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PubMed1978

Increased production and expression of tissue thromboplastin-like procoagulant activity in vitro by allogeneically stimulated human leukocytes.

Intravascular coagulation, thrombosis, and fibrin deposition often produce tissue damage in allogeneic inflammatory reactions such as allograft rejection. The mechanisms which initiate blood clotting in these reactions are poorly understood. We find that allogeneic stimulation of human leukocytes in vitro increases production and expression of tissue thromboplastin-like activity. In our experiments mixed leukocyte cultures (MLC) of cells from allogeneic (unrelated) donors produced and expressed more procoagulant activity than control cultures of cells from each donor alone. After 7 days, allogeneic MLC had 5- to 50-fold more total procoagulant activity than controls, as shown by assaying lysed whole cultures. Additionally, allogeneic MLC had 8- to 240-fold more procoagulant activity expressed on leukocyte surfaces and in culture supernates than controls after 7 days, as shown by assaying intact whole cultures and cell-free supernates. These increases were largely accounted for by gains in the amounts of procoagulant activity produced and expressed per cell in MLC as compared to controls. Controls and MLC produced and expressed considerable amounts of procoagulant activity during the 1st day of culture, and there were no differential effects of allogeneic stimulation on day 1. However, after day 1, the total amount of procoagulant activity produced and the amount expressed declined steadily in controls, nearly reaching preculture levels by day 7. In contrast, the total amount of procoagulant activity in allogeneic MLC remained high, and the amount of activity expressed on cell surfaces and in supernates increased severalfold by day 7. MLC of syngeneic (identical twin) cells produced and expressed the same amount of activity as controls over a 7-day period, whereas MLC of cells from each twin and an allogeneic donor produced and expressed more activity than controls (at least 9- and 35-fold more, respectively). Thus, increases of procoagulant activity production and expression were found only in MLC of genetically dissimilar cells. Therefore, these increases must have resulted from allogeneic stimulation.

Blood Coagulation↗

PubMed1985

Laboratory control of continuous intravenous heparin therapy with Howel time and activated partial thromboplastin time. A prospective, randomized and blind study.

Although heparin has been used extensively to treat Deep Venous Thrombosis (DVT) and arterial ischemia (AI), controversy still exists regarding optimal dosage and the need for monitoring. Different authors have employed various test with variable results. Others, however, persist in giving heparin without laboratory control. This study was made in order to compare, in a prospective, randomized and blind manner, two coagulation tests, namely: Howel Time (HT) and Activated Partial Thromboplastin Time (APTT), in controlling the dose of heparin given by continuous intravenous infusion in DVT and AI. Our results show no significant difference in complications and failures of the therapy with either test, although significantly higher doses of heparin were needed to maintain APTT within therapeutic range than those needed to keep HT within a similar range.

Aged↗

PubMed1992

Within-subject and between-subject biological variation of prothrombin time and activated partial thromboplastin time.

The within-subject and between-subject biological variation of prothrombin time (PT) and activated partial thromboplastin time (APTT) have been studied in a group of 19 women and 20 men over a period of 9 months. All measurements were performed with a mechanical coagulometer. The between-subject coefficients of variation were 6.8% for PT and 8.9% for APTT. The medians and the 90th percentiles of within-subject coefficients of variation were 1.7% and 5.9%, respectively, for PT, and 0.0% and 6.1% for APTT. These data were used to calculate the critical difference for significant change detection and the desirable between-day imprecision.

Adult↗

PubMed2003

Heparin in vitro sensitivity of the activated partial thromboplastin time in canine plasma depends on reagent.

The in vitro heparin sensitivity of 6 different commercial activated partial thromboplastin time (APTT) reagents was investigated based on artificial plasma samples prepared by addition of sodium heparin at different activities (0-1.5 IU/ml) to pooled normal canine plasma. Statistical analysis using 2-way analysis of variance was based on APTT ratios (APTT/mean APTT control). Significant differences between the APTT ratios of different APTT reagents (P < 0.00001) were found, which also depended on heparin activity (interaction between the factors; P < 0.00001). For example, mean APTT ratio at 0.7 IU/ml heparin varied between 1.2 and 2.5. The results of this study indicate that recommendations for the control of heparin therapy in dogs by APTT ratio should be reagent specific.

