Home About us Contact
|
Home About us Contact | ||
|
|
||
APTT
Selected AbstractsThe polysaccharide fucoidan inhibits microvascular thrombus formation independently from P- and l -selectin function in vivoEUROPEAN JOURNAL OF CLINICAL INVESTIGATION, Issue 9 2000Thorlacius Background Adhesion molecules of the selectin family (mainly P- and L-selectin) have been suggested to mediate interactions between platelets, leukocytes and endothelial cells in thrombus formation. The polysaccharide fucoidan has anticoagulative properties, but is also able to bind and block the function of the selectins. Here, we investigated in vivo (i) if fucoidan can prevent microvascular thrombus formation, and (ii) whether this is potentially mediated by the inhibition of P-and/or L-selectin. Materials and Methods For this purpose, we used intravital microscopy in the mouse cremaster microcirculation in which thrombosis was induced photochemically by light exposure to individual arterioles and venules after intravenous (i.v.) injection of FITC-dextran. Results We found that intravenous administration of fucoidan significantly prolonged the time required for complete occlusion in arterioles and venules by almost seven- and nine-fold, respectively. In contrast, treatment with monoclonal antibodies against P- and L-selectin had no effect on the development of microvascular thrombosis. Fucoidan and also the anti-P-selectin antibody completely inhibited baseline venular leukocyte rolling in the cremaster muscle, indicating that these treatment regimes abolished P-selectin function. Importantly, fucoidan and the anti-P-selectin antibody had no effect on systemic platelet and leukocyte counts. On the other hand, we found that fucoidan treatment significantly altered coagulation parameters, including prothrombin time (Quick percentage), activated partial thromboplastin time (APTT) and thrombin clotting time (TCT), which may explain the potent in vivo anticoagulative effect of fucoidan observed here. Conclusions Taken together, our novel findings suggest that fucoidan effectively prevents microvascular thrombus formation induced by endothelial damage in arterioles and venules in vivo. This protective effect of fucoidan is not attributable to inhibition of P- and L -selectin function but may instead be related to the anticoagulative capacity of fucoidan. [source] Influence of source of phospholipids for APTT-based factor IX assays and potential consequences for the diagnosis of mild haemophilia BHAEMOPHILIA, Issue 1 2009C. POUPLARD First page of article [source] Influence of factor VIII:C and factor IX activity in plasmas of haemophilic dogs on the activated partial thromboplastin time measured with two commercial reagentsHAEMOPHILIA, Issue 3 2000R. Mischke The present study is based on 145 plasma samples with a reduced activity of factor VIII:C (range: 0.009,0.62 IU mL,1) and 28 samples with a reduced factor IX activity (range: 0.035,0.55 IU mL,1). The samples were collected from dogs with haemophilia A (n=22) or haemophilia B (n=3), some of these during substitution therapy. For all samples the activated partial thromboplastin time (APTT) was measured with two commercial reagents containing kaolin as a contact activator. In each case, the deficiency of factor VIII:C or IX was reflected in abnormal results of the APTT. This was true for both reagents. A significant correlation (P < 0.001) was found between factor VIII:C activity and APTT (reagent 1, Pathromtin®; Spearman's rank correlation coefficient, rS=,0.731, reagent 2, PTT-Reagenz; rS=,0.875) as well as between factor IX activity and APTT (reagent 1, rS=,0.819; reagent 2, rS=,0.955]. In each case, the relationship between coagulation factor activity and APTT could be proven most precisely by geometric regression. The results of this study illustrate the applicability of commercial APTT test kits as a sensitive screening test of factor VIII:C and IX deficiencies in canine plasma. [source] Inadvertent postdialysis anticoagulation due to heparin line locksHEMODIALYSIS INTERNATIONAL, Issue 4 2007Ruth J. PEPPER Abstract Large-bore dual lumen in-dwelling venous catheters are used in hemodialysis. These catheters are usually locked with heparin after the treatment. This study addressed the underappreciated postdialysis coagulopathy that can result. Thirty-six patients were included: 7 dialyzed through arterio-venous fistulae, 29 through in-dwelling venous catheters. The latter group was further subdivided according to whether they received heparin or heparin-free dialysis. To assess the heparin lock, a full-dose heparin lock as well as a much weaker heparin lock and a citrate lock were used. To assess the coagulopathy, blood was taken 1 hr after dialysis. The activated partial thromboplastin time (APTT) and anti-Xa level was measured. Additionally, 6 venous catheters were removed and the amount of fluid expelled upon locking with saline was measured. Clotting from the patient group with arterio-venous fistulae was normal following dialysis. The patients with in-dwelling venous catheters and heparin locks had significantly deranged clotting; 6 out of 10 patients had abnormal APTT results. All patients with catheters, heparin-free dialysis, and heparin locks had deranged clotting (7 out of 7). The rate decreased significantly when heparinized saline was used as a lock. A subset of patients had a citrate lock rather than a heparin lock; the clotting results normalized in all but one patient. An in vitro study demonstrated immediate leakage of fluid from the end of the ports upon locking. Significant postdialysis anticoagulation can occur after dialysis, which can be attributed to the heparin line locks. This risk is considerably reduced when a citrate lock is used instead. [source] Effects of fibrinogen concentrate administration during severe hemorrhageACTA ANAESTHESIOLOGICA SCANDINAVICA, Issue 9 2010H. R. THORARINSDOTTIR Background: Fibrinogen concentrate has been shown to improve coagulation in dilutional coagulopathy in experimental studies, but clinical experience is still scarce. The aim of this study was to evaluate laboratory data and the clinical outcome of fibrinogen administration in patients suffering from severe hemorrhage. Materials and methods: A retrospective study over a 3-year observation period of consecutive patients who received a single dose of fibrinogen concentrate but not recombinant factor VIIa as part of their treatment of severe hemorrhage, defined as >6 U of packed red blood cells (PRBCs). Results: Thirty-seven patients were