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[Comparative studies of measuring condition of activated partial thromboplastin time and contact factors in experimental animals].

For establishing the optimal incubation time (OIT) for measurement of the activated partial thromboplastin time (APTT) in dogs, rabbits, guinea pigs, rats and mice, we determined the shortest clotting time of the plasma from each animal species and compared them with that of human plasma. The OIT for APTT determination was 15 to 30 sec in guinea pigs, rats and mice and 5 to 10 minutes in dogs and rabbits. The mouse APTT (about 30 sec) with the OIT thus determined was similar to human APTT, and relatively longer than APTT in other animal species (10-20 sec). To elucidate the mechanism of the species differences in OIT, we examined the plasma of each animal species for the activity of the contact factors such as factor XII, factor XI, high molecular weight kininogen (HMWK) and prekallikrein (PK) and their effect on the coagulation of contact factor-deficient plasma. The total activity of contact factors was higher in dogs and guinea pigs and lower in rabbits and mice than that in humans. Species difference with the factor XII, Factor XI and HMWK was noted in clotting time but not in OIT. These results suggest that the species difference in OIT for APTT is probably due to difference in activity of the plasma contact factors and in the mode of coagulation for each contact factor.

Animals↗

Correlation of activated clotting time and activated partial thromboplastin time to plasma heparin concentration.

STUDY OBJECTIVE: To determine the correlation between activated clotting time (ACT) or activated partial thromboplastin time (aPTT) and plasma heparin concentration. DESIGN: Two-phase prospective study. SETTING: University-affiliated community hospital. PATIENTS: Thirty patients receiving continuous-infusion intravenous heparin. INTERVENTIONS: Measurement of ACT, aPTT and plasma heparin concentrations. MEASUREMENTS AND MAIN RESULTS: Linear and log linear correlations were determined between clotting time tests and heparin concentrations. Linear correlations yielded r values of 0.58 for ACT (p=0.008) and 0.89 for aPTT (p=0.0001). Log linear correlations yielded r values of 0.60 for ACT (p=0.005) and 0.88 for aPTT (p=0.0001). A decision analysis was performed to determine possible consequences of dosage adjustments based on either test in relationship to the decision based on plasma heparin concentration. The decision analysis based on ACT disagreed with corresponding decisions based on plasma heparin concentration in 15 of 30 patients; 13 disagreements may have increased the risk of bleeding, and the other 2 may have increased the risk of thrombosis. Decisions based on aPTT disagreed with corresponding decisions based on plasma heparin concentration in 13 of 30 patients; 2 disagreements may have increased the risk of bleeding, and the other 11 may have increased the risk of thrombosis. CONCLUSION: There are significant statistical linear and log linear correlations between both clotting time tests and plasma heparin concentrations, with aPTT showing stronger correlation than ACT. However, decisions regarding heparin therapy based on ACT may increase a patient's risk of bleeding, whereas decisions based on aPTT may increase the risk of thrombus progression or rethrombosis.

Aged↗

Two instruments to determine activated partial thromboplastin time: implications for heparin monitoring.

STUDY OBJECTIVE: To measure the difference in therapeutic ranges of activated partial thromboplastin time (APTT) between two laboratory devices. DESIGN: Prospective, controlled laboratory study. SETTING: University-affiliated hospital. PATIENTS: Thirty inpatients receiving intravenous unfractionated heparin for treatment of myocardial infarction, unstable angina, deep venous thrombosis, or pulmonary embolism. INTERVENTIONS: Therapeutic APTT ranges were determined by a portable (whole blood assay) and a central laboratory device (plasma assay) based on heparin serum concentrations. They were compared with APTT ranges equivalent to 1.5-2.5 times the mean normal determination. MEASUREMENTS AND MAIN RESULTS: The central laboratory and portable devices produced therapeutic ranges of 61-93 and 56-73 seconds, respectively. Both differed from conventional therapeutic ratios of 1.5-2.5 times the mean normal (41-68 sec). Mean absolute APTT differences between instruments were statistically significant (12 +/- 20 sec, p<0.006), and 58% of paired APTT values differed by more than 10 seconds. CONCLUSION: A fixed APTT ratio as a goal for monitoring unfractionated heparin may result in significant underanticoagulation. Individual therapeutic APTT ranges must be reported for each instrument if more than one is used for heparin monitoring.

Adult↗

Monitoring unfractionated heparin therapy with antifactor Xa activity results in fewer monitoring tests and dosage changes than monitoring with the activated partial thromboplastin time.

