RESUMEN
BACKGROUND: In breast cancer patients routine thromboprophylaxis is not recommended but individualized risk assessment is encouraged. The incorporation of hypercoagulability biomarkers could increase the sensitivity of risk assessment models (RAM) to identify patients at VTE risk. To this aim we investigated the impact of cancer-related characteristics on hypercoagulability biomarkers. METHODS: Thrombin generation (TG) assessed with the Thrombogramme-Thrombinoscope®, levels of platelet derived microparticles (Pd-MP) assessed with flow cytometry, procoagulant phospholid dependent clotting time (PPL-ct) measured with a clotting assay and D-Dimers (were assessed in a cohort of 62 women with breast cancer and in 30 age matched healthy women. RESULTS: Patients showed significantly higher TG, Pd-MP, D-Dimers levels and shortened PPL-ct compared to the controls. The PPL-ct was inversely correlated with the levels of Pd-MP, which were increased in 97% of patients. TG and D-Dimers were increased in 76% and 59% of patients respectively. In any stage of the disease TG was significantly increased as compared to the controls. There was no significant difference of TG in patients with local, regional of metastatic stage. There was no significant difference in Pd-MP or Pd-MP/PS+ between the subgroups of patients with local or regional stage of cancer. Patients with metastatic disease had significantly higher levels of Pd-MP and Pd-MP/PS+ compared to those with regional stage. The D-Dimers increased in patients with metastatic stage. In patients on chemotherapy with less than 6 months since diagnosis TG was significantly higher compared to those on chemotherapy who diagnosed in interval > 6 months. Patients with metastatic disease had significantly higher levels of Pd-MP and D-Dimers compared to those with non-metastatic disease. CONCLUSION: In breast cancer patients the stage, the time elapsed since the diagnosis and the administration of chemotherapy are determinants of cellular and plasma hypercoagulability. The levels and the procoagulant activity of Pd-MP are interconnected with the biological activity and the overall burden of cancer. TG reflects the procoagulant properties of both breast cancer and chemotherapy in the initial period of cancer diagnosis. Thus the weighted incorporation of the biomarkers of cellular and plasma hypercoagulabilty in RAM for VTE might improve their predictive value.
Asunto(s)
Biomarcadores/sangre , Neoplasias de la Mama/tratamiento farmacológico , Neoplasias de la Mama/patología , Adulto , Anciano , Biomarcadores/metabolismo , Pruebas de Coagulación Sanguínea , Femenino , Productos de Degradación de Fibrina-Fibrinógeno/metabolismo , Humanos , Persona de Mediana Edad , Factores de Riesgo , Trombina/metabolismo , Trombofilia/sangre , Trombofilia/metabolismoRESUMEN
Low molecular weight heparins (LMWHs) and fondaparinux are widely used for prophylaxis and treatment of venous thromboembolic disease in cancer patients. However, the optimization of the antithrombotic treatment especially in patients with adenocarcinoma of the pancreas is a challenging issue. The understanding of the mechanism of action of the LMWHs and fondaparinux in cancer-induced hypercoagulability might help to optimize antithrombotic treatment. To this aim, we investigated the influence of BXPC3 pancreas adenocarcinoma cells on the antithrombotic activity of LMWHs and fondaparinux. Thrombin generation (TG) in normal platelet poor (PPP) and platelet rich plasma (PRP) spiked with clinically relevant concentrations of dalteparin, enoxaparin, nadroparin tinzaparin and fondaparinux was assessed with the Calibrated Automated Thrombogram assay. BXPC3 (5 cells/µl) were added to plasma. The mean rate index (MRI) of the propagation phase of TG and the endogenous thrombin potential (ETP) were analyzed. The IC50 of the studied compounds were determined and compared on the basis of anti-Xa and anti-IIa equivalent units. We demonstrate that the specific antithrombin (AT)-dependent anti-Xa activity of LMWHs and fondaparinux almost selectively inhibits the propagation phase of TG. The synergy between the anti-Xa and anti-IIa activities of LMWHs rather than the selective inhibition of FXa warrants abrogation of TG. The mean molecular weight and anti-Xa/anti-IIa ratio of the AT-dependent agents cannot predict the alteration of their capacity to inhibit TG. Tinzaparin was the most potent inhibitor of TG than the other LMWHs. Enoxaparin was more potent than nadroparin and dalteparin.
