RESUMO
Platelet biology owes to intravital studies not only a better understanding of platelets' role in primary hemostasis but also findings that platelets are important factors in inflammation and atherosclerosis. Researchers who enter the field of intravital platelet studies may be confused by the heterogeneity of experimental protocols utilized. On the one hand, there are a variety of stimuli used to activate platelet response, and on the other hand there are several approaches to measure the outcome of the activation. A number of possible combinations of activation factors with measurement approaches result in the aforementioned heterogeneity. The aim of this review is to present the most often used protocols in a systematic way depending on the stimulus used to activate platelets. By providing examples of studies performed with each of the protocols, we attempt to explain why a particular combination of stimuli and measurement method was applied to study a given aspect of platelet biology.
Assuntos
Plaquetas/fisiologia , Ativação Plaquetária/fisiologia , Testes de Função Plaquetária/métodos , Animais , Aterosclerose/sangue , Hemostasia/fisiologia , Humanos , Inflamação/sangue , Agregação Plaquetária/fisiologia , Testes de Função Plaquetária/tendências , Trombose/sangueRESUMO
Light transmission aggregometry (LTA) is still considered as the "gold standard" for platelet function assessment but, as acompletely manual technology, it is labour intensive. This challenge can be overcome by performing platelet aggregometry in anautomated method on a routine coagulation analyzer. We aimed to compare and correlate results obtained from a traditional manual LTA solution realized in our Reference Center with an optimized automated system using CE-marked agonist reagents. Platelet rich plasma from patients with suspected platelet disorders, von Willebrand disease or antiplatelet therapy have been assessed using a wide range of agonist concentrations. Results were expressed as Maximal Platelet Aggregation and correlation was analyzed using the Passing and Bablok regression test. Platelet aggregometry studies were performed in 49 samples. Maximal aggregation response with ADP (0.5-10 µM), collagen (2 mg/µL), ristocetin (1.2 mg/mL) and arachidonic acid (1 mM) agonists showed significant correlation between the two aggregometers (p< .001). We observed a more variable response using lowconcentrations of ADP (≤5 µM). Moreover, we also noted discrepancies with the low dose of ristocetin, showing excessive paradoxical agglutination with the CS-2500, suggesting that a lower ristocetin dose should be used with this system. These data show that CS-2500 has the advantages of a walk-away technology and the use of CE-marked reagents also permit the possibility of an easier certification.
Assuntos
Testes de Coagulação Sanguínea/métodos , Agregação Plaquetária/fisiologia , Testes de Função Plaquetária/métodos , Feminino , Humanos , MasculinoAssuntos
Estenose da Valva Aórtica/fisiopatologia , Hemostasia/fisiologia , Agregação Plaquetária/fisiologia , Fator de von Willebrand/fisiologia , Insuficiência da Valva Aórtica/fisiopatologia , Insuficiência da Valva Aórtica/cirurgia , Estenose da Valva Aórtica/cirurgia , Hemorreologia/fisiologia , Humanos , Complicações Pós-Operatórias/etiologia , Complicações Pós-Operatórias/fisiopatologia , Conformação Proteica , Substituição da Valva Aórtica Transcateter/efeitos adversos , Substituição da Valva Aórtica Transcateter/métodos , Fator de von Willebrand/químicaRESUMO
Nonsteroidal anti-inflammatory drugs (NSAIDs), including ibuprofen, are amongst the most commonly used medications and produce their anti-inflammatory and analgesic benefits by blocking cyclooxygenase (COX)-2. These drugs also have the potential to prevent and treat cancer and some members of the class including ibuprofen can produce anti-platelet effects. Despite their utility, all NSAIDs are associated with increased risk of cardiovascular side effects which our recent work suggests could be mediated by increased levels of the endogenous NO synthase (NOS) inhibitor asymmetric dimethylarginine (ADMA) leading to reduced endothelial NOS activity and associated endothelial cell dysfunction. ADMA is a cardiotoxic hormone and biomarker of cardiovascular risk whose effects can be prevented by l-arginine. The ibuprofen salt, ibuprofen arginate (Spididol®) was created to increase drug solubility but we have previously established that it not only effectively blocks COX-2 but also provides an arginine source able to reverse the effects of ADMA in vitro and in vivo. Here we have gone on to explore whether the formulation of ibuprofen with arginine influences the potency and efficacy of the parent molecule using a range of simple in vitro assays designed to test the effects of NSAIDs on (i) platelet aggregation and (iii) colon cancer cell killing. Our findings demonstrate that ibuprofen arginate retains these key functional effects of NSAIDs with similar or increased potency compared to ibuprofen sodium, further illustrating the potential of ibuprofen arginate as an efficacious drug with the possibility of improved cardiovascular safety.
