Clinical utility of thrombophilia, anticoagulant treatment, and maternal variables as predictors of placenta-mediated pregnancy complications: an extensive analysis.

Objective: The objective of this study is to analyze the usefulness of thrombophilia and antithrombotic drugs in combination with materno-fetal characteristics to generate a predictive model of placenta-mediated pregnancy complications (PMPC) for counseling treatment.Methods: A retrospective analysis was performed in women with singleton pregnancy that required a thrombophilia study, including 222 patients with unknown cause PMPC and 151 women with no complications at current pregnancy in Hospital Clínico Universitario, Lozano, Blesa, Zaragoza, Spain. Chi-squared and Mann-Whitney test were applied to analyze univariate risk factors. Multivariate analysis was performed using logistic regression model with candidate variables: maternal characteristics, obstetric history, thrombophilia, and treatment with low-molecular-weight heparin (LMWH) and/or with acid acetylsalicylic (ASA). The calibration, discrimination, and best cutoff point for the clinical application of the model was analyzed.Results: Maternal characteristics showed differences in median body mass index (BMI), odds ratio (OR): 0.4, smoking habit, OR: 8.5, and hypertension, OR: 11.4, appearing all of them as risk factors. In our study, a prior pregnancy that ended in a child alive was a protective factor OR: 0.02-0.4, and having a previous preterm child was a strong risk factor OR: 4.2. Thrombophilia was not a risk factor. Patients under LMWH treatment (15%) and/or ASA (6.2%) had better pregnancy outcomes, showing both as protective factors: ASA OR: 0.32 and LMWH OR: 0.16. The model has an AUC value of 0.847, with good calibration. A nomogram and an app is provided for this adjusted model with high discrimination ability in internal validation (AUC = 0.833). Our clinical utility analysis guide us to choose 40% as the best threshold probability.Conclusions: We found risk and protective factors associated with PMPC, but our data were not conclusive to demonstrate its relation with maternal thrombophilia. However, the challenger finding is the clinical utility of antithrombotic drugs as a protective factors in PMPC prevention. It is possible to identify patients with high risk of PMPC through a combined predictive model, for counseling treatment.

Molecular and clinical profile of von Willebrand disease in Spain (PCM-EVW-ES): comprehensive genetic analysis by next-generation sequencing of 480 patients.

Molecular diagnosis of patients with von Willebrand disease is pending in most populations due to the complexity and high cost of conventional molecular analyses. The need for molecular and clinical characterization of von Willebrand disease in Spain prompted the creation of a multicenter project (PCM-EVW-ES) that resulted in the largest prospective cohort study of patients with all types of von Willebrand disease. Molecular analysis of relevant regions of the VWF, including intronic and promoter regions, was achieved in the 556 individuals recruited via the development of a simple, innovative, relatively low-cost protocol based on microfluidic technology and next-generation sequencing. A total of 704 variants (237 different) were identified along VWF, 155 of which had not been previously recorded in the international mutation database. The potential pathogenic effect of these variants was assessed by in silico analysis. Furthermore, four short tandem repeats were analyzed in order to evaluate the ancestral origin of recurrent mutations. The outcome of genetic analysis allowed for the reclassification of 110 patients, identification of 37 asymptomatic carriers (important for genetic counseling) and re-inclusion of 43 patients previously excluded by phenotyping results. In total, 480 patients were definitively diagnosed. Candidate mutations were identified in all patients except 13 type 1 von Willebrand disease, yielding a high genotype-phenotype correlation. Our data reinforce the capital importance and usefulness of genetics in von Willebrand disease diagnostics. The progressive implementation of molecular study as the first-line test for routine diagnosis of this condition will lead to increasingly more personalized and effective care for this patient population.

Molecular and clinical profile of von Willebrand disease in Spain (PCM-EVW-ES): Proposal for a new diagnostic paradigm.

