Plasma F1 Research Articles

Daytime plasma cortisol and cortisol response to dexamethasone suppression are associated with a prothrombotic state in hypertension.

A prothrombotic state was demonstrated in patients with Cushing's syndrome and is involved in the development and progression of cardiovascular and renal damage in hypertensive patients. This study was designed to examine the relationships between cortisol secretion and the hemostatic and fibrinolytic systems in hypertension. In 149 middle-aged, nondiabetic, essential hypertensive patients free of cardiovascular and renal complications, we measured hemostatic markers that express the spontaneous activation of the coagulation and fibrinolytic systems and assessed daily cortisol levels (8 AM, 3 PM, 12 AM; area under the curve, AUC-cortisol) together with the cortisol response to dexamethasone overnight suppression (DST-cortisol). Plasma levels of D-dimer (D-dim), prothrombin fragment 1+2 (F1+2), and von Willebrand factor (vWF) were progressively and significantly higher across tertiles of AUC-cortisol and DST-cortisol, whereas no differences were observed in fibrinogen, tissue plasminogen activator, plasminogen activator inhibitor-1, antithrombin III, protein C, and protein S. D-dim, F1+2, and vWF were significantly and directly correlated with age and both AUC-cortisol and DST-cortisol. Multivariate regression analysis showed that both AUC-cortisol and DST-cortisol were related to plasma D-dim, F1+2, and vWF independently of age, body mass index, blood pressure, and renal function. Greater daily cortisol profile and cortisol response to overnight suppression are independently associated with a prothrombotic state in hypertensive patients and might contribute to the development of organ damage and higher risk of cardiovascular complications.

PD19-05 ELEVATED PROTHROMBIN FRAGMENT 1+2 IS A RISK FACTOR OF SEVERE ACUTE KIDNEY INJURY IN PATIENTS WITH UROLOGICAL SEPSIS

You have accessJournal of UrologyCME1 Apr 2023PD19-05 ELEVATED PROTHROMBIN FRAGMENT 1+2 IS A RISK FACTOR OF SEVERE ACUTE KIDNEY INJURY IN PATIENTS WITH UROLOGICAL SEPSIS Naoki Fujita, Masaki Momota, Osamu Soma, Daisuke Noro, Jotaro Mikami, Shingo Hatakeyama, Hiroyuki Ito, Takahiro Yoneyama, Yasuhiro Hashimoto, Kazuaki Yoshikawa, and Chikara Ohyama Naoki FujitaNaoki Fujita More articles by this author , Masaki MomotaMasaki Momota More articles by this author , Osamu SomaOsamu Soma More articles by this author , Daisuke NoroDaisuke Noro More articles by this author , Jotaro MikamiJotaro Mikami More articles by this author , Shingo HatakeyamaShingo Hatakeyama More articles by this author , Hiroyuki ItoHiroyuki Ito More articles by this author , Takahiro YoneyamaTakahiro Yoneyama More articles by this author , Yasuhiro HashimotoYasuhiro Hashimoto More articles by this author , Kazuaki YoshikawaKazuaki Yoshikawa More articles by this author , and Chikara OhyamaChikara Ohyama More articles by this author View All Author Informationhttps://doi.org/10.1097/JU.0000000000003285.05AboutPDF ToolsAdd to favoritesDownload CitationsTrack CitationsPermissionsReprints ShareFacebookLinked InTwitterEmail Abstract INTRODUCTION AND OBJECTIVE: Elevated prothrombin fragment 1+2 (F1+2), produced during conversion of prothrombin to thrombin, indicates excess thrombin production and serves as a molecular marker of activated coagulation. The aim of this study was to investigate the impact of elevated F1+2 on the development of severe acute kidney injury (AKI) in patients with urological sepsis. METHODS: We prospectively collected the data from 143 patients with urological sepsis between March 2017 and March 2019. AKI was defined according to the KDIGO criteria. Patients were divided into two groups: lower F1+2 (plasma F1+2 levels <395 pmol/L) and higher F1+2 groups (plasma F1+2 levels ≥395 pmol/L). Multivariable logistic regression analysis was performed to evaluate the impact of elevated F1+2 on the severe AKI development. We evaluated predictive ability of elevated F1+2 for the severe AKI development using the receiver operating characteristic curve and compared using net reclassification improvement (NRI) and integrated discrimination improvement (IDI). RESULTS: Median age was 78 years old in this cohort. Of 143 patients, 61 were classified as a higher F1+2 group and 36 developed to severe AKI (AKI stage 2-3). Rate of severe AKI development in the higher F1+2 group was significantly higher than that of the lower F1+2 group (Figure 1A; p=0.004). Plasma F1+2 levels in patients with severe AKI was significantly higher than that in patients with stage 0-1 AKI (Figure 1B; p=0.007). In multivariable analysis, plasma F1+2 level ≥395 pmol/L was selected as a significant independent risk factor for severe AKI development (Figure 1C; hazard ratio 3.067, p=0.006). The predictive ability of hydronephrosis plus elevated F1+2 levels for severe AKI development was significantly improved compared with hydronephrosis alone (Figure 1D). CONCLUSIONS: Elevated prothrombin F1+2 is a risk factor of severe AKI in patients with urological sepsis. Source of Funding: None © 2023 by American Urological Association Education and Research, Inc.FiguresReferencesRelatedDetails Volume 209Issue Supplement 4April 2023Page: e578 Advertisement Copyright & Permissions© 2023 by American Urological Association Education and Research, Inc.MetricsAuthor Information Naoki Fujita More articles by this author Masaki Momota More articles by this author Osamu Soma More articles by this author Daisuke Noro More articles by this author Jotaro Mikami More articles by this author Shingo Hatakeyama More articles by this author Hiroyuki Ito More articles by this author Takahiro Yoneyama More articles by this author Yasuhiro Hashimoto More articles by this author Kazuaki Yoshikawa More articles by this author Chikara Ohyama More articles by this author Expand All Advertisement PDF downloadLoading ...

