Serum Levels Of Thrombomodulin Research Articles

Serum levels of biomarkers related to severity staging of Raynaud’s phenomenon, neurosensory manifestations, and vibration exposure in patients with hand-arm vibration injury

Our aim was to explore possible relationships between serum levels of biomarkers in patients with hand-arm vibration injury in relation to the severity of the vascular, i.e., Raynaud’s phenomenon (RP), and neurosensory manifestations, the current exposure level, and the duration of exposure. This study was of case series design and involved 92 patients diagnosed with hand-arm vibration injury. Jonckheere’s trend test was used to assess any association between serum levels of biomarkers and RP as well as neurosensory manifestations, graded by the International Consensus Criteria. Generalized linear models with adjustment for possible confounders were also used for associations between serum levels of biomarkers and; (1) severity of RP recorded as the extent of finger blanching calculated with Griffin score, (2) vibration perception thresholds, (3) magnitude of current exposure as [A(8); (m/s2)] value, and (4) the duration of exposure in years. Serum levels of thrombomodulin, von Willebrand factor, calcitonin gene related peptide (CGRP), heat shock protein 27, and caspase-3 were positively associated with severity of RP. Serum levels of CGRP were positively associated with the neurosensory component. No associations with exposure were shown for these biomarkers. For Intercellular adhesion molecule 1 and monocyte chemoattractant protein 1, no associations were found with neither severity nor exposure. Levels of serum biomarkers associated with endothelial injury or dysfunction, inflammation, vasodilation, neuroprotection, and apoptosis were positively associated with the severity of hand-arm vibration injury.

Differential expression of serum TM, PAF, and CD62P in patients with autologous arteriovenous fistula and the correlation with vascular access function.

End-stage renal disease (ESRD) is the final stage of chronic kidney disease (CKD). We aimed to analyze the expression differences of serum thrombomodulin (TM), platelet-activating factor (PAF), and P-selectin (CD62P) in patients with autologous arteriovenous fistula (AVF) and the correlation with vascular access function. The case data were retrospectively analyzed. Moreover, 160 patients with AVF maintenance hemodialysis were selected as the AVF group, and 150 healthy participants were selected as the healthy control group. According to the function of vascular access, patients in the AVF group were divided into Group A (n = 50, after the first establishment of AVF), Group B (n = 64, normal vascular access function after hemodialysis treatment), and Group C (n = 46, vascular access failure). Pearson analysis was conducted to explore the correlation between serum TM, PAF, CD62P content, and vascular pathological examination indicators, to evaluate the value of TM, PAF, and CD62P levels in predicting vascular access failure in patients with AVF. The serum levels of TM, PAF, and CD62P were positively correlated with the expressions of CD68 and MCP-1, respectively (p < .001). Serum TM was positively correlated with the levels of PAF and CD62P (p < .001), and PAF was positively correlated with the levels of CD62P (p < .001), respectively. Serum levels of TM, PAF and CD62P were risk factors for vascular access failure in AVF patients (p < .05). The area under the curve of serum TM, PAF and CD62P levels in predicting vascular access failure in AVF patients was 0.879. The serum levels of TM, PAF, and CD62P in AVF patients were correlated with the vascular access function of AVF patients, which was very important for maintaining the stability of vascular access function, and had certain value in predicting vascular access failure/disorder in AVF patients, and could be popularized and applied.

Are serum thrombomodulin and interleukin-8 levels associated with disease severity and mortality in critically ill children with respiratory failure?

Thrombomodulin (TM) is found on endothelial cell surfaces and increases in response to endothelial injury of different organs. Interleukin (IL)-8 regulates pulmonary inflammation. TM and IL-8 are candidate biological markers of acute respiratory distress syndrome (ARDS). The aim of the present study was to compare TM and IL-8 levels in pediatric patients with and without ARDS who received respiratory support and to determine their relationships with prognosis. This was a prospective observational study of 55 patients who received respiratory support in the pediatric intensive care unit. Eighteen patients without active infection were defined as the control group. Two blood samples were taken for serum IL-8 and TM levels on the first and third days of respiratory support. The patient group had significantly higher IL-8 and TM levels than the control group [median IL-8: 102.7 (IQR: 180.42-189.47) vs. 45.4 (55.14-70.49) ng/L, p = 0.011; median TM: 6.9 (6.83-9.18) vs. 3.4 (3.62-5.05) ng/mL, p = 0.021]. Patients with ARDS had significantly higher marker levels on the first and third days than those who did not have ARDS. The TM and IL-8 levels of deceased patients were significantly higher than those of the survivors on the first day. In mortality prediction, the cut-off point for IL-8 was found to be >154.7 ng/L, which had sensitivity of 76.9% and specificity of 73.8%. The cut-off point for TM was >8.4 ng/mL, which had sensitivity of 76.9% and specificity of 66.7%. In our study, higher marker levels correlated with impaired oxygenation and higher mortality. Higher TM and IL-8 levels in ARDS might reflect the degree of vascular injury and inflammation.