Animals↗

PubMed1986

Effect of warfarin on the activated partial thromboplastin time.

Outpatients followed in an anticoagulation clinic were studied retrospectively to determine the effect of warfarin on the activated partial thromboplastin time (APTT). Twenty-nine patients were studied in part 1 of the trial to determine whether their APTT values were elevated when their prothrombin time (PT) was within 1.5 to 2.5 times the control PT. Part 2 was carried out using the data of 32 patients to determine whether a correlation existed between the degree of elevation of the patients' PT values due to warfarin and the concurrent degree of elevation of their APTT results. Baseline PT and APTT values and a minimum of three concurrent PT and APTT values determined during anticoagulation therapy with warfarin were used. Data were collected by chart review and review of Hematology Department records. Results indicated that a statistically significant difference was evident between the baseline APTT (30.79 sec) and the mean APTT (55.10 sec) when the PT was within the therapeutic range of 1.5 to 2.5 times the control while on warfarin therapy. Good linear correlation was evident (r = 0.821) between the degree of elevation of the PT and the degree of elevation of the APTT for the group. In most cases, a good linear correlation was also evident for individual patients. Routine ordering of concurrent APTT and PT tests for patients receiving warfarin therapy is not needed since the PT alone can be monitored under most circumstances.

Blood Coagulation Tests↗

PubMed1995

Activated clotting time versus activated partial thromboplastin time for therapeutic monitoring of heparin.

OBJECTIVE: To compare and contrast the activated partial thromboplastin time (aPTT) and the activated clotting time (ACT) for the therapeutic monitoring of heparin therapy. DATA SOURCES: Relevant articles were identified through an English-language MEDLINE search from 1966 to 1995. Additional sources were identified from the reference lists of these articles. STUDY SELECTION: Studies that investigated the use and limitations of the individual assays and those offering direct comparisons were chosen for review. DATA EXTRACTION: Features demonstrating clinical applications and limitations of the aPTT and the ACT were extracted. Where possible, data suggesting preferential application of either assay also were extracted. DATA SYNTHESIS: Both the aPTT and ACT are clinically useful for the monitoring of heparin therapy. The aPTT is used more frequently for routine monitoring; the ACT is used in specialized situations requiring large heparin doses. The ACT is typically performed at bedside and is capable of yielding results rapidly and perhaps at a lower cost than an aPTT performed by a central laboratory. Most practitioners are familiar with the central laboratory aPTT. A bedside aPTT device is available, but is not yet in widespread clinical use. Both assay techniques are subject to various limitations. CONCLUSIONS: The ACT is theoretically equally as useful as the aPTT for the routine monitoring of heparin therapy, but has not been well-studied. The ACT appears more useful in situations in which high serum concentrations of heparin are required. Further cost-effectiveness and clinical outcome studies directly comparing the ACT and the aPTT is specific clinical situations are needed.

Anticoagulants↗

PubMed2003

Differential effects of DX-9065a, argatroban, and synthetic pentasaccharide on tissue thromboplastin inhibition test and dilute Russell's viper venom test.

New synthetic direct and indirect factor Xa or factor IIa inhibitors are increasingly used for the prevention and treatment of thrombotic disorders, including patients suffering from antiphospholipid syndrome. In this study, the effects of the synthetic direct factor Xa inhibitor DX-9065a, the indirect synthetic heparinomimetic pentasaccharide, and the direct factor IIa inhibitor Argatroban were studied. These two widely used assays for the detection of lupus anticoagulant, namely the tissue thromboplastin inhibition (TTIT) and the dilute Russell viper venom tests (DRWT) proved useful. The drugs were added to a normal human plasma pool ranging in concentration from 0.04 to 10 microg/mL. Using the two tests named above, DX-9065a and Argatroban showed a dose-related prolongation of TTIT and DRWT in the concentration range from 0.04 to 5 micromol/mL, but the pentasaccharide only slightly prolonged the clotting times of these assays even at high concentrations. Argatroban had the more pronounced effect on both tests when compared with DX-9065a (p < 0.001). The most responsive assay for DX-9065a up to a concentration of 2.5 micromol/mL was the DRWT. For Argatroban both TTIT and DRWT were equally responsive. Patients whose plasma was tested for suspected lupus anticoagulant and who have been given DX-9065a or Argatroban may have false-positive results with the TTIT tests and DRWT. This effect should be considered during patient management. These results indicate that these assays could be used for the effective quantitation of the direct factor Xa or factor IIa inhibitors when suitable controls are used.