Assuntos
Apoptose/efeitos dos fármacos , Arginina/administração & dosagem , Sobrevivência Celular/efeitos dos fármacos , Neoplasias do Colo/tratamento farmacológico , Ibuprofeno/administração & dosagem , Agregação Plaquetária/efeitos dos fármacos , Agregação Plaquetária/fisiologia , Anti-Inflamatórios não Esteroides/administração & dosagem , Antineoplásicos/administração & dosagem , Células CACO-2 , Linhagem Celular Tumoral , Células Cultivadas , Neoplasias do Colo/patologia , Relação Dose-Resposta a Droga , Combinação de Medicamentos , Humanos , Inibidores da Agregação Plaquetária/administração & dosagem , Resultado do TratamentoRESUMO
BACKGROUND: Disseminated intravascular coagulopathy (DIC) relates to the consumption of coagulation factors and platelets with bleeding and micro thrombosis events. AIM: The aim of this study was to compare haemostasis parameters in critically ill patients with DIC versus patients without DIC, and in survivors versus non-survivors over time. Correlations between the DIC-score, the degree of organ failure and the haemostasis were assessed. METHOD: Patients admitted to the intensive care unit with a condition known to be associated with DIC and with an expected length of stay of >3 days were included. Routine laboratory tests, prothrombin time, activated partial thromboplastin time, platelet count, fibrinogen concentration and D-dimer were measured. Coagulation and platelet function were assessed with two point-of-care devices; Multiplate and ROTEM. DIC scores were calculated according to the International Society on Thrombosis and Haemostasis and Japanese Association for Acute Medicine. RESULTS: Blood was sampled on days 0-1, 2-3 and 4-10 from 136 patients with mixed diagnoses during 290 sampling events. The point-of-care assays indicated a hypocoagulative response (decreased platelet aggregation and reduced clot strength) in patients with DIC and, over time, in non-survivors compared to survivors. Patients with DIC as well as non-survivors had decreased fibrinolysis as shown by ROTEM. DIC scores were higher in non-survivors than in survivors. CONCLUSIONS: Patients with DIC displayed signs of a hypocoagulative response and impaired fibrinolysis, which was also evident over time in non-survivors. Patients with DIC had a higher mortality rate than non-DIC patients, and DIC scores were higher in non-survivors than in survivors.
Assuntos
Coagulação Intravascular Disseminada/diagnóstico , Coagulação Intravascular Disseminada/fisiopatologia , Hemostasia/fisiologia , Idoso , Estado Terminal , Feminino , Humanos , Masculino , Pessoa de Meia-Idade , Agregação Plaquetária/fisiologia , Sistemas Automatizados de Assistência Junto ao Leito , Estudos ProspectivosRESUMO
The objectives of this study were to establish feline references intervals for 3 commercial whole blood platelet function test analyzer systems: Multiplate analyzer (MP; Roche Diagnostics International Ltd., Rotkreuz, Switzerland), Platelet Function Analyzer-100 (PF: Siemens Canada, Mississauga, Ontario, Canada), and Plateletworks Combo-25 kit (PW; Helena Laboratories, Beaumont, TX). Venipuncture was performed on 55 healthy sedated cats, and platelet aggregation in response to adenosine diphosphate (ADP), collagen (COL), and arachidonic acid (AA; MP only) was assessed using citrated blood. For the MP analyzer, median (95% confidence intervals [CIs]) area under curve (Units) for ADP, COL, and AA agonists were 87 (11-176), 81 (32-129), and 91 (59-129), respectively. For the PF analyzer, median (95% CIs) closure time, using COL-ADP cartridges, was 69 (46-89) sec. For the PW assay, median (95% CIs) percent aggregations for ADP and COL agonists were 71 (18-92) and 49 (9-96), respectively, using impedance hematology analyzer platelet counts, and 94 (25-98) and 68 (14-119), respectively, using flow cytometry hematology analyzer platelet counts. There were low correlations between the PF analyzer (COL-ADP cartridge) and MP analyzer (COL agonist; ρ = 0.11), and between the PF analyzer (COL-ADP cartridge) and PW assay (COL agonist using impedance platelet counts; ρ = 0.14). The PW assay percent aggregations using impedance and flow cytometric platelet counts were correlated for both ADP (ρ = 0.64) and COL (ρ = 0.64) agonists. Platelet function testing using these tests are feasible in cats, but 95% CIs are wide, so single results may be difficult to interpret. Platelet counting by impedance or flow cytometry may be used for the PW assay but are not interchangeable.