Prognostic value of thrombomodulin molecule in COVID-19 patients

Background:The purpose of this study is to evaluate serum thrombomodulin(TM) levels in coronavirus disease 2019(COVID-19) patients and to investigate whether TM predicts the severity of COVID-19 and coagulopathy . Materials and methods:This study was conducted on 87 confirmed COVID-19 patients who were hospitalized due to symptomatic pneumonia.The patients were divided into two groups as intensive care unit(ICU) patients and non-ICU patients. Both groups were compared according to demographic features, comorbid conditions and measurement of the TM and other laboratory parameters of the patients. Results:Of 87 patients with COVID-19, 40(45%) were ICU patients and 47(55%) were non-ICU patients. 40(45%) of the patients were men.TM median value(IQR) was 1.22ng/mL (0.9–2.2) in all patients, 1.2ng/mL(0.8–2.3) in non-ICU patients and 1.3ng/mL (1.0–2.21) in ICU patients (p=0.485).As a receiver operating characteristic curve analysis the cut-off value of TM for predicting mortality in patients was 1.27 with 60% sensitivity and 54% specificity (AUC: 0.550, 95% CI: 0.384-0.716, P= 0.602).Also, we found significant cut-off values with respect to troponin, D-dimer, CRP, fibrinogen, procalcitonin, LDH, and ferritin(p&lt; 0.05). Conclusion:This study found no significant difference between TM levels in ICU versus non-ICU groups.We further showed that there is no relation between TM levels and mortality.

Variation in presepsin and thrombomodulin levels for predicting COVID-19 mortality

Coronavirus disease (COVID-19) has caused extensive mortality globally; therefore, biomarkers predicting the severity and prognosis of COVID-19 are essential. This study aimed to evaluate the application of presepsin (P-SEP) and thrombomodulin (TM), which are biomarkers of sepsis and endothelial dysfunction, respectively, in the prognosis of COVID-19. Serum P-SEP and TM levels from COVID-19 patients (n = 183) were measured. Disease severity was classified as mild, moderate I, moderate II, or severe based on hemoglobin oxygen saturation and the history of intensive care unit transfer or use of ventilation at admission. Patients in the severe group were further divided into survivors and non-survivors. P-SEP and TM levels were significantly higher in the severe group than those in the mild group, even after adjusting for creatinine values. In addition, TM levels were significantly higher in non-survivors than in survivors. Changes in the P-SEP levels at two time points with an interval of 4.1 ± 2.2 days were significantly different between the survivors and non-survivors. In conclusion, TM and continuous P-SEP measurements may be useful for predicting mortality in patients with COVID-19. Moreover, our data indicate that P-SEP and TM values after creatinine adjustment could be independent predictive markers, apart from renal function.

A-136 Usefulness of Presepsin and Thrombomodulin Measurements for Predicting COVID-19 Mortality