Anticoagulants↗

PubMed2004

A shortened activated partial thromboplastin time is associated with the risk of venous thromboembolism.

Hypercoagulability due to high coagulation factors XI, VIII, IX, II, and fibrinogen is recognized as a risk factor of venous thromboembolism (VTE). These factors are cumulatively explored by the activated partial thromboplastin time (APTT). To test the hypothesis that a short APTT increases the risk of VTE, a case-control study was carried out in 605 patients referred for thrombophilia testing after documented VTE and in 1290 controls. Median APTT ratio (coagulation time of test-to-reference plasma) values were 0.97 (range: 0.75-1.41) for patients and 1.00 (range: 0.72-1.33) for controls (P < .001). In patients who had an APTT ratio smaller than the fifth percentile of the distribution in controls, the odds ratio (OR) for VTE was 2.4 (95% confidence interval [CI]: 1.7-3.6) and was independent of inherited thrombophilic abnormalities. Further statistical analyses in 193 patients and 259 controls for whom factor VIII (FVIII) levels were available showed a decrease of the OR from 2.7 (95% CI: 1.4-5.3) to 2.1 (95% CI: 1.0-4.2), indicating that the risk was only partially mediated by high FVIII levels. In conclusion, hypercoagulability detected by a shortened APTT is independently associated with VTE. This inexpensive and simple test should be considered in the evaluation of the risk of VTE.

Adolescent↗

PubMed1994

In vitro reversal of heparin effect with heparinase: evaluation with whole blood prothrombin time and activated partial thromboplastin time in cardiac surgical patients.

This study was designed to evaluate the potential in vitro use of heparinase to eliminate functionally active heparin prior to performing whole blood (WB) prothrombin time (PT) and activated partial thromboplastin time (APTT) assays. A total of 250 U/kg of heparin for cardiopulmonary bypass (CPB) was administered to 30 cardiac surgical patients in three consecutive, divided doses (20, 80, and 150 U/kg) at 15-min intervals. Blood specimens were obtained prior to heparin administration (baseline) and 10 min after each heparin dose. After collection, blood specimens were fractionated into three aliquots of which the first was used for determination of heparin concentration. After gentle mixing, WB PT and APTT measurements were performed for heparinase (Aliquot 2)- and nonheparinase (Aliquot 3)-treated blood. With consecutive heparin doses of 20 and 80 U/kg, WB PT increased from a baseline of 12.3 +/- 0.1 s to 13.3 +/- 0.2 and 18.5 +/- 1.3 s, while WB APTT increased from a baseline of 28.3 +/- 1.1 s to 89.5 +/- 5.4 after the initial heparin dose (20 U/kg). When compared to baseline (no heparin) results, small, progressive increases in heparinase-treated WB PT (0.7 +/- 0.1, 1.5 +/- 0.1, 2.1 +/- 0.1 s) and APTT (2.3 +/- 0.3, 5.7 +/- 0.4, 9.5 +/- 0.5 s) were seen with increasing heparin concentration (0.23, 1.58, and 3.95 U/mL, respectively). Heparinase was highly effective in eliminating the anticoagulant effects of even large amounts of heparin in plasma from cardiac surgical patients.(ABSTRACT TRUNCATED AT 250 WORDS)

Adult↗

PubMed1989

Interaction between human tissue thromboplastin and human antithrombin III.