Assuntos
Plaquetas/fisiologia , Gatos/fisiologia , Testes de Função Plaquetária/veterinária , Animais , Área Sob a Curva , Feminino , Masculino , Agregação Plaquetária/fisiologia , Contagem de Plaquetas/veterinária , Testes de Função Plaquetária/instrumentação , Testes de Função Plaquetária/métodos , Valores de ReferênciaRESUMO
Assays measuring platelet aggregation (thrombus formation) at arterial shear rate mostly use collagen as only platelet-adhesive surface. Here we report a multi-surface and multi-parameter flow assay to characterize thrombus formation in whole blood from healthy subjects and patients with platelet function deficiencies. A systematic comparison is made of 52 adhesive surfaces with components activating the main platelet-adhesive receptors, and of eight output parameters reflecting distinct stages of thrombus formation. Three types of thrombus formation can be identified with a predicted hierarchy of the following receptors: glycoprotein (GP)VI, C-type lectin-like receptor-2 (CLEC-2)>GPIb>α6ß1, αIIbß3>α2ß1>CD36, α5ß1, αvß3. Application with patient blood reveals distinct abnormalities in thrombus formation in patients with severe combined immune deficiency, Glanzmann's thrombasthenia, Hermansky-Pudlak syndrome, May-Hegglin anomaly or grey platelet syndrome. We suggest this test may be useful for the diagnosis of patients with suspected bleeding disorders or a pro-thrombotic tendency.
Assuntos
Bioensaio/métodos , Trombose/diagnóstico , Trombose/metabolismo , Adulto , Biologia Computacional , Feminino , Humanos , Masculino , Pessoa de Meia-Idade , Agregação Plaquetária/fisiologia , Adulto JovemRESUMO
Patients with chronic myeloproliferative leukemia (CML) have frequent haemorrhage and/or thrombosis in their medical history. The mechanisms of these major and life-threatening complications remain unclear. Membrane organization influences many of the unique cellular functions and is strongly correlated, among other factors, to the membrane lipid composition; it may be evaluated by following up the membrane fluidity and aggregation properties of the platelet. In this study, we evaluated the platelet aggregation, the expression of platelet surface receptors, the membrane fluidity (as evaluated by fluorescence anisotropy) and its correlation to reactive oxygen species (ROS) production, in patients with chronic myeloid leukaemia (CML). It was found that the patients in accelerated and blastic phase of CML present an altered platelet aggregation response to all reagents except for ristocetin as compared with chronic phase group, which shows only epinephrine-altered response. We also found that BCR/ABL transcript leads to higher levels of ROS in accelerated and blastic CML phases. Patients without molecular remission have lower platelet membrane fluidity. We obtained a positive correlation between ROS level and membrane fluorescence anisotropy changes. The CD41 expression was decreased in CML patients and P selectin expression was found to be higher in these patients than in healthy volunteers. Platelets of CML patients have altered aggregation parameters in accelerated and blastic phases, in which BCR/ABL transcript level is increased. The increased level of ROS in CML patients without molecular remission is associated with a decrease in fluidity of platelet membrane and expression of CD41/CD61 receptors. These findings may contribute to understanding the mechanism of the altered platelet response reported in CML patients.
