RESUMEN
Several clinical, genetic and acquired risk factors for venous thromboembolism (VTE) have been identified. However, the molecular pathophysiology and mechanisms of disease progression remain poorly understood. This is reflected by uncertainties regarding the primary and secondary prevention of VTE and the optimal duration of antithrombotic therapy. A growing body of literature points to clinically relevant differences between VTE phenotypes (e.g. deep vein thrombosis (DVT) versus pulmonary embolism (PE), unprovoked versus provoked VTE). Extensive links to cardiovascular, inflammatory and immune-related morbidities are testament to the complexity of the disease. The GMP-VTE project is a prospective, multi-center cohort study on individuals with objectively confirmed VTE. Sequential data sampling was performed at the time of the acute event and during serial follow-up investigations. Various data levels (e.g. clinical, genetic, proteomic and platelet data) are available for multi-dimensional data analyses by means of advanced statistical, bioinformatic and machine learning methods. The GMP-VTE project comprises nâ¯=â¯663 individuals with acute VTE (mean age: 60.3⯱â¯15.9â¯years; female sex: 42.8%). In detail, 28.4% individuals (nâ¯=â¯188) had acute isolated DVT, whereas 71.6% subjects (nâ¯=â¯475) had PE with or without concomitant DVT. In the study sample, 28.9% (nâ¯=â¯129) of individuals with PE and 30.1% (nâ¯=â¯55) of individuals with isolated DVT had a recurrent VTE event at the time of study enrolment. The systems-oriented approach for the comprehensive dataset of the GMP-VTE project may generate new biological insights into the pathophysiology of VTE and refine our current understanding and management of VTE.
Asunto(s)
Tromboembolia Venosa/genética , Estudios de Cohortes , Femenino , Genotipo , Humanos , Masculino , Persona de Mediana Edad , Fenotipo , Estudios Prospectivos , Factores de RiesgoRESUMEN
Olive oil (OO) is the primary source of fat in the Mediterranean diet and has been associated with longevity and a lower incidence of chronic diseases, particularly CHD. Cardioprotective effects of OO consumption have been widely related with improved lipoprotein profile, endothelial function and inflammation, linked to health claims of oleic acid and phenolic content of OO. With CVD being a leading cause of death worldwide, a review of the potential mechanisms underpinning the impact of OO in the prevention of disease is warranted. The current body of evidence relies on mechanistic studies involving animal and cell-based models, epidemiological studies of OO intake and risk factor, small- and large-scale human interventions, and the emerging use of novel biomarker techniques associated with disease risk. Although model systems are important for mechanistic research nutrition, methodologies and experimental designs with strong translational value are still lacking. The present review critically appraises the available evidence to date, with particular focus on emerging novel biomarkers for disease risk assessment. New perspectives on OO research are outlined, especially those with scope to clarify key mechanisms by which OO consumption exerts health benefits. The use of urinary proteomic biomarkers, as highly specific disease biomarkers, is highlighted towards a higher translational approach involving OO in nutritional recommendations.
Asunto(s)
Biomarcadores/orina , Enfermedades Cardiovasculares/prevención & control , Aceite de Oliva/uso terapéutico , Animales , Humanos , Modelos Animales , Proteómica/métodos , Medición de Riesgo/métodosRESUMEN
The conditions of the cellular microenvironment in complex multicellular organisms fluctuate, enforcing permanent adaptation of cells at multiple regulatory levels. Covalent post-translational modifications of proteins provide the short-term response tools for cellular adjustment and growing evidence supports the possibility that protein tyrosine nitration is part of this cellular toolkit and not just a marker for oxidative damage. We have demonstrated that protein tyrosine nitration fulfils the major criteria for signalling and suggest that the normally highly regulated process may lead to disease upon excessive or inappropriate nitration.
Asunto(s)
Mitocondrias/metabolismo , Nitrógeno , Tirosina , Animales , Metabolismo Energético , Homeostasis , Óxido Nítrico/metabolismo , Nitrógeno/química , Nitrógeno/metabolismo , Estrés Oxidativo , Procesamiento Proteico-Postraduccional , Transducción de Señal/fisiología , Tirosina/química , Tirosina/metabolismoRESUMEN
As shown recently, in human skin fibroblasts both a constitutively expressed and the inducible nitric oxide synthase (NOS) isoform are present. To identify the NOS isoforms expressed under standard conditions in healthy human skin fibroblasts and fibroblasts with peroxisomal deficiencies (cell lines from patients suffering from X-chromosome linked Adrenoleukodystrophy (X-ALD) and the Zellweger Syndrome), we cultivated the cells in Dulbecco's modified Eagle's medium without inflammatory mediators. Our experiments clearly showed that human fibroblasts with and without peroxisomal deficiencies only contain the constitutively expressed endothelial nitric oxide synthase (eNOS) isoform and that the eNOS is tyrosine-phosphorylated. The inducible isoform (iNOS) could not be detected under standard conditions. Healthy human skin fibroblasts show a higher specific NOS activity than X-ALD and Zellweger cells (2.25 to 1.68 and 1.17 pmol L-citrulline/min/mg total cellular protein), although the content of eNOS protein does not differ significantly in these cell lines. However the tyrosine-phosphorylated portion of eNOS is significantly lower in X-ALD and Zellweger cells.
