Your browser doesn't support javascript.
loading
Mostrar: 20 | 50 | 100
Resultados 1 - 20 de 23
Filtrar
1.
Blood Cells Mol Dis ; 98: 102704, 2023 01.
Artículo en Inglés | MEDLINE | ID: mdl-36265282

RESUMEN

Lysosomal storage disorders (LSD) are a group of inherited metabolic diseases mainly caused by a deficiency of lysosomal hydrolases, resulting in a gradual accumulation of non-degraded substrates in different tissues causing the characteristic clinical manifestations of such disorders. Confirmatory tests of suspected LSD individuals include enzymatic and genetic testing. A well-oriented clinical suspicion can improve the cost-effectiveness of confirmatory tests and reduce the time expended to achieve the diagnosis. Thus, this work aims to retrospectively study the influence of clinical orientation on the diagnostic yield of enzymatic tests in LSD by retrieving clinical, biochemical, and genetic data obtained from subjects with suspicion of LSD. Our results suggest that the clinical manifestations at the time of diagnosis and the initial clinical suspicion can have a great impact on the diagnostic yield of enzymatic tests, and that clinical orientation performed in specialized clinical departments can contribute to improve it. In addition, the analysis of enzymatic tests as the first step in the diagnostic algorithm can correctly guide subsequent confirmatory genetic tests, in turn increasing their diagnostic yield. In summary, our results suggest that initial clinical suspicion plays a crucial role on the diagnostic yield of confirmatory enzymatic tests in LSD.


Asunto(s)
Enfermedades por Almacenamiento Lisosomal , Humanos , Hospitales , Enfermedades por Almacenamiento Lisosomal/diagnóstico , Enfermedades por Almacenamiento Lisosomal/genética , Enfermedades por Almacenamiento Lisosomal/metabolismo , Lisosomas/metabolismo , Estudios Retrospectivos
2.
J Clin Med ; 11(23)2022 Nov 30.
Artículo en Inglés | MEDLINE | ID: mdl-36498692

RESUMEN

Fetal echocardiography has limited prognostic ability in the evaluation of left-sided congenital heart defects (left heart defects). Cord blood cardiovascular biomarkers could improve the prognostic evaluation of left heart defects. A multicenter prospective cohort (2013−2019) including fetuses with left heart defects (aortic coarctation, aortic stenosis, hypoplastic left heart, and multilevel obstruction (complex left heart defects) subdivided according to their outcome (favorable vs. poor), and control fetuses were evaluated in the third trimester of pregnancy at three referral centers in Spain. Poor outcome was defined as univentricular palliation, heart transplant, or death. Cord blood concentrations of N-terminal precursor of B-type natriuretic peptide, Troponin I, transforming growth factor ß, placental growth factor, and soluble fms-like tyrosine kinase-1 were determined. A total of 45 fetuses with left heart defects (29 favorable and 16 poor outcomes) and 35 normal fetuses were included, with a median follow-up of 3.1 years (interquartile range 1.4−3.9). Left heart defects with favorable outcome showed markedly increased cord blood transforming growth factor ß (normal heart median 15.5 ng/mL (6.8−21.4) vs. favorable outcome 51.7 ng/mL (13.8−73.9) vs. poor outcome 25.1 ng/mL (6.9−39.0), p = 0.001) and decreased placental growth factor concentrations (normal heart 17.9 pg/mL (13.8−23.9) vs. favorable outcome 12.8 pg/mL (11.7−13.6) vs. poor outcome 11.0 pg/mL (8.8−15.4), p < 0.001). Poor outcome left heart defects had higher N-terminal precursor of B-type natriuretic peptide (normal heart 508.0 pg/mL (287.5−776.3) vs. favorable outcome 617.0 pg/mL (389.8−1087.8) vs. poor outcome 1450.0 pg/mL (919.0−1645.0), p = 0.001) and drastically reduced soluble fms-like tyrosine kinase-1 concentrations (normal heart 1929.7 pg/mL (1364.3−2715.8) vs. favorable outcome (1848.3 pg/mL (646.9−2313.6) vs. poor outcome 259.0 pg/mL (182.0−606.0), p < 0.001). Results showed that fetuses with left heart defects present a distinct cord blood biomarker profile according to their outcome.

3.
Hypertension ; 76(6): 1808-1816, 2020 12.
Artículo en Inglés | MEDLINE | ID: mdl-33012203

RESUMEN

Preeclampsia is caused by placental impairment with increased expression of sFlt-1 (soluble fms-like tyrosine kinase 1) and decreased PlGF (placental growth factor); it has been associated with cardiovascular morbidity and mortality later in life, but the underlying mechanism remains unknown. The aim of this study was to determine whether sFlt-1 and PlGF levels during preeclampsia are associated to long-term cardiovascular risk. We prospectively recruited 43 women with previous preeclampsia and 21 controls with uncomplicated pregnancies. Cardiovascular risk assessment ≈12 years later included maternal hemodynamic, cardiac function and structure, biomarker analysis, and carotid-intima thickness evaluation. Women with previous preeclampsia had higher prevalence of hypertensive disorders and dyslipidemia than controls. In addition, they had worse global longitudinal strain, thicker left ventricular septal and posterior walls, more myocardial mass and increased carotid intima-media thickness compared with controls. PlGF during pregnancy correlated positively with high-density lipoprotein (r=0.341; P=0.006), and negatively with global longitudinal strain (r=-0.581; P<0.001), carotid intima-media thickness (r=-0.251; P=0.045), and mean arterial blood pressure (r=-0.252; P=0.045), when adjusted by study group. sFlt correlated negatively with high-density lipoprotein (r=-0.372; P=0.002) and apolipoprotein A-1 (r=-0.257; P=0.040), and positively with carotid intima-media thickness (r=0.269; P=0.032) and left ventricular posterior wall thickness (r=0.368; P=0.003). The antiangiogenic state present in preeclampsia is related to greater prevalence of cardiovascular risk factors ≈12 years after delivery. The knowledge of altered angiogenic factors may help detect women with a higher risk for premature cardiovascular disease, who will require earlier follow-up after delivery.