The diagnosis of von Willebrand disease (VWD) remains difficult in a significant proportion of patients. A Spanish multicentre study investigated a cohort of 556 patients from 330 families who were analysed centrally. VWD was confirmed in 480. Next generation sequencing (NGS) of the whole coding VWF was carried out in all recruited patients, compared with the phenotype, and a final diagnosis established. A total of 238 different VWF mutations were found, 154 were not included in the Leiden Open Variation Database (LOVD). Of the patients, 463 were found to have VWF mutation/s. A good phenotypic/genotypic association was estimated in 96.5% of the patients. One hundred seventy-four patients had two or more mutations. Occasionally a predominant phenotype masked the presence of a second abnormality. One hundred sixteen patients presented with mutations that had previously been associated with increased von Willebrand factor (VWF) clearance. RIPA unavailability, central phenotypic results disagreement and difficult distinction between severe type 1 and type 3 VWD prevented a clear diagnosis in 70 patients. The NGS study facilitated an appropriate classification in 63 of them. The remaining seven patients presented with a VWF novel mutation pending further investigation. In five patients with a type 3 and two with a type 2A or 2B phenotype with no mutation, an acquired von Willebrand syndrome (AVWS) was suspected/confirmed. These data seem to support NGS as a first line efficient and faster paradigm in VWD diagnosis.

Hemostasis disorders caused by polymer coated iron oxide nanoparticles.

BACKGROUND: Superparamagnetic iron oxide nanoparticles (SPIONs) are inorganic nanomaterials gaining strong clinical interest due to their increasing number of biological and medical applications. The stabilization of SPIONs in a biocompatible stable suspension (bioferrofluid) is generally achieved by an adequate polymeric coating. As many applications using these materials are intended for clinical use through intravenous injection, it is of outmost importance to evaluate their hemostatic behaviour. OBJECTIVES: The aim of this work is to evaluate the hemocompatibility of selected polymer coated bioferrofluids and of their separated components by observing the effects of the bioferrofluid on: the coagulation process--by measuring the prothrombin time (PT) and activated partial thromboplastin time (aPTT)--, the complete blood count (CBC)--Erythrocytes, Leucocytes, Platelets, Hemoglobin and hematocrit--and the hemolysis. METHODS: A SPIONs/bioferrofluid model consisting of a magnetic core of iron oxide nanoparticles embedded within poly(4-vinyl pyridine) (P4VP) and all coated with polyethylene glycol (PEG) has been selected. RESULTS AND CONCLUSIONS: By increasing the concentration of the bioferrofluids an inhibitory effect on the intrinsic pathway of blood coagulation is observed, as indicated by significant increase in aPTT in vitro while PT values stay normal. The effect of the coating components on the inhibition of blood coagulation process shows that PEG has no effect on the process while the P4VP-g-PEG copolymer coating has a strong anticoagulant effect indicating that P4VP is at the origin of such effects. The studied bioferrofluids have no effect on the CBC neither they show in vitro hemolytic effect on blood.

[Research study on renal targeted chemotherapy with magnetic harpoons and intravenous administration of ferro-carbon nanoparticles]. / Estudio experimental sobre quimioterapia focalizada en riñón mediante arpón magnético y administración intravenosa de nanopartículas ferrocarbonosas.

OBJECTIVES: The use of nanoparticules for drug transport is one of the topics with priority interest within the field of biomedical research. Our objective is to show the initial results of an innovative method to focalize drug carrier ferro-carbon nanoparticules to solid organs. We obtained and characterized various types of ferrous magnetic nanoparticules and studied their behaviour in vitro and in vivo in laboratory animals with intrarenal magnetic targets laparoscopically implanted. METHODS: Using a plasma arch we obtained ferro-carbon nanoparticules with the ability to absorb and deliver doxorubicin, showing an excellent behaviour in in vitro rheological studies. Under general anesthesia and control we inserted a gold covered magnetic microharpoon in the left kidney of New Zealand rabbits. At the same time we injected intravenously different doses of various types of nanoparticules. The animals were sacrified ofter pre-established times and pathologic studies of their kidneys, spleens, livers, lungs and bone marrow were carried out. RESULTS: After selection of the most adequafe nanoparticules for our purposes, we ascertained significant differences in the distribution of nanoparticules in postmortem studies, with accumulation in the magnetic target and surrounding renal parenchyma. Nevertheless, the reticuloendothelial system retains a great amount of the injected dose. CONCLUSIONS: Although our magnetic focalization system is effective, nanoparticule temporary protection systems should be tested to allow us avoid the action of the immune system.

