between Visit 1 and Visit 3 showed no significant associations.Conclusions: By Visit 3 (the third trimester) women show significant increases in hemostatic activation, glycocalyx shedding and the release of another endothelial cell membrane protein. Increasing the levels of syndecan-1 in circulation may have a heparin-like effect on antithrombin activity while increasing soluble thrombomodulin levels may increase systemic protein C activation. However, the release of two potent anticoagulants from the endothelial surface can reduce the ability of the endothelial surface to help mediate and restrict thrombus growth at the site of injury. Future studies are needed to address whether there may be a prothrombotic contribution from the degraded endothelial surface that is offset by the release of anticoagulants into the circulatory system. DisclosuresNo relevant conflicts of interest to declare.

Damage-associated molecular patterns in intensive care unit patients with acute liver injuries: A prospective cohort study.

Acute liver injury (ALI) is frequently detected in an intensive care unit (ICU) and reportedly affects prognosis. Experimental animal studies suggested that increased extracellular histone and high morbidity group box-1 (HMGB1) levels might contribute to ALI development. Whether these damage-associated molecular patterns (DAMPs) play a crucial role in ALI remains unclear in the human clinical setting.We consecutively enrolled the patients admitted to our ICU. The patients with ALI were included in the analysis together with those without ALI by using frequency matching. Extracellular histone, HMGB1, soluble thrombomodulin (sTM), and interleukin-6 (IL-6) levels were measured in plasma collected at ICU admission. ALI was defined as an acute elevation in serum aminotransferase levels to >200 IU/L.A total of 805 patients were enrolled. Twenty ALI and forty non-ALI patients were analyzed. Plasma histone levels were significantly higher in the ALI group than in the non-ALI group, whereas HMGB1 levels were significantly lower in the ALI group. Furthermore, sTM was significantly increased in the ALI patients, whereas IL-6 levels were comparable between the groups. Multivariate logistic regression analysis demonstrated that histones were independently associated with ALI. There was no significant impact of ALI on in-hospital mortality.Extracellular histones showed an independent association with ALI. Histone elevation might be one of the possible pathogenic mechanisms in the development of ALI of ICU patients.

Circulating activated protein C levels are not increased in septic patients treated with recombinant human soluble thrombomodulin

BackgroundRecombinant human soluble thrombomodulin (rTM) has been used for the treatment of disseminated intravascular coagulation in Japan, and an international phase III clinical trial for rTM is currently in progress. rTM mainly exerts its anticoagulant effects through an activated protein C (APC)-dependent mechanism, but the circulating APC levels after rTM treatment have not been clarified. This prospective observational study investigated plasma APC levels after rTM treatment.MethodsPlasma levels of soluble thrombomodulin, thrombin-antithrombin complex (TAT), protein C, and APC were measured in eight septic patients treated with rTM. APC generation in vitro was assessed in the presence or absence of rTM.ResultsrTM significantly increased thrombin-mediated APC generation in vitro. In septic patients, soluble thrombomodulin levels were significantly increased during a 30–60-min period of rTM treatment and TAT levels were decreased. However, APC activity was not increased during the treatment period.ConclusionsPlasma APC activity is not increased in septic patients treated with rTM. It is possible that APC acts locally and does not circulate systemically.

Biomarkers of disseminated intravascular coagulation in pediatric intensive care unit in Thailand.

Disseminated intravascular coagulation (DIC) is a systemic activation of hemostatic system caused by several causes. Biomarkers including antithrombin (AT), protein C (PC), and thrombomodulin (TM) were reported as the additional markers for DIC in adults. This study aimed to determine the association between biomarkers among patients with overt DIC (ODIC) and nonovert DIC (NDIC) in children in PICU. We enrolled 103 subjects, aged 1month-18years, who were admitted to PICU at Chiang Mai University (CMU) Hospital >24hours with underlying conditions predisposing to DIC were enrolled. Biomarkers were tested after 24hours of admission. Subject who had NDIC on the 1st investigations would have other tests on days 3-5 of admission. The incidence of ODIC by the International Society on Thrombosis and Hemostasis (ISTH) DIC score was found 24%. The bleeding, thrombosis, and death were significantly higher in ODIC group (P<0.05). Mean levels of AT and PC in ODIC group were significantly different from NDIC one (66.9% vs 79.9%, P<0.001 and 46.1% vs 59.2%, P=0.004, respectively) while mean level of TM was not different between two groups. Adding AT to DIC score was better than the original score for predict mortality [area under curve (AUC)=0.662 vs AUC=0.65] and bleeding (AUC=0.751 vs AUC=0.732). ODIC is prevalent among critically ill children. Adverse outcomes were more commonly found in children with ODIC. AT and PC levels after 24hours of PICU admission seem to be the useful biomarkers for ODIC in PICU.

Does administration of eicosapentaenoic acid increase soluble thrombomodulin level in statin-treated patients with stable coronary artery disease?

Interventions targeting the serum eicosapentaenoic acid (EPA)/arachidonic acid (AA) ratio could be useful for the prevention of coronary artery disease (CAD). Few data exist regarding the effects of administration of EPA on the serum levels of soluble thrombomodulin (sTM) as a marker of endothelial damage, or on the relationship between the sTM and EPA/AA ratio in patients with CAD receiving statin treatment. We assigned stable CAD patients already receiving statin therapy to an EPA group (1800mg/day: n = 50) or control group (n = 50). A significant increase of the sTM level was observed in the EPA group as compared to that in the control group 0.40 (0.10/0.70) FU/mL vs. 0.20 (0/0.40) FU/mL, p = 0.004 at the 6-month follow-up examination. Multivariate regression analysis after adjustments for coronary risk factors and changes of the serum lipid levels identified an increased EPA/AA ratio as an independent predictor of increased serum sTM level (β = 0.244, p = 0.02). The results suggest that an increased sTM level caused by additional administration of EPA to statin might be associated with an increased EPA/AA ratio. The increase of the serum sTM after administration of EPA might reflect an increase of the TM expression on the endothelial surface rather than endothelial damage in CAD patients under statin treatment.Clinical Trial Registration Information UMIN ( http://www.umin.ac.jp/ ), Study ID: UMIN000010452.

