Clinical Experience with the Thrombostat 4000

1995 ◽  
Vol 21 (S 02) ◽  
pp. 1-10 ◽  
Author(s):  
Rasheed Alshameeri ◽  
Eberhard Mammen

Bleeding times are presently widely used to screen patients with primary hemostasis defects although their accuracy and reliability has been questioned by many investigators. Platelet aggregation studies are not suited for routine use. We investigated the performance characteristics of the Thrombostat 4000, a device that assesses primary hemostasis. Tests can be performed by adding ADP, epinephrine, CaCl2 or NaCl to the collagen onto which platelets adhere. It was found, using normal volunteers and patients, that ADP and epinephrine had acceptable reference ranges with coefficients of variance between 9–12% for within run and between runs. However, major differences were seen when different filter badges were used–a reflection of differences in collagen. Regular citrated blood, routinely drawn for coagulation studies, can be used; test performance can be delayed for up to five hours when the blood is kept at room temperature. The effects of aspirin on volunteers could be detected when epinephrine was used, but not with ADP. ADP addition allowed the detection of more patients with primary hemostasis defects than bleeding times, and epinephrine was as useful as ADP in detecting these abnormalities. The data suggest that the broadest spectrum of platelet defects (ASA use and platelet dysfunction) can be detected with epinephrine. Inconsistencies in collagen used for coating of the filters is a major drawback for the routine use of this device in screening primary hemostasis defects.

1987 ◽  
Author(s):  
D Declerck ◽  
F Vinckier ◽  
J Vermylen

Clinical experience suggests that acetylsalicylic acid prolongs bleeding after dental extraction, but objective evidence is lacking. We have studied the influence of platelet dysfunction on haemostasis following dental extraction in rabbits. Platelet defects were induced by oral administration of acetylsalicylic acid (ASA) or ticlopidine (T). Duration of bleeding was measured in following groups of Dutch rabbits:(A) control animals (n = 32),animals who received (B) ASA 100 mg/kg/day (n = 31), (C) ASA 300 mg/kg/day (n = 12)(D) T 60 mg/kg/day (n = 8) and (E) T 125 mg/kg/day (n = 12). Inhibition of thromboxane B2-formation was more than 99% in animals receiving ASA. ADP-induced platelet aggregation was inhibited in animals receiving T. Four front teeth were extracted on day 7 and tooth socket bleeding times were determined by inspecting the wounds at 5 min intervals. Oozing sockets were counted. The figure below shows the results.It is concluded that ASA or T prolong bleeding following dental extraction in rabbits. The effect appears to be dose-dependent for T. This model may be useful for evaluating haemostatic procedures applicable to patients with platelet disorders.


Blood ◽  
2010 ◽  
Vol 116 (21) ◽  
pp. 1436-1436
Author(s):  
Nina D’Abreo ◽  
Vatsala Kirtani ◽  
Jennifer L. Kujawa Eisenstein ◽  
Yelda Nouri ◽  
Matt Geller ◽  
...  