included, most of them suffering from severe hemorrhage following open heart surgery (68%). After a median fibrinogen dose of 2 g (range 1,6 g), an absolute increase in the plasma fibrinogen concentration of 0.6 g/l was observed (P<0.001). The activated partial thromboplastin time (APTT) decreased significantly (P<0.001), from 52 to 43 s and the prothrombin time (PT) decreased from 20 to 17 s, respectively. The transfusion requirement for PRBCs decreased from 6 to 3 U (P<0.01) in the 24 h after fibrinogen administration, but fresh-frozen plasma and platelet concentrate transfusions did not change significantly. Eight patients (22%) died in intensive care unit and the pre-operative fibrinogen concentration was not significantly different in these patients. Conclusion: Administration of fibrinogen for severe hemorrhage was associated with an increased fibrinogen concentration and a significant decrease in APTT, PT and the requirement for PRBCs. [source] Comparability of the results of PT,INR with local MNPT and APTTR with MNAPTT on different coagulation analyzers in ChinaINTERNATIONAL JOURNAL OF LABORATORY HEMATOLOGY, Issue 3 2009L. PENG Summary The prothrombin time (PT), International normalized ratio (INR) and activated partial thromboplastin time (APTT) are the most used coagulation tests in China, where more than one type of automated coagulation analyzer is often used in the clinical laboratory. The PT,INR results of 109 samples were compared with local mean normal PT (MNPT) and APTT ratio (APTTR) with mean normal APTT (MNAPTT) on two different coagulation analyzers in the same laboratory. The two different coagulation analyzers showed no significant difference (P > 0.05) in PT and INR determination, but there was a significant difference (P < 0.01) for APTT. The INR with local MNPT and APTTR with MNAPTT, obtained with the ACL Futura and CA 510, showed much better agreement; 98.8% (82/83) of bias for INR with local MNPT was less than 15% compared with 90.4% (75/83) of bias for INR; and 100% of bias for APTTR (62/62) was less than 15% compared with only 6.5% (4/62) of bias for APTT. Meanwhile, there was no significant difference (P = 0.865) for APTTR with MNAPTT compared with APTT (P = 0.002) between the ACL Futura and CA 510. In conclusion, these analyzers showed very poor agreement for both the PT and APTT, but the calculation of ratios significantly improved agreement. [source] Acquired haemophilia masked by warfarin therapyINTERNATIONAL JOURNAL OF LABORATORY HEMATOLOGY, Issue 1 2007C. M. VADIKOLIA Summary Acquired haemophilia is a rare phenomenon and prompt diagnosis is essential for successful treatment. Early laboratory detection could minimize its potentially devastating consequences and reduce mortality but when a masking element such as anticoagulant therapy is present, delay in diagnosis is not uncommon. A prolonged activated partial thromboplastin time (APTT) may be falsely attributed to warfarin alone, particularly when it is associated with oral anticoagulant overdose. We describe two patients on treatment with warfarin who presented with a bleeding diathesis and disproportionately prolonged APTT, which led to the diagnosis of antibodies directed against factor VIII. [source] Waveform analysis of clotting test optical profiles in the diagnosis and management of disseminated intravascular coagulation (DIC)INTERNATIONAL JOURNAL OF LABORATORY HEMATOLOGY, Issue 6 2002C. H. Toh Summary Transmittance waveform charts the changes in light transmittance on standard coagulation assays, such as the prothrombin time (PT) and activated partial thromboplastin time (APTT). Analysis and characterization of these data on photo-optical coagulation analysers provides additional qualitative and quantitative information to that obtained using the clotting time alone. The most thoroughly evaluated clinical application is that of the biphasic APTT waveform with disseminated intravascular coagulation (DIC). The degree of waveform abnormality correlates directly with the severity of haemostatic dysfunction and allows for both the prediction and monitoring from non-overt to overt DIC. As its performance is simple and rapid, this provides the means for targeting therapeutic intervention to an earlier stage of DIC. The recent identification that the mechanism underlying the biphasic waveform is a complex that exists in vivo between C reactive protein with very low density lipoprotein, provides potentially important insights into the molecular pathogenesis of DIC. Thus, in addition to the immediate clinical utility in diagnostic practice, it has important applications as a research tool. Preliminary experience in the application of this technology to the diagnosis and management of the haemophilias and the lupus anticoagulant syndrome has also provided evidence of the power and utility of waveform analysis in essentially simple clotting assays. [source] Biocompatibility investigation and urea removal from blood by urease-immobilized HEMA incorporated poly(ethyleneglycol dimethacrylate) microbeadsJOURNAL OF BIOMEDICAL MATERIALS RESEARCH, Issue 1 2003F. Ayhan Abstract The biocompatibility of modified and urease-immobilized poly(ethyleneglycol dimethacrylate/2-hydroxyethylmetacrylate) [poly(EGDMA/HEMA)] microbeads was tested through blood compatibility tests. Twelve percent HEMA incorporated nonporous particles of 105,125 ,m were used in the research. Hydroxyl groups on microbeads were chemically modified by following a three-step procedure that is composed of activation, spacer-arm incorporation (hexamethylene diamine) and, finally, glutaraldehyde bounding. Enzyme urease was immobilized on microbead surfaces, and adsorption of blood proteins in serum and plasma, blood coagulation time, and leukocyte and platelet adhesion were tested. Incubation of 1.5 cc of biological fluid with 100 mg of urease-immobilized poly(EGDMA/HEMA) microbeads at room temperature shows that protein adsorption on surfaces occurs, but protein content after treatment was in the range of healthy people. Adsorbed albumin and total globulin amounts per gram of microbeads is much greater than fibrinogen. Immobilization of urease reduced the protein adsorption and blood coagulation times compared with those of modified microbeads. Prothrombin time (PT) was not altered much, whereas poly(EGDMA/HEMA) microbeads induced a significant increase of activated partial thromboplastin time (APTT). The platelet and leukocyte adhesion slightly increased with the modification of poly(EGDMA/HEMA) and decreased with the introduction