Assuntos
Plaquetas/patologia , Leucemia Mielogênica Crônica BCR-ABL Positiva/sangue , Idoso , Plaquetas/metabolismo , Feminino , Humanos , Leucemia Mielogênica Crônica BCR-ABL Positiva/patologia , Masculino , Pessoa de Meia-Idade , Estadiamento de Neoplasias , Agregação Plaquetária/fisiologia , Prognóstico , Estudos Prospectivos , Espécies Reativas de Oxigênio/metabolismoAssuntos
Procedimentos Cirúrgicos do Sistema Digestório , Indicadores Básicos de Saúde , Hemorragia/sangue , Hemorragia/induzido quimicamente , Período Perioperatório , Inibidores da Agregação Plaquetária/efeitos adversos , Inibidores da Agregação Plaquetária/uso terapêutico , Agregação Plaquetária/fisiologia , Tromboembolia/prevenção & controle , Vísceras/cirurgia , Algoritmos , Aspirina/efeitos adversos , Aspirina/uso terapêutico , Doenças Cardiovasculares/sangue , Doenças Cardiovasculares/tratamento farmacológico , Clopidogrel , Substituição de Medicamentos , Quimioterapia Combinada , Hemorragia/mortalidade , Heparina/efeitos adversos , Heparina/uso terapêutico , Humanos , Agregação Plaquetária/efeitos dos fármacos , Complexo Glicoproteico GPIIb-IIIa de Plaquetas/antagonistas & inibidores , Medição de Risco , Tromboembolia/sangue , Tromboembolia/mortalidade , Ticlopidina/efeitos adversos , Ticlopidina/análogos & derivados , Ticlopidina/uso terapêuticoRESUMO
BACKGROUND: Early diagnosis of immune heparin-induced thrombocytopenia (HIT) is challenging. HemosIL® AcuStar HIT and heparin-induced multiple electrode aggregometry (HIMEA) were recently proposed as rapid diagnostic methods. OBJECTIVES: We conducted a study to assess performances of AcuStar HIT-IgG (PF4-H) and AcuStar HIT-Ab (PF4-H). The secondary objective was to compare the performances of the combination of Acustar HIT and HIMEA with standardised clinical diagnosis. METHODS: Sera of 104 suspected HIT patients were retrospectively tested with AcuStar HIT. HIMEA was performed on available sera (n=81). The clinical diagnosis was established by analysing in a standardized manner the patient's medical records. These tests were also compared with PF4-Enhanced®, LTA, and SRA in subsets of patients. Thresholds were determined using ROC curve analysis with clinical outcome as reference. RESULTS: Using the recommended thresholds (1.00AU), the negative predictive value (NPV) of HIT-IgG and HIT-Ab were 100.0% (95% CI: 95.9%-100.0% and 95.7%-100.0%). The positive predictive value (PPV) were 64.3% (95% CI: 35.1%-87.2.2%) and 45.0% (95% CI: 23.2%-68.6%), respectively. Using our thresholds (HIT-IgG: 2.89AU, HIT-Ab: 9.41AU), NPV of HIT-IgG and HIT-Ab were 100.0% (95% CI: 96.0%-100.0% and 96.1%-100.0%). PPV were 75.0% (95% CI: 42.7%-94.5%) and 81.8% (95% CI: 48.3%-97.7%), respectively. Of the 79 patients with a medium-high pretest probability score, 67 were negative using HIT-IgG (PF4-H) test at our thresholds. HIMEA was performed on HIT-IgG positive patients. Using this combination, only one patient on 79 was incorrectly diagnosed. CONCLUSION: Acustar HIT showed good performances to exclude the diagnosis of HIT. Combination with HIMEA improves PPV.