It is known that antithrombin III (ATIII) activity is inhibited by tissue thromboplastin (TP) in the presence of heparin. In our study on the mechanism of the inhibition, however, TP was found to inhibit ATIII activity even in the absence of heparin, indicating an interaction between ATIII and TP. We then studied effect of ATIII on the interaction between factor VII (FVII) and TP using FVII-depleted human plasma. When the mixture of FVII + ATIII + TP was incubated at 37 degrees C and mixed with FVII-depleted plasma, the interaction between FVII and TP was inhibited. Possible complex formation was examined by electrophoretic techniques. In the immunoelectrophoresis, ATIII shifted toward the cathode in the presence of TP and the substance which had changed the mobility of ATIII showed TP activity after zone electrophoresis. The results indicated that TP had shifted toward the anode due to ATIII while ATIII had shifted toward the cathode due to TP. In the immunoblotting analysis, ATIII was separated into several bands in the presence of TP. ATIII antigenicity was altered in the presence of both heparin and TP but not in the presence of TP alone. Our results strongly suggest that TP modulates ATIII activity in the initiation of the TP-mediated coagulation cascade and that in progressing coagulation ATIII participates in the inhibition of the coagulation cascade by blocking not only thrombin activity but also the interaction between TP and FVII.

Antithrombin III↗

PubMed1989

Inhibitory effects of ONO-3307 on various proteases and tissue thromboplastin in vitro and on experimental thrombosis in vivo.

The effect of ONO-3307 (4-sulfamoyl phenyl-4-guanidinobenzoate methanesulfonate), a new protease inhibitor, was studied on various proteases in vitro and in an experimental thrombosis model in vivo. ONO-3307 competitively inhibited trypsin, thrombin, plasma kallikrein, plasmin, pancreatic kallikrein and chymotrypsin; and their Ki values were 0.048 microM, 0.18 microM, 0.29 microM, 0.31 microM, 3.6 microM and 47 microM, respectively. In addition, ONO-3307 inhibited both elastase release from N-formyl-Met-Leu-Phe (fMLP)-stimulated leukocytes and tissue thromboplastin release from endotoxin-stimulated leukocytes. To examine the effects of ONO-3307 on disseminated intravascular coagulation (DIC), we developed an experimental thrombosis model. ONO-3307 (10 mg/kg/hr) completely inhibited the deposition of radioactive fibrin in kidney and lung. Gabexate mesilate (50 mg/kg/hr) was also effective in this model, but the effect of nafamostat mesilate was unclear. These results indicate that ONO-3307 exhibits a wide range of inhibitory effects on various proteases, and ONO-3307 may be useful for the treatment of protease-mediated diseases such as thrombosis and DIC.

Animals↗

PubMed2000

Short activated partial thromboplastin times are related to increased thrombin generation and an increased risk for thromboembolism.

This prospective study was designed to investigate whether patients with short activated partial thromboplastin times (aPTTs) have increased thrombin generation and are at increased risk for thromboembolism. During a 4-month period, routine coagulation specimens were screened for the presence of a short or normal aPTT, and, accordingly, 250 specimens were collected. Prothrombin fragment F1 + 2 (F1 + 2) was measured to evaluate thrombin activation, and a second aPTT was performed with a different reagent. Diagnoses were obtained from medical records after conclusion of sample collection. Five to 9 months later, patients were questioned on thromboembolic events during the previous 18 months by questionnaire and telephone interview. F1 + 2 and the incidence of venous thromboses were elevated significantly in the short aPTT group. Unexpectedly, patients with acute bleeding had short aPTTs, but 36% of these also had thromboembolic events during the 18 months proximal to blood collection. These findings were confirmed with the second aPTT reagent. Patients with short aPTTs have increased thrombin generation and are at increased risk for thromboembolism, mainly venous thromboses, despite the fact that a short aPTT can occur in the acute setting of bleeding.

Female↗

PubMed2001

Use of the activated partial thromboplastin time for heparin monitoring.