Asunto(s)
Anticoagulantes/uso terapéutico , Heparina de Bajo-Peso-Molecular/uso terapéutico , Neoplasias Pancreáticas/metabolismo , Polisacáridos/uso terapéutico , Trombina/antagonistas & inhibidores , Trombina/metabolismo , Anticoagulantes/farmacología , Fondaparinux , Heparina de Bajo-Peso-Molecular/farmacología , Humanos , Neoplasias Pancreáticas/tratamiento farmacológico , Polisacáridos/farmacología , Resultado del TratamientoRESUMEN
Platelets play a pivotal role in the regulation of both thrombosis and haemostasis. Functional testing of platelet response has been exclusively used in the diagnosis and management of bleeding disorders. Recent advances of light transmission aggregometry and development of more useful devices have demonstrated the clinical utility to enlarge platelet function testing in patients with cardiovascular disease. The ex vivo measurement of residual platelet response seems, with some assays, predictive of adverse clinical events. Still a debate, it represents an emerging area of interest for both the clinician and the basic scientist. Heparin-induced thrombocytopenia diagnosis is also difficult and a functional assay is now available for an easier and rapid method to rule out such a life-threatening situation. This review article will describe the available methods of measuring platelet response and will discuss both the limitations and emerging data supporting the role of platelet function studies in clinical practice.
Asunto(s)
Trastornos de la Coagulación Sanguínea/diagnóstico , Plaquetas/fisiología , Práctica Profesional , Trastornos de la Coagulación Sanguínea/fisiopatología , Trastornos de la Coagulación Sanguínea/terapia , Recolección de Muestras de Sangre/normas , Impedancia Eléctrica , Humanos , Mediciones Luminiscentes/métodos , Monitoreo Fisiológico/métodos , Fotometría/métodos , Agregación Plaquetaria/fisiología , Pruebas de Función Plaquetaria/métodos , Valor Predictivo de las PruebasRESUMEN
BACKGROUND: Patients with lung adenocarcinoma undergoing surgery are in high risk for VTE and receive routine post-operative thromboprophylaxis with LWMH. AIM: We investigated markers of hypercoagulability in patients with primary localized adenocarcinoma and the modifications induced by lobectomy and postoperative administration of enoxaparin. MATERIALS AND METHODS: Patients suffering from localised primary lung adenocarcinoma (n=15) scheduled for lobectomy were studied. The control group consisted of 15 healthy age and sex-matched individuals. Blood was collected before anaesthesia induction and after surgery, at several intervals until the 7th post-operative day. Samples were assessed for thrombin generation, phosphatidylserin expressing platelet derived microparticles expressing (Pd-MP/PS(+)), tissue factor activity (TFa), FVIIa and TFPI levels, procoagulant phospholipid dependent clotting time and anti-Xa activity. RESULTS: At baseline, patients showed increased thrombin generation and Pd-MP/PS(+). After lobectomy thrombin generation significantly decreased. Administration of enoxaparin attenuated thrombin generation. In about 50% of samples collected post-operatively an increase of thrombin generation occurred despite the presence of the expected anti-Xa activity in plasma. At the 7th post-operative day, 3 out of 15 patients showed a significant increase of thrombin generation. CONCLUSION: In patients with localized lung adenocarcinoma, hypercoagulability is characterized by high thrombin generation and increased concentration of Pd-MP/PS(+). Tumor mass resection is related with attenuation of thrombin generation, which is inhibited by postoperative thromboprophylaxis with enoxaparin. The response to enoxaparin is not predicted by the concentration of the anti-Xa activity in plasma. The assessment of thrombin generation during prophylaxis with enoxaparin allows to identify patients with high residual plasma hypercoagulability.
Asunto(s)
Adenocarcinoma/sangre , Adenocarcinoma/cirugía , Anticoagulantes/uso terapéutico , Enoxaparina/uso terapéutico , Neoplasias Pulmonares/sangre , Neoplasias Pulmonares/cirugía , Pulmón/cirugía , Trombofilia/tratamiento farmacológico , Adenocarcinoma/complicaciones , Adenocarcinoma del Pulmón , Anciano , Coagulación Sanguínea/efectos de los fármacos , Pruebas de Coagulación Sanguínea , Plaquetas/efectos de los fármacos , Plaquetas/patología , Micropartículas Derivadas de Células/patología , Inhibidores del Factor Xa , Femenino , Humanos , Neoplasias Pulmonares/complicaciones , Masculino , Persona de Mediana Edad , Periodo Posoperatorio , Trombina/análisis , Trombofilia/sangre , Trombofilia/complicaciones , Trombofilia/patología , Tromboplastina/análisisRESUMEN
Heparin-induced thrombocytopenia (HIT) is a potentially lethal adverse effect of heparin therapy. Accurate and rapid HIT laboratory diagnosis when HIT is suspected is crucial. The combination of an immunological assay with a functional test improves the accuracy of HIT, but functional assays are currently limited to a few laboratories. Multiplate® analyzer (Dynabyte, Munich, Germany) is a practical, semi-automated and easy-to-perform platelet aggregation assay. The aim of this study is to explore whether heparin-induced platelet aggregation in whole blood assessed by Multiplate® (Heparin-induced multiple electrode aggregometry, HIMEA) can replace platelet aggregation test (PAT) in platelet-rich plasma. For this purpose, HIMEA performance in HIT diagnosis was prospectively evaluated. HIMEA and PAT were compared to serotonin-release assay (SRA) in 200 well-characterized consecutive patients suspected for HIT. HIMEA was found to be more sensitive (81% vs. 76%) and more specific (99% vs. 96%) than PAT compared to SRA. Both tests showed a high negative predictive value while HIMEA had a better positive predictive value. HIMEA has overall better performance characteristics than PAT for the detection of HIT platelet-activating antibodies. The combination of an immunological assay with HIMEA could be a feasible option in non-specialized laboratories for HIT diagnosis optimization.