Assuntos
Heparina/efeitos adversos , Medições Luminescentes/métodos , Agregação Plaquetária/efeitos dos fármacos , Testes de Função Plaquetária/métodos , Trombocitopenia/induzido quimicamente , Trombocitopenia/diagnóstico , Adulto , Idoso , Automação , Estudos de Casos e Controles , Eletrodos , Ensaio de Imunoadsorção Enzimática , Feminino , Heparina/imunologia , Humanos , Medições Luminescentes/instrumentação , Masculino , Pessoa de Meia-Idade , Agregação Plaquetária/fisiologia , Estudos Retrospectivos , Trombocitopenia/sangue , Trombocitopenia/imunologiaRESUMO
BACKGROUND: Coagulopathy is common after cardiopulmonary bypass (CPB), and platelet dysfunction is frequently considered to be a major contributor to excessive bleeding. Exposure to hypothermia may exacerbate the platelet function defect. We assessed platelet function during and after deep hypothermia with multiple electrode aggregometry (Multiplate(®); Verum Diagnostica GmbH, Munich, Germany). METHODS: Twenty adult patients undergoing pulmonary endarterectomy for chronic pulmonary hypertension were cooled on CPB to 20°C and deep hypothermic arrest was used to facilitate surgery. We analyzed platelet aggregation in whole blood samples at 12 measuring points during and after the procedure. Platelet aggregation was stimulated via the thrombin receptor (TRAPtest) at the patient's actual body temperature (AUC-CT) and after rewarming the samples to 37°C (AUC-37). In addition, we tested samples at 2 time points after 2 minutes of in vitro incubation with 20 µg protamine (0.067 µg/µL). Results are expressed as area under the aggregation curve (AUC). RESULTS: Cooling resulted in a marked decrease of platelet aggregation to a minimum AUC-CT of 20.5 (95% confidence interval [CI] 8.9-32.1) at 20°C body temperature. AUC-CT was significantly different from baseline (92.8, 95% CI 82.5-103.1) for temperatures of ≤28°C (P < 0.001), whereas the change in AUC-37 only became significant at the lowest body temperature (59.4, 95% CI 41.3-77.4). After rewarming to 36°C, AUC-CT and AUC-37 had recovered to 67.6 (95% CI 53.9-81.3) and 71.7 (95% CI 52.5-90.8), respectively. The mean AUC-CT was significantly lower than the mean AUC-37 from cooling at 28°C to warming at 24°C inclusive, and the relationship with temperature during cooling was significantly different between AUC-CT and AUC-37 (regression coefficients 4.7 [95% CI 4.2-5.2] vs 1.3 [95% CI 0.7-1.9]; P < 0.0001). After administration of protamine, mean aggregation decreased significantly for both measurements by 38.2 (95% CI -27.9 to -48.5; P < 0.001) and 44.5 (95% CI -58.5 to -30.5; P < 0.001), respectively. Similarly, adding protamine in vitro resulted in a decrease of mean aggregation by 35.1 (95% CI -71.0 to 0.8; P = 0.055) when measured after administration of heparin, and 56.5 (95% CI -94.5 to -18.5; P = 0.005) at the end of CPB. CONCLUSION: Platelet aggregation, assessed by multiple electrode aggregometry (Multiplate), was severely affected during deep, whole-body hypothermia. This effect was partially reversible after rewarming, and was distinct from a general decline of platelet aggregation during CPB. Protamine also caused a significant decrease in platelet aggregation in vivo and in vitro.
Assuntos
Ponte Cardiopulmonar/métodos , Hipotermia Induzida/métodos , Agregação Plaquetária/efeitos dos fármacos , Sistemas Automatizados de Assistência Junto ao Leito , Protaminas/farmacologia , Adulto , Eletrodos , Humanos , Agregação Plaquetária/fisiologia , Testes de Função Plaquetária/métodosRESUMO
A non-invasive method for the study of platelet aggregation and formation of leukocyte-erythrocyte-platelet aggregates as well as certain hemostatic parameters is proposed. The method is based on the speckle-analysis of coherent light scattering from the surface of erythrocytes moving in an artificially isolated vessel segment. It was shown that light scattering index significantly correlated with ADP-, adrenalin-, or collagen-induced platelet aggregation, with the formation of leukocyte-erythrocyte or platelet-erythrocyte aggregates, and with the levels of fibrinogen, soluble fibrin-monomer complexes and related parameters. It is concluded that the proposed method for the study of hemostatic system can be used to roughly evaluate intensity of intravascular blood coagulation and probability of thrombosis.