The objectives of the present study were to evaluate the relationship between heparin concentration and activated partial thromboplastin time (aPTT) results, define a heparin concentration-derived therapeutic range for each aPTT instrument, compare aPTT- and heparin concentration-guided dosage adjustment decisions, and compare laboratory- and bedside aPTT-guided decisions. In phase 1, 102 blood samples were analyzed for bedside and laboratory aPTTs and heparin concentration (used to establish aPTT therapeutic range). In phase 2, 100 samples were analyzed in the same manner. Correlations for aPTT compared with heparin ranged from 0.36 to 0.82. Dosage adjustment decisions guided by the aPTT agreed with those based on heparin concentration 63% to 80% of the time. Laboratory and bedside aPTT dosage adjustment decisions agreed 59% to 68% of the time. The correlation of aPTT with heparin concentration and agreement between aPTT- and heparin-guided decisions vary with the aPTT instrument. Decisions guided by laboratory aPTT results often disagree with decisions guided by bedside aPTT results.

Aged↗

PubMed2002

Point-of-care testing for prothrombin time, but not activated partial thromboplastin time, correlates with laboratory methods in patients receiving aprotinin or epsilon-aminocaproic acid while undergoing cardiac surgery.

Point-of-care testing (POCT) of coagulation parameters can help optimize transfusion practice in cardiac surgery. Antifibrinolytic agents may interfere with the laboratory and/or POCT coagulation assays. This randomized controlled study compared coagulation parameters obtained from a whole blood POCT coagulation device with a typical laboratory instrument in cardiac surgery patients receiving aprotinin, epsilon-aminocaproic acid, or normal saline before undergoing cardiopulmonary bypass. Aliquots of arterial blood samples from 42 patients were collected perioperatively, and their prothrombin times (PTs) and activated partial thromboplastin times (aPTTs) were measured by POCT and laboratory instrumentation. Linear regression and error analyses were used for the method comparison. For PT, the POCT device compared favorably with the laboratory method. For aPTT, the POCT device did not compare well with the laboratory method. Treatment with antifibrinolytic agents does not interfere with determination of PT.

Aminocaproic Acid↗

PubMed2004

Biphasic activated partial thromboplastin time waveform and adverse events in non-intensive care unit patients.

The purpose of the present study was to determine whether the presence of a biphasic activated partial thromboplastin time (aPTT) waveform (BPW) is associated with adverse clinical outcomes among patients not in an intensive care unit (ICU). Consecutive patients from the emergency department or non-ICU inpatient floors with a BPW (n = 24) were enrolled prospectively, along with 24 matched control subjects with a normal aPTT waveform. Patients with a BPW had a significantly longer hospital stay (mean [median], 16.9 [11] vs 4.9 [2.5] days; P = .011), were more likely to have a positive microbial culture (16/24 [67%] vs 3/24 [13%]; P < .001), were transferred more often to an ICU (6/24 [25%] vs 0/24 [0%]; P = .010), and were more likely to receive an RBC transfusion (11/24 [46%] vs 5/24 [21%]; P = .047) or a fresh frozen plasma transfusion (5/24 [21%] vs 0/24 [0%]; P = .025). Emergency department patients with a BPW were more likely to be admitted (11/11 vs 5/11; P = .018). These results suggest that the BPW is associated with an increased rate of adverse events among non-ICU patients. Further study in this population is warranted.

Adult↗

PubMed2002

Correlation between activated clotting time and activated partial thromboplastin times.

OBJECTIVE: To evaluate the correlation between clotting time tests and heparin concentration, the correlation between activated clotting time (ACT) and activated partial thromboplastin time (aPTT) results, and to compare the clinical decisions based on ACT results with those based on aPTT results. METHODS: Retrospective evaluation of a large database containing heparin concentrations, ACT results (1 device), and aPTT results (3 different instruments: 2 bedside, 1 laboratory-based). Correlations between heparin concentrations and clotting time tests and between ACT results and aPTT results were determined. Clinical decisions regarding heparin dosage adjustments based on ACT results were compared with those based on aPTT results. RESULTS: Correlations between clotting time tests and heparin concentrations were r = 0.72 for ACT and r = 0.74-0.86 for the aPTT instruments. The laboratory-based aPTT had the highest correlation to heparin concentrations. The correlation between ACT and aPTT results ranged from r = 0.64-0.67. Heparin dosage adjustment decisions based on ACT results agreed with decisions based on aPTT results 59-63% of the time. CONCLUSIONS: The laboratory-based aPTT has a stronger correlation to heparin concentration than the bedside-based aPTT and ACT. The correlation between ACT and aPTT was similar among 3 different aPTT instruments. Decisions to adjust heparin therapy based on ACT results differed from decisions based on aPTT results more than one-third of the time.