Plasminogen activator inhibitor-1, thrombin-antithrombin, and prothrombin fragment F1+2 have higher diagnostic values than D-dimer for venous thromboembolism after TKA.

ObjectiveTo investigate the diagnostic values of D-dimer, plasminogen activator inhibitor-1 (PAI-1), thrombin–antithrombin (TAT), and prothrombin fragment F1 + 2 (F1 + 2) for predicting venous thromboembolism (VTE) after total knee arthroplasty (TKA).MethodsUltrasonography and CTPA were performed to diagnose VTE in 252 patients who underwent TKAs. Plasma D-dimer, PAI-1, TAT, and F1 + 2 levels were assessed 1–3 days prior to operation (T1), second hour (T2), first (T3), and third day (T4) after the operation. Receiver–operating characteristic curves (ROC) analysis was conducted and pairwise compared to evaluate the diagnostic value of those biomarkers.ResultsPlasma D-dimer levels differed between patients with and without VTE significantly on T4, PAI-1, TAT, and F1 + 2 levels differed on T3 and T4. The areas under ROC of D-dimer, PAI-1, TAT and F1 + 2 levels were 0.645, 0.773, 0.771 and 0.797, respectively. The most feasible cutoff values of D-dimer, PAI-1, TAT and F1 + 2 in predicting VTE after TKA were 2.24 ug/ml, 35.96 ng/ml, 13.36 ng/mg and 11.1 ng/ml, respectively. Pairwise comparison of ROC curves revealed that D-dimer level had the lowest diagnostic accuracy, whereas PAI-1, TAT and F1 + 2 level had similar diagnostic accuracy. There were significant differences in duration of tourniquet time and duration of anesthesia between patients with and without VTE.ConclusionAfter TKA, using 2.24ug/mL as the threshold value of D-dimer is more accurate than using 0.5ug/mL in the monitoring of VTE, PAI-1, TAT and F1 + 2 are more valuable than D-dimer in predicting VTE. Duration of tourniquet and duration of anesthesia are risk factors for the development of VTE.

Plasma fibrin clots of pulmonary embolism patients present increased amounts of factor XIII and alpha2-antiplasmin at 3 months' anticoagulation since the acute phase.

Fibrin cross-linking by coagulation factor (F)XIII leads to clot stabilization. Reduced plasma FXIII levels have been reported in acute pulmonary embolism (PE) patients. We investigated the impact of anticoagulant therapy on clot-bound amounts of FXIII and α2-antiplasmin and their associations with fibrin clot properties in patients with PE. Clots generated from plasma of 18 acute symptomatic patients on admission and after a 3-month treatment with rivaroxaban were assessed off anticoagulation using mass spectrometry. Plasma FXIII and α2-antiplasmin activity were determined at the 2 time points along with thrombin generation markers, plasma fibrin clot permeability (Ks), and clot lysis time (CLT). Following anticoagulant therapy, clot-bound FXIII increased from 2.97 (interquartile range, 1.98 - 4.08) to 4.66 (3.5 - 6.9) mg/g protein and α2-antiplasmin from 9.4 (7.2 - 10.6) to 11 (9.5 - 14) mg/g protein (both p < 0.0001). The two parameters showed positive correlation at baseline only (r = 0.63, p = 0.0056). Similarly to clot-bound amounts, plasma FXIII (+25.8%) and α2-antiplasmin activity (+12%) increased at 3 months. Plasma FXIII activity on admission, but not after 3 months since the index PE, was associated with amounts of clot-bound FXIII (r = 0.35, p = 0.043) and α2-antiplasmin (r = 0.47, p = 0.048). At baseline, clot-bound FXIII correlated with plasma F1+2 prothrombin fragments levels (r = 0.51, p = 0.03), while clot-bound α2-antiplasmin correlated with CLT (r = 0.43, p = 0.036). At 3 months associations of clot-bound FXIII and α2-antiplasmin were abolished. This study assessed for the first time changes in the fibrin clot composition following acute PE, suggesting an increase of clot-bound and plasma FXIII and α2-antiplasmin levels after 3 months.

