RESUMEN
Von Willebrand disease (VWD) is an inherited bleeding disorder caused by the quantitative or qualitative deficiency of von Willebrand factor (VWF). Replacement therapy with plasma-derived VWF/factor VIII (FVIII) concentrates is required in patients unresponsive to desmopressin. To assess the efficacy, safety and ease of use of a new, volume-reduced (VR) formulation of VWF/FVIII concentrate Haemate(®) P in patients requiring treatment for bleeding or prophylaxis for recurrent bleeding or for invasive procedures. Pharmacoeconomic variables were also recorded. Data were analysed using descriptive statistics. This was a multicentre, prospective, observational study. Consecutively enrolled patients received Haemate(®) P VR according to their needs, and were followed for 24 months. Of the 121 patients enrolled, 25.6% had type 3 VWD and more than 40% had severe disease. All patients were followed for 2 years, for a total of 521 visits. On-demand treatment was given to 61.9% of patients, secondary long-term prophylaxis to 25.6% and prophylaxis for surgery, dental or invasive procedures to 45.5%. The response to treatment was rated as good to excellent in >93-99% of interventions. The new formulation was well tolerated by all patients with no report of drug-related adverse events. The switch to volume-reduced Haemate(®) P was easy to perform and infusion duration was decreased twofold compared with the previous formulation. Volume-reduced Haemate(®) P was at least as effective and well-tolerated as the previous formulation.
Asunto(s)
Anticoagulantes/uso terapéutico , Factor VIII/uso terapéutico , Enfermedades de von Willebrand/tratamiento farmacológico , Factor de von Willebrand/uso terapéutico , Adolescente , Adulto , Anciano , Anticoagulantes/efectos adversos , Pérdida de Sangre Quirúrgica/prevención & control , Niño , Preescolar , Costo de Enfermedad , Sustitución de Medicamentos , Factor VIII/efectos adversos , Femenino , Hemorragia/prevención & control , Hospitalización/estadística & datos numéricos , Humanos , Italia , Masculino , Persona de Mediana Edad , Pasteurización , Estudios Prospectivos , Adulto Joven , Factor de von Willebrand/efectos adversosRESUMEN
AIM: The aim of this study was to determine, in patients with Turner syndrome (TS), the prevalence of thrombophilic disorders correlating with a higher risk of venous thromboembolism (VTE), to evaluate if thrombophilia is associated with the genetic features of these patients and whether screening before hormone replacement therapy (HRT) is advisable. PATIENTS AND METHODS: We examined 82 TS patients. In all patients we analyzed activated factor VIII:C, fibrinogen, antithrombin (AT), protein C (PC), protein S (PS), activated PC resistance, and homocysteine. For every patient, an investigation for mutations in prothrombin G20210A, factor V R506Q, methylenetetrahydropholate reductase (MTHFR) C 677T and A1298C was conducted. RESULTS: Low values of PC in 3 patients (3.70%), low values of PS in 12 (14.81%), and hyperhomocysteinemia in 4 (4.87%) were found; 52 girls (64.2%) presented hyperfibrinogenemia. Three patients were heterozygous for the prothrombin G20210A allele mutation (3.66%) and the factor V mutation was present in 4 patients (4.88%). No TS patient had a homozygous mutation. Mutations in the MTHFR gene were present in 62 girls, in 17 patients (20.7%) they were homozygous and in 45 patients (54.88%) heterozygous. CONCLUSIONS: Considering the increased risks with the association between VTE and the higher prevalence of PC and PS deficiencies, TT genotype mutations and high level of fibrinogen, it is advisable to perform a complete thrombophilia screening in TS patients before starting HRT.