Asunto(s)
Inductores de la Angiogénesis/metabolismo , Enfermedades Cardiovasculares/metabolismo , Factor de Crecimiento Placentario/metabolismo , Preeclampsia/metabolismo , Receptor 1 de Factores de Crecimiento Endotelial Vascular/metabolismo , Adulto , Enfermedades Cardiovasculares/diagnóstico , Grosor Intima-Media Carotídeo , Femenino , Factores de Riesgo de Enfermedad Cardiaca , Humanos , Hipertensión/diagnóstico , Hipertensión/metabolismo , Placenta/metabolismo , Preeclampsia/diagnóstico , Embarazo , Solubilidad , Factores de Tiempo
4.
Clin Immunol ; 191: 44-51, 2018 06.
Artículo en Inglés | MEDLINE | ID: mdl-29572183

RESUMEN

There is scarce literature about autoinflammation in syndromic patients. We describe a patient who, in addition to psychomotor and growth delay, presented with fevers, neutrophilic dermatosis, and recurrent orogenital ulcers. Comparative Genomic Hybridization (CGH) array permitted to identify a 13.13Mb deletion on chromosome 6, encompassing 53 genes, and including TNFAIP3 gene (A20). A20 is a potent inhibitor of the NF-kB signalling pathway and restricts inflammation via its deubiquitinase activity. Western blotting and immunoprecipitation assays showed decreased A20 expression and increased phosphorylation of p65 and IkBa. Patient's cells displayed increased levels of total K63-linked ubiquitin and increased levels of ubiquitinated RIP and NEMO after stimulation with TNF. We describe the molecular characterization of an autoinflammatory disease due to a large chromosomal deletion and review the phenotypes of patients with A20 haploinsufficiency. CGH arrays should be the first diagnostic method for comprehensive analysis of patients with syndromic features and immune dysregulation.


Asunto(s)
Deleción Cromosómica , Cromosomas Humanos Par 6 , Haploinsuficiencia/genética , Inflamación/etiología , Proteína 3 Inducida por el Factor de Necrosis Tumoral alfa/genética , Células Cultivadas , Niño , Hibridación Genómica Comparativa , Humanos , Masculino , FN-kappa B/fisiología
5.
J Transl Med ; 15(1): 43, 2017 02 21.
Artículo en Inglés | MEDLINE | ID: mdl-28222799

RESUMEN

BACKGROUND: Niemann-Pick disease type C (NP-C) is a rare, autosomal recessive neurodegenerative disease caused by mutations in either the NPC1 or NPC2 genes. The diagnosis of NP-C remains challenging due to the non-specific, heterogeneous nature of signs/symptoms. This study assessed the utility of plasma chitotriosidase (ChT) and Chemokine (C-C motif) ligand 18 (CCL18)/pulmonary and activation-regulated chemokine (PARC) in conjunction with the NP-C suspicion index (NP-C SI) for guiding confirmatory laboratory testing in patients with suspected NP-C. METHODS: In a prospective observational cohort study, incorporating a retrospective determination of NP-C SI scores, two different diagnostic approaches were applied in two separate groups of unrelated patients from 51 Spanish medical centers (n = 118 in both groups). From Jan 2010 to Apr 2012 (Period 1), patients with ≥2 clinical signs/symptoms of NP-C were considered 'suspected NP-C' cases, and NPC1/NPC2 sequencing, plasma chitotriosidase (ChT), CCL18/PARC and sphingomyelinase levels were assessed. Based on findings in Period 1, plasma ChT and CCL18/PARC, and NP-C SI prediction scores were determined in a second group of patients between May 2012 and Apr 2014 (Period 2), and NPC1 and NPC2 were sequenced only in those with elevated ChT and/or elevated CCL18/PARC and/or NP-C SI ≥70. Filipin staining and 7-ketocholesterol (7-KC) measurements were performed in all patients with NP-C gene mutations, where possible. RESULTS: In total across Periods 1 and 2, 10/236 (4%) patients had a confirmed diagnosis o NP-C based on gene sequencing (5/118 [4.2%] in each Period): all of these patients had two causal NPC1 mutations. Single mutant NPC1 alleles were detected in 8/236 (3%) patients, overall. Positive filipin staining results comprised three classical and five variant biochemical phenotypes. No NPC2 mutations were detected. All patients with NPC1 mutations had high ChT activity, high CCL18/PARC concentrations and/or NP-C SI scores ≥70. Plasma 7-KC was higher than control cut-off values in all patients with two NPC1 mutations, and in the majority of patients with single mutations. Family studies identified three further NP-C patients. CONCLUSION: This approach may be very useful for laboratories that do not have mass spectrometry facilities and therefore, they cannot use other NP-C biomarkers for diagnosis.