Asunto(s)
Anticoagulantes/efectos adversos , Heparina/efectos adversos , Trombocitopenia/inducido químicamente , Trombocitopenia/diagnóstico , Anticuerpos/inmunología , Ensayo de Inmunoadsorción Enzimática , Humanos , Agregación Plaquetaria , Factor Plaquetario 4/inmunología , Pruebas de Función Plaquetaria , Estudios Prospectivos , Sensibilidad y EspecificidadRESUMEN
We have previously demonstrated that matrix metalloproteinase-9 (MMP-9) is critical for breast cancer cell migration and is necessary but not sufficient for tubular network formation. Given the important angiogenic activity of vascular endothelial growth factor (VEGF), we investigate here its possible contribution in tubular network formation and its link with MMP-9. Exposure of resistant epithelial breast cancer cells (rMCF-7) to Avastin, a VEGF neutralising antibody, suppresses tubular network formation but not cell migration. However, their exposure to MMP-9 inhibitor markedly decreases both parameters. Besides, the addition of exogenous VEGF or MMP-9 alone or in combination to sensitive parental cells (sMCF-7) or rMCF-7 cells enhances tubular network formation by rMCF-7 cells but not by sMCF-7 cells. The evaluation of the expression levels of VEGF receptor (VEGFR) subtypes shows that sMCF-7 cells express only small quantities of VEGFR-2 and VEGFR-3 compared with rMCF-7 cells that express strong quantities. However, treatment of sMCF-7 cells by phorbol 12-myristate 13-acetate (PMA), a PKC activator, induces both tubular network formation and VEGFR-2/VEGFR-3 over-expressions. Interestingly, exposure of rMCF-7 cells or PMA-treated sMCF-7 cells to the specific inhibitors of VEGFR-2 and VEGFR-3 reduces markedly the tubular network formation. Together, our results demonstrate that the proteolytic enzyme MMP-9 promotes rMCF-7 cell migration and, consequently, tubular network formation through VEGFR-2/ VEGFR-3 activation. Understanding of mechanisms involved in vasculogenic mimicry and cell migration related to MMP-9 and VEGF may open new opportunities to improve cancer therapy.
Asunto(s)
Neoplasias de la Mama/patología , Metaloproteinasa 9 de la Matriz/genética , Receptor 2 de Factores de Crecimiento Endotelial Vascular/genética , Receptor 3 de Factores de Crecimiento Endotelial Vascular/genética , Adenocarcinoma/tratamiento farmacológico , Adenocarcinoma/genética , Adenocarcinoma/patología , Antibióticos Antineoplásicos/farmacología , Anticuerpos Monoclonales Humanizados/farmacología , Bevacizumab , Neoplasias de la Mama/tratamiento farmacológico , Neoplasias de la Mama/genética , Línea Celular Tumoral , Movimiento Celular , Doxorrubicina/farmacología , Resistencia a Antineoplásicos , Femenino , Regulación Neoplásica de la Expresión Génica , Humanos , Metaloproteinasa 9 de la Matriz/administración & dosificación , Acetato de Tetradecanoilforbol/farmacología , Factor A de Crecimiento Endotelial Vascular/administración & dosificación , Factor A de Crecimiento Endotelial Vascular/metabolismoRESUMEN
Sickle cell disease (SCD) is linked to hypercoagulability and is characterised by high concentrations of erythrocyte-derived microparticles (Ed-MPs). However, the impact of procoagulant cell-derived microparticles on the thrombin generation process remains unclear. We analysed the alterations of each phase of thrombin generation (TG) in relation to the concentration of erythrocyte- or platelet-derived microparticles (Ed-MPs and Pd-MPs) in a cohort of patients with steady-state SCD. We studied 92 steady-state SCD patients, 19 of which were under treatment with hydroxyurea, and 30 healthy age- and sex-matched individuals. TG was assessed by calibrated automated thrombogram. Ed-MP and Pd-MP expressing or not phosphatidylserine (PS) were determined by means of flow cytometry. Procoagulant phospholipid-dependent activity in the plasma was evaluated by the Procoag-PPL assay. Levels of thrombomodulin and haemoglobin in the plasma as well as red blood cell and reticulocyte counts were measured. SCD patients, independently of the administration of hydroxyurea, were marked by a significant acceleration in the propagation phase of TG which correlated with the Ed-MP/PS+ concentration. TG was significantly attenuated in hydroxyurea-treated patients. In conclusion, the acceleration of the propagation phase of TG, driven by Ed-MP/PS+, is a major functional alteration in blood coagulation in patients with steady-state SCD. Treatment with hydroxyurea, in addition to the regulation of haemolysis, lowers Ed-MPs and attenuates thrombin generation. The thrombogram could be a useful tool for the diagnosis of hypercoagulability and optimisation of the treatment in patients with SCD.