Asunto(s)
Trombofilia/epidemiología , Síndrome de Turner/epidemiología , Síndrome de Turner/genética , Adolescente , Adulto , Antitrombina III/metabolismo , Comorbilidad , Factor VIII/metabolismo , Femenino , Fibrinógeno/metabolismo , Genotipo , Homocisteína/metabolismo , Humanos , Metilenotetrahidrofolato Reductasa (NADPH2)/genética , Proteína C/metabolismo , Proteína S/metabolismo , Protrombina/genética , Tromboembolia/epidemiología , Tromboembolia/etiología , Trombofilia/complicaciones , Trombofilia/fisiopatología , Síndrome de Turner/fisiopatología , Adulto JovenRESUMEN
Hyperhomocysteinaemia represents an independent risk factor for atherosclerotic cardiovascular disease, stroke, peripheral arterial occlusive disease and venous thrombosis. Psoriasis is a chronic inflammatory skin disease associated with increased atherothrombosis and cardiovascular risk profile. The aim of this study is to investigate homocysteine, folic acid and vitamin B12 levels in a cohort of psoriatic patients and its relationship with the severity of the disease. A retrospective observational study in 98 patients with chronic plaque psoriasis and 98 healthy controls was performed. Total plasma homocysteine level, folic acid, vitamin B12 and PASI index were assessed in every patient. Patients with psoriasis had plasma homocysteine levels higher than controls (57% of cases and 25% of controls; p<0.0001). Folic acid and vitamin B12 plasma levels were lower in psoriatic patients than in controls (p = NS), lower levels of vitamin B12 were found in patients with hyperhomocysteinaemia compared to patients with a normal value of homocysteine (p = 0.0009). The severity of psoriasis assessed according to PASI (19.51+/-16.26) did not directly correlate either with higher levels of homocysteine or with vitamin B12 and folic acid plasma levels. In conclusion, a significantly higher prevalence of hyperhomocysteinaemia was found in psoriatic patients compared to healthy controls. A significant correlation between hyperhomocysteinaemia and lower vitamin B12 levels, but not folic acid, was evidenced. On the contrary, our data do not correlate the high level of homocysteine with higher PASI scores or psoriasis type, suggesting that homocysteine level can be considered an independent risk factor in psoriatic patients.
Asunto(s)
Ácido Fólico/sangre , Homocisteína/sangre , Psoriasis/sangre , Psoriasis/patología , Vitamina B 12/sangre , Femenino , Humanos , Hiperhomocisteinemia/sangre , Masculino , Persona de Mediana Edad , Estudios Retrospectivos , Factores de Riesgo , Piel/patologíaRESUMEN
OBJECTIVE: To investigate the prevalence of mitral valve prolapse (MVP) and abnormalities of haemostasis in children and adolescents with migraine with aura (MA) compared with peers affected by other idiopathic headaches. MATERIALS AND METHODS: We recruited 20 MA patients (10 men and 10 women; age range 8-17 years) and 20 sex- and age-matched subjects with other idiopathic headaches. Both groups underwent colour Doppler transthoracic echocardiography to detect MVP and the following laboratory work-up: plasma prothrombin time, activated partial thromboplastin time, thrombin time, fibrinogen, protein C, protein S, homocysteine, lupus anticoagulant, von Willebrand factor (vWF) ristocetin cofactor activity, immunoglobulins (Ig) G and M anticardiolipin antibodies (aCL). Factor V Leiden, factor II and methylenetetrahydrofolate reductase were investigated (we did not test the entire genes, but screened for specific point mutations). RESULTS: The prevalence of MVP was significantly higher in the MA subjects than in the patients affected by other idiopathic headaches (40% vs 10%; P < 0.05). Moreover, the MA patients showed a higher rate of above-normal IgM aCL titres (45% vs 10%; P < 0.05). Finally, in the group of patients with MVP we found a higher prevalence of aCL in those with MA compared with those affected by other idiopathic headaches. CONCLUSIONS: A proportion, at least, of the MA patients showed a more complex phenotype characterized by MVP and/or positive aCL titres. The pathogenetic role of these associations is obscure and larger studies are needed to confirm the usefulness of echocardiographic and laboratory investigations in this area and to identify possible new treatment approaches that might be explored in this group of MA patients.