Abstract Abstract 1436 Objectives: Myelodysplastic syndrome (MDS), immune thrombocytopenia purpura (ITP) and myeloproliferative disorders (MPD) are frequent diagnoses in a community hematology practice. Qualitative platelet defects are common features of these disorders but are difficult to characterize in daily practice as specialized technology is not readily available. The Verify Now® (VN) is a rapid, point-of service assay to measure platelet aggregation. VN assesses the degree of inhibition of GPIIb/IIIa mediated aggregation and uses minimal sample preparation. Its main application currently, is to determine the adequacy of anti-platelet therapy with aspirin (ASA), thienopyridines, such as clopidogrel (CPL), and GPIIb/IIIa inhibitors in patients with cardiovascular disease. (Verify Now for IIb/IIIa, aspirin, and P2Y12, Instructions for Use, Accumetrics) We used VN to detect platelet aggregation defects in patients with ITP, MDS and MPD and compared the to a platelet function analyzer PFA -100. (Francis JL; In Michelson AD, ed. Platelets, 2nded. San Diego: Elsevier/Academic Press, 2007:535-544) Methods: Subjects with MDS, ITP, and MPD were identified from our Hematology practice. Informed consent was obtained from all patients. Patients taking aspirin and /or clopidogrel were not excluded. Platelet function was determined concurrently using VN and PFA-100 using standard reference ranges. (Verify Now for IIb/IIIa, aspirin, and P2Y12, Instructions for Use, Accumetrics), (Mammen EF et al; Semin Throm Hemost. 1998; 24 (2):195-205). Results: Thirty three patients are enrolled in the study, 21 with ITP, 6 with MDS and 6 with MPD. Assay results are available for 30 patients. Platelet counts for all patients ranged from 2 to 1206 ×103/cmm, mean platelet count being 184 ×103/cmm (SD ± 215) and median count was 120 ×103/cmm. ITP: In 15 patients with ITP and platelet count ≥ 100 ×103/cmm, 11 were not on ASA/CPL. PFA-100 and VN each detected abnormalities in 4 patients but results were non-concordant in 2 patients. In 4 patients on ASA, VN detected ASA effect in all 4 whereas the PFA-100 detected 2. In 2 patients with platelet count < 100 ×103/cmm not on ASA/CPL, VN detected abnormalities in both, whereas PFA detected only 1. (Table 1) MDS: Two patients had counts ≥ 100 ×103/cmm. One was on ASA and both assays detected ASA effect. Neither assay detected an abnormality in the patient not on ASA. In 4 patients with counts <100 ×103/cmm, 3 were not on ASA/ CPL. Both assays detected abnormalities in all 3 and ASA effect in 1 patient on ASA. (Table 2) MPD: In 4 patients with counts < 400 ×103/cmm, not on ASA/CPL, both assays detected non-concordant abnormalities in 1 patient. Both assays detected CPL and ASA effect in 1 patient taking both drugs. 2 patients had count ≥400,000. In 1 patient taking CPL, VN detected CPL effect but PFA did not. In 1 patient on ASA, PFA detected abnormality but VN did not. (Table 3) Conclusion: In patients with MDS, ITP, and MPD, abnormal platelet function is common. These results show that VN may be as sensitive for detecting platelet dysfunction as PFA-100. Results were especially concordant in MDS patients. We plan to include 150 patients to confirm these results. We hope to define qualitative platelet defects in this population so that appropriate intervention can be recommended prior to invasive procedures. Disclosures: No relevant conflicts of interest to declare.


1973 ◽  
Vol 29 (01) ◽  
pp. 183-189
Author(s):  
C. A Praga ◽  
E. M Pogliani

SummaryTemperature represents a very important variable in ADP-induced platelet aggregation.When low doses of ADP ( < 1 (μM) are used to induce platelet aggregation, the length of the incubation period of PRP in the cuvette holder of the aggregometer, thermostatted at 37° C, is very critical. Samples of the same PRP previously kept at room temperature, were incubated for increasing periods of time in the cuvette of the aggregometer before adding ADP, and a significant decrease of aggregation, proportional to the length of incubation, was observed. Stirring of the PRP during the incubation period made these changes more evident.To measure the exact temperature of the PRP during incubation in the aggre- gometer, a thermocouple device was used. While the temperature of the cuvette holder was stable at 37° C, the PRP temperature itself increased exponentially, taking about ten minutes from the beginning of the incubation to reach the value of 37° C. The above results have a practical significance in the reproducibility of the platelet aggregation test in vitro and acquire particular value when the effect of inhibitors of ADP induced platelet aggregation is studied.Experiments carried out with three anti-aggregating agents (acetyl salicyclic acid, dipyridamole and metergoline) have shown that the incubation conditions which influence both the effect of the drugs on platelets and the ADP breakdown in plasma must be strictly controlled.