Estudio experimental sobre quimioterapia focalizada en riñón mediante arpón magnético y administración intravenosa de nanopartículas ferrocarbonosas / Research study on renal targeted chemotherapy with magnetic harpoons and intravenous administration of ferro-carbon nanoparticles

OBJETIVOS: El empleo de nanopartículas en el transporte de fármacos es actualmente uno de los temas de interés prioritario dentro del campo de la investigación biomédica. Nuestro objetivo es mostrar los resultados iniciales de un método inédito para focalizar en órganos sólidos nanopartículas ferro carbonosas quimioportadoras. Hemos obtenido y caracterizado diversos tipos de nanopartículas ferromagnéticas, y hemos estudiado su comportamiento in vitro e in vivo en animales de experimentación con dianas magnéticas intrarrenales implantadas laparoscópicamente. METODOS: Obtenemos mediante arco de plasma nanopartículas ferro carbonosas capaces de absorber y desorber doxorrubicina y mostrar excelente comportamiento en estudios reológicos in vitro. Bajo anestesia general insertamos mediante control laparoscópico en el riñón izquierdo de conejos de raza neozelandesa un microarpón magnético encapsulado en oro. En el mismo acto inyectamos por vía intravenosa diferentes dosis de distintos tipos de nanopartículas. Los diversos lotes de animales se sacrifican tras diferentes tiempos y se analizan histológicamente ambos riñones, bazo, hígado, pulmones y médula ósea. RESULTADOS: Tras haber seleccionado las nanopartículas más adecuadas para nuestros fines, hemos constatado en los estudios post-mortem diferencias significativas en la distribución de las nanopartículas, con cúmulos de las mismas en la diana magnética y en el parénquima renal circundante. No obstante, el sistema retículoendotelial retiene gran parte de la carga inyectada. CONCLUSIONES: Aunque nuestro sistema de focalización magnética resulta efectivo, deben ensayarse sistemas de blindaje temporal de las nanopartículas que les permita eludir la acción del sistema inmune (AU)

OBJECTIVES: The use of nanoparticules for drug transport is one of the topics with priority interest within the field of biomedical research. Our objective is to show the initial results of an innovative method to focalize drug carrier ferro-carbon nanoparticules to solid organs. We obtained and characterized various types of ferrous magnetic nanoparticules and studied their behaviour in vitro and in vivo in laboratory animals with intrarenal magnetic targets laparoscopically implanted. METHODS: Using a plasma arch we obtained ferro-carbon nanoparticules with the ability to absorb and deliver doxorubicin, showing an excellent behaviour in in vitro rheological studies. Under general anesthesia and control we inserted a gold covered magnetic microharpoon in the left kidney of New Zealand rabbits. At the same time we injected intravenously different doses of various types of nanoparticules. The animals were sacrified ofter pre-established times and pathologic studies of their kidneys, spleens, livers, lungs and bone marrow were carried out. RESULTS: After selection of the most adequafe nanoparticules for our purposes, we ascertained significant differences in the distribution of nanoparticules in postmortem studies, with accumulation in the magnetic target and surrounding renal parenchyma. Nevertheless, the reticuloendothelial system retains a great amount of the injected dose. CONCLUSIONS: Although our magnetic focalization system is effective, nanoparticule temporary protection systems should be tested to allow us avoid the action of the immune system (AU)

Estudio de la posible trombogenicidad de los medios de contraste no iónico a las 24 horas de su empleo / Study of possible thrombogenicity of nonionic contrast media at 24 hours from time of use

Objetivo: Investigar la posible trombogenicidad de los medios de contraste radiológicos no iónicos, mediante la determinación de las modificaciones en los valores de tres marcadores de hipercoagulabilidad, transcurridas 24 h desde su empleo. Material y método: Se estudiaron 16 pacientes a los que se les practicó coronariografía diagnóstica por sospecha clínica de cardiopatía isquémica. Analizamos las variaciones existentes en los valores de tres marcadores de hipercoagulabilidad, el fragmento 1+2 de la protrombina, el complejo trombina-antitrombina y el D-dímero, tras la realización de la coronariografía con un medio de contraste radiológico no iónico. Para ello, realizamos dos determinaciones, una previa a la exploración, y otra a las 24 h. También se realizaron dos estudios de coagulación convencionales, coincidentes con cada toma, para comprobar que no existían alteraciones en la hemostasia detectables mediante los procedimientos analíticos habituales. Como herramienta estadística se empleó el test de la t de Student, con un intervalo de confianza del 95 por ciento. Resultados: No se produjeron modificaciones significativas ni en los parámetros de los estudios de coagulación convencionales, ni en los valores de ninguno de los marcadores de hipercoagulabilidad estudiados a las 24 h de la realización de la coronariografía. Conclusiones: En nuestra serie, el empleo de medios de contraste radiológico no iónico no desencadena un estado de hipercoagulabilidad subclínico a las 24 h de su uso, y, por tanto, no existe un incremento en el riesgo de producción de fenómenos trombóticos por su utilización pasado ese período (AU)