Abstract Background Coronavirus disease (COVID-19) is a novel infectious disease developed into a pandemic since early 2020, resulting in extensive morbidity and mortality globally. Therefore, it is important to identify biomarkers for predicting the severity and mortality of COVID-19. We focused on presepsin (P-SEP) and thrombomodulin (TM), which are biomarkers of sepsis and endothelial dysfunction, respectively. Some reports have already suggested that these biomarkers can be associated with COVID-19, however, most of them assessed blood P-SEP and TM levels using only a single day measurement. In addition, since both P-SEP and TM are excreted by the kidney, renal dysfunction should be considered when evaluating these levels. In this study, to evaluate the usefulness of P-SEP and TM for the prognosis and prediction of COVID-19 severity, we investigated the changes in these levels after admission with or without adjustment for creatinine (CRE) level in COVID-19 patients. Methods Samples were residual sera obtained from COVID-19 patients (n = 674) admitted to the TMDU Hospital. Serum P-SEP and TM levels were measured using the STACIA clinical assay system. Disease severity was classified as mild, moderate, or severe based on hemoglobin oxygen saturation and the history of intensive care unit transfer or use of ventilation at admission. Patients in the severe group were further classified into survivors and non-survivors. This study was approved by the institutional research ethics committee (No. M2020-110). Results Since patients who received recombinant human TM treatment had abnormally high TM levels (&amp;gt; 1000 U/mL), data after administration were excluded from the analysis. P-SEP and TM levels during hospitalization were significantly correlated with CRE levels. In the correlation between P-SEP and CRE, the slope of the severe group was different from that of the non-severe group. When compared the data extracted within 5 days after admission (n = 183) among the three severity groups, CRE levels showed no significant difference among them. On the other hand, P-SEP and TM levels at admission were significantly higher in the severe group than those in the mild group, even after adjusting for CRE levels. In addition, there was no significant difference in the P-SEP levels between survivors and non-survivors, whereas TM levels were significantly higher in non-survivors than in survivors. Changes in the P-SEP levels at two time points (interval: 4.1 ± 2.2 days) were significantly different between survivors and non-survivors. The second measurement of TM levels were elevated in both groups from the first measurement at admission. Conclusion This study suggested that the elevation of P-SEP and TM levels in severe COVID-19 patients might not only fluctuate with renal dysfunction, but might reflect severity. TM and continuous P-SEP measurements are available for predicting mortality in patients with COVID-19. Moreover, P-SEP and TM values after CRE adjustment would be independent predictive markers, apart from renal function.

Potential mechanisms of resistance to current anti-thrombotic strategies in Multiple Myeloma.

Multiple Myeloma (MM) is a common haematological malignancy that is associated with a high rate of venous thromboembolism (VTE) with almost 10% of patients suffering thrombosis during their disease course. Recent studies have shown that, despite current thromboprophylaxis strategies, VTE rates in MM remain disappointingly high. The pathophysiology behind this consistently high rate of VTE is likely multifactorial. A number of factors such as anti-thrombin deficiency or raised coagulation Factor VIII levels may confer resistance to heparin in these patients, however, the optimal method of clinically evaluating this is unclear at present, though some groups have attempted its characterisation with thrombin generation testing (TGT). In addition to testing for heparin resistance, TGT in patients with MM has shown markedly varied abnormalities in both endogenous thrombin potential and serum thrombomodulin levels. Apart from these thrombin-mediated processes, other mechanisms potentially contributing to thromboprophylaxis failure include activated protein C resistance, endothelial toxicity secondary to chemotherapy agents, tissue factor abnormalities and the effect of immunoglobulins/“M-proteins” on both the endothelium and on fibrin fibre polymerisation. It thus appears clear that there are a multitude of factors contributing to the prothrombotic milieu seen in MM and further work is necessitated to elucidate which factors may directly affect and inhibit response to anticoagulation and which factors are contributing in a broader fashion to the hypercoagulability phenotype observed in these patients so that effective thromboprophylaxis strategies can be employed.

Serum thrombomodulin level in neonates with septic shock

ObjectiveThe aim was to assess the potential role of serum thrombomodulin level as a biochemical diagnostic marker in neonates with septic shock.BackgroundNeonatal septic shock is a significant cause of morbidity and mortality. Thrombomodulin is expressed on vascular endothelial cells and is found in the body in a bound form and a soluble form.Patients and methodsThis prospective cohort study was done on 120 neonates who were divided into two groups: patients' group included 80 neonates with septic shock, and control group included 40 healthy neonates. Detailed history, thorough clinical examination, and laboratory investigations including level of serum soluble thrombomodulin were done.ResultsNeonates with septic shock had statistically significant higher level of serum soluble thrombomodulin than controls. Serum soluble thrombomodulin levels were significantly higher in patients with gram-positive bacteria in comparison with gram-negative ones. Nonsurviving septic neonates had statistically significant increase in serum soluble thrombomodulin levels than survived septic neonates. There was a statistically significant positive correlation between serum soluble thrombomodulin levels and quantitative C-reactive protein (CRP) values. The cutoff value of 20 of serum soluble thrombomodulin levels had a sensitivity of 91.5%, a specificity of 84.7%, positive predicted value of 91.9%, negative predicted value of 88.3% with accuracy 92.94% in the diagnosis of septic shock. Serum soluble thrombomodulin showed a higher accuracy in the diagnosis of septic shock than quantitative CRP values.ConclusionSerum soluble thrombomodulin level has a more sensitivity diagnostic role in septic shock than quantitative CRP value. Higher serum soluble thrombomodulin levels predict the prognosis of septic shock.