of urease. When blood samples were treated with urease-immobilized microbeads, BUN values of patients were lowered to almost acceptable amounts. © 2002 Wiley Periodicals, Inc. J Biomed Mater Res Part B: Appl Biomater 64B: 13,18, 2003 [source] Use of a new silica clotting time for diagnosing lupus anticoagulant in patients who meet the clinical criteria for antiphospholipid syndromeJOURNAL OF CLINICAL LABORATORY ANALYSIS, Issue 1 2006Panagiotis Grypiotis Abstract The silica clotting time (SCT) is a phospholipid-dependent coagulation assay used for the laboratory diagnosis of lupus anticoagulant (LA) antibodies. The sensitivity and specificity of a new commercial SCT for identifying LA in patients who meet the clinical criteria for antiphospholipid syndrome (APS), and its association with thrombotic events were evaluated here. Forty-five patients who met the clinical criteria for APS according to the Sapporo International Consensus Statement were examined. Sixty-nine patients who did not meet the clinical criteria for APS, and 20 blood donors were used as controls. Plasma samples from the patients and controls were tested for LA using a new commercial SCT with low and high synthetic phospholipid concentrations. The results were compared with those obtained by diluted Russell's viper venom time (dRVVT) and activated partial thromboplastin time (APTT). SCT's sensitivity for identifying LA in patients who met the clinical criteria of APS was higher compared to APTT and dRVVT (53.3% vs. 31.1% and 31.1%), and the specificities of these assays were 96.6%, 100%, and 98.9%, respectively. When dRVVT was combined with SCT, and dRVVT was combined with APTT their sensitivities were 57.7% and 48.8%, and their specificities were 96.6% and 98.9%, respectively. A stepwise logistic regression analysis indicated that the combination of dRVVT with SCT was associated with total thrombotic events (odds ratio (OR)=11.5, 95% confidence interval (CI)=1.25,106.3, P=0.031) as well as with venous thrombosis (OR=4.09, 95% CI=1.16,14.43, P=0.028). According to our results, SCT is the most sensitive assay for identifying LA in patients who meet the clinical criteria for APS. Moreover, the highest sensitivity was reached with a combination of SCT and dRVVT. The method's association with total thrombotic events and venous thrombosis was in fact significant. J. Clin. Lab. Anal. 20:15,18, 2006. © 2006 Wiley-Liss, Inc. [source] Effects of pentoxifylline on coagulation profile and disseminated intravascular coagulation incidence in Egyptian septic neonatesJOURNAL OF CLINICAL PHARMACY & THERAPEUTICS, Issue 3 2010M. Adel Msc Summary Background and objectives:, Neonatal sepsis is frequently associated with pathological activation of the coagulation system, leading to microcirculatory derangement and multiple organ dysfunction syndrome (MODS). The key role in the pathogenesis of sepsis has been attributed to proinflammatory cytokines. These trigger the development of disseminated intravascular coagulation (DIC) via the tissue factor-dependent pathway of coagulation. Pentoxifylline (PTX), a methylxanthine derivative that is used in peripheral vascular disease, has the potential to modify inflammatory response. The current work was designed to evaluate the potential protective effects of PTX against sepsis-induced microcirculatory derangement in Egyptian neonates. Methods:, A double-blind placebo-controlled quasi-randomized design was used. Thirty-seven neonates with sepsis were randomly allocated into two groups. Seventeen patients were given PTX (5 mg/kg/h for 6 h; for 6 successive days). Twenty patients received equivalent volume of normal saline and represented the placebo group. Prothrombin time (PT), Activated partial thromboplastin time (APTT), fibrinogen, d -dimer, C-reactive protein (CRP), complete blood count (CBC), also hemodynamic parameters comprising arterial blood pressure, heart rate, capillary refill and urinary output were assessed in both groups before and after treatment. Results:, Coagulation parameters in the two groups showed no significant differences. However, a higher incidence of DIC was observed in the placebo group neonates. PTX significantly lowered the percentage of bleeding (P = 0·0128) and less frequent use of FFP was observed in the PTX group (35·53% in PTX group vs. 80% in placebo group, P = 0·003). Incidence of MODS was significantly lower (P = 0·037) and hospital stay duration of survivors was significantly shorter (P = 0·044) in the PTX treated-infants. Conclusion:, Pentoxifylline protects against sepsis-induced microcirculatory derangement in neonates. It significantly lowered the incidence of bleeding and MODS and shortened the length of hospital stay. [source] Prolonged activated partial thromboplastin time in thromboprophylaxis with unfractionated heparin in patients undergoing cesarean sectionJOURNAL OF OBSTETRICS AND GYNAECOLOGY RESEARCH (ELECTRONIC), Issue 1 2010Shigeki Matsubara Abstract Aim:, Hemorrhage is an important complication of heparin-thromboprophylaxis after surgery. We attempted to clarify the incidence rate of prolonged activated partial thromboplastin time (APTT), representative of hemorrhagic tendency, in Japanese women who received thromboprophylaxis with unfractionated subcutaneous heparin administration after cesarean section (CS). We also determined factors which affected postoperative APTT. Methods:, We studied 280 women who were administered thromboprophylaxis with unfractionated subcutaneous heparin 5000 IU two times per day after CS. Postoperative APTT under heparin was measured and the incidence of its prolongation was determined. Preoperative APTT, blood loss during surgery, postoperative hematocrit, postoperative serum total protein level, and postpartum body weight were measured, and their correlation with postoperative APTT was determined. Results:, Preoperative and postoperative APTT values were 28.3 (26.7,30.3) and 33.8 (31.0,37.5) seconds for median (interquartile range), respectively. Overall, 7.1% of patients showed ,45 s postoperative APTT. Two patients (0.7%) showed ,60 s APTT, one of whom suffered subcutaneous hemorrhage around the abdominal incision with complete healing. There were no other hemorrhagic complications. Preoperative APTT positively, and postpartum body weight inversely, correlated with postoperative APTT. The amount of blood loss, postoperative hematocrit, and postoperative serum total protein level did not correlate with postoperative APTT. No discernible deep vein thrombosis or pulmonary embolism