Estudi citogenètic de líquid amniòtic i vellositats corials: valor de la implementació del cribatge combinat de primer trimestre de gestació en el diagnòstic prenatal d’anomalies cromosòmiques

La implementació del cribatge combinat de primer trimestre de gestació ha repercutit en els estudis citogenètics, disminuïnt el nombre de proves invasives i augmentant la taxa de detecció d’anomalies cromosòmiques. Es presenten els resultats citogenètics de mostres de líquid amniòtic i de vellositats corials realitzats del 1999 al 2010 al servei laboratori d’Hematologia de l’Hospital Germans Trias i Pujol per valorar la implementació d’aquest cribatge.

Prenatal exposure to traffic-related air pollution and ultrasound measures of fetal growth in the INMA Sabadell cohort

Background: Few studies have used longitudinal ultrasound measurements to assess the effect of traffic-related air pollution on fetal growth.Objective: We examined the relationship between exposure to nitrogen dioxide (NO2) and aromatic hydrocarbons [benzene, toluene, ethylbenzene, m/p-xylene, and o-xylene (BTEX)] on fetal growth assessed by 1,692 ultrasound measurements among 562 pregnant women from the Sabadell cohort of the Spanish INMA (Environment and Childhood) study.Methods: We used temporally adjusted land-use regression models to estimate exposures to NO2 and BTEX. We fitted mixed-effects models to estimate longitudinal growth curves for femur length (FL), head circumference (HC), abdominal circumference (AC), biparietal diameter (BPD), and estimated fetal weight (EFW). Unconditional and conditional SD scores were calculated at 12, 20, and 32 weeks of gestation. Sensitivity analyses were performed considering time–activity patterns during pregnancy.Results: Exposure to BTEX from early pregnancy was negatively associated with growth in BPD during weeks 20–32. None of the other fetal growth parameters were associated with exposure to air pollution during pregnancy. When considering only women who spent 2 hr/day in nonresidential outdoor locations, effect estimates were stronger and statistically significant for the association between NO2 and growth in HC during weeks 12–20 and growth in AC, BPD, and EFW during weeks 20–32.Conclusions: Our results lend some support to an effect of exposure to traffic-related air pollutants from early pregnancy on fetal growth during mid-pregnancy.

The Plesiomonas shigelloides wbO1 gene cluster and the role of O1-antigen LPS in pathogenicity

The Plesiomonas shigelloides 302-73 strain (serotype O1) wb gene cluster encodes 15 proteins which are consistent with the chemical structure of the O1-antigen lypopolysaccharide (LPS) previously described for this strain. The P. shigelloides O1-antigen LPS export uses the Wzy-dependent pathway as correspond to heteropolysaccharides structures. By the isolation of two mutants lacking this O1-antigen LPS, we could establish that the presence of the O1-antigen LPS is crucial for to survive in serum mainly to become resistant to complement. Also, it is an important factor in the bacterial adhesion and invasion to some eukaryotic cells, and in the ability to form biofilms. This is the first report on the genetics from a P. shigelloides O-antigen LPS cluster (wb) not shared by Shigella like P. shigelloides O17, the only one reported until now.

Aspectos éticos de la información y asesoramiento en el diagnóstico prenatal de anomalías congénitas.

En el transcurso de los últimos años, el incremento de la tecnología ha modificado la atención prenatal. La posibilidad de aplicación de pruebas y estudios ha hecho posible que el diagnóstico prenatal de defectos congénitos se convierta en un objetivo del control prenatal. Ronal Dworkin defiende el principio de autonomía procreativa, según el cual todas aquellas decisiones que afedan a la propia reproducción son individuales y fundamentales. Sin embargo, las decisiones que se toman en el ámbito procreativo se inscriben en el seno de una relación asistencial que se establece a lo largo del tiempo. En este artículo se pretende realizar un breve repaso de los principios filosóficos que fundamentan el principio de autonomía, para en un segundo tiempo realizar una reflexión sobre la aplicación de este concepto de autonomía a la toma de decisiones en el caso que nos ocupa: el diagnóstico prenatal de anomalías congénitas

Aspectos éticos de la información y asesoramiento en el diagnóstico prenatal de anomalías congénitas.