Assuntos
Plaquetas/metabolismo , Agregação Eritrocítica/fisiologia , Eritrócitos/metabolismo , Hemostasia/fisiologia , Leucócitos/metabolismo , Agregação Plaquetária/fisiologia , Trombose/diagnóstico , Adolescente , Adulto , Idoso , Análise Química do Sangue/métodos , Coagulação Sanguínea , Criança , Pré-Escolar , Feminino , Humanos , Lactente , Recém-Nascido , Masculino , Pessoa de Meia-Idade , Trombose/sangue , Adulto JovemRESUMO
The quality of platelet aggregation and dense granule deficiency testing is important for diagnosing platelet function disorders. After a successful pilot exercise on diagnosing platelet dense granule deficiency by electron microscopy (EM), the North American Specialized Coagulation Laboratory Association (NASCOLA) has launched regular external quality assurance (EQA) for dense granule EM, as well as for the interpretation of platelet aggregation findings. EQA records were analyzed to assess performance. For EM EQA, between 2009 and 2011, there was excellent performance in distinguishing normal from dense granule-deficient samples and good (>70%) agreement on classifying most electron dense structures in platelets. For aggregation EQA, some normal variants were misclassified and overall case interpretations were more acceptable for rare disorders than for common findings. NASCOLA experiences with these EQAs indicate that there is a need to improve the quality of platelet disorder evaluations. For aggregometry interpretations, deficits in performance could be addressed by translating guideline recommendations into practice.
Assuntos
Transtornos Plaquetários/sangue , Transtornos Plaquetários/diagnóstico , Plaquetas/patologia , Grânulos Citoplasmáticos/patologia , Testes de Função Plaquetária/métodos , Garantia da Qualidade dos Cuidados de Saúde/métodos , Plaquetas/ultraestrutura , Grânulos Citoplasmáticos/ultraestrutura , Coleta de Dados , Humanos , Microscopia Eletrônica , Agregação Plaquetária/fisiologia , Testes de Função Plaquetária/normas , Controle de QualidadeRESUMO
OBJECTIVES: Evaluation of aspirin (ASA) responsiveness with platelet function tests varies by the choice of blood mixture and functional test and cut off values for defining the the treatment used. Addition to that we also aimed to determine agreement between three tests and to research whether there is any necessity to measure baseline platelet activity. METHODS: The study group comprised of 52 patients with multiple risk factors receiving primary prophylaxis of ASA (100 mg/day). For each patient inhibition of platelet aggregation with aspirin was determined using three different whole blood tests: Multiplate electrical impedance aggregometry, Verify Now Aspirin, and collagen-epinephrine closure time PFA-100. Platelet aggregation was assessed with multiplate electrical impedance aggregometry,and was defined as the area under curve (AUC,AUxmin). Maximal 6,4 microM collagen-induced AUC were used to quantify platelet aggregation due to ASA. The ASA response was defined as >30 % reduction in basal platelet aggregation with multiplate electrical impedance aggregometry. Collagen induced platelet aggregation at the Verify Now Aspirin assay quantitated the ASA-induced platelet inhibition as aspirin reaction units (ARU). According to manufacturer insert ARU>550 indicates aspirin resistance. ASA platelet function studies were assessed twice at baseline (pre-aspirin), and after 7 day(post-aspirin) were performed. RESULTS: After ASA intake none of the patients was found aspirin resistant with PFA-100. (CEPI-CT (129+/-36 vs 289+/-18 ). None of the patients was found aspirin resistant with PFA-100. As>30 % reduction in basal platelet aggregation with multiplate electrical impedance aggregometry is selected all of the patients have been stratified as responders.(COL TEST 688+/-230 vs 169+/-131 AU) None of the patients with Verify Now Aspirin found resistance to ASA(594+/-62 vs 446+/-43).Prior to ASA intake 15 of all patients with VN(501+/-16) and 2 of all patients with multiplate electrical impedance aggregometry (223+/-40 AUC )aggregation levels below the cut off label before ingestion of ASA.None of the patients was above the cut off label with PFA -100 (129+/-36). CONCLUSIONS: Verify Now ASA assay, multiplate electrical impedance aggregometry and PFA-100 seem to be reliable tests in reflecting ASA effect on platelets. Cut off labels for the defining the responsiveness given by manufacturer may show significant interindividual variability with Verify Now ASA assay and multiplate electrical impedance aggregometry, and these test may show platelet inhibition despite the absence of ASA intake. Consideration of the pretreatment values may eliminate the risk of overestimation in the assessment of platelet inhibition by ASA.