Novel fibrin-fibronectin matrix accelerates mice skin wound healing

Plasma fibrinogen (F1) and fibronectin (pFN) polymerize to form a fibrin clot that is both a hemostatic and provisional matrix for wound healing. About 90% of plasma F1 has a homodimeric pair of γ chains (γγF1), and 10% has a heterodimeric pair of γ and more acidic γ′ chains (γγ′F1). We have synthesized a novel fibrin matrix exclusively from a 1:1 (molar ratio) complex of γγ′F1 and pFN in the presence of highly active thrombin and recombinant Factor XIII (rFXIIIa). In this matrix, the fibrin nanofibers were decorated with pFN nanoclusters (termed γγ′F1:pFN fibrin). In contrast, fibrin made from 1:1 mixture of γγF1 and pFN formed a sporadic distribution of “pFN droplets” (termed γγF1+pFN fibrin). The γγ′F1:pFN fibrin enhanced the adhesion of primary human umbilical vein endothelium cells (HUVECs) relative to the γγF1+FN fibrin. Three dimensional (3D) culturing showed that the γγ′F1:pFN complex fibrin matrix enhanced the proliferation of both HUVECs and primary human fibroblasts. HUVECs in the 3D γγ′F1:pFN fibrin exhibited a starkly enhanced vascular morphogenesis while an apoptotic growth profile was observed in the γγF1+pFN fibrin. Relative to γγF1+pFN fibrin, mouse dermal wounds that were sealed by γγ′F1:pFN fibrin exhibited accelerated and enhanced healing. This study suggests that a 3D pFN presentation on a fibrin matrix promotes wound healing.

Further Evidence for Hypercoagulability in Women With Ovarian Endometriomas.

Our previous studies have shown that platelets play a crucial role in the development of endometriosis, and women with endometriosis appear to be in a state of hypercoagulability. However, a recent study could only replicate part of our previous finding, casting doubts on this notion. We further investigated this question through a cross-sectional study by measuring additional coagulation factors in women with and without endometriosis. To this end, we conducted a cross-sectional study of 100 women with laparoscopically and pathologically diagnosed ovarian endometriomas (OMA) and another 100 women without endometriosis. The platelet count; platelet activation rate; maximum platelet aggregation rate; plasma levels of D-dimer, fibrinogen, fibrin degradation products (FDPs), plasma soluble P-selectin (sP-sel), and prothrombin fragment 1+2 (F1+2); prothrombin time; thrombin time (TT); and activated partial thromboplastin time were measured before surgery and 3 months after surgery, and their clinical data were recorded. These measurements were also performed in control patients. We found that, compared with controls, women with OMA had a significantly higher platelet activation rate and platelet aggregation rate, elevated plasma D-dimer, fibrinogen, FDPs, sP-sel, and F1+2 levels as well as shortened TT. Remarkably, TT was prolonged, and all the other coagulation measurements, except plasma fibrinogen level, were significantly reduced 3 months after surgical removal of endometriotic lesions. Thus, our study provides another piece of evidence that endometriosis is a hypercoagulable disease, and anticoagulation therapy may hold promises in treating endometriosis.

Prothrombin fragment F1+2 in plasma and urine during total hip arthroplasty

ObjectiveTo study the time course and correlation of prothrombin fragment (F1+2) in plasma and urine during THA. MethodF1+2 concentrations were recorded at four time-points during THA. ResultsF1+2 increased during surgery. In contrast to urine, plasma F1+2 was temporarily reduced the first postoperative day. At day six, plasma F1+2 was significantly elevated indicating on-going thrombin activation, while urine F1+2 was normalized. ConclusionThe response in coagulation to operative trauma can be recorded with F1+2 both in plasma and urine. We were not able to demonstrate any correlation of F1+2 in plasma and urine later in the postoperative period.

Effect of rivaroxaban on prothrombin fragment 1 + 2 compared with warfarin in patients with acute cardioembolic stroke: Insight from its serial measurement

IntroductionPatients with intracerebral hemorrhage during rivaroxaban treatment have small hematoma and favorable outcomes compared with those with warfarin. We investigated its possible mechanism, focusing on prothrombin fragment 1+2 (F1+2), a marker of thrombin generation. Materials and methodsIn 65 patients with acute cardioembolic stroke (median 77years), rivaroxaban was initiated at 5days after the onset. Plasma F1+2 level (normal range, 69–229pmol/L), prothrombin time (PT), and rivaroxaban concentration evaluated by anti-Xa activity were serially measured. ResultsMedian plasma F1+2 was 276 (IQR, 195–454) pmol/L before starting rivaroxaban, and significantly decreased to 196 (141–267) and 192 (151–248) on 7 and 28days after rivaroxaban, respectively (both p<0.05). Serial measurements of PT and rivaroxaban concentration at trough, 2, 4, and 6h after taking rivaroxaban showed a positive correlation (R2=0.69, p<0.01). PT at 4h after rivaroxaban was significantly prolonged compared with trough (16.6 versus 11.5s, p<0.0001). F1+2 at 4h was also decreased compared with trough (160 (123–245.5) versus 196 (141–266.5), p=0.04), but no patients showed F1+2 below the normal range at 4h. In other 34 patients with warfarin treatment (77years), median PT-INR and F1+2 were 2.06 (1.75–2.50) and 75 (48–111) (p<0.0001 versus 4h after rivaroxaban). Notably, of those with PT-INR≥2.0 (18/34), 12 (12/18, 67%) showed F1+2 below the normal range. ConclusionsRivaroxaban retains a normal thrombin generation even at its peak level with prolonged PT, whereas warfarin at therapeutic levels inhibits thrombin generation. This may partly explain different outcomes in patients complicated with bleeding events.