En el transcurso de los últimos años, el incremento de la tecnología ha modificado la atención prenatal. La posibilidad de aplicación de pruebas y estudios ha hecho posible que el diagnóstico prenatal de defectos congénitos se convierta en un objetivo del control prenatal. Ronal Dworkin defiende el principio de autonomía procreativa, según el cual todas aquellas decisiones que afedan a la propia reproducción son individuales y fundamentales. Sin embargo, las decisiones que se toman en el ámbito procreativo se inscriben en el seno de una relación asistencial que se establece a lo largo del tiempo. En este artículo se pretende realizar un breve repaso de los principios filosóficos que fundamentan el principio de autonomía, para en un segundo tiempo realizar una reflexión sobre la aplicación de este concepto de autonomía a la toma de decisiones en el caso que nos ocupa: el diagnóstico prenatal de anomalías congénitas

Infección prenatal

Protocolos terapeuticos. Infección prenatal. Riesgo de infección prenatal. La infección prenatal requiere un alto índice de sospecha, ya que no siempre, los antecedentes se hallan presentes bien porque faltan o bien porque hayan pasado desapercibidos. Dentro del concepto de infección prenatal se encuentran las englobadas en el acrónimo Torches (toxoplasmosis, rubeola, citomegalovirosis, herpes o sífilis) )...

Increasing the scalability and the speedup of a fish school distributed simulator

El present treball fa un anàlisi i desenvolupament sobre les millores en la velocitat i en l’escalabilitat d'un simulador distribuït de grups de peixos. Aquests resultats s’han obtingut fent servir una nova estratègia de comunicació per als processos lògics (LPs) i canvis en l'algoritme de selecció de veïns que s'aplica a cadascun dels peixos en cada pas de simulació. L’idea proposada permet que cada procés lògic anticipi futures necessitats de dades pels seus veïns reduint el temps de comunicació al limitar la quantitat de missatges intercanviats entre els LPs. El nou algoritme de selecció dels veïns es va desenvolupar amb l'objectiu d'evitar treball innecessari permetent la disminució de les instruccions executades en cada pas de simulació i per cadascun del peixos simulats reduint de forma significativa el temps de simulació.

Implicació de la inflamació i la resposta immunitària en la lesió induïda per la isquèmia

El sistema nerviós central (SNC) i el sistema immunitari (SI) estan estretament connectats. Es produeixen nombroses alteracions en el sistema immunitari després de la isquèmia i la inflamació es reconeix com una de les principals causes de la progressió de la lesió isquèmica. És molt important determinar el paper de les diferents cèl•lules implicades en la resposta immunitària i inflamatòria després de la isquèmia i el perfil de citocines que s’alliberen. A partir de l’estudi de les diferents poblacions de leucòcits en sang circulant després de la isquèmia, hem determinat que la subpoblació de monòcits (CD43high/CD11bhigh) augmenta a 48h de manera proporcional al volum d’infart. Aquesta població està formada per dos subtipus descrits de monòcits, els no clàssics (CD43high/Ly6C-) i els intermedis (CD43high/Ly6Cdim), i sembla expressar un perfil de citocines anti-inflamatòries així com una major capacitat fagocítica. Per altra banda, observem la presència de CD43 en el cervell i la seva degradació a 4 dies després de la isquèmia. També s’observa l’aparició de la fracció soluble del CD43 en el parènquima cerebral després del trencament de la barrera hematoencefàlica. Addicionalment, hem estudiat com s’alteren els canvis a nivell immunològic en ratolins deficients en CD69 i hem observat una pitjor progressió del volum d’infart en els animals CD69KO. A més a més hem volgut esbrinar el paper dels limfòcits utilitzant ratolins RAG (-/-) que tenen infarts més petits que els WT, però quan aquests careixen de CD69, tenen infarts significativament més grans. En l’estudi del procés inflamatori en la isquèmica, hem treballat amb ratolins deficient en ApoE i IL10. Pel que fa als ratolins ApoE (-/-), observem que tenen un volum d’infart més gran a les 24h i que es manté fins als 4 dies, i proposem que NFkB pot tenir un paper molt rellevant en aquest procés. Pel que fa a la IL-10, els animals deficients en aquesta citocina presenten un volum d’infart major i una expressió de citocines proinflamatòries més alt. A més a més, els ratolins IL10 KO presenten uns nivells de IL-12 més elevats de manera basal, i proposem que això és degut a la falta de la IL-10 per a inhibir la via.