Value of Thrombomodulin as a Marker for Sepsis in Critically Ill Children.

Pediatric sepsis is altered organ function in critically ill children and a main etiology of mortality for children. Therefore, the authors aimed to assess the role of serum thrombomodulin as valuable biomarker in the diagnosis and prognosis of sepsis in acutely ill pediatrics in the intensive unit. This prospective clinical study conducted on 140 acutely ill patients admitted to the Pediatric Intensive Care Unit (PICU) of Menoufia University Hospital and 50 apparently healthy controls from October 2018 through September 2019. All included children were subjected to clinical examination and the Pediatric Risk of Mortality (PRISM) and Pediatric Index of Mortality II (PIM II) scores were calculated. Serum thrombomodulin was measured for both patients and the control group upon admission. The children were followed for a period of 30 d. Serum thrombomodulin level was increased among all the patients and those with systemic inflammatory response syndrome (SIRS), sepsis and severe sepsis compared with controls (p < 0.001). Furthermore, serum thrombomodulin was higher in patients who died than who survived (p =0.005). Thrombomodulin had area under Receiver Operating Characteristic Curve (AUC) =0.915 for predicting sepsis, whereas C-reactive protein had AUC = 0.789. According to the prognosis, thrombomodulin had AUC = 0.711 for predicting mortality whereas PRISM and PIM scores had AUC = (0.918, 0.960) respectively. Serum thrombomodulin is a promising marker for pediatric sepsis. The data showed that serum thrombomodulin had a valuable role in diagnosis of sepsis early in critically ill pediatrics.

Low Vitamin D Serum Level Is Associated with HDL-C Dyslipidemia and Increased Serum Thrombomodulin Levels of Insulin-Resistant Individuals.

BackgroundInsulin-resistant individuals are known to have dyslipidemia and are predicted to be at high risk of cardiovascular events. Vitamin D deficiency was shown to be associated with dyslipidemia; however, the type of dyslipidemia associated with vitamin D deficiency in insulin-resistant individuals is not determined. Furthermore, there is evidence linking insulin resistance with low-grade inflammation suggesting levels of pro-inflammatory cytokines to be increased in insulin-resistant states.ObjectiveThis study was performed to evaluate the impact of vitamin D deficiency, defined as serum level of 25(OH)D below 20 ng/mL, on lipid profile and inflammatory markers such as interleukin (IL-6) and IL-8, as well as soluble thrombomodulin (TM) in the serum of insulin-resistant individuals.MethodsA total of 4114 individuals had simultaneous serum 25(OH)D, insulin, and lipid panel testing during 2013 as part of the United Arab Emirates National Diabetes and Lifestyle (UAEDIAB) study. Multivariate logistic regression analysis was used to assess the association between serum level of 25(OH)D and lipid profile in insulin-sensitive versus -resistant individuals. The lipid panel was stratified into high total cholesterol (TC: >6.2 mmol/L), high low-density lipoprotein-cholesterol (LDL-C: >2.59 mmol/L), high triglycerides (TG: >2.3 mmol/L), and low high-density lipoprotein-cholesterol (HDL-C: <1.55 mmol/L) dyslipidemia. Furthermore, the immunomodulatory and vasculoprotective effects of 25(OH)D were assessed by measuring the levels of IL-6, IL-8, and soluble TM in serum using ELISA.ResultsMore than half of the 4114 individuals were insulin resistant (n=2760, 67%) and around one-fifth of them were vitamin D-deficient (n=796, 19%). After adjusting for age, gender, body mass index, smoking, ethnicity, and educational level, the only dyslipidemia associated with vitamin D-deficient-insulin-resistant individuals (OR 2.09 [95]; P=0.009) was lower HDL-C. Furthermore, deficient 25(OH)D individuals with low HDL-C levels had higher circulatory IL-6 and IL-8 levels, and higher serum soluble TM compared to individuals with sufficient 25(OH)D and normal lipid profiles (median, IL-6 pg/mL 0.82 vs 1.71, P=0.001; median, IL-8 pg/mL 51.31 vs 145.6, P=0.003; and median, soluble TM ng/mL 5.19 vs 7.38, P<0.0001; in sufficient vs deficient groups, respectively).ConclusionThe results of our study showed that in insulin-resistant individuals, vitamin D deficiency status is associated with HDL-C dyslipidemia and higher serum inflammatory and endothelial damage markers.