1987 ◽  
Author(s):  
M Taki ◽  
M Inagaki ◽  
T Miura ◽  
N Saito ◽  
T Meguro ◽  
...  

It has been reported recently that DDAVP might be an useful tool in the therapy and prevention of bleeding in patients with congenital afibrinogenemia (CA).To study the mechanism of its efficacy, changes in the platelet functions of a patient with CA were examined prior to, and one hour after, the infusion of DDAVP (0.4 μg/Kg). A patient with Glanzmann's thrombasthenia (GT) was also examined, to allow a study of the role of platelet membrane glycoprotein IIb/IIIa (GP IIb/IIIa), a deficient platelet in GT, in the resulting effects of the drug. When both patients were infused with DDAVP, the level of plasma von Willebrand factor (vWF) increased two- to fourfold, accompanied by an enhancement of ristocetin-induced platelet agglutination. The level of plasma fibrinogen was never changed.The prolonged bleeding time observed was markedly improved only in the CA patient, remaining unchanged in the GT patient, after the infusion of DDAVP. This indicates that DDAVP is effective in diminishing the bleeding tendency in CA, but not in GT. Among the platelet functions tested, only the platelet retention rate on glass beads, ADP-induced platelet aggregation and collagen-induced platelet aggregation improved in CA, each remaining unchanged in GT. In particular, collagen-induced platelet aggregation was markedly improved in the CA patient. However, the platelet adhesion to collagen (50 μg/ml)-Sepharose remained normal, both before and after the infusion of DDAVP in CA.These results suggest that an increase in the plasma vWF level and the existence of platelet membrane GPIIb/IIIa may be necessary for the improvement of primary hemostasis, after the infusion of DDAVP. The vWF-mediated platelet aggregation by collagen or ADP may produce this effect in the CA patient.


1994 ◽  
Vol 19 (3) ◽  
pp. 301-302 ◽  
Author(s):  
B. BARTON ◽  
J. M. KLEINERT

Caffeine has been shown to increase mean blood pressure, but studies documenting the effect of caffeine on digits are lacking. We evaluated the effect of caffeine on digital blood pressure and pulse volume in normal volunteers. In the first part of the study, 24 subjects were given water containing either 200 mg of caffeine or placebo. Bilateral brachial and middle finger digital pressures were measured at room temperature before ingestion and at 30 and 60 minutes after ingestion. In the second part of the study, pulse volume recordings (PVRs) were obtained in 24 subjects at the level of the distal phalanx of the middle finger of one hand immediately prior to beverage ingestion and at 10 minute intervals for 90 minutes. Differences in mean digital systolic pressure, digital/brachial index, or PVR amplitude between the control and caffeine groups were not statistically significant. The administration of caffeine was found not to alter the haemodynamics of digital blood flow or digital pressure in this population.


Blood ◽  
2009 ◽  
Vol 114 (2) ◽  
pp. 261-263 ◽  
Author(s):  
Alfonso Quintás-Cardama ◽  
Xin Han ◽  
Hagop Kantarjian ◽  
Jorge Cortes

Abstract Dasatinib is associated with increased risk of bleeding among patients with chronic myeloid leukemia, even in the absence of thrombocytopenia, suggesting the presence of a hemostatic defect. We tested platelet aggregation in 91 patients with chronic myeloid leukemia in chronic phase either off-therapy (n = 4) or receiving dasatinib (n = 27), bosutinib (n = 32), imatinib (n = 19), or nilotinib (n = 9). All but 3 patients simultaneously receiving imatinib and warfarin had normal coagulation studies. All 4 patients off therapy had normal platelet aggregation. Impaired platelet aggregation on stimulation with arachidonic acid, epinephrine, or both was observed in 70%, 85%, and 59% of patients on dasatinib, respectively. Eighty-five percent of patients on bosutinib, 100% on nilotinib, and 33% on imatinib had normal platelet aggregation. Dasatinib 400 nM induced rapid and marked prolongation of closure time to collagen/epinephrine in normal whole blood on the PFA-100 system. In conclusion, dasatinib and, to some extent, imatinib produce abnormalities in platelet aggregometry testing.