Study the association of adiponectin with inflammation and hypercoagulability in case of type 2 diabetic subjects with renal dysfunction

AbstractAdiponectin is a 30-kDa polypeptide chiefly secreted by adipose tissue and plays a pivotal role in cardiovascular complication and kidney disease. The aim of this study was to find if there was any association of adiponectin with inflammation and hypercoagulability in case of type 2 diabetic subjects with renal dysfunction.From May 2012 to January 2014, 205 outpatients with normo, micro, and macroalbuminuria having type 2 diabetes (T2DM) were included in this study.We found a U-shaped association of adiponectin with albumin excretion. Macroalbuminuria showed a positive correlation with adiponectin (p < 0.0001). Plasma F1+2 was strongly associated with degree of renal failure (p < 0.001). CRP, IL6, TNFα, and pentosidine status showed a linear increase from normoalbuminuria and microalbuminuria to macroalbuminuria (p < 0.001, p < 0.0001).Monitoring adiponectin, pasma F1+2, inflammatory and oxidative stress markers provide a predictive value for presence of macrovascular complications in patients wit...

Sub-Clinical Activation of Thrombosis May Predict Survival in Cancer Patients

Background: Approximately 10% of newly diagnosed cancer patients will develop thromboembolic disease (TED) in the first year following diagnosis. We hypothesize that a subgroup of cancer patients at diagnosis will have subclinical changes in their coagulation system that will indicate increased risk for subsequent TED and/or predict increased adverse cancer outcome.Methods: We performed a prospective, non-interventional study of recently diagnosed cancer patients who were scheduled to start chemotherapy. We obtained a single plasma sample at the time of diagnosis and prior to therapy, and measured prothrombin 1.2 fragment (F1.2), a measure of thrombin activation, and plasma microparticle levels (MP), a measure of platelet, endothelial, and other cell activation. We reviewed subsequent clinical outcomes including survival and incidence of venous thrombosis. We accrued 22 patients with mean age 63 years (range 39 to 89; 20 males, 2 females), 12 with lung, 6 with colorectal, and 4 with head and neck cancer. Mean follow-up was 24 months (range 3 to 57). Plasma F1.2 and MP were assayed using commercially available ELISA kits.Results:T1.2 concentrations were significantly higher in the deceased subgroup (table 1; p<0.002), and in patients with venous thrombosis (table 2; p<0.05). MPs were not informative. Sensitivity and specificity for prediction of survival and venous thrombosis (table 3) were determined by comparing patients in highest quartile of levels with lower quartiles combined and by using threshold value of mean +2.5 SD’s determined by measuring levels in 8 healthy controls.Table 1Survival: F1.2 and MP (Mann-Whitney)Deceased Patients (N=7)Alive Patients (N=15)Mean +/- SDMedianMean +/- SDMedianp-valueF1.2 (pM)436+/-214355121+/-117105<0.002MP (nM)10+/-1038.8+/-2.82.8Not SignificantTable 2Venous Thrombosis: F1.2 and MP (Mann-Whitney)Thrombosis (N=4)No Thrombosis (N=18)Mean +/- SDMedianMean +/- SDMedianp-valueF1.2 (pM)368+/-267313189+/-191163<0.05MP (nM)5.9+/-5.33.99.8+/-15.62.7Not SignificantTable 3Sensitivity/Specificity Analysis>mean + 2.5 SD’sHighest QuartileMPT1.2MPT1.2Death SENSITIVITY43%43%43%71%Death SPECIFICITY87%100%87%93%Thrombosis SENSITIVITY25%25%25%50%Thrombosis SPECIFICITY78%89%78%78%Conclusion: T1.2 levels measured at time of diagnosis of cancer correlated with increased mortality and development of venous thrombosis. MP levels were not informative, possibly due to the limited size of the study population. Additional studies are needed to elucidate whether these biomarkers may be useful in identifying a higher risk population for death and/or venous thrombosis, and to help guide enhanced therapy for such higher risk patients. DisclosuresNo relevant conflicts of interest to declare.

The hypercoagulable status in common Mediterranean β-thalassaemia mutations trait.