Asunto(s)
Quimiocinas CC/sangre , Hexosaminidasas/sangre , Enfermedad de Niemann-Pick Tipo C/sangre , Enfermedad de Niemann-Pick Tipo C/diagnóstico , Adolescente , Adulto , Niño , Preescolar , Simulación por Computador , Demografía , Familia , Femenino , Filipina , Humanos , Lactante , Masculino , Persona de Mediana Edad , Mutación , Enfermedad de Niemann-Pick Tipo C/enzimología , Oxiesteroles , Estudios Prospectivos , Adulto Joven
6.
Placenta ; 42: 44-50, 2016 Jun.
Artículo en Inglés | MEDLINE | ID: mdl-27238713

RESUMEN

OBJECTIVE: To explore in women with late-onset preeclampsia (PE) the association between maternal levels of angiogenic/antiangiogenic factors in the first trimester of pregnancy and histological findings attributable to placental underperfusion (PUP). METHODS: A nested case-control cohort study was conducted in 73 women with pregnancies complicated by late-onset PE (>34 weeks at delivery) matched with controls. First trimester uterine artery Doppler (UtA); maternal levels of placental growth factor (PlGF) and soluble fms-like tyrosine kinase-1 (sFlt-1) were retrieved. Placentas were histologically evaluated using a hierarchical and standardized classification system. One-way ANOVA with linear polynomial contrast or linear-by-linear association test was performed to test the hypothesis of a linear association across study groups (controls, PE without PUP and PE with PUP). RESULTS: In 54 (74%) placentas, 89 placental histological findings qualifying for PUP were found. Across study groups, significant values were observed in maternal levels of decreased PlGF (MoM values: 1.53, 1.41 and 1.37; p < 0.001), increased sFlt-1 (MoM values: 3.11, 3.11 and 3.22; p = 0.002), increased sFlt-1/PlGF ratio (MoM values: 2.3, 2.3 and 2.44; p < 0.001), abnormal UtA Doppler (MoM values: 1, 1.26 and 1.32; p < 0.001), and worse perinatal outcomes in terms of gestational age at delivery, cesarean section for not reassuring fetal status, birth weight and neonatal acidosis. DISCUSSION: In late-onset PE an imbalance of circulating angiogenic and anti-angiogenic factors already present at 8-10 weeks of pregnancy was associated with histological findings reflecting placental insufficiency. An early first trimester screening by angiogenic factors might help to identify patients with placental involvement among late-onset PE cases. CONCLUSION: In late-onset preeclampsia, first-trimester uterine Doppler and circulating levels of angiogenic/antiangiogenic factors are associated with placental underperfusion.


Asunto(s)
Factor de Crecimiento Placentario/sangre , Insuficiencia Placentaria/diagnóstico , Preeclampsia/diagnóstico , Receptor 1 de Factores de Crecimiento Endotelial Vascular/sangre , Adulto , Biomarcadores/sangre , Estudios de Casos y Controles , Femenino , Humanos , Recién Nacido , Insuficiencia Placentaria/sangre , Insuficiencia Placentaria/diagnóstico por imagen , Insuficiencia Placentaria/patología , Preeclampsia/sangre , Preeclampsia/diagnóstico por imagen , Preeclampsia/patología , Embarazo , Primer Trimestre del Embarazo , Ultrasonografía Doppler en Color , Ultrasonografía Prenatal , Arteria Uterina/diagnóstico por imagen
7.
Biol Chem ; 397(3): 269-79, 2016 Mar.
Artículo en Inglés | MEDLINE | ID: mdl-26756094

RESUMEN

The pathogenic basis of abnormal placentation and dysfunction in preeclampsia (PE) is highly complex and incompletely understood. Secretory sphyngomyelinase activity (S-ASM) was analyzed in plasma samples from 158 pregnant women developing PE and 112 healthy pregnant controls. Serum PlGF, sFlt-1, s-Endoglin and sVCAM were measured. Results showed S-ASM activity to be higher in women who later developed PE than in those with uncomplicated pregnancies (40.6% and 28.8% higher in the late- and early-onset groups, respectively). Plasma S-ASM activity correlated significantly with circulating markers of endothelial damage in the late-PE group (endoglin and sVCAM-1), with plasma cholesterol and total lipid levels. However, these significant associations were not observed in the early-PE or control groups. This work provides the first evidence of significantly elevated circulating S-ASM activity in the first trimester of pregnancy in women who go on to develop PE; thus, it may be deduced that the circulating form of ASM is biologically active in PE and could contribute to promoting endothelial dysfunction and cardiovascular programming. Plasma S-ASM measurement may have clinical relevance as a further potential biomarker contributing to the earliest identification of women at risk of developing preeclampsia.


Asunto(s)
Preeclampsia/sangre , Esfingomielina Fosfodiesterasa/metabolismo , Adulto , Antígenos CD/sangre , Biomarcadores/sangre , Estudios de Casos y Controles , Endoglina , Femenino , Humanos , Lípidos/sangre , Proteínas de la Membrana/sangre , Preeclampsia/epidemiología , Embarazo , Primer Trimestre del Embarazo , Receptores de Superficie Celular/sangre , Esfingomielina Fosfodiesterasa/sangre , Molécula 1 de Adhesión Celular Vascular/sangre , Receptor 1 de Factores de Crecimiento Endotelial Vascular/sangre
8.
Fetal Diagn Ther ; 40(1): 13-20, 2016.
Artículo en Inglés | MEDLINE | ID: mdl-26421431

RESUMEN

OBJECTIVE: To evaluate in normally growing fetuses at routine 32-36 weeks scan the performance of maternal angiogenic factors, Doppler and ultrasound indices in predicting smallness for gestational age (SGA) at birth. METHODS: A cohort of 1,000 singleton pregnancies with normal estimated fetal weight (EFW, ≥10th centile) at 32-36 weeks scan was included. At inclusion, Doppler indices (mean uterine artery pulsatility index [mUtA-PI], cerebroplacental ratio and normalized umbilical vein blood flow by EFW (ml/min/kg) were evaluated, and blood samples were collected and frozen. Nested in this cohort, maternal circulating placental growth factor (PlGF) and soluble fms-like tyrosine kinase-1 (sFlt-1) were assayed by enzyme-linked immunosorbent assay in all cases with a birth weight <10th centile by customized standards and in an equivalent number of controls (birth weight ≥10th centile). RESULTS: 160 cases were included (80 SGA and 80 controls). EFW (2,128 vs. 2,279 g, p < 0.001), mUtA-PI z-values (-0.25 vs. -0.65, p = 0.034) and sFlt-1/PlGF ratio (11.10 vs. 6.74, p < 0.005) were lower in SGA. The combination of sFlt-1/PlGF ratio and EFW resulted in a 66.3% detection rate for subsequent SGA, with 20% of false-positives. Fetal Doppler indices were not predictive of SGA. CONCLUSIONS: In normally growing fetuses, maternal angiogenic factors add to ultrasound parameters in predicting subsequent SGA at birth. This supports further research to investigate composite scores in order to improve the definition and identification of fetal growth restriction.