occurred. Conclusion:, Although 7.1% of women under heparin-thromboprophylaxis showed a prolonged APTT that was 150% of the preoperative APTT, serious side effects were not observed. Subcutaneous administration of unfractionated heparin, if checking APTT prolongation 1 day after surgery, may be safe method of thromboprophylaxis after CS. [source] A further face of the partial thromboplastin time APTTJOURNAL OF THROMBOSIS AND HAEMOSTASIS, Issue 5 2005C. H. TOH No abstract is available for this article. [source] Thrombin generation for the control of heparin treatment, comparison with the activated partial thromboplastin timeJOURNAL OF THROMBOSIS AND HAEMOSTASIS, Issue 8 2004R. Al Dieri Summary., Heparin can be quantified with antifactor Xa and IIa tests (aXa, aIIa) but the anticoagulant power of heparin depends upon plasma properties as well as upon heparin concentrations and thus differs between subjects. Measuring the effect, as with the activated partial thromboplastin time (APTT) therefore is clinically more relevant. Here we investigate the use of the endogenous thrombin potential (ETP) for this purpose. In 12 volunteers 9000 IU of four heparins of different mol. wt distributions were injected. Samples were taken at 11 time points between 0 and 24 h. With the exception of the 0 and 24-h time points, heparin could be demonstrated by its aIIa and aXa activity in virtually all samples. The APTT showed the effect of this heparin in 34% of the samples; the ETP in 80%. This is partly due to the wide margins of the normal values, caused by large interindividual variation [coefficient of variation (CV) approximately 12% for the APTT, approximately 17% for the ETP]. The intraindividual variation is much smaller (CV approximately 4% for the APTT, approximately 5% for the ETP). Relative to the baseline value of the individual, the heparin effect was recognized by the APTT in 55% of the cases and by the ETP in 98%. There were no large differences between the different types of heparin. [source] The inhibition of blood coagulation by heparins of different molecular weight is caused by a common functional motif,the C-domainJOURNAL OF THROMBOSIS AND HAEMOSTASIS, Issue 5 2003R. Al Dieri Summary.,Background:,Heparins in clinical use differ considerably as to mode of preparation, molecular weight distribution and pharmacodynamic properties. Objectives:,Find a common basis for their anticoagulant action. Methods:,In 50 fractions of virtually single molecular weight (Mr), prepared from unfractionated heparin (UFH) and four low-molecular-weight heparins (LMWH), we determined: (i) the molar concentration of material (HAM) containing the antithrombin binding pentasaccharide (A-domain); (ii) the specific catalytic activity in thrombin and factor Xa inactivation; (iii) the capacity to inhibit thrombin generation (TG) and prolong the activated partial thromboplastin time (APTT). We also calculated the molar concentration of A-domain with 12 sugar units at its non-reducing end, i.e. the structure that carries antithrombin activity (C-domain). Results:,The antithrombin activity and the effects on TG and APTT are primarily determined by the concentration of C-domain and independent of the source material (UFH or LMWH) or Mr. High Mr fractions (>15 000) are less active, probably through interaction with non-antithrombin plasma proteins. Anti-factor Xa activity is proportional to the concentration of A-domain, it is Ca2+ - and Mr-dependent and does not determine the effect on TG and APTT. Conclusion:,For any type of heparin, the capacity to inhibit the coagulation process in plasma is primarily determined by the concentration of C-domain, i.e. the AT-binding pentasaccharide with 12 or more sugar units at its non-reducing end. [source] Prognostic Factors for Mortality and Thromboembolism in Canine Immune-Mediated Hemolytic Anemia: A Retrospective Study of 72 DogsJOURNAL OF VETERINARY INTERNAL MEDICINE, Issue 5 2002Anthony P. Carr Medical records of 72 dogs diagnosed with immune-mediated hemolytic anemia (IMHA) were reviewed to find risk factors for the disease, for mortality, and for thromboembolism. Coagulation data of 32 patients were evaluated for mortality or thromboembolism risk factors. Cocker Spaniels were at increased risk for IMHA (P= .012). Timing of vaccination was not associated with development of IMHA. PCV ranged from 5 to 33%, with a mean of 16 ± 5%. Autoagglutination was present in 42% of the dogs. Platelet counts (n = 60) varied from 3,000 to 793,000/,L (mean, 160,117 ± 133,571; median, 144,000). Thrombocytopenia (platelet count, <200,000/,L) was present in 70% of the dogs, with severe thrombocytopenia (platelet count, <50,000/,L) being present in 22%. One-step prothrombin time (OSPT) was prolonged in 28% of the dogs tested, and activated partial thromboplastin time (APTT) was prolonged in 47% of the dogs tested. Fibrin(ogen) degradation products (FDPs) were detected in 16 of 28 dogs tested (57%). Disseminated intravascular coagulation (DIC) was diagnosed in 10 of 31 (32%) dogs and was suspected in 8 dogs. Thromboemboli were found in 20 of 25 dogs given postmortem examinations. Mortality rate was 58%. Thrombocytopenia (P= .008) and serum bilirubin concentration of >5 mg/dL (P= .015) were risk factors for mortality, and hypoalbuminemia approached significance (P= .053). Severe thrombocytopenia (P= .046), serum bilirubin concentration of >5 mg/dL (P= .038), and hypoalbuminemia (P= .016) were risk factors for thromboembolism. On evaluation of continuous data, decreased platelet count (P= .057), increased bilirubin (P= .062), and decreased albumin (P= .054) approached significance for decreased survival. A higher risk for thrombosis was found with increased alkaline phosphatase (ALKP) (P= .042), increased bilirubin (P= .047), and decreased albumin (P= .012). [source] Screening coagulation testing using the APTT: Which reagent to choose?,AMERICAN JOURNAL OF HEMATOLOGY, Issue 9 2010Francine R. Dembitzer No abstract is available for this article. [source] The effect of aprotinin on transfusion requirements in pediatric orthotopic liver transplantationPEDIATRIC TRANSPLANTATION, Issue 2 2003Toni M. Rentoul Abstract: The use of aprotinin to reduce blood loss by inhibiting fibrinolysis thereby decreasing transfusion requirements during orthotopic liver transplantation (OLT), is well-documented in adults. We set out to test the hypothesis that the prophylactic use of aprotinin reduced blood product requirements during pediatric OLT. A retrospective study was performed, reviewing data from 24 OLTs performed