Asunto(s)
Anemia de Células Falciformes/complicaciones , Coagulación Sanguínea , Micropartículas Derivadas de Células/metabolismo , Eritrocitos/metabolismo , Trombina/metabolismo , Trombofilia/etiología , Adolescente , Adulto , Anemia de Células Falciformes/sangre , Anemia de Células Falciformes/tratamiento farmacológico , Antidrepanocíticos/uso terapéutico , Pruebas de Coagulación Sanguínea , Estudios de Casos y Controles , Micropartículas Derivadas de Células/efectos de los fármacos , Eritrocitos/efectos de los fármacos , Femenino , Citometría de Flujo , Hemoglobinas/metabolismo , Humanos , Hidroxiurea/uso terapéutico , Masculino , Paris , Fosfatidilserinas/sangre , Trombomodulina/sangre , Trombofilia/sangre , Factores de Tiempo , Adulto JovenAsunto(s)
Anticoagulantes/farmacología , Coagulación Sanguínea/efectos de los fármacos , Morfolinas/farmacología , Tiempo de Tromboplastina Parcial , Tiempo de Protrombina , Tiofenos/farmacología , Biomarcadores/sangre , Factores de Coagulación Sanguínea/metabolismo , Relación Dosis-Respuesta a Droga , Humanos , Valor Predictivo de las Pruebas , Reproducibilidad de los Resultados , Rivaroxabán , Factores de TiempoRESUMEN
Cancer histology influences the risk of venous thromboembolism and tissue factor (TF) is the key molecule in cancer-induced hypercoagulability. We investigated the relation between TF expression by pancreatic and breast cancer cells (BXPC3 and MCF7 respectively) and their capacity to trigger in vitro thrombin generation in normal human plasma. Flow cytometry and Western blot analysis for TF expression were performed using murine IgG1 monoclonal antibody against human TF. Real-time PCR for TFmRNA was also performed. Activity of TF expressed by cancer cells was measured with a specific chromogenic assay. Thrombin generation in PPP was assessed using calibrated automated thrombogram. Cancer cells were added to platelet poor plasma from healthy volunteers. In separate experiments cells were incubated with the anti-TF antibody at concentration that completely neutralized the activity of recombinant human TF on thrombin generation. BXPC3 cells expressed significantly higher amounts of functional TF as compared to MCF7 cells. Incubation of BXPC3 and MCF7 cells with PPP resulted in acceleration of the initiation phase of thrombin generation. BXPC3 cells manifested higher procoagulant potential than MCF7 cells. The incubation of BXPC3 or MCF7 cells with the anti-TF monoclonal antibody which resulted in reversal of their effect on thrombin generation. The present study establishes a link between the amount of TF expressed by cancer cells with their procoagulant activity. Both studied types of cancer cells trigger thrombin generation but they have different procoagulant potential. The procoagulant activity of BXPC3 and MCF7 cells is related to the amount of TF expressed. Kinetic parameters of thrombogram are the most relevant for the detection of the TF-dependent procoagulant activity of cancer cells. TF expression is one of the mechanisms by which cancer cells manifest their procoagulant potential but it is not the unique one. The present experimental model will allow the characterization the procoagulant fingerprint of cell lines from the same or different histological types of cancer.
Asunto(s)
Neoplasias de la Mama/metabolismo , Neoplasias Pancreáticas/metabolismo , Trombina/metabolismo , Tromboplastina/biosíntesis , Tromboembolia Venosa/metabolismo , Neoplasias de la Mama/genética , Neoplasias de la Mama/patología , Línea Celular Tumoral , Femenino , Citometría de Flujo , Humanos , Células MCF-7 , Neoplasias Pancreáticas/genética , Neoplasias Pancreáticas/patología , ARN Mensajero/biosíntesis , ARN Mensajero/genética , Trombina/genética , Tromboplastina/genética , Tromboplastina/metabolismo , Tromboembolia Venosa/genética , Tromboembolia Venosa/patologíaRESUMEN
In this study, we evaluate the relationships between aspirin nonresponsiveness and the cyclooxygenase-1 (Cox-1) gene C50T polymorphism in stable coronary artery disease (CAD) in Tunisian patients. One hundred twenty-five stable CAD patients were included. The Cox-1 gene C50T polymorphism was determined by the polymerase chain reaction/restriction fragment length polymorphism method. Aspirin response was evaluated by measuring the collagen epinephrine closer time and the urinary dehydro-thromboxane B2 excretion. According to the collagen epinephrine closer time values, the frequency of the -50T allele was not significantly different in bad responders when compared with good responders (36.8% vs. 15.7%; p=0.1). Similarly, the presence of the -50T mutant allele was not statistically different comparing bad and good responders according to the urinary 11-dehydro-thromboxane B2 excretion concentration (60% vs. 40%; p=0.43). Our study did not demonstrate any association between the Cox-1 gene C50T polymorphism and aspirin nonresponsiveness status in stable CAD patients.