Association between GIS-based exposure to urban air pollution during pregnancy and birth weight in the INMA Sabadell Cohort

Background: There is growing evidence that traffic-related air pollution reduces birth weight. Improving exposure assessment is a key issue to advance in this research area.Objective: We investigated the effect of prenatal exposure to traffic-related air pollution via geographic information system (GIS) models on birth weight in 570 newborns from the INMA (Environment and Childhood) Sabadell cohort.Methods: We estimated pregnancy and trimester-specific exposures to nitrogen dioxide and aromatic hydrocarbons [benzene, toluene, ethylbenzene, m/p-xylene, and o-xylene (BTEX)] by using temporally adjusted land-use regression (LUR) models. We built models for NO2 and BTEX using four and three 1-week measurement campaigns, respectively, at 57 locations. We assessed the relationship between prenatal air pollution exposure and birth weight with linear regression models. We performed sensitivity analyses considering time spent at home and time spent in nonresidential outdoor environments during pregnancy.Results: In the overall cohort, neither NO2 nor BTEX exposure was significantly associated with birth weight in any of the exposure periods. When considering only women who spent < 2 hr/day in nonresidential outdoor environments, the estimated reductions in birth weight associated with an interquartile range increase in BTEX exposure levels were 77 g [95% confidence interval (CI), 7–146 g] and 102 g (95% CI, 28–176 g) for exposures during the whole pregnancy and the second trimester, respectively. The effects of NO2 exposure were less clear in this subset.Conclusions: The association of BTEX with reduced birth weight underscores the negative role of vehicle exhaust pollutants in reproductive health. Time–activity patterns during pregnancy complement GIS-based models in exposure assessment.

La meiosi

La unitat didàctica s’inicia amb una seqüència per emfatitzar la importància del llenguatge en ciències, contextualitzada en el desxiframent dels jeroglífics egipcis i en el mètode hipotètic-deductiu que s’utilitzà per a desxifrar-los. Aquest, serà el fil conductor per introduir la meiosi, ara emmarcada en un cas de Síndrome de Down, determinat en el cariotip de l’embrió d’una parella que rep el diagnòstic prenatal d’un equip investigador. Amb activitats proposades durant les sis sessions l’alumne haurà d’ aprendre a identificar evidències, a justificar proves a partir de continguts teòrics, i a formular hipòtesis a partir d’un cas; a explicitar el procés amb recursos simbòlics i visuals; a comprendre’n la funcionalitat i a caracteritzar-ne les fases. S’utilitzen eines de regulació tals com el treball cooperatiu i la coavaluació, així com pautes i bases d’orientació. També es treballen les idees prèvies i l’atenció a la diversitat present a l’aula.