Biomarkers associating endothelial dysregulation in pediatric-onset systemic lupus erythematous

Background/purposeEndothelium is a key element in the regulation of vascular homeostasis and its alteration can lead to the development of vascular diseases. Systemic lupus erythematosus (SLE) is a systemic autoimmune disease with potential extensive vascular lesions, involving skin vessels, renal glomeruli, cardiovascular system, brain, lung alveoli, gastrointestinal tract vessels and more. We aimed to assess endothelial dysregulation related biomarkers in pediatric-onset SLE (pSLE) patient serum and elucidate its correlation with their clinical features, laboratory parameters, and the overall disease activity.MethodsDisease activities were evaluated by SLE disease activity index (SLEDAI). Patient characteristics were obtained by retrospective chart review. Six biomarkers associated with endothelial dysregulation, including Angiopoietin-1 (Ang-1), Angiopoietin-2 (Ang-2), Tie2, Vascular endothelial growth factor (VEGF), thrombomodulin, and a disintegrin-like and metalloprotease with thrombospondin type 1 motif (ADAMTS13) were tested through enzyme-linked immunosorbent assay (ELISA) measurement.ResultsThis study comprised 118 pSLE patients. Data from 40 age-matched healthy controls were also obtained. The mean diagnostic age was 13 ± 4.12 years-old and 90.7% are females. Serum levels of VEGF, Tie2, thrombomodulin were significantly higher while serum ADAMTS13 was lower in active pSLE patients when compared to those with inactive diseases (all p < 0.05). In organ specific association, serum thrombomodulin level was higher in pSLE patient with renal involvement, and serum ADAMTS13 levels was negatively associated with neurological involvement (p < 0.05). A cutoff of thrombomodulin at 3333.6 pg/ml best correlated renal involvement. (AUC = 0.752, p < 0.01).ConclusionEndothelial dysregulation associating proteins seems to be potent biomarkers for pSLE activity as well as organ involvement in pSLE patients. These biomarkers may be beneficial in understanding of the vascular pathogenesis and disease monitoring.

Serum thrombomodulin as a metastatic and prognostic marker in soft tissue sarcomas.

Thrombomodulin (TM) has multiple biological functions and modulates not only anti-coagulation, but also cell proliferation, adhesion, and anti-inflammation activities. The main function of TM is to activate the anticoagulant pathway of protein C. Soluble TM is related to metastasis by its inactivation of thrombin. To clarify the correlation between serum TM levels and clinicopathological parameters. The plasma TM levels (FU/ml) of 135 primary soft tissue tumors (benign, 67; soft tissue sarcoma (STS), 68) were measured before biopsy or treatment. TM levels were analyzed and compared to various clinicopathological parameters. Log-rank test and Cox proportional analysis were used to evaluate recurrence-free survival, metastasis-free survival, and overall survival. STS tumors had significantly higher TM values (15.9) than benign tumors (13.7) (p= 0.0138). 5-year MFS was 81.1% in low TM and 40.0% in high TM (p= 0.00671), and 5-year OS was 85.5% in low and 52.5% in high TM in grades 1-3 (p= 0.0673). In multivariate COX proportional analysis, high-TM showed a significant difference (MFS: HR 4.37, p= 0.0147; OS: HR 3.60, p= 0.0557) in grades 1-3. We demonstrated that a high level of soluble TM has the potential to be a significant predictor of metastasis and poor prognosis in STS patients. TM is a candidate molecular marker for high metastatic potential and can be clinically useful for guiding therapeutic strategy.

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.