over a 4-yr period. Six patients did not receive aprotinin (group 1), while 18 (group 2) received a weight-based dose of aprotinin. Both groups were comparable with respect to demographics, baseline characteristics and surgical variables except for a significantly more prolonged activated partial thromboplastin time (APTT) in the aprotinin group (p = 0.015). Despite the fact that median values for transfused volumes of red blood cells (78.3 vs. 36.7 mL/kg) and fresh frozen plasma (51.9 vs. 23.7 mL/kg) were more than halved in the aprotinin group, there was no statistical difference demonstrated. The failure to reach statistical significance can probably be explained by the small number in group 1 and a high level of scatter. All patients in group 1 required intraoperative transfusion of RBC and fresh frozen plasma (FFP) while two patients in group 2 did not require RBC and seven received no FFP. There were four patients in group 1 and 17 in group 2 who did not receive platelets while five in group 1 and 12 in group 2 did not receive cryoprecipitate. The differences between the groups in avoidance of these blood products did not reach statistical significance. There was little difference between groups with respect to albumin and crystalloid requirements. No statistical difference was demonstrated in intraoperative hematologic profiles between the two groups except during the anhepatic phase of surgery when there was a statistically significant more prolonged prothrombin time (p = 0.04) and a greater international normalized ratio (p = 0.027) in group 2. [source] Removal of lipopolysaccharide and reactive oxygen species using sialic acid immobilized polysulfone dialyzerPOLYMERS FOR ADVANCED TECHNOLOGIES, Issue 12 2009Jung-Jhih Chang Abstract Sialic acid (N -acetylneuraminic acid, NANA) was covalently immobilized onto the surface of a polysulfone (PSF) hollow fiber membrane. Prior to the immobilization, the surface of PSF was treated with ozone, followed by grafting with acrylic acid, and then the esterification of NANA. The surface concentration of NANA was determined by 2-thiobarbituric acid (TBA) test. Hemocompatibility, the capability of suppressing oxidative stress, and clearance of lipopolysaccharide (LPS) from the resulting hollow fiber membrane were evaluated. The results show that by immobilizing NANA onto PSF hollow fiber, the adhesion of platelet was reduced, while both APTT and PT were little affected. Furthermore, oxidative stress was suppressed by NANA-immobilized PSF hollow fibers. The level of LPS was also greatly reduced. Copyright © 2009 John Wiley & Sons, Ltd. [source] Protein adsorption and platelet adhesion of polysulfone membrane immobilized with chitosan and heparin conjugatePOLYMERS FOR ADVANCED TECHNOLOGIES, Issue 2 2003M.-C. Yang Abstract Polysulfone (PSF) membranes were treated with ozone to introduce peroxides, and then grafted with either acrylic acid or chitosan, followed by the immobilization of heparin. The effect of spacer arm on blood compatibility was investigated using three chitosans of different molecular weight [1170 (water soluble), 160,000, and 400,000] and similar degrees of deacetylation (75%). The hydrophilicity was evaluated by measuring the contact angle of water. Blood compatibility was evaluated using the activated partial thromboplastin time (APTT) as well as the adhesion of platelets. The protein affinity was determined by the absorption of human serum albumin (HSA) and human plasma fibrinogen (HPF). The results show that by the coupling of chitosan, the amount of heparin immobilized can be increased by four times. Water contact angle (from 78,° to 41,°) decreased with the increase of the amount of heparin immobilized, showing increased wettability. The heparinized PSF membrane showed longer APTT and decreasing platelet adhesion, compared to that of unmodified PSF membrane. The adsorption of HSA and HPF were reduced to 17 and 6%, respectively. This suggests that longer spacer binding to heparin can increase the opportunity of anti-coagulation on contacting blood. These results demonstrated that the hydrophilicity and blood compatibility of PSF membrane could be improved by chitosan and heparin conjugate. Copyright © 2003 John Wiley & Sons, Ltd. [source] Sustained and therapeutic levels of human factor IX in hemophilia B mice implanted with microcapsules: key role of encapsulated cellsTHE JOURNAL OF GENE MEDICINE, Issue 3 2006Jianping Wen Abstract Background A gene therapy delivery system based on microcapsules enclosing recombinant cells engineered to secrete a therapeutic protein was explored in this study. In order to prevent immune rejection of the delivered cells, they were enclosed in non-antigenic biocompatible alginate microcapsules prior to being implanted intraperitoneally into mice. We have shown that encapsulated C2C12 myoblasts can temporarily deliver therapeutic levels of factor IX (FIX) in mice, but the C2C12 myoblasts elicited an immune response to FIX. In this study we report the use of mouse fetal G8 myoblasts secreting hFIX in hemophilia mice. Methods Mouse G8 myoblasts were transduced with MFG-FIX vector. A pool of recombinant G8 myoblasts secreting ,1500 ng hFIX/106 cells/24 h in vitro were enclosed in biocompatible alginate microcapsules and implanted intraperitoneally into immunocompetent C57BL/6 and hemophilic mice. Results Circulating levels of hFIX in treated mice reached ,400 ng/ml for at least 120 days (end of experiment). Interestingly, mice treated with encapsulated G8 myoblasts did not develop anti-hFIX antibodies. Activated partial thromboplastin time (APTT) of plasmas obtained from treated hemophilic mice was reduced from 107 to 82 sec on day 60 post-treatment, and whole blood clotting time (WBCT) was also corrected from 7,9 min before treatment to 3,5 min following microcapsule implantation. Further, mice were protected against bleeding following major trauma. Thus, the FIX delivery in vivo was biologically active. Conclusions Our findings suggest that the type of cells encapsulated play a key role in the generation of immune responses against the transgene. Further, a judicious selection of encapsulated cells is critical for achieving sustained gene expression. Our findings support the feasibility of encapsulated G8 myoblasts as a gene therapy approach for hemophilia B. Copyright © 2005 John Wiley & Sons, Ltd. [source] Validation of a bedside activated clotting time test (Hemochron® Jr II Signature) with low dose heparin therapyANAESTHESIA, Issue 4 2009L. Racioppi Summary The purpose