The coagulation activation in β-thalassaemia is multifactorial and most likely a consequence of the exposure of phosphatidylserine (PS) on RBCs surface. The degree of PS exposure and procoagulant activity of RBCs in β-thalassaemia trait (BTT) subjects carrying common Mediterranean mutations were assessed. Eighty BTT subjects carrying common Mediterranean mutations (β+, n=53 and β0 , n=27) and sixty healthy subjects served as controls were studied. Plasma prothrombin fragment 1+2 (F1+2), percentage of PS expression on RBCs membrane, clotting times of modified thromboplastin generation test (MTGT) and modified partial thromboplastin with kaolin (MPTTK) were estimated. The percentage of annexin V positive RBCs and plasma F1+2 had a significant increase and MTGT had a significant decrease in BTT subjects versus controls and in β0 group versus β+ group. MPTTK was significantly shorter in BTT subjects than controls, but no significant deference between BTT subjects. The percentage of annexin V positive RBCs showed a significant negative correlation with haemoglobin level, MTGT and MPTTK, and a significant positive correlation with plasma F1+2. BTT subjects may have a risk of hypercoagulable state particularly in β0 genotype. Measurement of PS exposure on RBCs and the plasma F1+2 is useful to evaluate hypercoagulability state.

Urinary prothrombin fragment 1+2 in patients with venous thrombosis and myocardial infarction

Patients with venous-thromboembolism (VTE) and myocardial infarction (MI) have elevated prothrombin fragment 1+2 (F1+2) levels. In patients with postoperative VTE, urinary F1+2 (uF1+2) was higher than in individuals without VTE. To explore the relationship between plasma and uF1+2 we performed a pilot study in patients with thrombotic events and healthy controls. In 40 patients with VTE or MI, and 25 age- and sex-matched healthy controls, F1+2 and D-dimer levels were measured in urine and plasma within 48h after diagnosis. In addition, in all subjects renal function was assessed. Plasma and uF1+2 levels were positively correlated. Compared to controls, patients with VTE had higher levels of both plasma F1+2 (271 vs 160pmolL(-1), p<0.05) and uF1+2 levels (38 vs 28pmolL(-1)), the latter, however, was not statistically significant. Patients with acute MI had similar F1+2 levels as controls in both plasma and urine. Differences in urinary F1+2 levels could not be attributed to differences in concentrations of creatinine or albumin in spot urine samples. Overall, D-dimer and F1+2 levels in urine were extremely low in all groups.

A Phase 1 Study of Prasugrel in Subjects with Sickle Cell Disease: Effects on In Vivo Markers of Platelet Activation and of Coagulation

Abstract 1049▪▪This icon denotes a clinically relevant abstract Introduction:Evidence suggests that platelets are activated in sickle cell disease (SCD) and this appears to increase further during painful crises caused by vascular occlusions from sickled red blood cells. Antiplatelet therapy may be useful in reducing the frequency and severity of acute pain episodes and reducing the risk of thrombotic complications. Prasugrel, an ADP receptor antagonist, irreversibly inhibits the P2Y12 ADP receptor, blocking ADP-stimulated platelet activation and aggregation and reducing downstream procoagulant activities. Here we present the first evaluation of prasugrel's effects on markers of in vivo platelet activation and of coagulation in subjects with SCD. Methods:Twenty-six adult subjects were enrolled and 25 completed the study: 12 with SCD and 13 well-matched healthy controls. Subjects were examined before and after 12±2 days of treatment with oral prasugrel (5.0 mg/day for subjects weighing <60 kg and 7.5 mg/day for subjects weighing ≥60 kg). Markers of platelet activation and coagulation included whole-blood platelet-monocyte and -neutrophil aggregates, and whole blood platelet-associated P-selectin and platelet CD40L, all measured by flow cytometry and presented as percent (%) of marker positive cells. Plasma soluble (s) P-selectin, CD40L, and plasma prothrombin fragment 1.2 (F1.2) were evaluated by ELISA. Results:Results from the biomarkers are presented in the table. Prior to prasugrel administration (baseline), subjects with SCD had significantly higher levels of the following biomarkers compared to healthy subjects: Platelet-monocyte aggregates, platelet-neutrophil aggregates, platelet CD40L, and plasma F1.2. In addition, subjects with SCD had numerically higher values of sCD40L, as well as platelet-associated and sP-selectin.Prasugrel treatment resulted in numerical decreases in levels of all biomarkers (with the exception of platelet-associated CD40L for control subjects), most notably in SCD subjects with elevated baseline levels.Prasugrel was safe and well tolerated with no serious adverse events observed during the study. No subject discontinued the study due to an adverse event (AE) and the majority of AEs were mild. No subjects with SCD reported any bleeding-related AEs. Conclusion:In this study, compared to healthy controls, baseline elevation of several platelet-activation and coagulation markers among adult subjects with SCD is consistent with that seen in previous studies of both children and adults with SCD. The decrease in platelet activation biomarkers following 12 days of prasugrel treatment in subjects with SCD suggests prasugrel interrupts SCD-related platelet activation in vivo and raises the possibility that prasugrel may modulate the frequency and/or severity of painful crises associated with SCD. These data support additional studies of the safety and efficacy of prasugrel in the treatment of vascular complications associated with SCD.BiomarkersTimeControl (Mean ± SD)SCD (Mean ± SD)Platelet-monocyte aggregates (%)Baseline50.7 ± 17.978.0 ± 25.6*Day 1240.4 ± 23.562.2 ± 24.3**Platelet-neutrophil aggregates (%)Baseline17.7 ± 4.027.5 ± 15.2*Day 1216.1 ± 8.225.9 ± 17.8CD40L (%)Baseline13.0 ± 4.617.3 ± 4.0*Day 1217.8 ± 4.8**16.4 ± 3.8P-selectin (% CD62P)Baseline10.3 ± 6.415.8 ± 7.5Day 125.4 ± 2.1**12.0 ± 12.7sCD40L (pg/mL)Baseline266.6 ± 184.2610.2 ± 644.2Day 12155.6 ± 83.8**419.4 ± 640.2sP-selectin (ng/mL)Baseline29.5 ± 11.137.6 ± 9.2*Day 1226.7 ± 8.032.8 ± 12.3F1.2 (nmol/L)Baseline205.8 ± 291.91777.0 ± 2487.4*Day 12135.6 ± 57.6403.0 ± 265.5*p<.05 for between group differences at baseline by t-tests**p<.05 for within-group changes from baseline by t-tests Disclosures:Jakubowski:Eli Lilly and Company: Employment, Equity Ownership. Off Label Use: This abstract discusses prasugrel treatment in patients with sickle cell disease. Please see USPI for most up-to-date information. Zhou:Eli Lilly and Company: Employment, Equity Ownership. Small:Eli Lilly and Company: Employment, Equity Ownership. Winters:Eli Lilly and Company: Employment, Equity Ownership. Lachno:Eli Lilly and Company: Employment, Equity Ownership. Frelinger:Takeda: Research Funding; Daiichi Sankyo Company, Ltd. and Eli Lilly and Company: Consultancy, Research Funding; GLSynthesis: Research Funding. Howard:Daiichi Sankyo Company, Ltd. and Eli Lilly and Company: Research Funding. Payne:Eli Lilly and Compnay: Employment, Equity Ownership.