STUDY OBJECTIVE: To determine how much more costly it is to monitor unfractionated heparin (UFH) therapy by antifactor Xa heparin activity (HA) than by activated partial thromboplastin time (aPTT). DESIGN: Prospective, randomized, unmasked, cohort, single-center study. SETTING: A 625-bed, adults-only, private teaching hospital. PATIENTS: Two hundred sixty-eight patients with a variety of indications for UFH therapy. INTERVENTIONS: Patients were treated with UFH based on ideal weight (75 U/kg bolus, 20 U/kg initial infusion) and monitored by either HA or aPTT, MEASUREMENTS AND MAIN RESULTS: After adjusting for gender, groups were equivalent in patient characteristics and UFH dosage. The HA group had fewer monitoring tests and dosage changes/24 hours than the aPTT group. These reductions neutralized much of the increased cost of the HA assay itself. CONCLUSION: Monitoring UFH therapy over 96 hours with an HA assay costs $4.37 more than monitoring with aPTT. This modest increase may be acceptable given other advantages of the HA assay.

Aged↗

Challenge and rechallenge: drotrecogin alfa (activated)-induced prolongation of activated partial thromboplastin time in a patient with severe sepsis.

An 81-year-old woman with ischemic bowel underwent laparotomy with small-bowel resection and developed septic shock. She required broadspectrum antibiotics, norepinephrine, and mechanical ventilation. The patient received drotrecogin alfa (activated) 24 microg/kg/hour for a total of 67.5 hours. Coagulation parameters were monitored during her therapy. Significant increases in activated partial thromboplastin time (aPTT) during infusion led to two temporary discontinuations of the drug. Coagulation parameters decreased when the drug was held and increased with each rechallenge. The patient survived the episode and was discharged on postoperative day 27. Medical records of 26 other patients who received drotrecogin alfa (activated) at our institution from November 2001-August 2003 were reviewed retrospectively for coagulation parameters and bleeding rate. Of the 26 patients, nine (35%) were treatment compliant (>90% of the 96-hr course). Coagulopathy and bleeding resulted in early discontinuation in four (15%) and six (23%) patients, respectively. An increase in aPTT from baseline to during infusion of drotrecogin alfa (activated) was noted in 14 patients with complete data (p=0.56). A decrease in median platelet count from baseline to during infusion was noted in the six patients who bled during therapy (p=0.01). Two of these patients had platelet counts less than 30x10(3)/mm3 during administration. Drotrecogin alfa (activated) should be considered an anticoagulant. In postmarketing reports, clinically significant bleeding occurred more frequently than was noted in a large, randomized, multicenter trial. Patients receiving drotrecogin alfa (activated) should be closely monitored for prolongation of coagulation parameters. Temporary discontinuation of the drug should be considered when international normalized ratio is greater than 3.0, platelet count is less than 15x10(3)/mm3, and aPTT is greater than 100 seconds.

APACHE↗

Prothrombin, activated partial thromboplastin, and proteins induced by vitamin K absence or antagonists clotting times in 20 hyperthyroid cats before and after methimazole treatment.

The effect of daily doses of 5-15 mg of methimazole on the platelet count, prothrombin time (PT), activated partial thromboplastin time (APTT), and proteins induced by vitamin K absence or antagonists (PIVKA) clotting time in 20 hyperthyroid cats was determined. No significant (P > .05) difference was found in median platelet count. PT, APTT, or PIVKA clotting time before treatment compared to median values at 2-6 weeks or > or =7-12 weeks of methimazole treatment. No cat had a prolonged APTT at any time. At 2-6 weeks of methimazole treatment, 1 cat each developed thrombocytopenia or prolonged PIVKA clotting time despite initially normal values. Three cats had abnormal coagulation tests (prolonged PT [n = 1] and PIVKA clotting time [n = 3]) before treatment that fluctuated during treatment. Excluding the 3 cats that had abnormal PIVKA clotting time before treatment, prolonged PIVKA clotting time developed in 6% (1/17; 95% confidence interval, 0-28%) cats treated with methimazole for 2-6 weeks. Seemingly. doses of methimazole commonly used to treat hyperthyroidism in cats do not cause alteration in PT and APTT, and only rarely prolong PIVKA clotting time. Nevertheless, abnormal PIVKA clotting time may explain bleeding tendencies unassociated with thrombocytopenia in methimazole-treated hyperthyroid cats.

Animals↗

The suitability of the Australasian reference thromboplastin for use on an automated centrifugal analyser.