Asunto(s)
Inductores de la Angiogénesis/metabolismo , Retardo del Crecimiento Fetal/diagnóstico por imagen , Recién Nacido de Bajo Peso , Biomarcadores/metabolismo , Estudios de Casos y Controles , Estudios de Cohortes , Femenino , Humanos , Recién Nacido , Modelos Logísticos , Embarazo , Tercer Trimestre del Embarazo , Curva ROC , Ultrasonografía Prenatal
9.
PLoS One ; 10(8): e0135873, 2015.
Artículo en Inglés | MEDLINE | ID: mdl-26287674

RESUMEN

Nonsense mutations are quite prevalent in inherited diseases. Readthrough drugs could provide a therapeutic option for any disease caused by this type of mutation. Geneticin (G418) and gentamicin were among the first to be described. Novel compounds have been generated, but only a few have shown improved results. PTC124 is the only compound to have reached clinical trials. Here we first investigated the readthrough effects of gentamicin on fibroblasts from one patient with Sanfilippo B, one with Sanfilippo C, and one with Maroteaux-Lamy. We found that ARSB activity (Maroteaux-Lamy case) resulted in an increase of 2-3 folds and that the amount of this enzyme within the lysosomes was also increased, after treatment. Since the other two cases (Sanfilippo B and Sanfilippo C) did not respond to gentamicin, the treatments were extended with the use of geneticin and five non-aminoglycoside (PTC124, RTC13, RTC14, BZ6 and BZ16) readthrough compounds (RTCs). No recovery was observed at the enzyme activity level. However, mRNA recovery was observed in both cases, nearly a two-fold increase for Sanfilippo B fibroblasts with G418 and around 1.5 fold increase for Sanfilippo C cells with RTC14 and PTC124. Afterwards, some of the products were assessed through in vitro analyses for seven mutations in genes responsible for those diseases and, also, for Niemann-Pick A/B. Using the coupled transcription/translation system (TNT), the best results were obtained for SMPD1 mutations with G418, reaching a 35% recovery at 0.25 µg/ml, for the p.W168X mutation. The use of COS cells transfected with mutant cDNAs gave positive results for most of the mutations with some of the drugs, although to a different extent. The higher enzyme activity recovery, of around two-fold increase, was found for gentamicin on the ARSB p.W146X mutation. Our results are promising and consistent with those of other groups. Further studies of novel compounds are necessary to find those with more consistent efficacy and fewer toxic effects.


Asunto(s)
Codón de Terminación/genética , Gentamicinas/uso terapéutico , Mucopolisacaridosis III/genética , Mucopolisacaridosis VI/genética , Animales , Células COS , Línea Celular , Chlorocebus aethiops , Codón sin Sentido/efectos de los fármacos , Codón sin Sentido/genética , Codón de Terminación/efectos de los fármacos , Fibroblastos/citología , Humanos , Lisosomas/metabolismo , Mucopolisacaridosis III/tratamiento farmacológico , Mucopolisacaridosis VI/tratamiento farmacológico , Enfermedad de Niemann-Pick Tipo A/tratamiento farmacológico , Enfermedad de Niemann-Pick Tipo A/genética , Enfermedad de Niemann-Pick Tipo B/tratamiento farmacológico , Enfermedad de Niemann-Pick Tipo B/genética , ARN Mensajero/genética
10.
Oxid Med Cell Longev ; 2015: 509241, 2015.
Artículo en Inglés | MEDLINE | ID: mdl-25852816

RESUMEN

Down's syndrome (DS) is characterized by a complex phenotype associated with chronic oxidative stress and mitochondrial dysfunction. Overexpression of genes on chromosome-21 is thought to underlie the pathogenesis of the major phenotypic features of DS, such as premature aging. Using cultured fibroblasts with trisomy 21 (T21F), this study aimed to ascertain whether an imbalance exists in activities, mRNA, and protein expression of the antioxidant enzymes SOD1, SOD2, glutathione-peroxidase, and catalase during the cell replication process in vitro. T21F had high SOD1 expression and activity which led to an interenzymatic imbalance in the antioxidant defense system, accentuated with replicative senescence. Intracellular ROS production and oxidized protein levels were significantly higher in T21F compared with control cells; furthermore, a significant decline in intracellular ATP content was detected in T21F. Cell senescence was found to appear prematurely in DS cells as shown by SA-ß-Gal assay and p21 assessment, though not apoptosis, as neither p53 nor the proapoptotic proteins cytochrome c and caspase 9 were altered in T21F. These novel findings would point to a deleterious role of oxidatively modified molecules in early cell senescence of T21F, thereby linking replicative and stress-induced senescence in cultured cells to premature aging in DS.