of this study was to validate a device designed to measure activated clotting time in low-range heparin plasma concentrations (ACT-LR) prospectively during the post-operative period of vascular surgery. Measurement of ACT-LR and activated partial thromboplastin time (APTT) were performed before heparinisation (T0) and at the end of surgery (T1). ACT-LR(T1) and ,ACT-LR (defined as ACT-LR(T1) , ACT-LR(T0)) were evaluated as diagnostic tests for excessive anticoagulation, defined by APTT more than twice the laboratory's normal, by Bland-Altman method and receiver operating characteristic (ROC) curves. In 103 patients, mean (SD) ACT-LR was 137 (33) s at T0 and 176 (39) s at T1. Bland-Altman graph did not show a good agreement between APTT and ACT-LR. Areas under ROC curves were 0.82 (95% CI: 0.75,0.89) and 0.87 (95% CI: 0.80,0.93) for ACT-LR(T1) and ,ACT-LR, respectively. Using a threshold of 32 s for ,ACT-LR, test sensitivity was 87% (95% CI: 81,93%), specificity was 85% (95% CI: 78,92%), positive predictive value was 90% (95% CI: 84,96%) and negative predictive value was 81% (95% CI: 73,86%). While ,ACT-LR may have some potential in evaluating excessive anticoagulation in vascular surgery, the poor correlation between ACT-LR and APTT does not support its routine use. [source] Activated coagulation times in normal cats and dogs using MAX-ACTTM tubesAUSTRALIAN VETERINARY JOURNAL, Issue 7 2009AM See Objective, To establish reference values for activated coagulation time (ACT) in normal cats and dogs, by visual assessment of clot formation using the MAX-ACTTM tube. Subjects, We recruited 43 cats and 50 dogs for the study; 11 cats and 4 dogs were excluded from the statistical analysis because of abnormalities on clinical examination or laboratory testing including anaemia, prolonged prothrombin time (PT) or activated partial thromboplastin time (APTT), or insufficient plasma volume for comprehensive laboratory coagulation testing. Procedure, Blood samples were collected via direct venipuncture for MAX-ACT, packed cell volume/total solids, manual platelet estimation and PT/APTT measurement. Blood (0.5 mL) was mixed gently in the MAX-ACT tube at 37°C for 30 s, then assessed for clot formation every 5 to 10 s by tipping the tube gently on its side and monitoring for magnet movement. The endpoint was defined as the magnet lodging in the clot. The technique was tested with 10 dogs by collecting two blood samples from the same needle insertion and running a MAX-ACT on each simultaneously. Results, In normal cats the mean MAX-ACT was 66 s (range 55,85 s). In normal dogs the mean was 71 s (range 55,80 s). There was no statistical difference between the first and second samples collected from the same needle insertion. Conclusions and Clinical Relevance, In both cats and dogs, a MAX-ACT result >85 s should be considered abnormal and further coagulation testing should be performed. Additionally, failure to discard the first few drops of the sample does not appear to significantly affect results. [source] Population modelling of the effect of inogatran, at thrombin inhibitor, on ex vivo coagulation time (APTT) in healthy subjects and patients with coronary artery diseaseBRITISH JOURNAL OF CLINICAL PHARMACOLOGY, Issue 1 2001Marie Cullberg Aims, The purpose of this study was to characterize the relationship between the degree of anticoagulation, assessed by APTT, and the plasma concentration of inogatran in healthy subjects and in patients with coronary artery disease. Methods, Data from five phase I studies in 78 healthy males and two phase II multicentre studies in 948 patients of both sexes with unstable angina pectoris or non-Q-wave myocardial infarction were evaluated. A total of 3296 pairs of concentration-APTT samples were obtained before, during, and after intravenous infusions of inogatran. Mixed effects modelling was used for population pharmacodynamic analysis of the drug effect and for describing the variability in baseline APTT. Results, The population mean baseline APTT was 29 s, but large variations between individuals (s.d. 3.6 s) were observed. The variability between studies (1.3 s) and centres (1.8 s) were of less importance, though statistically significant. APTT increased in a nonlinear manner with increasing inogatran concentration and the relationship was well described by a combined linear and Emax model. A significant part of the overall variability could be ascribed to the APTT reagent and equipment used at the different study centres. These method-dependent differences were compensated for by including the lower limit of the normal reference range as a covariate, affecting both baseline and Emax, in the model. For the typical healthy subject and patient, the method-corrected population mean parameters were: APTTbaseline 35 and 31 s, slope 8.0 and 5.8 s l µmol,1, Emax 36 and 34 s, and EC50 0.54 and 0.72 µmol l,1, respectively. The model predicted plasma concentration needed to double the APTT from the baseline value was 1.25 and 1.45 µmol l,1 in the healthy volunteer and patient, respectively. Conclusions, The nonlinear relationship between APTT and inogatran concentration in plasma was well described by a combined linear and Emax model. Pooling of data was made possible by incorporating a centre-specific characteristic of the assay method in the model. Patients had lower baseline APTT and appeared to have less pronounced effect of inogatran than young healthy subjects. [source] The management of heparin-induced thrombocytopeniaBRITISH JOURNAL OF HAEMATOLOGY, Issue 3 2006David Keeling Abstract The Haemostasis and Thrombosis Task Force of the British Committee for Standards in Haematology has produced a concise practical guideline to highlight the key issues in the management of heparin-induced thrombocytopenia (HIT) for the practicing physician in the UK. The guideline is evidence-based and levels of evidence are included in the body of the article. All patients who are to receive heparin of any sort should have a platelet count on the day of starting treatment. For patients who have been exposed to heparin in the last 100 d, a baseline platelet count and a platelet count 24 h after starting heparin should be obtained. For all patients receiving unfractionated heparin (UFH), alternate day platelet counts should be performed from days 4 to 14. For surgical and medical patients receiving low-molecular-weight heparin (LMWH) platelet counts should be performed every 2,4 d from days 4 to 14. Obstetric patients receiving treatment doses of LMWH should have platelet