1999 ◽  
Vol 82 (10) ◽  
pp. 1250-1254 ◽  
Author(s):  
Olga Panes ◽  
Blanca Muñoz ◽  
Edgar Pais ◽  
Rodrigo Tagle ◽  
Fernando González ◽  
...  

Summary Background: A defect in platelet function is the main determinant of the prolonged bleeding time in chronic renal failure (CRF). We previously reported a significant correlation between platelet abnormalities and elevated plasma markers of plasmin and thrombin generation. Our aim was to explore the effect of inhibiting both plasmin action with tranexamic acid (TA) and thrombin production with low molecular weight heparin (LMWH), on the bleeding time (BT) and platelet function in patients with CRF. Methods: 37 patients with CRF (mean creatinine 8.6 ± 4.4 mg/dl) under conservative treatment, with prolonged BT, entered this study and received TA during 6 days, with (n = 24) and without LMWH (n = 13). BT, platelet aggregation/secretion, platelet granule contents, von Willebrand factor and parameters of coagulation and fibrinolysis were recorded before and at the end of treatment. Results: The BT was shortened in 26/37 (67%) patients. This effect was associated with significant improvement of platelet aggregation and secretion, with decrease to a normal range of fibrin/fibrinogen degradation products, mild increase in plasmin-antiplasmin complexes and pronounced reduction of circulating plasminogen. No differences were seen among patients with or without LMWH. No serious side effects or complications were observed. Interpretation: These findings indicate that the activation of fibrinolysis plays a significant role in the defect of primary hemostasis in patients with CRF. Inhibition of plasmin activity with TA shortens the BT and improves platelet function in the majority of patients with severe disease.


1999 ◽  
Vol 67 (6) ◽  
pp. 2763-2768 ◽  
Author(s):  
Masaaki Iwaki ◽  
Kazunari Kamachi ◽  
Nikolaus Heveker ◽  
Toshifumi Konda

ABSTRACT The effect of Bordetella pertussis adenylate cyclase toxin (ACT) on platelet aggregation was investigated. This cell-invasive adenylate cyclase completely suppressed ADP (10 μM)-induced aggregation of rabbit platelets at 3 μg/ml and strongly suppressed thrombin (0.2 U/ml)-induced aggregation at 10 μg/ml. The suppression was accompanied by marked increase in platelet intracellular cyclic AMP (cAMP) content and was diminished by the anti-ACT monoclonal antibody B7E11. A catalytically inactive point mutant of ACT did not show the suppressive effect. Since an increase of cAMP content is a known cause of platelet dysfunction, these results indicate that the observed platelet inactivation was due to the catalytic activity of ACT through increase of intracellular cAMP.


2021 ◽  
Author(s):  
Elizabeth Fisher ◽  
Christian James ◽  
Diana Mosca ◽  
Bart J Currie ◽  
Anna P Ralph