Asunto(s)
Trastornos de la Coagulación Sanguínea/epidemiología , Migraña con Aura/epidemiología , Prolapso de la Válvula Mitral/epidemiología , Adolescente , Trastornos de la Coagulación Sanguínea/genética , Pruebas de Coagulación Sanguínea , Niño , Comorbilidad , Estudios Transversales , Ecocardiografía Doppler , Femenino , Tamización de Portadores Genéticos , Homocigoto , Humanos , Incidencia , Masculino , Migraña con Aura/genética , Prolapso de la Válvula Mitral/genética , Cefalea de Tipo Tensional/epidemiologíaRESUMEN
Xenopus laevis oocytes codify a G-protein-activated inward rectifier potassium channel (GIRK5 or Kir3.5). Coinjection of other GIRKs, the muscarinic m2 receptor, or Gbetagamma protein cRNAs is required to observe functional GIRKx-GIRK5 heteromultimers in oocytes. Studies with GIRK2 isoforms have shown that the size of the amino or carboxyl terminus plays a crucial role on giving functional K(+) channels. In this work we studied the properties of a GIRK5 with 25 amino acids deleted toward its amino-terminal domain. Injection of GIRK5-Delta25 cRNA alone displayed large basal and transient inward rectifying currents in oocytes. The instantaneous currents reached a stationary level after a long duration voltage pulse (10 s). For this relaxation, fast (tau(1)) and slow (tau(2)) time constants were estimated at different voltages. Recovery from inactivation followed a monoexponential function (tau=0.95+/-0.07 s). By contrast with other inward rectifier channels, blockade of GIRK5-Delta25 by extracellular Ba(2+) was voltage-independent (K(d)=102+/-2 microM), suggesting the presence of a Ba(2+) site at the external channel vestibule. To confirm this hypothesis, the Ba(2+) sensitivity of two charged mutants GIRK5-Delta25(N129E) and GIRK5-Delta25(K157E) at each of the external loops was determined. GIRK5-Delta25(N129E) and GIRK5-Delta25(K157E) showed a 100-fold and 2-fold higher affinity to Ba(2+), respectively, supporting the existence of this Ba(2+) binding site.
Asunto(s)
Oocitos/metabolismo , Canales de Potasio de Rectificación Interna , Canales de Potasio/metabolismo , Animales , Bario/química , Bario/metabolismo , Compuestos de Bario/farmacología , Sitios de Unión , Cationes Bivalentes , Cloruros/farmacología , Electrofisiología , Canales de Potasio Rectificados Internamente Asociados a la Proteína G , Mutación , Bloqueadores de los Canales de Potasio , Canales de Potasio/genética , Isoformas de Proteínas/metabolismo , Proteínas Recombinantes/metabolismo , Electricidad Estática , Transfección , Xenopus laevisRESUMEN
The purpose of the present study was to analyze the long-term regulation of renal bumetanide-sensitive Na+-K+-2Cl- cotransporter and thiazide-sensitive Na+-Cl- cotransporter gene expression during changes in NaCl and water metabolism. Male Wistar rats exposed to high or low NaCl intake, saline loading, dehydration, water loading, and furosemide administration during 7 days were studied. Control groups had access to regular food and tap water. Rats were kept in metabolic cages for 4 days before and during the experiment to determine daily urinary electrolyte excretion and osmolarity. At the end of the experiment, creatinine clearance and serum electrolyte levels were also measured. Kidneys were excised and macroscopically subdivided into cortex and outer and inner medulla. Total RNA was extracted from each individual cortex or outer medulla by use of the guanidine/cesium chloride method. The Na+-K+-2Cl- cotransporter expression in outer medulla total RNA was assessed by nonradioactive Northern blot analysis and the Na+-Cl- cotransporter expression in renal cortex total RNA was assessed by semiquantitative polymerase chain reaction. Experimental maneuvers were adequately tolerated, and all groups developed the appropriate renal response to each challenge. However, the level of expression of both cotransporters did not change in any model, except for a 2.8-fold increase in the Na+-Cl- cotransporter expression during dehydration. We conclude that nephron adaptation to 7-day modifications in NaCl and water metabolism does not include changes in the amount of electroneutral sodium-coupled cotransporter gene expression at the mRNA level.