Asunto(s)
Antioxidantes/metabolismo , Senescencia Celular , Fibroblastos/metabolismo , Especies Reactivas de Oxígeno/metabolismo , Adenosina Trifosfato/metabolismo , Caspasa 9/metabolismo , Catalasa/metabolismo , Supervivencia Celular , Células Cultivadas , Citocromos c/metabolismo , Síndrome de Down/metabolismo , Síndrome de Down/patología , Feto/citología , Fibroblastos/citología , Glutatión Peroxidasa/genética , Glutatión Peroxidasa/metabolismo , Humanos , Mitocondrias/metabolismo , Estrés Oxidativo , Superóxido Dismutasa/genética , Superóxido Dismutasa/metabolismo , Superóxido Dismutasa-1 , Trisomía/genética , Proteína p53 Supresora de Tumor/metabolismo
11.
Proteins ; 83(1): 91-104, 2015 Jan.
Artículo en Inglés | MEDLINE | ID: mdl-25382311

RESUMEN

Loss-of-function mutations of the enzyme alpha-galactosidase A (GLA) causes Fabry disease (FD), that is a rare and potentially fatal disease. Identification of these pathological mutations by sequencing is important because it allows an early treatment of the disease. However, before taking any treatment decision, if the mutation identified is unknown, we first need to establish if it is pathological or not. General bioinformatic tools (PolyPhen-2, SIFT, Condel, etc.) can be used for this purpose, but their performance is still limited. Here we present a new tool, specifically derived for the assessment of GLA mutations. We first compared mutations of this enzyme known to cause FD with neutral sequence variants, using several structure and sequence properties. Then, we used these properties to develop a family of prediction methods adapted to different quality requirements. Trained and tested on a set of known Fabry mutations, our methods have a performance (Matthews correlation: 0.56-0.72) comparable or better than that of the more complex method, Polyphen-2 (Matthews correlation: 0.61), and better than those of SIFT (Matthews correl.: 0.54) and Condel (Matthews correl.: 0.51). This result is validated in an independent set of 65 pathological mutations, for which our method displayed the best success rate (91.0%, 87.7%, and 73.8%, for our method, PolyPhen-2 and SIFT, respectively). These data confirmed that our specific approach can effectively contribute to the identification of pathological mutations in GLA, and therefore enhance the use of sequence information in the identification of undiagnosed Fabry patients.


Asunto(s)
Enfermedad de Fabry/enzimología , Enfermedad de Fabry/genética , Mutación/genética , alfa-Galactosidasa/genética , Secuencia de Aminoácidos , Secuencia Conservada , Enfermedad de Fabry/patología , Humanos , Datos de Secuencia Molecular , Programas Informáticos , Relación Estructura-Actividad , alfa-Galactosidasa/química
12.
Prenat Diagn ; 35(2): 183-91, 2015 Feb.
Artículo en Inglés | MEDLINE | ID: mdl-25346181

RESUMEN

OBJECTIVE: The aim of this article is to develop the best first-trimester screening model for preeclampsia (PE) based on maternal characteristics, biophysical parameters, and angiogenic factors in a low-risk population. METHODS: A prospective cohort of 9462 pregnancies undergoing first-trimester screening is used. Logistic regression predictive models were developed for early and late PE (cut-off of 34 weeks' gestation at delivery). Data included the a priori risk (maternal characteristics), mean arterial pressure (MAP), and uterine artery (UtA) Doppler (11-13 weeks) in all cases. Plasma levels (8-11 weeks) of human chorionic gonadotrophin, pregnancy-associated plasma protein A, placental growth factor (PlGF), and soluble Fms-like tyrosine kinase-1 (sFlt-1) were analyzed using a nested case-control study design. RESULTS: The best model for early PE (n = 57, 0.6%) included a priori risk, MAP, UtA Doppler, PlGF, and sFlt-1 achieving detection rates of 87.7% and 91.2% for 5% and 10% false-positive rates, respectively (AUC: 0.98 [95% CI: 0.97-0.99]). For late PE (n = 246, 2.6%), the best model included the a priori risk, MAP, UtA Doppler, PlGF, and sFlt-1 achieving detection rates of 68.3% and 76.4% at 5% and 10% of false-positive rates, respectively (AUC: 0.87 [95% CI: 0.84-0.90]). CONCLUSION: Preeclampsia can be predicted with high accuracy in general obstetric populations with a low risk for PE, by combined algorithms. Angiogenic factors substantially improved the prediction.


Asunto(s)
Tamizaje Masivo , Preeclampsia/diagnóstico , Adulto , Presión Sanguínea , Estudios de Casos y Controles , Gonadotropina Coriónica Humana de Subunidad beta/sangre , Femenino , Humanos , Análisis Multivariante , Factor de Crecimiento Placentario , Valor Predictivo de las Pruebas , Embarazo , Proteínas Gestacionales/sangre , Primer Trimestre del Embarazo , Proteína Plasmática A Asociada al Embarazo/metabolismo , Ultrasonografía , Arteria Uterina/diagnóstico por imagen , Receptor 1 de Factores de Crecimiento Endotelial Vascular/sangre
13.
Fetal Diagn Ther ; 35(4): 258-66, 2014.
Artículo en Inglés | MEDLINE | ID: mdl-24714555

RESUMEN

OBJECTIVE: To explore the predictive role of angiogenic factors for the prediction of early and late preeclampsia (PE) in the first trimester. METHODS: A nested case-control study, within a cohort of 5,759 pregnancies, including 28 cases of early, 84 of late PE (cut-off 34 weeks) and 84 controls. Maternal characteristics, mean blood pressure (MAP), uterine artery (UtA) Doppler (11-13 weeks), vascular endothelial growth factor, placental growth factor (PlGF), soluble Fms-like tyrosine kinase-1 (sFlt-1) and soluble endoglin (8-11 weeks) were measured/recorded. All parameters were normalized by logarithmic transformation; logistic regression analysis was used to predict PE. RESULTS: For early PE, significant contributions were chronic hypertension, previous PE, MAP, UtA Doppler, PlGF and sFlt-1. A model including these predictors achieved detection rates (DR) of 77.8 and 88.9% for 5 and 10% false-positive rates (FPR), respectively (AUC 0.958; 95% CI 0.920-0.996). For late PE, significant contributions were provided by body mass index, previous PE, UtA Doppler, PlGF and sFlt-1. The model including these factors achieved DR of 51.2 and 69% at 5 and 10% FPR, respectively (AUC 0.888; 95% CI 0.840-0.936). CONCLUSIONS: Among angiogenic factors, not only PlGF but also sFlt-1 substantially improve the prediction for early and late PE. The data need confirmation in larger studies.