Asunto(s)
Aspirina/farmacología , Plaquetas/efectos de los fármacos , Enfermedad de la Arteria Coronaria/tratamiento farmacológico , Ciclooxigenasa 1/genética , Resistencia a Medicamentos , Polimorfismo Genético , Aspirina/uso terapéutico , Enfermedad de la Arteria Coronaria/genética , Humanos , Reacción en Cadena de la Polimerasa , Polimorfismo de Longitud del Fragmento de Restricción , Tromboxano B2/análogos & derivados , Tromboxano B2/orina , TúnezRESUMEN
The aim of this pilot study was to compare the effect of two different regimens of aspirin dosage on platelet of coronary artery disease (CAD) diabetic patients. Twenty-five CAD diabetic patients were included. Initially, all patients received aspirin 100 mg/day for 10 days. At day 10, aspirin antiplatelet effect was determined by measuring the collagen/epinephrine closure time (CT) 2 h after the last aspirin dosage and the next morning at 8 a.m.. The aspirin regimen was modified to 100 mg twice daily for patients showing a non-optimal platelet-inhibitory effect (CT < 298 s at 8 a.m.). Persistent high platelet reactivity (HPR) was defined by a CT < 160 s. During the 100 mg/day aspirin regimen, the prevalence of HPR at 8 a.m. was 48%, and only 7 patients (28%) had showed an optimal platelet-inhibitory effect. Bridging to the twice-daily regimen, the HPR was significantly reduced (p=0.025), and the optimal platelet-inhibitory effect was reached for 3 other patients. Our results showed that 100 mg aspirin twice-daily dosing rather than a once-daily dose significantly improves the aspirin effect on platelet of CAD diabetic patients. However, large prospective studies were needed to confirm whether this strategy will be clinically relevant and safe.
Asunto(s)
Aspirina/administración & dosificación , Enfermedad de la Arteria Coronaria/tratamiento farmacológico , Angiopatías Diabéticas/tratamiento farmacológico , Adulto , Anciano , Aspirina/farmacología , Aspirina/uso terapéutico , Plaquetas/efectos de los fármacos , Quimioprevención/métodos , Diabetes Mellitus/tratamiento farmacológico , Esquema de Medicación , Humanos , Persona de Mediana Edad , Proyectos Piloto , Inhibidores de Agregación Plaquetaria/administración & dosificación , Resultado del TratamientoRESUMEN
Platelet glycoprotein IIb/IIIa is a membrane receptor which plays a key role in coronary artery disease and thrombotic events. However, there is a considerable controversy regarding the clinical impact of glycoprotein IIIa platelet antigen 1 (PlA1)/platelet antigen 2 (PlA2) polymorphism as a risk factor for myocardial infarction. To evaluate the association between glycoprotein IIIa PlA1/PlA2 polymorphism and 1-year cardiovascular events occurrence in aspirin-treated patients with stable coronary artery disease. We prospectively included 188 postacute coronary syndrome patients (183 men) aged 59 ± 10 years and receiving aspirin (250 mg/day). The clinical outcome at 1 year was the composite end point of nonfatal myocardial infarction, stroke, recurrent unstable angina or cardiac death. Genotyping for PlA1/PlA2 polymorphism was conducted using PCR and restriction fragment length polymorphism analysis. The genotype distribution of glycoprotein IIIa PlA1/PlA2 polymorphism was PlA1/PlA1, 55.3%; PlA1/PlA2, 39.3% and PlA2/PlA2, 4%. Incidence of composite end point in homozygous PlA1/PlA1 carriers was significantly higher than in PlA2/PlA2 and PlA1/PlA2 patients [14.4 vs. 3.6% odds ratio 4.5 (1.2-16.6, 95% confidence interval); P = 0.012]. Multivariate analysis identified three strong predictive factors of cardiac death: age more than 65 years [odds ratio = 6.8, (1.4-34, 95% confidence interval); P = 0.018], ventricular ejection fraction less than 50% [odds ratio = 8.6, (1.7-42.6, 95% confidence interval); P = 0.008] and homozygous PlA1/PlA1 genotype [odds ratio = 8.8, (1.0-78.6, 95% confidence interval); P = 0.014]. Our results demonstrated that glycoprotein IIIa PlA1/PlA1 genotype carriers have a significantly increased risks of acute vascular ischemic events associated with a poor prognosis at 1 year. These postacute coronary syndrome patients might require an optimized secondary antithrombotic prophylaxis strategy.