Asunto(s)
Bumetanida/farmacología , Proteínas Portadoras/efectos de los fármacos , Diuréticos/farmacología , Riñón/metabolismo , ARN Mensajero/metabolismo , Cloruro de Sodio/metabolismo , Simportadores , Animales , Benzotiadiazinas , Proteínas Portadoras/metabolismo , Transporte Iónico/efectos de los fármacos , Riñón/efectos de los fármacos , Masculino , Ratas , Ratas Wistar , Inhibidores de los Simportadores del Cloruro de Sodio/farmacología , Simportadores del Cloruro de Sodio , Simportadores de Cloruro de Sodio-Potasio , Agua/metabolismoRESUMEN
PURPOSE: To assess the natural history and risk factors for thrombosis in a large cohort of unselected patients with antiphospholipid antibodies. PATIENTS AND METHODS: Three hundred sixty consecutive patients (118 males, 242 females, median age 39 years [range 2 to 78]) fulfilling the currently accepted criteria for diagnosis of lupus anticoagulant (LAC) (n = 326) and/or raised immunoglobulin G anticardiolipin antibodies (IgG ACA) (n = 185) were collected from 16 Italian institutions and prospectively observed for a median of 3.9 years (range 0.5 to 5). Main endpoints were the occurrence of arterial or venous thrombosis, the outcome of pregnancies, and any severe complications leading to hospitalization or death. RESULTS: Thirty-four patients developed a thrombotic complication, with a total incidence of 2.5% patient-years. Multivariate logistic regression analysis identified two independent risk factors for thrombotic events: a previous thrombosis (RR 4.9; 95% CI, 1.76 to 13.7; P < 0.005) and IgG ACA titer above 40 units (RR 3.66; 95% CI, 1.24 to 10.8; P < 0.01). A total of 28 pregnancies were observed in 25 women and 11 (39%) were abortive. Adverse pregnancy outcomes were significantly more frequent in women with a history of miscarriage or vascular occlusion (9/16, 56%) than in asymptomatic women (2/12, 17%) (P = 0.035). Four patients developed non-Hodgkin's lymphoma during the follow-up. Eighteen patients died. Vascular events and hematological malignancies represented the most frequent causes of death (n = 5 for each). CONCLUSIONS: The present study shows that: (a) previous thrombosis and ACA titer > 40 U are independent predictors of thrombosis; (b) history of miscarriage or vascular disease is significantly associated with adverse pregnancy outcome; (c) hematological malignancies can develop during follow-up in patients with antiphospholipid antibodies.
Asunto(s)
Anticuerpos Antifosfolípidos/análisis , Trombosis/etiología , Adolescente , Adulto , Anciano , Anticuerpos Anticardiolipina/análisis , Anticoagulantes/uso terapéutico , Causas de Muerte , Niño , Preescolar , Estudios de Cohortes , Femenino , Estudios de Seguimiento , Humanos , Inmunoglobulina G/análisis , Incidencia , Modelos Logísticos , Masculino , Persona de Mediana Edad , Embarazo , Estudios Prospectivos , Factores de Riesgo , Trombosis/epidemiología , Trombosis/inmunología , Factores de Tiempo , Warfarina/uso terapéuticoRESUMEN
PURPOSE: To study the effect of aminoguanidine (AMG), an inhibitor of nitric oxide production, on the ocular infection of Balb/c mice with herpes simplex virus (HSV) type 1 strain F and HSV-2 strain G. METHODS: Animals were treated with different amounts of AMG (0.5, 0.1, and 0.05 mg/mouse) by topical application in the eye from postinfection (PI) days -2 through +5, considering 0 the day of infection. At different PI days, development of herpetic keratitis was evaluated in treated and control mice. RESULTS: Treated animals showed a dose-dependent increase in ocular disease after viral infection, compared with control animals. Viral titers in ocular washings were higher in AMG-treated mice (PI day 2, HSV-1: AMG 0.5 mg, 1.3 x 10(3) plaque-forming units (PFU)/ml; control, 0. 22 x 10(2) PFU/ml, P < 0.025). At PI day 3, control corneas had only scattered inflammatory cells, whereas those from treated animals showed a conspicuous infiltrate consisting primarily of neutrophils. Viral titers were also higher in brains of treated mice. These animals died earlier and in a greater proportion than control animals (percentage of mortality, PI day 12, HSV-1: AMG 0.5 mg, 40% +/- 4%; control, 18% +/- 3%, P < 0.05). CONCLUSIONS: These data indicate an inhibitory effect of nitric oxide on HSV ocular infection.