Comparison of perioperative serum thrombomodulin levels in patients underwent off-pump coronary artery bypass grafting with and without cardiogenic shock

Objective: To observe the perioperative changes of serum thrombomodulin in patients with and without cardiogenic shock undergoing off-pump coronary artery bypass grafting surgery. Methods: A total of 66 patients with coronary artery disease who underwent off-pump coronary artery bypass grafting surgery between June and December 2015 in our hospital were included in this study.The patients were divided into non-cardiogenic shock group (n=51) and shock group (n=15) according to the absence or presence of cardiogenic shock.The clinical data of the two groups were analyzed.Arterial blood samples were collected immediately after anesthesia, and at 4, 8, 16, 24, 48 and 72 hours after surgery.The levels of serum thrombomodulin were tested with enzyme-linked immunosorbent assay. Results: (1)The prevalence of diabetes was significantly higher (64.7%(33/51) vs. 20.0%(3/15), P<0.01), while prevalence of myocardial infarction was significantly lower (41.2%(21/51)vs. 100%(15/15), P<0.01) in non-cardiogenic shock group than in cardiogenic shock group.(2)The peak plasma creatinine level, troponin I level and creatine kinase MB level were significantly lower in the non-cardiogenic shock group than in the cardiogenic shock group ((88.5±36.7) μmol/L vs. (122.6±71.1) μmol/L, 1.3(0.2, 2.7) μg/L vs. 16.4(5.8, 23.4) μg/L and (18.8±4.7) μg/L vs.(49.3±15.9) μg/L, respectively, all P<0.05). (3)Mechanical ventilation time was significantly longer (11.5 (9.0, 18.0) hours vs. 20.0 (8.5, 82.5) hours, P=0.02), and frequency of intra-aortic balloon pump use (0 vs. 100%, P<0.01) was significantly higher in the cardiogenic shock group than in the non-cardiogenic shock group.(4) Prevalence of atrial fibrillation (5.9%(3/51) vs. 80.0%(12/15)) and myocardial infarction (19.6%(10/51)vs. 93.3%(14/15)) that occurred during and after surgery was significantly lower in the non-cardiogenic shock group than in the shock group(all P<0.01). (5)The serum thrombomodulin concentration was similar among various time point in the non-cardiogenic shock group: 3.30(2.68, 7.44), 4.09(2.95, 7.18), 4.35(2.68, 8.22), 3.50(2.95, 8.00), 3.41(2.60, 5.97), 3.30(2.65, 5.42)and 3.94(2.82, 5.60) μg/L, taken immediately after anesthesia, and at 4, 8, 16, 24, 48 and 72 hours after surgery respectively, and was 2.44(1.97, 2.67), 2.21(1.93, 2.83), 2.64(2.29, 2.67), 2.84(2.26, 2.94), 3.35(2.43, 4.05), 2.76(2.73, 2.97) and 3.81(2.96, 5.96)μg/L respectively, in the cardiac shock group (all P>0.05). The serum thrombomodulin levels at 4 and 8 hours after surgery were higher in the non-cardiogenic shock group than cardiogenic shock group(all P<0.05). Conclusion: Compared with the non-cardiogenic shock group, the peak serum thrombomodulin level appears later and recovers slower in cardiogenic shock patients who underwent off-pump coronary artery bypass grafting surgery.

Increased serum thrombomodulin level is associated with disease severity and mortality in pediatric sepsis.

BackgroundEndothelial dysfunction plays an important role in the pathophysiology of sepsis. As previously reported, the serum thrombomodulin is elevated in diseases associated with endothelial injury.ObjectiveThe aim of this study was to investigate the association of serum thrombomodulin level in different pediatric sepsis syndromes and evaluate the relationship with disease severity and mortality.MethodsWe prospectively collected cases of sepsis treated in a pediatric intensive care unit from June 2012 to July 2015 at Chang Gung Children’s Hospital in Taoyuan, Taiwan. Clinical characteristics and serum thrombomodulin levels were analyzed.ResultsIncreased serum thrombomodulin levels on days 1 and 3 of the diagnosis of sepsis were found in different pediatric sepsis syndromes. Patients with septic shock had significantly increased serum thrombomodulin levels on days 1 and 3 [day 1: median, 6.9 mU/ml (interquartile range (IQR): 5.8–12.8) and day 3: median, 5.8 mU/ml (IQR: 4.6–10.8)] compared to healthy controls [median, 3.4 mU/ml (IQR: 2.3–4.2)] (p = <0.001 and 0.001, respectively) and those with sepsis [day 1: median, 2.9 mU/ml (IQR: 1.8–4.7) and day 3: median, 3 mU/ml (IQR: 1.5–3.5)] and severe sepsis [day 1: median, 3.3 mU/ml (IQR: 1.3–8.6) and day 3: median, 4.4 mU/ml (IQR: 0.5–6)] (p = <0.001 and 0.001, respectively). There was also a significant positive correlation between serum thrombomodulin level on day 1 and day 1 PRISM-II, PELOD, P-MOD and DIC scores. The patients who died had significantly higher serum thrombomodulin levels on days 1 and 3 [day 1: median, 9.9 mU/ml (IQR: 6.2–15.6) and day 3: median, 10.4 mU/ml (IQR: 9.2–11.7)] than the survivors [day 1; median, 4.4 mU/ml (IQR: 2.2–7.5) and day 3: [median, 3.5 mU/ml (IQR: 1.6–5.7)] (p = 0.046 and 0.012, respectively).ConclusionIncreased serum thrombomodulin levels were found in different pediatric sepsis syndromes and correlated with disease severity and mortality.