Changes in the enterocyte cytoskeleton in newborn rats exposed to ethanol in utero

BACKGROUND: Cytoskeletal changes after longterm exposure to ethanol have been described in a number of cell types in adult rat and humans. These changes can play a key part in the impairment of nutrient assimilation and postnatal growth retardation after prenatal damage of the intestinal epithelium produced by ethanol intake. AIMS: To determine, in the newborn rat, which cytoskeletal proteins are affected by longterm ethanol exposure in utero and to what extent. ANIMALS: The offspring of two experimental groups of female Wistar rats: ethanol treated group receiving up to 25% (w/v) of ethanol in the drinking fluid and control group receiving water as drinking fluid. METHODS: Single and double electron microscopy immunolocalisation and label density estimation of cytoskeletal proteins on sections of proximal small intestine incubated with monoclonal antibodies against actin, alpha-tubulin, cytokeratin (polypeptides 1, 5, 6, 7, 8, 10, 11, and 18), and with a polyclonal antibody anti-beta 1,4-galactosyl transferase as trans golgi (TG) or trans golgi network (TGN) marker, or both. SDS-PAGE technique was also performed on cytoskeletal enriched fractions from small intestine. Western blotting analysis was carried out by incubation with the same antibodies used for immunolocalisation. RESULTS: Intestinal epithelium of newborn rats from the ethanol treated group showed an overexpression of cytoskeletal polypeptides ranging from 39 to 54 kDa, affecting actin and some cytokeratins, but not tubulin. Furthermore, a cytokeratin related polypeptide of 28-29 kDa was identified together with an increase in free ubiquitin in the same group. It was noteworthy that actin and cytokeratin were abnormally located in the TG or the TGN, or both. CONCLUSIONS: Longterm exposure to ethanol in utero causes severe dysfunction in the cytoskeleton of the developing intestinal epithelium. Actin and cytokeratins, which are involved in cytoskeleton anchoring to plasma membrane and cell adhesion, are particularly affected, showing overexpression, impaired proteolysis, and mislocalisation.

Defective TNF-alpha-mediated hepatocellular apoptosis and liver damage in acidic sphingomyelinase knockout mice

This study addressed the contribution of acidic sphingomyelinase (ASMase) in TNF-alpha-mediated hepatocellular apoptosis. Cultured hepatocytes depleted of mitochondrial glutathione (mGSH) became sensitive to TNF-alpha, undergoing a time-dependent apoptotic cell death preceded by mitochondrial membrane depolarization, cytochrome c release, and caspase activation. Cyclosporin A treatment rescued mGSH-depleted hepatocytes from TNF-alpha-induced cell death. In contrast, mGSH-depleted hepatocytes deficient in ASMase were resistant to TNF-alpha-mediated cell death but sensitive to exogenous ASMase. Furthermore, although in vivo administration of TNF-alpha or LPS to galactosamine-pretreated ASMase(+/+) mice caused liver damage, ASMase(-/-) mice exhibited minimal hepatocellular injury. To analyze the requirement of ASMase, we assessed the effect of glucosylceramide synthetase inhibition on TNF-alpha-mediated apoptosis. This approach, which blunted glycosphingolipid generation by TNF-alpha, protected mGSH-depleted ASMase(+/+) hepatocytes from TNF-alpha despite enhancement of TNF-alpha-stimulated ceramide formation. To further test the involvement of glycosphingolipids, we focused on ganglioside GD3 (GD3) because of its emerging role in apoptosis through interaction with mitochondria. Analysis of the cellular redistribution of GD3 by laser scanning confocal microscopy revealed the targeting of GD3 to mitochondria in ASMase(+/+) but not in ASMase(-/-) hepatocytes. However, treatment of ASMase(-/-) hepatocytes with exogenous ASMase induced the colocalization of GD3 and mitochondria. Thus, ASMase contributes to TNF-alpha-induced hepatocellular apoptosis by promoting the mitochondrial targeting of glycosphingolipids.