Asunto(s)
Córnea/efectos de los fármacos , Inhibidores Enzimáticos/uso terapéutico , Guanidinas/uso terapéutico , Queratitis Herpética/tratamiento farmacológico , Óxido Nítrico Sintasa/antagonistas & inhibidores , Administración Tópica , Animales , Córnea/enzimología , Córnea/virología , ADN Viral/análisis , Progresión de la Enfermedad , Relación Dosis-Respuesta a Droga , Herpesvirus Humano 1/aislamiento & purificación , Herpesvirus Humano 1/fisiología , Herpesvirus Humano 2/aislamiento & purificación , Herpesvirus Humano 2/fisiología , Queratitis Herpética/patología , Queratitis Herpética/virología , Ratones , Ratones Endogámicos BALB C , Óxido Nítrico Sintasa/genética , Óxido Nítrico Sintasa/metabolismo , Soluciones Oftálmicas , Reacción en Cadena de la Polimerasa de Transcriptasa Inversa , Ganglio del Trigémino/virologíaRESUMEN
The immediate effect of cigarette-smoking on ADP-induced platelet aggregation and on platelet adhesiveness was investigated in 12 normal subjects aged 20 to 40, in 10 normal subjects aged 43 to 72 and in 10 patients with cerebrovascular disease aged 45 to 75. All the subjects were heavy smokers (more than 20 cigarettes a day). After smoking 2 cigarettes a significant increase in ADP aggregation and platelet adhesiveness was found in the group of young heavy smokers, while in the old subjects with or without cerebrovascular disease the increase in platelet activity was never significant. These data were discussed and some hypotheses for this higher reactivity of platelets from young people were suggested.
Asunto(s)
Plaquetas/efectos de los fármacos , Fumar , Adenosina Difosfato , Adulto , Anciano , Envejecimiento , Trastornos Cerebrovasculares/sangre , Humanos , Persona de Mediana Edad , Adhesividad Plaquetaria/efectos de los fármacos , Agregación Plaquetaria/efectos de los fármacosRESUMEN
The authors report a long-term evaluation of the effects of chemical and surgical synovectomies performed in two groups of patients which were comparable as regards number of haemarthrosis per year and joint mobility. The results obtained indicated that both synovectomy and synoviorthesis were effective in reducing the bleeding episodes; however, while synovectomy stopped almost completely the recurrence of haemarthrosis, synoviorthesis in 3 patients younger than 12 years did not modify it and in one case it even increased the bleeding tendency. Moreover, the occurrence of arthritic pain was observed in 6 knees treated by synoviorthesis and only in 3 knees treated by synovectomy. Finally, the joint function was significantly reduced in the synoviorthesis group as compared to the synovectomy group. In conclusion, synovectomy gave better results than synoviorthesis but the latter treatment has a number of economical and technical advantages and should therefore be considered in patients older than 12 year of age with good joint function and without evident radiological changes, in whom full dose replacement therapy has had no effect on the recurrence of haemarthrosis.
Asunto(s)
Hemartrosis/complicaciones , Hemofilia A/complicaciones , Sinovectomía , Adolescente , Adulto , Artritis Reumatoide/terapia , Niño , Hemartrosis/tratamiento farmacológico , Hemartrosis/cirugía , Hemofilia A/tratamiento farmacológico , Humanos , Cuidados a Largo Plazo , Tetróxido de Osmio/uso terapéutico , RecurrenciaRESUMEN
Some haemostatic parameters have been evaluated in a group of rigorously selected patients with maturity-onset diabetes mellitus without thromboembolic complications and in apparently normal subjects of the same age before and after the venous occlusion test (VOT). In basal conditions diabetics had higher levels of AT III as biological activity and higher fibrinolytic and antifibrinolytic activities than controls. After VOT, F VIII R:Ag increased significantly in both groups, more markedly in controls than in diabetics, while F VIII: C showed no modification. Also AT III R:Ag increased after the test, but such variation was significant only in diabetics; on the contrary, the biological activity of AT III was always significantly decreased after the test. After VOT there were also in both groups highly significant increases in the fibrinolytic and antifibrinolytic activities. Finally, HbA1c levels directly correlated with AT III as biological activity before VOT, but with no other parameter either before or after the test. These data suggest the existence in patients with diabetes mellitus without thromboembolic complications of an activated protective mechanism against intravascular clotting.