Asunto(s)
Preeclampsia/diagnóstico , Proteínas Gestacionales/sangre , Primer Trimestre del Embarazo/sangre , Receptor 1 de Factores de Crecimiento Endotelial Vascular/sangre , Adulto , Antígenos CD/sangre , Biomarcadores/sangre , Presión Sanguínea , Estudios de Casos y Controles , Estudios de Cohortes , Diagnóstico Precoz , Endoglina , Femenino , Humanos , Modelos Logísticos , Análisis Multivariante , Factor de Crecimiento Placentario , Preeclampsia/sangre , Valor Predictivo de las Pruebas , Embarazo , Resultado del Embarazo , Receptores de Superficie Celular/sangre , Medición de Riesgo , Factor A de Crecimiento Endotelial Vascular/sangre
14.
Stroke ; 45(2): 432-7, 2014 Feb.
Artículo en Inglés | MEDLINE | ID: mdl-24335228

RESUMEN

BACKGROUND AND PURPOSE: Fluorescent molecular peroxidation products (FMPPs) are considered potential markers of molecular oxidative damage and may provoke increased permeability and disruption of the blood-brain barrier. This study aimed to determine the value of FMPPs as a biomarker to predict neurological worsening related to early hemorrhagic transformation. METHODS: Baseline FMPP levels were measured in 186 consecutive acute ischemic stroke patients before tissue plasminogen activator treatment was administered. A serial FMPP profile (baseline before tissue plasminogen activator treatment, and 1, 2, 12, and 24 hours from treatment) was determined in a subset of 100 patients. Computed tomographic scans were performed at admission and repeated at 24 to 48 hours or after neurological worsening occurred. Symptomatic intracranial hemorrhage was defined as blood at any site in the brain associated with neurological deterioration. RESULTS: Patients who worsened had higher median FMPP levels compared with those who did not (59.68 [48.63-85.73] versus 44.87 [36.37-58.90] Uf/mL; P=0.035) at baseline. After logistic regression multivariate analysis, FMPP >48.2 Uf/mL together with age, hypertension, and systolic blood pressure remained baseline predictors of worsening at 48 hours. Moreover, baseline FMPP determination helped to distinguish between patients who worsened and those who did not (Integrated Discrimination Improvement index, 5.7%; P=0.0004). Finally, within patients who had worsened at 48 hours, those with symptomatic intracranial hemorrhage had higher FMPP levels (P=0.038). CONCLUSIONS: FMPPs might be a valuable biomarker of poor early neurological outcome and be related to the appearance of symptomatic intracranial hemorrhage in tissue plasminogen activator-treated patients, one of the most feared neurological complications after thrombolytic treatment of acute ischemic stroke.


Asunto(s)
Biomarcadores/sangre , Hemorragia Cerebral/etiología , Enfermedades del Sistema Nervioso/etiología , Peróxidos/sangre , Accidente Cerebrovascular/complicaciones , Accidente Cerebrovascular/tratamiento farmacológico , Terapia Trombolítica/efectos adversos , Anciano , Análisis de Varianza , Hemorragia Cerebral/diagnóstico por imagen , Hemorragia Cerebral/epidemiología , Progresión de la Enfermedad , Determinación de Punto Final , Femenino , Fibrinolíticos/efectos adversos , Fibrinolíticos/uso terapéutico , Fluorescencia , Humanos , Masculino , Estrés Oxidativo , Pronóstico , Activador de Tejido Plasminógeno/efectos adversos , Activador de Tejido Plasminógeno/uso terapéutico , Tomografía Computarizada por Rayos X
15.
Int J Stroke ; 9(7): 910-6, 2014 Oct.
Artículo en Inglés | MEDLINE | ID: mdl-24148838

RESUMEN

BACKGROUND AND AIMS: Chitotriosidase, a component of innate immunity, constitutes a sensitive parameter of macrophage activation and its elevated plasma activity reflects an inflammatory response. Given the deleterious effects of inflammation in brain ischemia, we aimed to assess the prognostic value of chitotriosidase activity in acute stroke patients. METHODS: The study comprised 159 acute stroke patients and 51 age-matched controls. Plasma chitotriosidase activity was serially determined by fluorometric assay. Short-term neurological outcome was determined at 48 h and functional outcome at three-months. Predictors of neurological and functional outcome were determined via multivariate analysis, and the additional predictive value of chitotriosidase was tested with the Integrated Discrimination Index and the Net Reclassification Improvement. RESULTS: Stroke patients showed increased levels of baseline chitotriosidase activity compared to controls [114·2 (74·65-182·95) nmol/ml/h vs. 54·4 (32·7-76·4); P < 0·0001]. Chitotriosidase activity (<118·75) was found to be an independent predictor of neurological improvement at 48 h (odds ratio: 3·25; 95% confidence interval: 1·54-6·85; P=0·002), and the addition of plasma chitotriosidase activity showed a better prediction of improvement at 48 h (Integrated Discrimination Index=5·7%, Net Reclassification Improvement=11·6%, P<0·05) over the predictive model constituted only with clinical information. Although patients disabled at three-months showed higher baseline chitotriosidase levels, it was not an independent predictor of long-term disability. CONCLUSIONS: Baseline chitotriosidase activity in acute stroke patients treated with tissue plasminogen activator (tPA) may constitute a prognostic predictor of short-term outcome, adding a moderate additional predictive value. Our results underline the deleterious role of inflammation in acute stroke patients.