The Australasian Reference Thromboplastin (ART) is now produced in a low turbidity form (ART-LT) for use in photo-electrical coagulometers. This study was designed to assess the suitability of ART and ART-LT for use on a fully automated centrifugal analyser, the ACL300 Research (ACL). These reagents were compared to Thromborel S (TRL), the reagent currently in successful use in our laboratory. It was found that regular ART was an unsuitable reagent for the ACL due to poor daily quality control testing (QC) precision at an INR of 4.5. The ACL also had the effect of significantly altering the ISI of ART from its recommended value as determined for manual testing. ART-LT was found to be an acceptable reagent on the ACL even though its QC precision was lower than that of TRL. The reagent was stable throughout its 8 wk shelf-life. The ACL significantly altered the ISI (International Sensitivity Index) of ART-LT. This may no longer be a disadvantage, as ART-LT is now provided with an ISI specifically for its use on the ACL.

Blood Coagulation Tests↗

The production of tumour necrosis factor, tissue thromboplastin, lactoferrin and cathepsin C during lipopolysaccharide stimulation in whole blood.

The release of tumour necrosis factor (TNF), lactoferrin (LF) and cathepsin C (CC) into plasma and production of thromboplastin (TPL) in monocytes were studied in lipopolysaccharide (LPS) stimulated heparinized whole blood from 10 healthy donors. The influence of dextran 70, haemaccel and methylprednisolone on levels of these parameters were examined. TNF concentration in plasma 5 min after the addition of LPS (0 h) was 250 pg/ml (median), 520 pg/ml after 1 h and 1300 pg/ml after 3 h. The addition of dextran 70 to the blood in addition to LPS at the same intervals gave significantly higher values of 740 pg/ml and 1800 pg/ml after 1 h and 3 h respectively. Unstimulated cells had no TPL but after 1 h with LPS, the TPL activity in incubated cells was 2.3 mU/10(6) monocytes and after 3 h, 2.7 mU/10(6) monocytes. LPS induced the secretion of LF from granulocytes (PMN) and the levels 5 min after the addition of LPS (0 h) were 2.1 mg/l (control 0.2 mg/l) and after 1 h, 5.3 mg/l (control 1.3 mg/l) in plasma after LPS stimulation. Haemaccel enhanced the LPS-induced generation of TPL in monocytes and production of CC. The LPS-induced secretion of LF was, to a small extent, influenced by the three reagents tested. Methylprednisolone (1 mmol/l) reduced the production and appearance of TNF in plasma and the generation of TPL activity in monocytes. This model for stimulating heparinized whole blood is suitable for examination of the production and appearance of cellular factors and the influence of drugs on this production.

Adult↗

Effects on leukocyte function by arthroplasty. Thromboplastin activity and oxygen-derived free radicals studied in rheumatoid arthritis and arthrosis.

We examined thromboplastin activity (TA) of monocytes and release of oxygen-derived free radicals (ODRFs) from monocytes and granulocytes before and after implantation of a hip or a knee prosthesis in 7 patients with rheumatoid arthritis and in 8 patients with arthrosis. Monocyte TA rose threefold on the first postoperative day in the rheumatoid patients, but was unaltered postoperatively in the arthrosis patients. Granulocyte chemiluminescence doubled in the arthrosis group on the second postoperative day, but was unaltered in the rheumatoid patients. Monocyte chemiluminescence was not influenced by the operation. Thus, leukocytes from the rheumatoid patients responded differently from surgical trauma when compared with leukocytes from the arthrosis patients. This difference may have an impact postoperatively.

Arthritis, Rheumatoid↗

Change in tissue thromboplastin content of brain following trauma.

BACKGROUND: Tissue thromboplastin (TTP) is an integral membrane protein contributing to coagulopathy after trauma of brain, which is a rich source of TTP. AIMS: A study was undertaken to establish the TTP content of various areas of normal brain and estimate the changes in TTP activity of brain in response to varying degrees of trauma. MATERIALS AND METHODS: Samples from different areas of brain of ten cadavers were used as controls and they were compared with contused brain tissue obtained after surgery in 25 head injury (HI) patients of varying severity. RESULTS: In the study group, the TTP activity of the frontal, parietal, and temporal lobes after HI was significantly raised in contrast to that of the control group. The TTP activity was also significantly higher in the severe HI patients than those having moderate HI. The mode of injury and the time lapse after HI had no significant bearing on the TTP activity. Subjects above 40 years of age demonstrated a higher mean TTP activity after HI, though it was not statistically significant. CONCLUSION: The study provides quantitative data on TTP activity of normal brain and highlights the role of TTP in coagulopathy following HI through its increased activity after HI, more so in the severe HI group.