Postprandial changes in the phospholipid composition of circulating microparticles are not associated with coagulation activation

IntroductionEvidence is present that the phospholipid composition of circulating cell-derived microparticles (MP) affects coagulation in vivo, and that postprandial metabolic alterations may be associated with hypercoagulable state. Our objective was to investigate whether postprandial metabolic responses affect the phospholipid composition of MP, and whether such changes are associated with coagulation activation. Materials and MethodsTwelve healthy males were studied twice and randomly received two consecutive meals or remained fasted. Blood was collected before and at 2, 4, 6 and 8h following breakfast. Plasma concentrations of prothrombin-F1+2 and thrombin-antithrombin-complexes were measured. Numbers and cellular origin of MP were determined by flowcytometry. The phospholipid composition of MP was determined by hpTLC. In vitro procoagulant activity of MP was studied by fibrin generation. ResultsDuring the meal visit, plasma glucose, triglyceride and insulin levels increased, compared to baseline and the fasting visit (all P<0.05). Postprandially, the total numbers of MP increased in time compared to the fasting visit (P<0.05). Erythrocyte-derived MP increased (6-fold) during the meal visit, but remained constant on the fasting day (P<0.001). On the meal versus fasting day circulating MP contained increased phosphatidylcholine (P<0.05) and decreased sphingomyelin (P<0.05) amounts. The amount of phosphatidylserine did not change. Concentrations of plasma F1+2 and thrombin-antithrombin were similar on both days, as was the ability of MP to generate fibrin in vitro. ConclusionAlthough numbers, cellular origin and phospholipid composition of MP alter during exposure to two consecutive meals in healthy subjects, this does not lead to changes in the coagulation activation in vivo.

Activation of coagulation in bullous pemphigoid and other eosinophil-related inflammatory skin diseases

Bullous pemphigoid (BP) is a skin disease caused by autoantibodies to hemidesmosomal proteins BP180 and BP230, with eosinophils participating in blister formation. Tissue factor (TF), the initiator of coagulation, is embodied within the eosinophil granules and exposed upon activation. We evaluated the coagulation activation in patients with BP (63), chronic urticaria (CU; 20), atopic dermatitis (AD; 14), cutaneous drug reactions (CDRs; six), psoriasis (20), dermatitis herpetiformis (DH; four) and primary cutaneous T cell lymphoma (CTCL; five), and in 40 healthy controls. Prothrombin fragment F1+2 and d-dimer (coagulation markers) were measured by enzyme-linked immunosorbent assay (ELISA) in all plasma samples and BP blister fluid. Skin TF expression was evaluated immunohistochemically in the patients and 20 controls. F1+2 and d-dimer levels were higher in BP plasma than in control plasma (P = 0·0001 for both), and dramatically high in blister fluid; both correlated positively with disease severity, esinophil counts and anti-BP180 antibodies (P = 0·006-0·0001). Plasma F1+2 and d-dimer levels were higher in the CU, AD and CDR patients than in controls (P = 0·0001 for all), but normal in the psoriasis, DH and CTCL patients. Skin TF was expressed in the BP (P = 0·0001), CU (P = 0·0001), AD (P = 0·001) and CDR patients (P = 0·01), but not in the psoriasis, DH or CTCL patients. Co-localization confocal microscopy studies confirmed eosinophils as the source of TF in 10 BP patients. The coagulation cascade is activated in BP and other eosinophil-mediated skin disorders, but not in non-eosinophil driven conditions. This hypercoagulability may contribute to inflammation, tissue damage and, possibly, thrombotic risk.