Effects of alloantibodies to human leukocyte antigen on endothelial expression and serum levels of thrombomodulin

Rationale Thrombomodulin (TBM) is an anticoagulant and anti-inflammatory transmembrane protein expressed on endothelial cells. Donor-specific alloantibodies, particularly those against human leukocyte antigen (HLA) class II, are associated with microvascular endothelial damage in solid allografts. Objective Our aim was to characterize the effects of anti-HLA antibodies on endothelial expression of TBM, and in particular, the differential effects of anti-HLA class I compared with those of anti-HLA class II. Methods We used human glomerular microvascular endothelial cells to examine TBM expression on anti-HLA-treated cells, and we tested sera from transplant recipients for soluble TBM. Results We found that whereas membrane TBM expression increased in a dose-dependent manner in the presence of anti-HLA class I antibodies, treatment with anti-HLA class II led to minimal TBM expression on the endothelial surface but to a cytosolic accumulation. Platelet adhesion studies confirmed the functional impact of anti-HLA class II. Quantitative densitometry of the membrane lysates further suggested that anti-HLA class II impairs TBM glycosylation. Furthermore, we found a significant association between the presence of circulating anti-HLA class II antibodies in transplant recipients and low serum levels of TBM. Conclusion These results indicate that ligation of anti-HLA class I and II antibodies produces different effects on the endothelial expression of TBM and on serum levels of TBM in transplant recipients. Anti-HLA class II antibodies may be associated with a prothrombotic state, which could explain the higher occurrence of microangiopathic lesions in the allograft and the poor outcomes observed in patients with these alloantibodies.

Label-Free and Sensitive Detection of Thrombomodulin, a Marker of Endothelial Cell Injury, Using Quartz Crystal Microbalance.

Thrombomodulin (TM), an integral glycoprotein on the surface of endothelial cells, can be released during endothelial cell injury and the levels of serum TM are regarded as an important parameter of activity in vasculitides in vivo. Quantitative detection of TM and investigation on the release of soluble thrombomodulin (sTM) by the injured HUVEC-C cells using quartz crystal microbalance (QCM) were achieved in this work. Anti-antibody (AAb) and bovine serum albumin (BSA) were bound on gold nanoparticles (GNPs) to construct BSA-GNPs-AAb nanocomposites and they were characterized by transmission electron microscope, UV-vis, and infrared spectrophotometry, respectively. The capture of the nanocomposites on the TM antibody modified electrode, which was tested by scanning electron microscope, could result in a great decrease of the resonant frequency (f0). This binding was effectively inhibited by the beforehand immobilized TM proteins on the electrode surface due to the strong steric hindrance effect. It led to the decrease of the frequency changing extent. The relative frequency-shift was found to be proportional to the logarithm of the TM concentration from 10 to 5000 ng mL(-1) with a detection limit of 2 ng mL(-1). By analyzing the growth medium used for cell incubation, the release of sTM by the injured HUVEC-C cells in the presence of H2O2 was confirmed. The sTM amount in the growth medium was increased with the enhancement of contact time of the cells with H2O2, proving that sTM may serve as a specific marker of endothelial cell injury.

The investigation of relationship between joint findings and serum angiogenic and inflammatory factor levels in severe hemophilia A patients.