Asunto(s)
Hexosaminidasas/sangre , Accidente Cerebrovascular/sangre , Anciano , Biomarcadores/sangre , Femenino , Humanos , Masculino , Análisis Multivariante , Pronóstico , Accidente Cerebrovascular/diagnóstico , Accidente Cerebrovascular/terapia , Resultado del Tratamiento
16.
Immunobiology ; 217(1): 44-53, 2012 Jan.
Artículo en Inglés | MEDLINE | ID: mdl-21903293

RESUMEN

Several soft-tissue dermal fillers have been reported to provoke immunogenicity and may cause adverse reactions despite claims regarding their safety. This study aimed to assess biomaterial-induced macrophage activation, cell-mediated immune response and oxidative stress in 169 patients with dermal bioimplants. To this end, we analysed plasma concentrations of myeloperoxidase (MPO), the chitinase-like proteins chitotriosidase and YKL-40 and molecular oxidative damage. The present study shows, for the first time, that the components of innate immunity: chitotriosidase and YKL-40, are significantly higher in patients with certain bioimplants and these markers of monocyte/macrophage activation rose progressively as adverse reactions (AR) evolved. Plasma MPO levels increased 4-fold in filler users with AR and 3-fold in those without. Analysis by filler type showed subjects injected with calcium hydroxylapatite, methacrylate, acrylamides and silicone to have values significantly above those of non-filler subjects for at least two plasma biomarkers, probably because the afore-mentioned biomaterials are permanent and prone to trigger AR in the long term. By contrast, hyaluronic acid alone elicited little immune response. Plasma concentrations of markers of oxidative damage to lipids and proteins were found to be significantly higher in users of four of the nine dermal fillers studied. These diffusible products of molecular peroxidation would stem from the reaction catalysed by MPO that generates potent oxidants, leading to cell oxidative damage which, in turn, may exert deleterious effects on the organism. Overall, the results of this study on the effects of a range of dermal fillers point to chronic activation of the immune response mediated by macrophages and PMNs. The increases in plasma of MPO, chitotriosidase and YKL-40 proteins and products of macromolecular peroxidation suggests that these molecules could serve as blood-based biochemical markers and alert to the risk of chronic immune system activation and development of adverse events that may arise from the use of certain bioimplants.


Asunto(s)
Adipoquinas/inmunología , Materiales Biocompatibles/administración & dosificación , Hexosaminidasas/inmunología , Inmunidad Celular/efectos de los fármacos , Lectinas/inmunología , Activación de Macrófagos/efectos de los fármacos , Macrófagos/efectos de los fármacos , Peroxidasa/inmunología , Adipoquinas/sangre , Administración Cutánea , Adulto , Anciano , Materiales Biocompatibles/efectos adversos , Materiales Biocompatibles/metabolismo , Materiales Biocompatibles/uso terapéutico , Biomarcadores/sangre , Proteína 1 Similar a Quitinasa-3 , Femenino , Hexosaminidasas/sangre , Humanos , Ácido Hialurónico/administración & dosificación , Ácido Hialurónico/uso terapéutico , Lectinas/sangre , Peroxidación de Lípido/efectos de los fármacos , Peroxidación de Lípido/inmunología , Activación de Macrófagos/inmunología , Macrófagos/inmunología , Masculino , Persona de Mediana Edad , Oxidación-Reducción/efectos de los fármacos , Estrés Oxidativo/efectos de los fármacos , Estrés Oxidativo/inmunología , Peroxidasa/sangre , Prótesis e Implantes/efectos adversos
17.
Biotechnol Prog ; 27(5): 1206-17, 2011.
Artículo en Inglés | MEDLINE | ID: mdl-21618456

RESUMEN

Successful production of recombinant proteins (r-proteins) by transient gene expression (TGE) depends on several parameters (including producer cells, culture conditions, transfection procedure, or expression vector) that should be optimized when producing any recombinant product. In this work, TGE-based production of human α-galactosidase A (GLA) is described. Producer cells, expression vectors, and parameters influencing cell metabolism after transfection have been tested. The enzyme is secreted, has the right molecular weight, and is enzymatically active. Productivities of up to 30-40 mg/L have been achieved, with a simple, fast procedure. A 6 × His tag allows enzyme purification in a single step, rendering a highly pure product. We propose a TGE-based protocol able to produce up to several milligrams per liter of highly pure, active GLA in a time as short as a few days. By this, enough amounts of engineered versions of the enzyme can be easily produced to be tested in vitro or in preclinical trials.


Asunto(s)
alfa-Galactosidasa/biosíntesis , Animales , Línea Celular , Cromatografía de Afinidad , Cromatografía en Gel , Cricetinae , Humanos , Espectrometría de Masas , Proteínas Recombinantes/biosíntesis , Proteínas Recombinantes/genética , Proteínas Recombinantes/aislamiento & purificación , Transfección , alfa-Galactosidasa/genética , alfa-Galactosidasa/aislamiento & purificación
18.
Stroke ; 41(4): 653-60, 2010 Apr.
Artículo en Inglés | MEDLINE | ID: mdl-20185784