BACKGROUND Acute Rheumatic Fever (ARF) is a critically important condition for which there is no diagnostic test. Diagnosis requires the use of a set of criteria comprising clinical, laboratory, electrocardiographic and echocardiographic findings. The complexity of the algorithm and the fact that clinicians lack familiarity with ARF, make ARF diagnosis ideally suited to an electronic decision support tool. We developed an ARF Diagnosis Calculator to assist clinicians in diagnosing ARF and correctly assigning categories of ‘possible, ‘probable’ or ‘definite’ ARF. OBJECTIVE To evaluate the acceptability and accuracy of the ARF Diagnosis Calculator as perceived by clinicians in Northern Australia where ARF rates are high, and test performance against a ‘gold standard’. METHODS Three strategies were used to provide triangulation of data. Users of the calculator employed at Top End Health Service, Northern Territory, Australia were invited to participate in an online survey about the calculator, and clinicians with ARF expertise were invited to participate in semi-structured interviews. Qualitative data were analysed using inductive analysis. Performance of the calculator in correctly assigning a diagnosis of possible, probable or definite ARF, or not ARF, was assessed using clinical data from 35 patients presenting with suspected ARF. Diagnoses obtained from the calculator were compared using the Kappa statistic with those obtained from a panel of expert clinicians, considered the ‘gold standard’. Findings were shared with developers of the calculator and changes were incorporated. RESULTS Survey responses were available from 23 Top End Health Service medical practitioners, and interview data were available from five expert clinicians. Using a 6-point Likert scale, participants highly recommended the ARF Diagnosis Calculator (median score 6, IQR 1) and found it easy to use (median 5, IQR 1). Participants believed the calculator helped them diagnose ARF (median 5, IQR 1). Valued features included educational content and laboratory test reference ranges. Criticisms included: too many pop-up messages to be clicked through; that it is less helpful in remote areas which lack access to investigation results; and the need for more clarity about actively excluding alternative diagnoses to avoid false-positive ARF diagnoses. Importantly, clinicians with ARF expertise noted that electronic decision making is not a substitute for clinical experience. There was high agreement between the ARF Diagnosis Calculator and the ‘gold standard’ ARF diagnostic process (κ=0.767, 95% CI: 0.568-0.967). However, incorrect assignment of diagnosis occurred in 4/35 (11%) patients highlighting the greater accuracy of expert clinical input for ambiguous presentations. Sixteen changes were incorporated into a revised version of the calculator. CONCLUSIONS The ARF Diagnosis Calculator is an easy-to-use, accessible tool, but it does not replace clinical expertise. Effective resources to support clinicians in diagnosing and managing ARF are critically important for improving the quality of care of ARF.


Blood ◽  
1974 ◽  
Vol 44 (3) ◽  
pp. 411-425 ◽  
Author(s):  
V. J. McPherson ◽  
M. B. Zucker ◽  
N. M. Friedberg ◽  
P. L. Rifkin

Abstract Plasma of normal heparinized blood contained 0.284 µM ± SD 0.097 (ADP + ATP) with an ATP:ADP ratio of 2.5:1. Plasma from thrombocytopenic blood contained only 0.106 µM ± 0.073 (ADP + ATP). Blood with normal platelet retention released 0.234 µM ± 0.187 (ADP + ATP) during passage through a glass bead column, with an ATP:ADP ratio of 1.6:1. Significantly less was released in blood with low retention, i.e., samples from patients with von Willebrand’s disease, thrombasthenia, or thrombocytopenia, and some samples from normal individuals. Thus, nucleotides in the plasma of pre- and postcolumn blood appear to be derived from platelets; their release within glass bead columns is closely associated with normal platelet retention. Since release occurred at room temperature and was not prevented by acetylsalicylic acid or accompanied by measurable release of 14C-serotonin, the classic release reaction may not have been responsible. The low retention in platelet-rich plasma was variably increased by adding 0.5 µM ADP, an increase at least partly due to trapping of preformed aggregates. Retention in undisturbed blood was markedly inhibited by creatine phosphokinase with creatine phosphate (CPK-CP) and moderately inhibited by apyrase I (ATPase:ADPase 0.8:1) at an ADP-removing activity between 1 and 5 U/ml, indicating that ADP is essential for retention. At less than 1 U/ml, both apyrase I and II (ATPase: ADPase 2.8:1) enhanced retention in undisturbed blood, but CPK-CP was still inhibitory. These results suggest that enhancement is due to conversion of released ATP to ADP, as shown to occur in studies of platelet aggregation with ATP and ADP. At less than 1 U/ml, all three enzymes protected against the inhibitory effect of disturbance; this protection was marked with apyrase II, moderate with apyrase I and slight with CPK-CP. These observations provide additional evidence that ADP is responsible for the low retention caused by disturbance of the blood.


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