Asunto(s)
Enfermedad de la Arteria Coronaria/genética , Integrina beta3/genética , Polimorfismo Genético , Factores de Edad , Anciano , Aspirina/uso terapéutico , Enfermedad de la Arteria Coronaria/diagnóstico , Determinación de Punto Final , Femenino , Genotipo , Humanos , Masculino , Persona de Mediana Edad , Pronóstico , Estudios Prospectivos , Factores de Riesgo , Volumen Sistólico , Resultado del TratamientoRESUMEN
Matrix metalloproteinase-9 (MMP-9) strongly influences tumor development and metastasis. Using resistant (rMCF-7) and sensitive (sMCF-7) breast cancer lines we investigated the role of MMP-9 in cell migration (CM) and tubular network (TN) formation, two processes implied in tumor growth and metastasis. Our data demonstrate that MMP-9 which is critical for CM is necessary but not sufficient for TN formation and suggest a link between MDR1/P-gp and constitutive MMP-9. Both TN formation and CM are dependent on PKC and ERK1/2 pathways. This study reinforces the logic of combining therefore MMP inhibitors in cancer therapy, especially in patients with chemoresistance and invasion/metastasis.
Asunto(s)
Neoplasias de la Mama/patología , Metaloproteinasa 9 de la Matriz/fisiología , Proteína Quinasa 1 Activada por Mitógenos/fisiología , Proteína Quinasa 3 Activada por Mitógenos/fisiología , Proteína Quinasa C/fisiología , Transducción de Señal , Neoplasias de la Mama/tratamiento farmacológico , Línea Celular Tumoral , Movimiento Celular/efectos de los fármacos , Resistencia a Antineoplásicos , Femenino , Flavonoides/farmacología , Humanos , Acetato de Tetradecanoilforbol/farmacologíaRESUMEN
Vitamin K antagonists (VKA) treatment starts with co-administration of low-molecular-weight heparin (LMWH). The anticoagulation induced by the two drugs is still not well determined. In the present study we used thrombin generation assay to evaluate the hypo-coagulation induced by treatment with VKA and by the combination of VKA with LMWH. Tissue factor triggered thrombin generation in platelet-poor plasma was assessed in samples from 15 healthy volunteers, 97 samples from patients treated with VKA and 41 samples from patients receiving enoxaparin and VKA. Patients were classified according to international normalised ratio (INR) level (<2, 2-3 and >3). In plasma samples from patients treated with VKA having INR 2-3 the inhibition of thrombin generation reached 50% compared to controls. In samples with INR>3 this inhibition was 80%. In samples from patients receiving both LMWH and VKA, thrombin generation was significantly decreased compared to the controls and VKA group. In samples with an INR 2-3 obtained from patients treated with LMWH and VKA, the inhibition of thrombin generation was similar to that observed in samples with an INR>3 obtained from VKA treated patients. Thrombin generation assay is sensitive to detect the global the anticoagulant effect produced by the association of LMWH and VKA. For equal INR dual anticoagulant treatment induces significantly more profound inhibition of thrombin generation compared to treatment with VKA alone. The clinical relevance of this observation merits to be studied in prospective studies in patients with defined indications of anticoagulant therapy.
Asunto(s)
Anticoagulantes/farmacología , Antifibrinolíticos/antagonistas & inhibidores , Heparina de Bajo-Peso-Molecular/farmacología , Trombina/biosíntesis , Vitamina K/antagonistas & inhibidores , Adolescente , Adulto , Anciano , Anciano de 80 o más Años , Pruebas de Coagulación Sanguínea , Sinergismo Farmacológico , Femenino , Humanos , Técnicas In Vitro , Masculino , Persona de Mediana Edad , Tromboplastina/metabolismo , Adulto JovenRESUMEN
Cancer is linked with hypercoagulability and risk of thrombosis and this close association was recognized by Armand Trousseau in 1865. The relation between cancer and blood coagulation is reciprocal: cancer induces a hypercoagulable state and predisposes to thrombosis and activation of platelets, blood coagulation and fibrinolysis interfere with tumor cell biology, tumor growth, angiogenesis and metastatic process. In the present article, we analyze the clinical trials which assessed the influence of anticoagulant treatment on the survival of patients with cancer. The available data show that low-molecular-weight heparins (LMWHs) tend to be more effective and safer than vitamin K antagonists (VKA) in improving survival in patients with cancer. The beneficial effect of anticoagulation with either LMWHs or VKA is not universal for all patients with cancer. There are some histological types of cancers which, at early stages, appear to be more sensitive than others to the effect of anticoagulant treatment. The available clinical trials, although limited, are encouraging for the beneficial effect of anticoagulant treatment on the survival of cancer patients. More clinical trials are needed, targeting groups of patients homogenous regarding the type of cancer, the stage of the disease and life expectancy. The forthcoming clinical trials have to address some issues regarding the optimal dose, the timing and the duration of treatment with LMWH in relation to chemotherapy or other anticancer therapies.