Asunto(s)
Diabetes Mellitus/sangre , Coagulación Intravascular Diseminada/prevención & control , Fibrinólisis , Anciano , Antígenos/análisis , Antitrombina III/análisis , Antitrombina III/inmunología , Constricción , Factor VIII/análisis , Factor VIII/inmunología , Femenino , Hemoglobina A/análisis , Humanos , Masculino , Persona de Mediana Edad , Activadores Plasminogénicos/sangre , Venas , Factor de von WillebrandRESUMEN
Blood coagulation abnormalities induced by administration of E. coli L-asparaginase were investigated in 25 patients with acute lymphoblastic leukemia treated according to the GIMEMA ALL 0288 trial. Dosage of L-asparaginase was relatively low (6,000 U/m2/day for 7 days total dose 42,000 U/m2) as compared to the conventional dosages (120,000-140,000 U/m2 over 10-14 days). A significant decrease in fibronogen, plasminogen, alpha2-antiplasmin and antithrombin III was observed from day IV of L-asparaginase and it was maximum on day VIII, with return to the baseline levels on day XV. Protein C levels had only a borderline reduction, while no modification of protein S or factor VII was observed. Two of the patients investigated developed thrombosis. The presence of a prothrombotic state induced even by this low dosage of E. coli L-asparaginase was suggested by a significant increase of sensitive markers of hypercoagulability such as fibrinopeptide A, thrombin-antithrombin complexes, and prothrombin fragment F1 + 2.
Asunto(s)
Asparaginasa/efectos adversos , Trastornos de la Coagulación Sanguínea/inducido químicamente , Escherichia coli/enzimología , Leucemia-Linfoma Linfoblástico de Células Precursoras/tratamiento farmacológico , Trombina/biosíntesis , Adolescente , Adulto , Asparaginasa/administración & dosificación , Factores de Coagulación Sanguínea/metabolismo , Femenino , Humanos , Incidencia , Masculino , Persona de Mediana Edad , Leucemia-Linfoma Linfoblástico de Células Precursoras/sangre , Sensibilidad y Especificidad , Tromboflebitis/inducido químicamente , Tromboflebitis/epidemiologíaRESUMEN
Whereas in vitro studies showed that plasmin may induce both inhibition and activation of platelets, in vivo and ex vivo investigations suggested that thrombolytic agents are responsible for platelet stimulation. To gain further information on this topic, ex vivo platelet function was studied in 24 subjects with acute myocardial infarction treated with streptokinase or recombinant tissue-type plasminogen activator (rt-PA). Ten patients with acute myocardial infarction who did not receive thrombolytic treatment were also investigated. The data shows that at the end of thrombolytic infusion, the maximal extent of platelet aggregation and adenosine triphosphate release was reduced in treated patients compared with that in untreated ones. In subjects treated with streptokinase, the defect in platelet aggregation derived from both cellular and plasmatic defects. Plasmatic beta-thromboglobulin concentration was significantly reduced after streptokinase, but unchanged after rt-PA. Three days after thrombolytic treatment, platelet aggregation of patients receiving streptokinase or rt-PA was not significantly different from that of untreated subjects. A similar defect in platelet function was obtained in vitro, incubating normal platelet-rich plasma with pharmacologic concentrations of streptokinase. Again, platelet function defect derived from both cellular and plasmatic damages. It cannot be excluded that platelet activation occurs in patients with acute myocardial infarction during the very early phases of thrombolytic treatment. However, it is suggested that a transient defect in platelet function follows both streptokinase and rt-PA infusion.
Asunto(s)
Plaquetas/efectos de los fármacos , Infarto del Miocardio/sangre , Infarto del Miocardio/tratamiento farmacológico , Estreptoquinasa/uso terapéutico , Terapia Trombolítica , Activador de Tejido Plasminógeno/uso terapéutico , Adenosina Trifosfato/sangre , Plaquetas/fisiología , Humanos , Técnicas In Vitro , Agregación Plaquetaria/efectos de los fármacos , Factores de TiempoRESUMEN
Various pathogenic factors have been proposed to explain the abnormal hemostasis observed in AL amyloidosis. Since imbalance between clotting factors and inhibitors could play a pathogenic role in both hemorrhagic and thrombotic manifestations, we investigated the thrombin-antithrombin pathway in 35 patients with AL amyloidosis. Ten patients suffered from bleeding while 3 patients experienced deep venous thrombosis. Thrombin time was prolonged in 29 subjects, the mean values of antithrombin III activity (ATIII Act) were significantly lower than those of antithrombin III antigen (ATIII Ag) with loss of relationship between these two different techniques of ATIII detection, normally observed in healthy controls. In 19 patients increased levels of thrombin-antithrombin (TAT) complexes were present. Crossed immunoelectrophoresis of ATIII, performed in presence of heparin, evidenced ATIII forms with reduced binding capacity to heparin and TAT complexes of various electrophoretic mobilities. In conclusion, the impairment of the thrombin-antithrombin pathway, in association with the low ATIII biological activity, might play a pathogenic role in the hypercoagulable state reported in AL amyloidosis, despite the higher frequency of bleeding manifestations.