counts performed every 2,4 d from days 4 to 14. Obstetric patients receiving prophylactic LMWH are at low risk and do not need routine platelet monitoring. If the platelet count falls by 50% or more, or falls below the laboratory normal range and/or the patient develops new thrombosis or skin allergy between days 4 and 14 of heparin administration HIT should be considered and a clinical assessment made. If the pretest probability of HIT is high, heparin should be stopped and an alternative anticoagulant started at full dosage unless there are significant contraindications while laboratory tests are performed. Platelet activation assays using washed platelets have a higher sensitivity than platelet aggregation assays but are technically demanding and their use should be restricted to laboratories experienced in the technique. Non-expert laboratories should use an antigen-based assay of high sensitivity. Only IgG class antibodies need to be measured. Useful information is gained by reporting the actual optical density, inhibition by high concentrations of heparin, and the cut-off value for a positive test rather than simply reporting the test as positive or negative. In making a diagnosis of HIT the clinician's estimate of the pretest probability of HIT together with the type of assay used and its quantitative result (enzyme-linked immunosorbent assay, ELISA, only) should be used to determine the overall probability of HIT. Clinical decisions should be made following consideration of the risks and benefits of treatment with an alternative anticoagulant. For patients with strongly suspected or confirmed HIT, heparin should be stopped and full-dose anticoagulation with an alternative, such as lepirudin or danaparoid, commenced (in the absence of a significant contraindication). Warfarin should not be used until the platelet count has recovered. When introduced in combination with warfarin, an alternative anticoagulant must be continued until the International Normalised Ratio (INR) is therapeutic for two consecutive days. Platelets should not be given for prophylaxis. Lepirudin, at doses adjusted to achieve an activated partial thromboplastin time (APTT) ratio of 1·5,2·5, reduces the risk of reaching the composite endpoint of limb amputation, death or new thrombosis in patients with HIT and HIT with thrombosis (HITT). The risk of major haemorrhage is directly related to the APTT ratio, lepirudin levels and serum creatinine levels. The patient's renal function needs to be taken into careful consideration before treatment with lepirudin is commenced. Severe anaphylaxis occurs rarely in recipients of lepirudin and is more common in previously exposed patients. Danaparoid in a high-dose regimen is equivalent to lepirudin in the treatment of HIT and HITT. Danaparoid at prophylactic doses is not recommended for the treatment of HIT or HITT. Patients with previous HIT who are antibody negative (usually so after >100 d) who require cardiac surgery should receive intraoperative UFH in preference to other anticoagulants that are less validated for this purpose. Pre- and postoperative anticoagulation should be with an anticoagulant other than UFH or LMWH. Patients with recent or active HIT should have the need for surgery reviewed and delayed until the patient is antibody negative if possible. They should then proceed as above. If deemed appropriate early surgery should be carried out with an alternative anticoagulant. We recommend discussion of these complex cases requiring surgery with an experienced centre. The diagnosis must be clearly recorded in the patient's medical record. [source] Comparison of unfractioned and low molecular weight heparin for prophylaxis of coagulopathies in 52 horses with colic: a randomised double-blind clinical trialEQUINE VETERINARY JOURNAL, Issue 5 2003K. FEIGE Summary Reasons for performing study: Unfractioned heparin (UFH) is widely used for prophylaxis of coagulation disorders, especially in colic-affected horses. However, it is accompanied by certain side effects. Objectives: To compare the efficacy and side effects of unfractioned and low molecular weight heparin (LMWH) in horses with colic. Methods: The study was carried out as a randomised, double-blind, controlled clinical trial. Fifty-two horses with colic were treated subcutaneously with either UFH (heparin calcium, 150 iu/kg bwt initially, followed by 125 iu/kg bwt q. 12 h for 3 days and then 100 iu/kg bwt q. 12 h) or LMWH (dalteparin, 50 iu/kg bwt q. 24 h). All horses underwent daily physical examination including assessment of jugular veins, local reaction to heparin injections, haematological evaluation and coagulation profiles over up to 9 days. Results: The type of heparin used did not affect the general behaviour and condition. There were significantly more jugular vein changes in horses treated with UFH. Packed cell volume decreased significantly within the first few days of UFH treatment, but did not change significantly in horses treated with LMWH. Activated partial thromboplastin time (aPTT) and thrombin time (TT) were prolonged in horses treated with UFH but not in those treated with LMWH. Conclusions: It was concluded that, in comparison to UFH, LMWH has markedly fewer side effects in horses. Potential relevance: Therefore, LMWH is recommended for prophylaxis of coagulation disorders in colic patients. [source] Influence of temperature and time before centrifugation of specimens for routine coagulation testingINTERNATIONAL JOURNAL OF LABORATORY HEMATOLOGY, Issue 4 2009G. L. SALVAGNO Summary The accurate standardization of the preanalytical phase is of pivotal importance for achieving reliable results of coagulation tests. Because information on the suitable storage conditions for coagulation testing is controversial, we aimed at investigating the sample stability with regard to the temperature and time before centrifugation. The activated partial thromboplastin time (aPTT), prothrombin time (PT), fibrinogen and D-dimer were assayed in specimens collected from 26 consecutive patients on antivitamin K therapy on the ACL TOP analyzer. Three primary 3.6-ml siliconized evacuated tubes containing 0.109 mol/l buffered trisodium citrate were sequentially collected from each patient. These three tubes were mixed, pooled and divided into seven identical aliquots. The first aliquot was immediately centrifuged according to the standard protocol [1500 g for 15 min at room temperature (RT)] and analyzed. The other aliquots were left for 3, 6 and 24 h, respectively, at RT or 4 °C, and then