Asunto(s)
Anticoagulantes/uso terapéutico , Ensayos Clínicos como Asunto/estadística & datos numéricos , Neoplasias/tratamiento farmacológico , Trombofilia/tratamiento farmacológico , Trombosis/prevención & control , Animales , Anticoagulantes/efectos adversos , Método Doble Ciego , Ensayos de Selección de Medicamentos Antitumorales , Fibrinolíticos/efectos adversos , Fibrinolíticos/uso terapéutico , Hemorragia/inducido químicamente , Heparina/efectos adversos , Heparina/uso terapéutico , Heparina de Bajo-Peso-Molecular/efectos adversos , Heparina de Bajo-Peso-Molecular/uso terapéutico , Humanos , Metaanálisis como Asunto , Ratones , Estudios Multicéntricos como Asunto/estadística & datos numéricos , Metástasis de la Neoplasia , Neoplasias/sangre , Neoplasias/mortalidad , Neoplasias/patología , Neovascularización Patológica/tratamiento farmacológico , Ensayos Clínicos Controlados Aleatorios como Asunto/estadística & datos numéricos , Proyectos de Investigación , Análisis de Supervivencia , Trombofilia/sangre , Trombofilia/etiología , Trombosis/sangre , Trombosis/etiología , Trombosis/mortalidad , Vitamina K/antagonistas & inhibidores , Warfarina/efectos adversos , Warfarina/uso terapéuticoRESUMEN
Thromboelastography analysis providing a global assessment of coagulation is gaining new interest in clinical practice. MinimalTF triggered whole blood thromboelastography provides a valuable tool for studying the kinetics of clot formation (expressed by the parameters R, K and alpha-angle) and the physical characteristics of the clot, such as its firmness and the elastic modulus shear (expressed by the parameters maximal amplitude MA and G). We studied the influence of fibrin polymerization and platelet functional status on each parameter of thromboelastographic trace obtained by minimalTF activation inWB by employing increasing concentrations of a fibrin polymerization inhibitors (the tetrapeptide Gly-Pro-Arg-Pro-OH.AcOH; Pefabloc-FG) and an inhibitor of actin polymerization (Cytochalasin D). Pefabloc-FG at concentrations higher than 5 mg/ml prolonged the R and K times and decreased the alpha-angle in a concentration-dependent manner but it did not modify MA and G parameters. At the concentration of 5 mg/ml, Pefabloc-FG completely inhibited clot formation. Cytochalasin D had no effect on R time but decreased the alpha-angle, MA and G parameters by reaching a plateau at the concentration of 5 microM. The effect of cytochalasin D was more pronounced on MA and G than on the alpha-angle. A combination of both Pefabloc-FG (0.5 mg/ml) and cytochalasin D (50 microM) significantly decreased alpha-angle compared to control as well as their single effect. However, G value was dramatically reduced in the presence of cytochalasin D exposure, without any additional effect when both inhibitors were combined. This study confirms the importance of fibrin polymerisation on the kinetics of thrombus formation and demonstrates the close association between the quality of the thrombus and the functional status of platelets. Normal platelet contractile forces are of major importance for the maximum amplitude of TEG which is related to the strength and elastic modulus of the thrombus.
Asunto(s)
Coagulación Sanguínea , Plaquetas/fisiología , Fibrina/metabolismo , Tromboelastografía , Fenómenos Biomecánicos , Ácido Cítrico , Citocalasina D/farmacología , Relación Dosis-Respuesta a Droga , Fibrina/antagonistas & inhibidores , Humanos , Oligopéptidos/farmacología , Sulfonas/farmacología , TrombosisRESUMEN
The in vitro closure time (CT), determined by the Platelet Function Analyzer (PFA-100), is used to monitor patients treated with aspirin. A relatively high percentage of in vitro aspirin resistance was reported despite an adequate inhibition of platelet response to arachidonic acid and we investigated whether high plasma levels of von Willebrand factor ristocetin cofactor activity (vWF:RCo) may contribute to this profile. Platelet aggregation test, CT [collagen adrenaline (CEPI-CT) and collagen adenosine 5'-diphosphate (ADP) (CADP-CT)], and vWF:RCo levels were evaluated in 55 consecutive patients receiving aspirin (75-250 mg/d) versus 32 untreated control subjects. All the aspirin-treated patients showed platelet aggregation responses that reflected the aspirin intake. However, CT data analysis enabled aspirin good-responder (GR) and aspirin bad-responder (BR) patients to be identified. All GR group subjects (n = 27), had a CEPI-CT and a CADP-CT longer than 300 s and 96 s respectively. The BR group (n = 28) had CEPI-CT values below 200 s and all CADP-CT were in the normal range (77 +/- 19 s). Interestingly, the BR plasma vWF:RCo levels were significantly higher (159 +/- 43%) than those of the GR group (121 +/- 34%) (P < 0.01), which were similar to control values (114 +/- 31%). A negative correlation between vWF:RCo and CT values was established. We demonstrate that in vitro aspirin-resistance, revealed by PFA-100 CT prolongation failure, is correlated to increased plasmatic vWF:RCo levels, reinforcing its particular importance in PFA-100 cartridges performance.