Endothelial dysfunction in patients with eosinophilic granulomatosis with polyangiitis

Eosinophilic granulomatosis with polyangiitis (EGPA) is a rare form of vasculitis associated with asthma and eosinophilia. Endothelial dysfunction has been well documented in other types of vasculitis but not in EGPA. Thirty patients (10 men and 20 women) diagnosed with EGPA and remaining in a remission, and 58 controls (24 men and 34 women) matched for age, sex, and body mass index, were enrolled in the study. We assessed each participants for typical risk factors of cardiovascular diseases and measured serum levels of vascular cell adhesion molecule-1 (VCAM-1), interleukin 6 (IL-6), and thrombomodulin. We also measured flow-mediated dilatation (FMD) of the brachial artery and intima-media thickness (IMT) of the common carotid artery using ultrasonography. Patients with EGPA had 20% higher serum level of VCAM-1 (p < 0.001) and 41.9% of thrombomodulin (p < 0.001). They also had 38.8% lower relative increase of FMD (FMD%) (p < 0.001), indicating endothelial dysfunction. These differences remained significant also after adjustment for potential confounders. Laboratory and ultrasonographic parameters of endothelial injury were correlated to the markers of inflammation and impaired kidney function. Determinants of lower FMD% in a simple regression model were pack-years of smoking (β = − 0.3 [95% confidence interval (CI) − 0.5 to − 0.1]), serum level of IL-6 (β = − 0.36 [95% CI − 0.62 to − 0.1]), and thrombomodulin (β = − 0.34 [95% CI − 0.6 to − 0.08]). EGPA patients are characterized by inflammatory endothelial injury that is likely related to the pathogenesis of the disease. Proper immunosuppressive treatment is the best method to prevent atherosclerosis and future cardiovascular events, the patients may also benefit from additional preventive interventions.

Decreased antithrombin activity in the early phase of trauma is strongly associated with extravascular leakage, but not with antithrombin consumption: a prospective observational study

BackgroundWe conducted a prospective observational study for investigating coagulofibrinolytic changes and mechanisms of antithrombin (AT) alternations in trauma.MethodsTrauma patients hospitalized for more than seven days were analyzed for coagulofibrinolytic biomarkers. The patients were stratified into two groups according to AT activity level on admission (day 0), comprising normal AT and low AT patients.ResultsThirty-nine patients (median Injury Severity Score 20) exhibited initial coagulatory activation and triphasic fibrinolytic changes. AT activity did not show a negative linear correlation with levels of thrombin-antithrombin complex (TAT), a marker of coagulation activity and AT consumption, but was strongly correlated with levels of albumin (Alb), an index of vascular permeability, on day 0 (r = 0.702, p < 0.001). Furthermore, Alb was one of the independent predictors for AT on day 0. IL-6 on day 0 and thrombomodulin (TM) levels during the study period, reflecting systemic inflammation and endothelial cell injury, respectively, were significantly higher in the lower AT group (n = 10) than in the normal group (n = 29) (IL-6, p = 0.004; TM, p = 0.017). On days 2 and 4, TAT levels in the lower AT group were significantly higher than in the normal group.ConclusionsTrauma caused clear triphasic coagulofibrinolytic changes. Decreased AT in the later phase might lead to a prolonged hypercoagulation. AT reduction in the initial phase of trauma is strongly associated with extravascular leakage as suggested by the association of Alb depletion with IL-6 and TM elevation, but not with AT consumption.

Evaluation of thrombomodulin and pentraxin-3 as diagnostic biomarkers in calves with sepsis

Early diagnosis and treatment of sepsis in patients are crucial for their survival and can help reduce mortality rates. Novel biomarkers, such as thrombomodulin and pentraxin-3, have been used as diagnostic, prognostic and mortality indicators in patients with sepsis. Plasma thrombomodulin is a vascular endothelial membrane-bound glycoprotein and pentraxin-3 is an acute-phase protein. In the present study, thrombomodulin and pentraxin-3 levels were determined in calves with sepsis, to determine their diagnostic values as well as usefulness as indicators of health status. To this end, 20 neonatal calves with sepsis (G1) and ten healthy neonatal calves (G2) were used. Additionally, group G1 was also divided into two groups consisting of surviving (G1-S; n = 9) and non-surviving calves (G1-NS; n = 11). A single blood sample was collected from all the calves and the prepared serum samples were used to measure thrombomodulin and pentraxin-3 levels using bovine-specific ELISA kits. The serum concentrations of thrombomodulin and pentraxin-3 were found to be significantly higher (P &amp;lt; 0.01) in the G1 group than in G2. Thrombomodulin and pentraxin-3 levels were also found to be higher in the G1-NS group than in G1-S but the difference was not significant. We conclude that thrombomodulin and pentraxin-3 may have some diagnostic value in calves with sepsis. Furthermore, these findings may also help in understanding the pathogenesis of sepsis in neonatal calves. Further studies are required to determine the importance of thrombomodulin and pentraxin-3 as diagnostic and prognostic biomarkers in calves with sepsis and to evaluate the concentrations of these biomarkers also in other disease states.