Circulating tissue factor and microparticles are not increased in patients with deep vein thrombosis

Circulating tissue factor (TF) is associated with inflammation and may contribute to thrombotic events. Aim of this study was to analyze circulating TF activity and proinflammatory cytokines in patients with deep venous thrombosis. Forty-eight patients with deep vein thrombosis and 45 control subjects were included. Venous blood samples were obtained at diagnosis for analysis of TF activity, TF antigen, prothrombin fragment F1 + 2, microparticles (expressing phosphatidylserine and supporting FXa generation), Interleukin (IL)-1beta, IL-6, IL-8, IL-10, IL-12 and tumor-necrosis-factor-alpha (TNF). TF antigen, activity and microparticles were similar in both groups: In contrast, a significant increase in plasma IL-6, IL-8 and F1 + 2 levels was found in thrombosis. This increase in IL-6 and IL-8 as well as F1 + 2 was not correlated with the extent of thrombosis, predisposing factors or onset of symptoms. Circulating TF and microparticles are not elevated in deep venous thrombosis. The increase in IL-6, IL-8 and F1 + 2 during thrombosis was not proportional to the extent or predisposing risk factors.

Additional impact of morning haemostatic risk factors and morning blood pressure surge on stroke risk in older Japanese hypertensive patients

Stroke events occur most frequently in the morning hours. Impaired haemostatic activity and morning blood pressure (BP) surge, defined as the morning BP increase from sleep, have individually been associated with stroke risk in general or hypertensive populations. However, their combined impact on the risk of a stroke remains unknown. A total of 514 hypertensive patients aged > 50 years (mean 72.3 years; 37% men) underwent 24 h BP monitoring, measurement of haemostatic risk factors [plasma fibrinogen, plasminogen activator inhibitor-1 (PAI-1), and prothrombin fragment 1+2(F1+2)], and brain MRI at baseline. The incidence of stroke was prospectively ascertained. During an average of 41 months (1751 person-years), there were 43 stroke events (ischaemic, 30; haemorrhagic, 5; undefined, 8). On multivariable analysis adjusted for confounding factors, the hazard ratio [HR (95% confidence interval (CI)] for stroke in the highest vs. lower quartiles of PAI-1 was 2.5 (1.3-4.6), that for F1+2 was 2.6 (1.4-5.0), and that for the morning BP surge was 1.2 (1.1-1.4; all P< 0.01). In particular, the ratio was substantially higher in cases with the highest quartile of both PAI-1 and F1+2 levels compared with those with the lower quartiles of both parameters (HR: 8.2; 95% CI: 3.7-18.2; P< 0.001). Among the patients with the highest quartile of the morning BP surge (n= 128), the multivariable HR (95% CI) for the highest vs. lower quartiles of PAI-1 was 3.4 (1.3-9.1) and that for F1+2 was 3.3 (1.3-8.7) (both P< 0.05). High levels of plasma PAI-1 and F1+2, as well as an excessive morning BP surge, are independently and additively associated with an increased risk of stroke in older hypertensive patients.

Measurements of Prothrombin Fragment 1.2 In Urine and Blood to Assess Hypercoagulability In Preeclampsia