Adolescent↗

Fatal dural sinus thrombosis associated with heterozygous factor V Leiden and a short activated partial thromboplastin time.

Inherited thrombophilia is a risk factor for dural sinus thrombosis (DST). To our knowledge, this is the first description with autopsy findings of a patient with DST associated with heterozygous factor V Leiden and a short activated partial thromboplastin time (aPTT). A 51-year-old woman presented with a 3-day history of headache, nausea, right-sided weakness, and focal motor seizure; she died 3 days after admission. At autopsy, a gross examination showed hemorrhage of bilateral parietal lobes and left primary motor cortex, uncal and tonsillar herniation, and pulmonary embolus of the right upper lobe. A microscopic examination of the brain showed an organizing thrombus in the superior sagittal sinus, diffuse cerebral edema, and extensive venous congestion. Laboratory studies showed heterozygous factor V Leiden by polymerase chain reaction and a very short aPTT of 17 seconds (reference range, 22-30 seconds). The combination of a heterozygous factor V Leiden mutation and a short aPTT may have contributed to the fatal DST in this patient.

Factor V↗

Diagnostic uses of the activated partial thromboplastin time and prothrombin time.

The activated partial thromboplastin time (APTT) and prothrombin time (PT) have three principal uses. In screening for coagulation disorders (or increased risk of postoperative hemorrhage), the tests add no information to the preoperative care of patients without clinical findings indicative of increased bleeding risk. Furthermore, the prevalence of asymptomatic congenital coagulopathies is so low that false-positive test results greatly outnumber true-positive results. Thus, clinicians may use clinical assessment to screen and should reserve coagulation tests to investigate patients with abnormal findings. In evaluating abnormal bleeding, these tests are sufficiently sensitive that if both are negative, further investigation of the coagulation system is obviated. If one or both tests are positive, the pattern of results directs further attention to limited segments of the coagulation sequence. In monitoring anticoagulation therapy, the APTT and PT tests appear to contribute to the safety and effectiveness of heparin and warfarin therapies, respectively.

Anticoagulants↗

Comparison of three prothrombin time and activated partial thromboplastin time reagent systems on the MLA 700 coagulation analyzer.

OBJECTIVE: To determine the difference in prothrombin time (PT) and activated partial thromboplastin time (aPTT) results among three reagent systems using a single analyzer instrument. DESIGN: Convenience sample of 100 patient specimens tested in duplicate with three reagent systems: Baxter-Dade, Pacific Hemostasis, and Organon Teknika. SETTING: A tertiary hospital that services other institutions within a three-state area. PATIENTS: Patients were divided into four groups: (1) normal preoperative patients who received no anticoagulants, (2) patients who received warfarin for at least the week immediately before the study, (3) patients who received heparin on the day of the testing, and (4) patients with severe liver disease accompanied by abnormal liver function tests. MAIN OUTCOME MEASURE: Coefficients of correlation of Baxter-Dade results versus the other two systems. RESULTS: PT values were significantly different in normal samples and in warfarin-treated patients. aPTT values were significantly different for normal patients and, for the Organon system only, for heparin-treated patients. When expressed as international normalized ratio (INR) values, taking reagent sensitivity into consideration, the results correlated well. Problems with precipitation when using Organon's system limited its practical utility. CONCLUSION: Compatibility between a reagent system and analyzer instrument should be verified by the manufacturer of the instrument. Use of the INR format produced more accurate and comparable results, allowing safer and more effective dosage adjustments. Laboratories should convert PT and aPTT results to the INR format routinely.

Adult↗

Comparison of six activated partial thromboplastin time reagents: intrinsic system factors' sensitivity and responsiveness.

Six activated partial thromboplastin reagents from four manufacturers were evaluated for precision, sensitivity, and dose responsiveness to Factors VIII, IX, and XII. Normal and abnormal APTT samples were prepared. Precision was assessed using 10 replicates on three consecutive days. Factor sensitivity and responsiveness were measured using a log-log curve of percent activity versus the upper limit of normal of prepared samples. All reagents had acceptable precision (C.V. less than 5%) in the normal range, but three performed unacceptably in the abnormal range. Two of the reagents were determined to be too sensitive to certain factors, giving an abnormal APTT result when the factor activity was in the normal range. Sensitivity and dose responsiveness to factors should be considered as two separate characteristics of an APTT reagent.

Adult↗

[A new system of analysis for partial thromboplastin time. Comparison with other blood coagulation tests in liver diseases].