Despite the use of primary prophylactic Factor VIII replacement in severe hemophilia A patients, bleeding into joints cannot be prevented completely and early diagnosis and treatment of the joint bleedings are important for prevention of permanent joint damage. Recent studies have shown that neoangiogenesis plays important role in development of synovitis after recurrent joint bleedings. This study aimed to investigate the relationship between joint findings and levels of serum angiogenic and inflammatory factors in severe hemophilia A patients.The patient groups consisted of 10 severe hemophilia A patients with acute joint bleeding and 25 severe hemophilia A patients without acute joint bleeding. They were all inhibitor negative. The control group consisted of 22 healthy male children. Complete blood cell count analysis, C-reactive protein (CRP), serum ferritin, lactic acid, and ELISA-based detection of vascular endothelial growth factor (VEGF), intercellular adhesion molecule-1, thrombomodulin, macrophage migration inhibitory factor (MIF), and endostatin were performed from peripheral blood of patient and the control groups. CRP and MIF levels were detected significantly higher in hemophilia patients with acute joint bleeding than patients without acute joint bleeding. There was a positive correlation between serum thrombomodulin, VEGF, and MIF levels. In this study, we demonstrated that serum CRP and MIF levels increases in acute bleeding period regardless of the presence of previous joint damage in children with severe hemophilia. CRP elevation may be a useful and rapid marker for acute bleeding in these patients.

Single Nucleotide Polymorphisms Within The Thrombomodulin Gene (THBD) Predict Risk Of Non-Relapse Mortality In Patients With Graft-Versus-Host Disease

Purpose Acute graft-versus-host disease (GVHD) is a major complication after allogeneic stem cell transplantation (allo-SCT). With the current treatment approaches focussing on escalating immunosuppression, mortality rates of therapy-refractory disease courses are approx. 80-90%, underlining the medical need for novel therapeutic concepts. The pathophysiology of refractory GVHD is incompletely understood, however, recent evidence points towards a crucial role for endothelial damage. We have previously shown that steroid-refractory GVHD associates with increasing serum thrombomodulin (TM) levels 1, 2 and loss of endothelial TM expression 3, a constellation that indicates endothelial stress. However prior to allo-SCT, baseline TM levels did not predict steroid-refractory GVHD or survival. We hypothesized that endothelial vulnerability caused by single nucleotide polymorphisms within the TM gene (THBD) is a risk factor for refractory GVHD predicting outcome after allo-SCT. Patients and Methods Seven SNPs within the THBD gene were studied (rs1962.TC, rs1042579.TC, rs1042580.AG, rs3176123.TG, rs3176124GA, rs3176126.GA and rs3176134.CT) in a training cohort of 465 allografted patients. The relevant genotypes were then re-assessed in an independent validation cohort (n=386). Results Allele frequencies of seven THBD SNPs allowed meaningful statistical correlation with outcome after allo-SCT. An increased risk of non-relapse mortality (NRM) was associated with three SNPs: rs1962.CC, rs1042579.TT (455V) and rs1042580.GG. When patients were divided into risk cohorts (one vs. no high-risk SNP), a strong correlation with NRM was observed in both cohorts (training cohort: p=0.002, HR 2.32 CI 1.36-3.95; validation cohort: p=0.007, HR 2.13, CI 1.23-3.70) (Figure 1). NRM was predicted by THBD SNPs in particular for patients who later develop GVHD (validation cohort: p=0.00056, HR 3.03 CI 5.68-1.61, training cohort: p=0.012, HR 2.38 CI 1.21-4.69) but not in patients without GVHD. In contrast, THBD SNPs did not predict incidence of acute GVHD. Multivariate analyses adjusting for clinical variables confirmed the independent effect of THBD SNPs on NRM in both cohorts. Conclusions Our results demonstrate that THBD-SNPs predict mortality of GVHD patients without influencing GVHD incidence. This supports our hypothesis that endothelial vulnerability contributes to therapy refractory courses of the disease. Rather than focussing on the escalation of combinations of (endothelial cell toxic) immunosuppressive drugs, therapeutic approaches aiming at endothelial protection should be evaluated. 1. Dietrich S, Falk CS, Benner A, et al. Endothelial vulnerability and endothelial damage are associated with risk of graft-versus-host disease and response to steroid treatment. Biol Blood Marrow Transplant 2013; 19(1): 22-7. 2. Luft T, Dietrich S, Falk C, et al. Steroid-refractory GVHD: T-cell attack within a vulnerable endothelial system. Blood 2011; 118(6): 1685-92. 3. Andrulis M, Dietrich S, Longerich T, et al. Loss of endothelial thrombomodulin predicts response to steroid therapy and survival in acute intestinal graft-versus-host disease. Haematologica 2012; 97(11): 1674-7. Disclosures: Dreger: Riemser Pharma : Consultancy, Honoraria, Research Funding.