Anticoagulants↗

PubMedDate not supplied

Changes in prothrombin and activated partial thromboplastin time during replacement therapy with human recombinant growth hormone in growth hormone deficient adults.

BACKGROUND: In rodents, Growth Hormone (GH) has been shown to stimulate coagulation parameters, including Prothrombin Time (PT), activated Partial Thromboplastin Time (aPTT) and vitamin K dependent coagulation factors. However, there are no reports on the influence of GH replacement therapy on global coagulation tests in Growth Hormone Deficiency (GHD). OBJECTIVE: The aim of this study was to investigate the effects of GH administration on basic coagulation parameters: PT, aPTT and fibrinogen concentrations in adult GHD patients before and during one year of GH replacement. DESIGN: Twenty-one adult patients with severe GHD (mean age +/- SE: 38.6 +/- 2.8 years) were included in this hospital based, prospective, interventional study. All patients were treated with rhGH for 12 months (GH dose: 0.4 mg/day for male and 0.6 mg/day for female patients). IGF-1 concentrations were determined using RIA-INEP kits. Basic coagulation tests, i.e. aPTT and fibrinogen concentrations, were measured before and after 3, 6 and 12 months of treatment with rhGH. Control values were obtained from fourteen "healthy" subjects matched by age, sex and body mass index (BMI). RESULTS: At baseline, we observed no significant differences in PT, aPTT and fibrinogen values between GHD and healthy subjects. IGF-1 concentrations increased significantly within 3 months of GH therapy (8.2 +/- 1.5 vs. 24.2 +/- 2.9 nmol/l, p <0.05) and remained stable thereafter. A significant increase in PT values, which was more pronounced in female subjects, was noted after 6 and 12 months of treatment with GH. aPTT values increased significantly after 12 months of treatment only in male patients (28.8 +/- 4.6 vs. 39.7 +/- 2.1 s.; p <0.05). No significant changes in fibrinogen concentrations were found during the study. CONCLUSIONS: Twelve months of GH replacement therapy led to a significant increase in PT and aPTT values in adult GHD patients, while fibrinogen concentrations did not change. Changes in PT were more pronounced in female GHD patients, while an increase in aPTT values was observed only in male patients with GHD. The clinical significance of these changes needs further evaluation.

Adult↗

PubMed1998

Evaluation of the activated partial thromboplastin time (APTT) sensitivity to heparin using five commercial reagents: implications for therapeutic monitoring.

Heparin is an effective drug for prevention and treatment of thromboembolic conditions. Although several biological assays have been proposed for monitoring unfractionated heparin therapy, the measurement of the activated partial thromboplastin time (APTT) is the most widely employed test, and the overall risk of thromboembolic episodes was markedly reduced by maintaining APTT ratios above 1.5. However, the adjustment of the heparin therapy on the basis of APTT presents several questions which are still unresolved. Major discrepancies were found in APTTs performed using different reagents in both ex vivo and in vitro heparinized samples and occasionally with different lots of the same reagents; poor correlation was observed between APTT values and plasma heparin concentrations. In order to gain further insights into this phenomenon, we analysed the sensitivity to heparin of five commercial reagents for APTT measurement in 19 ex vivo heparinized samples. Differences were observed; correlation coefficients ranged from 0.820 to 0.985 and slopes of linear regressions from 0.26 to 1.14. Moreover, unsatisfactory correlations were obtained when APTT ratios were compared with heparin plasma concentrations in the same patients' samples. In the heparin therapeutic range of 0.35 - 0.70 U/ml, reagent-dependent differences were observed in the corresponding APTT values. These results point out a critical role of the assay methodology in monitoring heparin therapy by APTT. We suggest that reference materials and methods should be urgently identified, a universally agreed scale for reporting results should be established and reference ranges for the unfractionated heparin therapy should be reconsidered taking on account the reagent employed.

Case-Control Studies↗