A new analysis system for the partial thromboplastin test (Cephotest) has been applied to 40 subjects (normal, suffering from nutritional toxic hepatopathy, chronic hepatopathy and cirrhosis of the liver). The test was then compared with other blood clotting tests (Howell's test, PTT test, Quick's test, Normotest and Thrombotest) and proved to have very good standardization and sensitivity.

Adult↗

Properties of human tissue thromboplastins from brain, lung, arteries, and placenta.

Properties of purified and delipidated human tissue thromboplastins (TTPs) from brain (BTTP), lung (LTTP), arteries (ATTP) and placenta (PTTP) were studied. The extinction coefficients were closely similar with a mean value of 8.3 +/- 0.01 (SD). The molecular weights were 200,000 +/- 3,000 (SD) BTTP), 90,000 +/- 2,000 (SD) (LTTP), 110,000 +/- 3,000 (SD) (ATTP) and 250,000 +/- 4,000 (SD) (PTTP). The maximum activity of each delipidated TTP after relipidation was obtained when phospholipid-delipidated TTP ratio was 2.0, and the maximum specific activity was 440 units per mg BTTP, 270 units per mg LTTP, 80 units per mg ATTP and 600 units per mg PTTP. Ouchterlony analysis with anti-delipidated PTTP antibody showed the reaction of partial identity between delipidated TTPs and the antibody. Studies of the reactivity of I-125-delipidated TTPs with the antibody gave the following average values for % bound I-125-TTPs in 2 hours: 3.6 +/- 0.2(SD) % (I-125-BTTP), 11.0 +/- 0.3 (SD) % (I-125-LTTP), 4.4 +/- 0.2 (SD) % (I-125-ATTP) and 13.7 +/- 0.3 (SD) % (I-125-PTTP). It was also found that the antibody markedly neutralized the coagulant activity of saline extracts of brain, lung and placenta. These results indicate that delipidated BTTP, LTTP, ATTP and PTTP are different while being similar in some aspects and that complete TTPs are not coated with lipids in a manner inaccessible to anti-delipidated PTTP antibody.

Antibodies↗

[Suggestions and propositions to resolve some issues for standardization of prothrombin time and activated partial thromboplastin time].

Prothrombin time (PT) and activated partial thromboplastin time (APTT), popularized as a routine assay for screening blood coagulation disorders and monitoring anticoagulant therapy, still involve some issues regarding standardization. In this lecture, we present propositions to resolve these problems in respective laboratory. Although international normalized ratio (INR) calculated by international sensitivity index (ISI) of PT reagent seems to improve discrepancy of sensitivity between reagents, local calibration of sensitivity of PT reagent in respective laboratories (local SI) is reasonable to make INR/ISI system more useful. However, local calibration of reagent is not easy by WHO recommended method in a small size laboratory. By using AK calibrant (IMMUNO AG), one of calibration plasma for INR, we investigated its possibility to calibrate local SI in four different reagents, compared with the recommended methodology. The results led the following process to determine reagent and calibrate local SI for practical use of INR/ISI system. (a) Use PT reagent of which ISI is close to 1.0 if possible, and utilize manufacture's ISI as is for INR. (b) Select PT reagent labeled specific ISI for an instrument as the same as used in the lab., and use the manufacture's ISI as is, if impossible to choose small ISI reagent. (c) If use a reagent of which ISI is close to 2.0 and shown no specific ISI for used detector, adjustment of local SI by commercial calibration plasma is recommended when unavailable warfarinized patient plasma. In APTT, we attempted to evaluate sensitivity between five different APTT reagents with a patient model by hemophilia A plasma contained various FVIII: C. This model reflected difference of sensitivity between reagents in results. Because standardization of APTT is not improved in this point, certification of APTT pattern in each laboratories with patient models is required for not only monitoring of heparinization, but also screening of typical coagulation disorders such as hemophilia and von Willebrand disease.

Humans↗

[Test for detection of activated partial thromboplastin time using ellagic acid].

A simple and sensitive method for estimation of activated partial thromboplastin time (APTT) is developed, making use a complex reagent containing the activator (plant phospholipids) and contact factor (ellagic acid). The test requires additionally only 0.025 M CaCl2. The test is more sensitive to the presence of heparin in the blood and to insufficiency of blood clotting factors VIII and IX than the reagents containing insoluble substances (kaolin and animal phosphatides). Addition of soluble ellagic acid into reagent for APTT estimation allows studies on optic coagulometers.

Blood Coagulation↗