centrifuged and analyzed. Test results were compared with those obtained on the reference specimen. Statistically significant prolongations were observed for aPTT in all the samples. Such differences exceeded the analytical quality specifications for desirable bias in the samples stored for 24 h. A significant reduction, yet comprised within the desirable bias, was observed for PT and fibrinogen in uncentrifuged specimens stored at RT for 3 and 6 h. No significant biases could be recorded in D-dimer. In conclusion, a 6-h storage of uncentrifuged specimens at either RT or 4 °C may still be suitable to achieve results of routine coagulation testing comprised within the analytical quality specifications for desirable bias. [source] An evaluation of monitoring possibilities of argatroban using rotational thromboelastometry and activated partial thromboplastin timeACTA ANAESTHESIOLOGICA SCANDINAVICA, Issue 1 2010M. ENGSTRÖM Background: Rotational thrombelastometry/thrombelastography with ROTEM® and TEG® is becoming available bedside in an increasing number of intensive care units, where many patients with heparin-induced thrombocytopenia (HIT) are treated. The study has been performed in an effort to find out whether ROTEM® could be an alternative to activated partial thromboplastin time (aPTT) when argatroban is used for anticoagulation. Methods: Argatroban was added in vitro to a series of citrated whole-blood samples from 10 healthy volunteers to obtain whole-blood concentrations of 0, 0.125, 0.25, 0.5, 1.0, 2.0, 4.0 and 8.0 mg/l. ROTEM® and whole-blood aPTT analyses were performed at each argatroban concentration. Correlation analyses were performed using the Spearman correlation analysis. Results: There was a significant and strong correlation between argatroban concentrations and clotting time (CT in ROTEM® analysis with INTEM) (P<0.0001 and r=0.98). Also, the ROTEM® time to maximum clot formation velocity (MAXV-t) appeared to have a very strong and highly significant correlation to argatroban concentrations (P<0.0001 and r=0.95). When we studied the correlation between aPTT and CT, we found a highly significant and strong correlation between these two analyses (P<0.0001 and r=0.97), especially so in the clinically relevant therapeutic range up to 100 s aPTT prolongation for HIT patients. Conclusion: A significant and strong correlation was found between argatroban concentrations and several ROTEM® parameters. Rotational thrombelastometry/thrombelastography has a potential role in increasing the safety of argatroban anticoagulation in critically ill patients. [source] Hemostatic Defect in Baboons Autotransfused Treated Plasma to Simulate Shed BloodJOURNAL OF CARDIAC SURGERY, Issue 6 2006C. Robert Valeri M.D. This study was done to determine how autotransfusion of nontreated plasma and plasma treated with urokinase with and without aprotinin affected hemostasis in healthy baboons. Methods: A 500-mL volume of blood was collected from the baboon, a 250-mL volume of plasma was isolated, and the RBCs were reinfused. Three baboons were autotransfused untreated plasma. Four baboons received plasma that had been treated with 3000 IU/mL urokinase at +37°C for 30 minutes. Eight baboons received plasma that had been treated first with urokinase 3000 IU/mL at +37°C for 30 minutes and then with aprotinin (1000 KIU/mL). Bleeding time, fibrinogen degradation products (FDP), D-dimer, and alpha-2 antiplasmin levels were measured. Results: During the 4-hour period following autotransfusion of the urokinase-aprotinin-treated plasma, the levels of D-dimer and FDP were significantly higher and fibrinogen and alpha-2 antiplasmin levels were significantly lower than those levels seen after the autotransfusion of nontreated plasma. FDP and D-dimer levels showed significant positive correlations with prothrombin time (PT) and activated partial thromboplastin time (aPTT). A significant negative correlation was observed between thrombin time (TT) and fibrinogen level. A significant positive correlation was observed between bleeding time and D-dimer level and a significant negative correlation between the bleeding time and the fibrinogen level. Conclusions: The infusion of a volume of urokinase or urokinase-aprotinin treated autologous plasma equivalent to 15% of the blood volume was not associated with a bleeding diathesis in healthy baboons. [source] Recombinant activated factor VII efficacy and safety in a model of bleeding and thrombosis in hypothermic rabbits: a blind studyJOURNAL OF THROMBOSIS AND HAEMOSTASIS, Issue 2 2007A. GODIER Summary.,Background:,Recombinant activated factor VII (rFVIIa) is increasingly used to secure hemostasis in hemorrhagic situations in trauma and surgical patients. Hypothermia is often observed in these clinical settings. Objective:,To study the efficacy and safety of rFVIIa in hypothermia in a rabbit model of bleeding and thrombosis. Methods:,Sixty-nine rabbits were anesthetized, ventilated and monitored for blood pressure, temperature and carotid flow. The Folts model was used: a stenosis (75%) and an injury were carried out on the carotid artery, inducing thrombosis. Blood flow decreased as thrombus size increased until the pressure gradient was such that the thrombus was released and local arterial blood flow was suddenly restored. This is known as a cyclic flow reduction (CFR). After counting baseline CFRs during a 20-min period (P1), rabbits were randomized blindly to one of four groups: normothermic (NT) placebo or rFVIIa (150 ,g kg,1), hypothermic (HT) (34 °C) placebo or rFVIIa. Then CFRs were recorded over a second period (P2). At the end of the experiment, a hepato-splenic section was performed and the amount of blood loss was recorded. After each period, the following were measured: ear immersion bleeding time (BT), hemoglobin, platelet count, prothrombin time (PT), activated partial thromboplastin time (aPTT) and fibrinogen. Results:,Hypothermia increased BT and blood loss. These effects were reversed by rFVIIa. In NT rabbits, rFVIIa shortened BT but did not reduce blood loss. rFVIIa-treated rabbits bled similarly regardless of temperature. The incidence of CFRs was higher in treated than placebo animals regardless of temperature. rFVIIa decreased PT and aPTT without modifying platelet count or fibrinogen level. Conclusion:,Hemostatic efficacy of rFVIIa was maintained in hypothermia. However, the number of CFRs was higher in the rFVIIa-treated group than in the placebo groups, whether for NT or HT rabbits. [source] | |||||||||||||||||||||||||||||||