AbstractAbstract 3174 Objective:Pregnancy is an acquired hypercoagulable condition as coagulation is shifted towards a more procoagulant state. Prothrombin fragment 1.2 (F1.2) is a direct measure of thrombin generation and thus of activation of the haemostatic system. We aimed to investigate the relationship between plasma F1.2 and urinary elimination in pregnancies complicated by PE compared to uncomplicated normotensive pregnancies and to establish whether levels of urinary F1.2 reflects thrombin generation de facto or is influenced or a result of proteinuria. Methods:In 62 pregnancies with PE and 10 randomly selected normotensive pregnancies, blood samples were collected at inclusion and at gestational weeks 17, 23, 29, 33, 37, 39 and 41, and urine samples were collected every two weeks from inclusion until delivery or in a subgroup of patients only once before delivery. Levels of plasma and urinary F1.2, albumin and creatinine and plasma D-dimer were measured and standardized data collected on pregnancy, delivery, placenta and neonatal outcome. F1.2 in urine and plasma were analyzed by enzyme immunoassay Enzygnost® F1.2 (Dade Behring Marburg GmbH, Germany). All measurements were performed in duplicate and a reference curve established for each series. Creatinine was analyzed by enzymatic reaction IDMS (Isotope Dilution Mass Spectroscopy) by Ortho Clinical (Johnson & Johnson Nordic AB, Sweden), albumin was analyzed by direct immunoassay by Ortho Clinical (Johnson & Johnson Nordic AB, Sweden), and finally D-dimer analyzed by BCSXP (Siemens 2750-DK, reagens Auto Dimer, Biopool, UK). Results:In all pregnancies, levels of urinary F1.2 and plasma F1.2 increased throughout pregnancy until delivery especially in pregnancies complicated by PE (Figure 1). This difference between PE and controls was more pronounced at gestational weeks 30 and 37. D-dimer demonstrated a small increase with gestational age in all pregnancies with non-significant higher measurements in the second and third trimesters of women with PE. Creatinine was fairly constant in all pregnancies. In PE, albuminuria was significantly increased at 30 and 37 gestational weeks, coinciding with declining plasma levels. There was no significant association between levels of urinary F1.2 and albuminuria (Figure 2). [Display omitted] [Display omitted] Conclusion:In all pregnancies, levels of uF1.2 increased with gestational age with an even further increase at 37 gestational weeks. The increase in urinary F1.2 was even more pronounced in pregnancies with PE, emphasising the hypercoagulable character of this disease. Furthermore, the levels of uF1.2 reflected plasma generation of F1.2 and thus the amount of thrombin generated. F1.2 was eliminated in urine irrespective of the co-existence of proteinuria and urinary F1.2 may be a more reliable and sensitive parameter for assessing haemostatic activation than plasma levels although the exact role of uF1.2 warrants further studies. Disclosures:Lassen:BMS: Consultancy, Membership on an entity's Board of Directors or advisory committees; Pfizer: Consultancy, Membership on an entity's Board of Directors or advisory committees; Bayer: Consultancy, Speakers Bureau; Sanofi Aventis: Consultancy; Astellas Pharma: Consultancy; GSK: Consultancy.

Relationship between disseminated intravascular coagulation and levels of plasma thrombinogen segment 1+2, D-dimer, and thrombomodulin in patients with multiple injuries

To explore the relationship between disseminated intravascular coagulation (DIC) and levels of plasma thrombinogen segment 1+2 (F1+2), D-dimer (D-D), and thrombomodulin (TM) in patients with severe multiple injuries. In this study, 66 patients (49 males and 17 females, aged 15-74 years, mean=38.4 years) with multiple injuries, who were admitted to our hospital within 24 hours after injury with no personal or family history of hematopathy or coagulopathy, were divided into a minor injury group (ISS<16, n=21) and a major injury group (ISS>or=16, n=45) according to the injury severity. The patients in the major injury group were divided into a subgroup complicated with DIC (DIC subgroup, n=12) and a subgroup complicated with no DIC (non-DIC subgroup, n=33). Ten healthy people (7 males and 3 females, aged 22-61 years, mean=36.5 years+/-9.0 years), who received somatoscopy and diagnosed as healthy, served as the control group. Venous blood samples were collected once in the control group and 1, 3 and 7 days after trauma in the injury groups. The F1+2 and TM concentrations were determined by enzyme linked immunosorbent assay (ELISA), and D-D concentrations were measured by automated latex enhanced immunoassay. F1+2, D-D and TM levels were higher in the minor and major injury groups than in the control group. They were markedly higher in the major injury group than in the minor injury group. In the non-DIC subgroup, F1+2 levels declined gradually while D-D and TM levels declined continuously. In the DIC subgroup, F1+2 and D-D levels remained elevated while TM levels exhibited an early rise and subsequent decrease. Plasma F1+2, D-D and TM levels were higher in the DIC patients than in the non-DIC patients. Injury-induced increases in F1+2, D-D and TM plasma levels had significant positive correlation with each other at each time point. Besides being related to trauma severity, F1+2, D-D and TM levels correlate closely with the occurrence of posttraumatic DIC. Therefore, changes in plasma F1+2, D-D and TM levels may predict the occurrence of DIC.

Coagulation activation in autoimmune bullous diseases

The main autoimmune blistering skin disorders are pemphigus vulgaris (PV) and bullous pemphigoid (BP). They differ in the inflammatory infiltrate, which is more intense in BP. Inflammation is known to activate coagulation in several disorders. Local and systemic activation of coagulation was evaluated in BP and PV. We studied 20 BP patients (10 active and 10 remittent), 23 PV patients (13 active and 10 remittent) and 10 healthy subjects. The coagulation markers prothrombin fragment F1+2 and D-dimer were measured by enzyme-immunoassays in plasma. The presence of tissue factor (TF), the main initiator of blood coagulation, was evaluated immunohistochemically in skin specimens from 10 patients with active PV, 10 patients with active BP and 10 controls. Plasma F1+2 and D-dimer levels were significantly high in active BP (P = 0.001), whereas in active PV the levels were normal. During remission, F1+2 and D-dimer plasma levels were normal in both BP and PV. TF immunoreactivity was found in active BP but neither in active PV nor in normal skin. TF reactivity scores were higher in active BP than in controls or active PV (P = 0.0001). No difference in TF scores was found between active PV and controls. BP is associated with coagulation activation, which is lacking in PV. This suggests that BP but not PV patients have an increased thrombotic risk. The observation that thrombotic complications occur more frequently in BP than in PV further supports this view.