Asunto(s)
Amiloidosis/metabolismo , Antitrombina III/metabolismo , Péptido Hidrolasas/metabolismo , Adulto , Anciano , Amiloide/sangre , Amiloide/orina , Amiloidosis/complicaciones , Trastornos de las Proteínas de Coagulación/fisiopatología , Femenino , Heparina/metabolismo , Humanos , Inmunoelectroforesis , Masculino , Persona de Mediana Edad , Unión Proteica , Tiempo de TrombinaRESUMEN
In a previous study we found a correlation between metastatic potential and platelet aggregating activity in sublines of a benzopyrene-induced murine fibrosarcoma ( mFS6 ); the purpose of the present work was to elucidate the role of thromboxane biosynthesis by platelets and/or by neoplastic cells in the activation of platelets in this system. The cells of the more malignant subline induced higher aggregation and TxB2 production than those of the non metastasizing one. The supernatants of aggregating cell suspensions contained very few TxB2; furthermore, preincubation of platelets with ASA or Apyrase resulted in inhibition of aggregation and TxB2 production, while preincubation of the cells was ineffective; these results suggest the platelet origin of the measured TxB2 and indicate that platelet-derived ADP plays an important role in their activation, while the production of ADP by the cells does not seem to be relevant in this model. The involvement of platelet prostaglandin biosynthesis pathway in neoplastic cell induced platelet activation could play an important role in the development of platelet-dependent tumour metastasis.
Asunto(s)
Plaquetas/metabolismo , Transformación Celular Neoplásica , Fibrosarcoma/metabolismo , Agregación Plaquetaria , Tromboxano B2/biosíntesis , Tromboxanos/biosíntesis , Animales , Apirasa/farmacología , Aspirina/farmacología , Línea Celular , Transformación Celular Neoplásica/efectos de los fármacos , Humanos , Ratones , Agregación Plaquetaria/efectos de los fármacosRESUMEN
The physiological relevance of calcium in many vital processes requires that its concentration in extracellular fluids be kept within a narrow range. The near-constancy of this parameter emphasizes the remarkable sensitivity of cells sensing changes in extracellular calcium concentration to minimal fluctuations (< 2%) and the level of sophistication of the homeostatic system (1). The identification of a cell surface, Ca2+ (polyvalent cation)-sensing receptor (CaR), has shed considerable light on the molecular aspects of hypercalcemia on cell function (2). Activation of the receptor by calcium triggers an intracellular cascade of second messengers producing a variety of biological effects, many of which have yet to be understood. This suggests, for the first time, that Ca2+ can exert its effects in a hormone-like fashion without crossing the plasma membrane. The demonstration that inherited genetic disorders of Ca2+ homeostasis are associated with mutations that reduce or enhance responsiveness of the receptor to extracellular Ca2+ concentration clearly proposes CaR as the main regulator of divalent mineral ion excretion (3). This hypothesis is confirmed by the assessment of the presence of the receptor in all regions involved in Ca2+ homeostasis (e.g., parathyroid glands, kidney, calcitonin-secreting C cells, bone-derived cell lines, and intestine) (1,4-8). Recently, the receptor has also been found in regions not normally involved in mineral ion metabolism, such as the brain, eye, stomach, and pancreas (9-13). This clearly indicates a much broader relevance of CaR in the maintenance of local ionic homeostasis and, possibly, in the involvement in vital processes such as the regulation of cell fate.