Asunto(s)
Aspirina/administración & dosificación , Inhibidores de Agregación Plaquetaria/administración & dosificación , Factor de von Willebrand/fisiología , Adulto , Anciano , Enfermedades Cardiovasculares/sangre , Enfermedades Cardiovasculares/prevención & control , Resistencia a Medicamentos , Femenino , Humanos , Masculino , Persona de Mediana Edad , Pruebas de Función Plaquetaria/métodos , Estudios Prospectivos , Factor de von Willebrand/análisisRESUMEN
A novel C-type lectin protein (CLP), lebecetin, was purified to homogeneity from the venom of Macrovipera lebetina by gel filtration on a Sephadex G75 column and ion exchange chromatography on Mono S column. Lebecetin is a basic protein with a pHi=9.9 and migrates in SDS-PAGE as a single band or two distinct bands under nonreducing and reducing conditions, respectively. These results are further confirmed by MALDI-TOF mass spectrometry that indicates a molecular mass of 29779 Da for native lebecetin and molecular masses of 15015 and 16296 Da for alpha and beta subunits, respectively. The N-terminal amino acid sequences of lebecetin subunits show a high degree of similarity with those of C-type lectin-like proteins. In addition, functional studies showed that lebecetin has a potent inhibitory effect on platelet aggregation induced by thrombin in a concentration-dependent manner. In contrast, no inhibitory effect is observed when platelets are exposed to thromboxane A2 (TxA2) mimetic (U46619) or arachidonic acid. Moreover, there was no effect either on blood coagulation or A, B and O washed human erythrocytes agglutination. Furthermore, flow cytometric analysis revealed that fluoro-isothiocyanate (FITC)-labelled lebecetin bound to human formalin fixed platelets in a saturable and concentration manner and this binding was specifically prevented by anti-glycoprotein Ib (GPIb) mAb. These observations suggest that lebecetin is a C-type lectin-like protein that selectively binds to platelet GPIb.
Asunto(s)
Venenos de Crotálidos/química , Lectinas Tipo C/metabolismo , Inhibidores de Agregación Plaquetaria/metabolismo , Venenos de Víboras/metabolismo , Viperidae/metabolismo , Secuencia de Aminoácidos , Animales , Plaquetas/metabolismo , Femenino , Humanos , Lectinas Tipo C/química , Lectinas Tipo C/genética , Lectinas Tipo C/aislamiento & purificación , Masculino , Datos de Secuencia Molecular , Agregación Plaquetaria/fisiología , Inhibidores de Agregación Plaquetaria/química , Inhibidores de Agregación Plaquetaria/aislamiento & purificación , Subunidades de Proteína/química , Subunidades de Proteína/metabolismo , Alineación de Secuencia , Trombina/metabolismo , Venenos de Víboras/química , Venenos de Víboras/genética , Venenos de Víboras/aislamiento & purificaciónRESUMEN
Prostaglandin H synthase 2 (PGHS-2), a highly inducible isoenzyme, is responsible for overproduction of the prostaglandins (PGs) in inflammatory sites. We established that among fish oil polyunsaturated fatty acids (PUFAs), eicosapentaenoic acid (EPA), but not docosahexaenoic acid (DHA), greatly decreased interleukin-1beta (IL-1beta)-induced PGHS-2 expression in human pulmonary microvascular endothelial cells (HPMECs). Lipoxygenase products 12 (S)-hydroperoxyeicosapentaenoic acid ((S)-HpEPE), 15 (S)-HpEPE and leukotriene (LT) D5 reproduced similar inhibitory effect, suggesting that they may be the intermediate metabolites responsible for PGHS-2 down-regulation by EPA. Accordingly, the EPA effect is prevented by nordihydroguaiaretic acid (NDGA) and by REV 5901, nonspecific and specific 5-lipoxygenase inhibitors, respectively. Besides, inhibition of cyclooxygenase activity by ibuprofen, indomethacin or aspirin was not able to prevent this effect. Moreover, cyclooxygenase metabolites of EPA (PGs D3, E3 and I3) markedly potentiate IL-1beta-induced PGHS-2 expression, probably by increasing intracellular cAMP levels. Peroxisome proliferator-activated receptors (PPARs) are known to be activated by fatty acids (FAs) such as EPA. We found here that HPMECs express only weak amounts of PPARalpha and PPARgamma whose activation by synthetic agonists, Wy-14,643 and ciglitazone, does not cause any inhibition of IL-1beta-induced PGHS-2 expression. This finding ruled out the involvement of PPARs in the EPA inhibitory effect. In addition, we established that EPA, which failed to inhibit nuclear factor-kappaB (NF-kappaB) activation, suppressed p38 mitogen-activated protein kinase (MAPK) phosphorylation in stimulated HPMECs. Our data demonstrate that EPA, unlike DHA, down-regulates PGHS-2 expression in HPMECs probably through its 5-lipoxygenase-dependent metabolites and advocates a beneficial role for this FA in limiting inflammatory response.