Assuntos
Aspirina/uso terapêutico , Plaquetas/fisiologia , Agregação Plaquetária/fisiologia , Testes de Função Plaquetária/instrumentação , Testes de Função Plaquetária/métodos , Aspirina/administração & dosagem , Aspirina/farmacologia , Bioensaio/instrumentação , Plaquetas/efeitos dos fármacos , Técnicas de Laboratório Clínico/instrumentação , Colágeno/farmacologia , Impedância Elétrica , Epinefrina/farmacologia , Hemostasia/efeitos dos fármacos , Hemostasia/fisiologia , Humanos , Agregação Plaquetária/efeitos dos fármacos , Contagem de Plaquetas/instrumentação , Risco , Fatores de RiscoRESUMO
BACKGROUND: In this study, we explored whether antiplatelet medications impair whole blood impedance aggregometry after cardiac surgery and cardiopulmonary bypass (CPB) compared with classical light transmission aggregometry (LTA). METHODS: Multiplate (M) assays measuring changes in electrical resistance as aggregation units over time, and LTA assays (% aggregation) induced by collagen (COL), adenosine diphosphate (ADP), or arachidonic acid were performed simultaneously using arterial blood samples obtained before induction of anesthesia, 15 min and 3 h after neutralization of heparin in 70 consecutive patients scheduled for elective coronary artery bypass grafting. Patients in Group A (n = 48) discontinued intake of antiplatelet drugs for at least 7 days and served as controls, patients in Group B (n = 11) received aspirin 100 mg/d and those in Group C (n = 11) aspirin 100 mg/d and clopidogrel 75 mg/d (dual antiplatelet therapy) until the day before surgery. RESULTS: In patients without antiplatelet therapy, 15 min and 3 h after protamine a significant decrease in platelet aggregation was observed with all three agonists and both aggregation methods. In patients receiving aspirin alone, LTA-COL, LTA-ADP and M-ADP changed significantly over time, and ADP assays of both aggregation methods showed a significant decrease in platelet aggregation 15 min after protamine in patients receiving dual antiplatelet therapy. When calculating the areas under the receiver-operating characteristic curves for discrimination of antiplatelet agents, LTA-COL was able to discriminate between controls and patients receiving aspirin or dual antiplatelet therapy 15 min and 3 h after CPB and the M-ADP assay was able to discriminate between controls and patients receiving dual antiplatelet therapy 3 h after protamine. CONCLUSION: Whole blood and classical LTA performed with all commonly used agonists enable detection of CPB-induced changes in platelet aggregation in patients not taking antiplatelet medication, whereas in patients receiving antiplatelet therapy, ADP-induced antiplatelet assays are preferable for detecting CPB-induced impairment of platelet aggregation.
Assuntos
Plaquetas/fisiologia , Ponte Cardiopulmonar/efeitos adversos , Agregação Plaquetária/fisiologia , Testes de Função Plaquetária , Idoso , Anestesia , Aspirina/uso terapêutico , Perda Sanguínea Cirúrgica , Clopidogrel , Feminino , Humanos , Masculino , Pessoa de Meia-Idade , Projetos Piloto , Inibidores da Agregação Plaquetária/uso terapêutico , Testes de Função Plaquetária/métodos , Curva ROC , Ticlopidina/análogos & derivados , Ticlopidina/uso terapêuticoRESUMO
Light transmission aggregometry (LTA) is the gold standard for the study of patients with defects of platelet function. Use of LTA in clinical practice for predicting the risk of thrombosis or monitoring the pharmacologic effects of antiplatelet agents should be discouraged, because not only is the monitoring of treatment with antiplatelet agents (with any laboratory test) not indicated at present, but also the lack of standardization of the technique for LTA makes it additionally unsuitable for this purpose. The need for standardization of LTA has recently been emphasized by the results of four surveys, which showed that there is a wide variation in the methodology used worldwide. A modification of the traditional LTA is the lumiaggregometer, which measures platelet secretion in parallel with platelet aggregation. This technique is probably preferable to traditional LTA in the diagnostic workup of patients with inherited defects of platelet function, because it is more sensitive to the most common disorders, which are characterized by abnormalities of platelet secretion. LTA (or lumiaggregometry) is useful as a first screening test of patients with the clinical suspicion of defects of platelet function because it helps to provide an interim diagnostic hypothesis, which can then be confirmed or discounted using appropriate and specific tests.