Asunto(s)
Calcio/metabolismo , Receptores de Superficie Celular/fisiología , Animales , Calcio/fisiología , Homeostasis/fisiología , Humanos , Especificidad de Órganos/fisiología , Receptores Sensibles al Calcio , Receptores de Superficie Celular/metabolismoRESUMEN
BACKGROUND: The basolateral isoform of the Na(+):K(+):2Cl(-) cotransporter is expressed in several epithelial and non-epithelial cells, in which it is involved in ion secretion processes and in cell volume regulation. In humans, this cotransporter has been implicated in the development of primary hypertension. The major goal of the present study was to characterize the effect of protein kinase C activation on the function of the Na(+):K(+):2Cl(-) cotransporter isoform present in Xenopus laevis oocytes. METHODS: Oocytes were surgically harvested from adult female Xenopus laevis frogs, defolliculated by incubation in frog ringer containing collagenase B (2 mg/mL) under vigorous shaking, and by hand under the microscope. Only stage V-VI oocytes were used in the study. After overnight incubation in regular frog Ringer, oocytes were switched to a Cl(-)-free ringer for at least 12 h before beginning uptake experiments. The function of the Na(+):K(+):2Cl(-) cotransporter was determined by assessing tracer 22Na(+) uptake in the control group as well as under several experimental conditions, such as changes in extracellular osmolarity, absence of one of the cotransported ions, or the presence of drugs such as the specific cotransporter inhibitor bumetanide, phorbol esters (TPA, PDBu, or 4alphaPDD), and the PKC inhibitor bisindolylmaleimide I. At the end of the uptake period, tracer Na(+) uptake was counted by liquid scintillation of each individual oocyte previously dissolved in SDS. RESULTS: Xenopus oocytes exhibited a bumetanide-sensitive Na(+):K(+):2Cl(-) cotransporter in the plasma membrane activated by hypertonicity and inhibited by hypotonicity. The bumetanide-sensitive fraction of Na(+) uptake was significantly reduced by the addition of phorbol esters TPA or PDBu to the uptake media. This inhibitory effect of PKC activators was dose- and time-dependent. Phorbol ester 4alphaPDD, which cannot activate PKC, exhibited no effect on Na(+):K(+):2Cl(-) cotransporter function. In addition, pretreatment of oocytes with the PKC inhibitor bisindolylmaleimide I partially abolished TPA-induced reduction in the cotransporter function. CONCLUSION: In defolliculated Xenopus laevis oocytes, phorbol esters reduce the function of the Na(+):K(+):2Cl(-) cotransporter by a mechanism that includes the activation of an endogenous PKC.
Asunto(s)
Proteínas Portadoras/antagonistas & inhibidores , Oocitos/metabolismo , Proteína Quinasa C/metabolismo , Animales , Proteínas Portadoras/metabolismo , Activación Enzimática , Femenino , Simportadores de Cloruro de Sodio-Potasio , Xenopus laevisRESUMEN
Nephron tubular epithelium possesses the capacity of adaptation to any salt ingestion condition. The mechanism of adaptation is due in part to an increase in the activity of Na(+):K(+):ATPase at the basolateral membrane. The goal of the present study was to analyze the long-term regulation of the Na(+):K(+):ATPase alpha(1)-subunit mRNA expression during changes in NaCl metabolism. Male Wistar rats given a normal, high, or low NaCl diet, and intraperitoneal administration of the loop diuretic furosemide from 12 h to 7 days were studied. Rats were kept in metabolic cages 4 days before and throughout the study to determine daily urinary electrolyte excretion and osmolarity. At the end of each experimental period, creatinine clearance and serum electrolytes were also measured. Total RNA was extracted from each individual cortex or outer medulla and from pooled inner medullas using the guanidine/cesium chloride method. Na(+):K(+):ATPase alpha(1)-subunit mRNA expression was assessed by nonradioactive dot-blot analysis. Experimental maneuvers were well tolerated and all groups developed the appropriate renal response to each experimental condition. Urinary sodium excretion was significantly higher in rats administered a high sodium diet or furosemide and lower in rats treated with a low sodium diet after 7 days of treatment. Glomerular filtration rate was similar among all groups. However, the level of expression of the Na(+):K(+):ATPase alpha(1)-subunit did not change in any model. Nephron adaptation to the modification in NaCl intake or furosemide administration over 7 days did not include changes in Na(+):K(+):ATPase alpha(1)-subunit mRNA levels.