Asunto(s)
Adrenoleucodistrofia/enzimología , Óxido Nítrico Sintasa/metabolismo , Trastorno Peroxisomal/enzimología , Piel/enzimología , Síndrome de Zellweger/enzimología , Adrenoleucodistrofia/patología , Línea Celular , Células Cultivadas , Fibroblastos/citología , Fibroblastos/enzimología , Fibroblastos/patología , Humanos , Óxido Nítrico Sintasa/análisis , Óxido Nítrico Sintasa de Tipo II , Óxido Nítrico Sintasa de Tipo III , Trastorno Peroxisomal/patología , Fosfotirosina/metabolismo , Proteínas/metabolismo , Valores de Referencia , Piel/citología , Piel/patología , Fracciones Subcelulares/enzimología , Síndrome de Zellweger/patologíaRESUMEN
The role of nitric oxide, a compound involved in neurotransmission and regulation of cerebral blood flow, in cerebral ischemia is still not fully elucidated yet. Although well studied in adult systems of cerebral ischemia/hypoxia, information on nitric oxide in perinatal asphyxia is limited and, in particular, no direct evidence for its generation has been provided. We therefore decided to study nitric oxide generation in brain of asphyctic rat pups by biophysical and biochemical methods. We used a simple, non-invasive rat model resembling the clinical situation in perinatal asphyxia: rat pups delivered by Caesarean section were placed into a water bath at 37 degrees C still in patent membranes for various asphyctic periods (up to 20 min). Brain pH, cerebral blood flow, neuronal nitrix oxide synthase messenger RNA (by northern and dot blot analysis), immunoreactive protein (by western blot analysis) and nitric oxide synthase activity were determined; generation of nitric oxide was evaluated directly by electron paramagnetic resonance spectroscopy. Neuronal nitric oxide synthase messenger RNA activity and nitric oxide generation were unaffected, whereas neuronal nitric oxide synthase-immunoreactive protein of 150,000 mol. wt was decreased and of 136,000 mol. wt was increased with the length of the asphyctic period. This is the first report on direct evidence for the generation of nitric oxide in perinatal asphyxia and we demonstrate that nitric oxide production remains unaffected even by 20 min of asphyxia, at a time-point when cerebral blood flow was increased four-fold and severe acidosis was present. However, it was found that levels of immunoreactive neuronal nitric oxide synthase of 136,000 mol. wt were increased paralleling the length of asphyxia. Levels of the 150,000 mol. wt immunoreactive neuronal nitric oxide synthase protein decreased, suggesting a different regulation pattern. Thus, the present biochemical and biophysical results form the basis for further investigations on nitric oxide in perinatal asphyxia.
Asunto(s)
Asfixia Neonatal/metabolismo , Química Encefálica , Hipoxia Fetal/metabolismo , Proteínas del Tejido Nervioso/fisiología , Óxido Nítrico Sintasa/fisiología , Óxido Nítrico/fisiología , Animales , Espectroscopía de Resonancia por Spin del Electrón , Femenino , Humanos , Recién Nacido , Neuronas/enzimología , Óxido Nítrico/análisis , Embarazo , Ratas , Ratas Sprague-Dawley , Factores de TiempoRESUMEN
Thyroid hormone abnormalities are strongly associated with Down Syndrome (DS) with elevated thyroid stimulating hormone (TSH) levels as the most consistent finding. Using subtractive hybridization for gene hunting we found significant overexpression of mRNA levels for the TSH-receptor (TSH-R) in brain of a fetus with DS. Based upon this observation we determined TSH-R protein levels in five brain regions of patients with DS (n=8), Alzheimer disease (AD, n=8) and controls (C, n=8). Western blots revealed significantly elevated immunoreactive TSH-R protein(s) 40 kD and 61 kD in temporal and frontal cortex of patients with DS and, unexpectedly, in AD. Levels for the 40 kD protein in temporal cortex were 1.00+/-0.036 (arbitrary units+/-SD) in C, 1.35+/-0.143 in DS, 1.52+/-0.128 in AD; in frontal cortex: 1.00+/-0.046 in C, 1.10+/-0.03 in DS, 1.10+/-0.038 in AD. Levels for the 61 kD protein in temporal cortex were 1.01+/-0.015 in C, 1.47+/-0.013 in DS, 1.623+/-0.026 in AD; in frontal cortex: 1.02+/-0.020 in C, 1.18 +/-0.123 in DS, 1.48+/-0.020 in AD. These results show that elevated brain immunoreactive TSH-R is not specific for DS and maybe reflecting apoptosis, a hallmark of both neurodegenerative disorders, as it is well-documented that the thyroid hormone system is involved in the control of programmed cell death.