Assuntos
Trifosfato de Adenosina/sangue , Transtornos Plaquetários/diagnóstico , Plaquetas/metabolismo , Agregação Plaquetária/fisiologia , Transtornos Plaquetários/sangue , Humanos , Testes de Função Plaquetária/métodosRESUMO
Platelet function testing is essential for the diagnosis of congenital/acquired bleeding disorders and may be useful for the prediction of surgical bleeding. Nowadays there is also much interest in monitoring the efficacy of anti-platelet therapy and measuring platelet hyper-function. However, this often presents clinical laboratories with significant challenges as platelet function tests are complex, poorly standardized, time consuming and quality assurance is not straightforward. There are also few comprehensive modern guidelines available and many recent published surveys have revealed poor standardization between laboratories. Up until the late 1980's the traditional clinical platelet function tests that were available were the bleeding time (BT), light transmission (LTA) and whole blood aggregometry (WBA) and various biochemical assays. These were also usually performed within specialized research and clinical laboratories. Since the last BCSH guidelines were published in 1988 a variety of new platelet function tests have become available. These include flow cytometry and an ever increasing choice of new commercial instruments. Although the potential clinical utility of the new assays is emerging some have not yet entered into routine clinical practice. It is encouraging that a number of standardization committees (e. g. CLSI, BCSH and ISTH Platelet Physiology SSC) are now beginning to produce new platelet function testing guidelines and this will hopefully improve clinical practice, quality assurance and result in less variability between different laboratories.
Assuntos
Plaquetas/fisiologia , Agregação Plaquetária/fisiologia , Testes de Função Plaquetária/métodos , Tempo de Sangramento , Plaquetas/citologia , Desenho de Equipamento , Citometria de Fluxo , Humanos , Testes de Função Plaquetária/instrumentação , Testes de Função Plaquetária/normas , Resistência ao CisalhamentoRESUMO
The main aim of the study is to estimate a state of thrombocytic and coagulation haemostasis of elderly and senile patients with ischemic heart disease through using a combination of magnitotherapy and hyperbaric oxygenation. 108 patients aged between 70 and 85 years are recruited so far. The range of diagnoses varies from ischemic heart disease, stable angina of II functional class. The studies indicators include thrombocytes aggregation and coagulation which are provided before and after treatment. It has been shown that the results are more positive when magnitotherapy and hyperbaric oxygenation are included in the treatment. That is effected in positive dynamic of thrombocytes spontaneous and induced aggregation, coagulation haemostasis indicators and fibrinolytic activity, which characterize improvement of cardiovascular system functioning.
Assuntos
Envelhecimento/sangue , Coagulação Sanguínea , Oxigenoterapia Hiperbárica , Magnetoterapia , Isquemia Miocárdica/terapia , Agregação Plaquetária , Idoso , Idoso de 80 Anos ou mais , Coagulação Sanguínea/fisiologia , Feminino , Humanos , Masculino , Isquemia Miocárdica/sangue , Agregação Plaquetária/fisiologia , Resultado do TratamentoRESUMO
To treat acute lung failure, an intravenous membrane gas exchange device, the Hattler Catheter, is currently under development. Several methods were employed to evaluate the biocompatibility of the device during preclinical testing in bovines, and potential coatings for the fibers comprising the device were screened for their effectiveness in reducing thrombus deposition in vitro. Flow cytometric analysis demonstrated that the device had the capacity to activate platelets as evidenced by significant increases in circulating platelet microaggregates and activated platelets. Thrombus was observed on 20 +/- 6% of the surface area of devices implanted for up to 53 h. Adding aspirin to the antithrombotic therapy permitted two devices to remain implanted up to 96 h with reduced platelet activation and only 3% of the surface covered with thrombus. The application of heparin-based coatings significantly reduced thrombus deposition in vitro. The results suggest that with the use of appropriate antithrombotic therapies and surface coatings the Hattler Catheter might successfully provide support for acute lung failure without thrombotic complications.