RESUMEN

BACKGROUND AND PURPOSE: Animal models of transient ischemia suggest that oxygen-derived free radicals produced on reperfusion of ischemic brain could constitute the main cause of reperfusion injury. We aimed to determine the presence and role of lipid peroxidation and protein oxidation-related molecules after tissue plasminogen activator-induced recanalization in human stroke. METHODS: A total of 160 patients with strokes involving the middle cerebral artery and treated with tissue plasminogen activator and 60 healthy controls were included. Blood samples, transcranial Doppler recordings, and National Institutes of Health Stroke Scale scores were obtained at baseline (pretreatment), 1 hour and 2 hours after tissue plasminogen activator bolus, and 12 hours and 24 hours after stroke onset. The main lipid peroxidation end-product malondialdehyde, advanced oxidation protein products, and plasma concentrations of myeloperoxidase were assessed. RESULTS: At baseline, all oxidative stress biomarkers were higher than in control subjects (P<0.01 for all comparisons). Malondialdehyde remained high compared with controls during the study period, whereas myeloperoxidase concentrations were significantly raised at baseline, 1 hour after tissue plasminogen activator administration, and 12 hours after stroke onset. Malondialdehyde concentrations correlated with stroke severity and were associated with outcome and with hemorrhagic complications. Regarding recanalization, among those patients with middle cerebral artery recanalization by the end of tissue plasminogen activator infusion (44%) or anytime thereafter, no peaking of any of the studied molecules could be identified. CONCLUSIONS: Our study showed that systemic oxidative damage to lipids and proteins had already occurred at baseline in stroke. In contrast to animal studies, a relationship between free radical-mediated oxidative damage to lipids or proteins and reperfusion injury after arterial recanalization could not be established.


Asunto(s)
Fibrinolíticos/uso terapéutico , Estrés Oxidativo , Reperfusión , Accidente Cerebrovascular/tratamiento farmacológico , Terapia Trombolítica , Activador de Tejido Plasminógeno/uso terapéutico , Anciano , Animales , Biomarcadores/metabolismo , Hemorragia Cerebral/etiología , Hemorragia Cerebral/fisiopatología , Femenino , Humanos , Masculino , Malondialdehído/metabolismo , Peroxidasa/metabolismo , Especies Reactivas de Oxígeno/metabolismo , Accidente Cerebrovascular/complicaciones , Terapia Trombolítica/efectos adversos
19.
Atherosclerosis ; 195(1): 57-65, 2007 Nov.
Artículo en Inglés | MEDLINE | ID: mdl-17222414

RESUMEN

We examined the effect of chronic social stress, similar to that endured by humans, on lipid metabolism of mice. The activity of the lipoprotein lipase (LPL) enzyme increased in adrenals, while in plasma it diminished significantly. Hepatic lipase (HL) was strongly affected in liver and adrenal glands, increasing four-fold and three-fold, respectively. At the same time, scavenger receptor class-B type-I (SR-BI), which are considered the high-density lipoprotein (HDL) receptor in the liver, increased significantly. Although the adrenals do not synthesise HL, the increase in HL may facilitate the uptake of HDL cholesterol for the synthesis of corticoids, which increase significantly following chronic stress. The volume of adrenal glands in control animals was significantly higher than in stressed animals (1.23+/-0.12 mm3 versus 0.29+/-0.06 mm3, p<0.001), corresponding with the weight difference of these organs. Medulla volume was also different in the two groups (0.27+/-0.10 mm3 versus 0.04+/-0.02 mm3, p<0.05). Despite this, corticosterone in plasma was significantly higher in stressed animals. Our results shows, for the first time, the effect of chronic social stress on lipid metabolism in general, and in particular on the SR-BI receptor and HL, which is directly involved in cholesterol reverse transport.


Asunto(s)
Regulación de la Expresión Génica , Metabolismo de los Lípidos , Receptores Depuradores de Clase B/metabolismo , Corticoesteroides/metabolismo , Glándulas Suprarrenales/metabolismo , Animales , Apolipoproteínas/metabolismo , Colesterol/metabolismo , Lipoproteína Lipasa/metabolismo , Hígado/enzimología , Hígado/metabolismo , Masculino , Ratones , Ratones Endogámicos C57BL , Estrés Fisiológico , Sustancias Reactivas al Ácido Tiobarbitúrico
20.
Anal Biochem ; 343(2): 277-82, 2005 Aug 15.
Artículo en Inglés | MEDLINE | ID: mdl-15993372

RESUMEN

This paper describes the development of a procedure for measuring total lipid, triacylglyceride, and cholesterol content using small amounts of tissue (50 mg for liver samples and 10 mg for adipose tissue samples). In the first step, total lipids are extracted with an organic solvent mixture of hexane and isopropanol. In the second step, extracted lipids are emulsified by sonication in a buffer containing 1,4-piperazinediethanesulfonic acid (28.75 mM), magnesium chloride.6H(2)O (57.76 mM), free fatty acids-bovine serum albumin (8.76 microM), and sodium dodecyl sulfate (0.1%). Lipid concentrations from the resulting emulsion can then be assessed using commercial enzymatic kits. This method is also suitable for determining direct homogenate triacylglyceride and total cholesterol content in cases where a lipid percentage is not needed. The proposed method increases the yield of lipid recovery from small tissue samples and allows the measurement of both triacylglyceride and cholesterol content from a single starting sample. The methodology described here is the first to accomplish simultaneous determination of both parameters and is potentially useful for animal small tissue samples, particularly human biopsy samples.


Asunto(s)
Técnicas de Química Analítica/métodos , Colesterol/análisis , Lípidos/análisis , Triglicéridos/análisis , Tejido Adiposo/química , Animales , Tampones (Química) , Emulsiones/química , Hígado/química , Masculino , Ratones , Ratones Endogámicos C57BL , Polisorbatos/química , Ratas , Ratas Wistar , Reproducibilidad de los Resultados , Especificidad de la Especie
SELECCIÓN DE REFERENCIAS
DETALLE DE LA BÚSQUEDA
...