Temporal changes in the expression and distribution of adhesion molecules during liver development and regeneration

We have compared by immunocytochemistry and immunoblotting the expression and distribution of adhesion molecules participating in cell-matrix and cell-cell interactions during embryonic development and regeneration of rat liver. Fibronectin and the fibronectin receptor, integrin alpha 5 beta 1, were distributed pericellularly and expressed at a steady level during development from the 16th day of gestation and in neonate and adult liver. AGp110, a nonintegrin fibronectin receptor was first detected on the 17th day of gestation in a similar, nonpolarized distribution on parenchymal cell surfaces. At that stage of development haemopoiesis is at a peak in rat liver and fibronectin and receptors alpha 5 beta 1 and AGp110 were prominent on the surface of blood cell precursors. During the last 2 d of gestation (20th and 21st day) hepatocytes assembled around lumina. AGp110 was initially depolarized on the surface of these acinar cells but then confined to the lumen and to newly-formed bile canaliculi. At birth, a marked increase occurred in the canalicular expression of AGp110 and in the branching of the canalicular network. Simultaneously, there was enhanced expression of ZO-1, a protein component of tight junctions. On the second day postpartum, presence of AGp110 and of protein constituents of desmosomes and intermediate junctions, DGI and E-cadherin, respectively, was notably enhanced in cellular fractions insoluble in nonionic detergents, presumably signifying linkage of AGp110 with the cytoskeleton and assembly of desmosomal and intermediate junctions. During liver regeneration after partial hepatectomy, AGp110 remained confined to apical surfaces, indicating a preservation of basic polarity in parenchymal cells. A decrease in the extent and continuity of the canalicular network occurred in proliferating parenchyma, starting 24 h after resection in areas close to the terminal afferent blood supply of portal veins and spreading to the rest of the liver within the next 24 h. Distinct acinar structures, similar to the ones in prenatal liver, appeared at 72 h after hepatectomy. Restoration of the normal branching of the biliary tree commenced at 72 h. At 7 d postoperatively acinar formation declined and one-cell-thick hepatic plates, as in normal liver, were observed.

Changes in the enterocyte cytoskeleton in newborn rats exposed to ethanol in utero

BACKGROUND: Cytoskeletal changes after longterm exposure to ethanol have been described in a number of cell types in adult rat and humans. These changes can play a key part in the impairment of nutrient assimilation and postnatal growth retardation after prenatal damage of the intestinal epithelium produced by ethanol intake. AIMS: To determine, in the newborn rat, which cytoskeletal proteins are affected by longterm ethanol exposure in utero and to what extent. ANIMALS: The offspring of two experimental groups of female Wistar rats: ethanol treated group receiving up to 25% (w/v) of ethanol in the drinking fluid and control group receiving water as drinking fluid. METHODS: Single and double electron microscopy immunolocalisation and label density estimation of cytoskeletal proteins on sections of proximal small intestine incubated with monoclonal antibodies against actin, alpha-tubulin, cytokeratin (polypeptides 1, 5, 6, 7, 8, 10, 11, and 18), and with a polyclonal antibody anti-beta 1,4-galactosyl transferase as trans golgi (TG) or trans golgi network (TGN) marker, or both. SDS-PAGE technique was also performed on cytoskeletal enriched fractions from small intestine. Western blotting analysis was carried out by incubation with the same antibodies used for immunolocalisation. RESULTS: Intestinal epithelium of newborn rats from the ethanol treated group showed an overexpression of cytoskeletal polypeptides ranging from 39 to 54 kDa, affecting actin and some cytokeratins, but not tubulin. Furthermore, a cytokeratin related polypeptide of 28-29 kDa was identified together with an increase in free ubiquitin in the same group. It was noteworthy that actin and cytokeratin were abnormally located in the TG or the TGN, or both. CONCLUSIONS: Longterm exposure to ethanol in utero causes severe dysfunction in the cytoskeleton of the developing intestinal epithelium. Actin and cytokeratins, which are involved in cytoskeleton anchoring to plasma membrane and cell adhesion, are particularly affected, showing overexpression, impaired proteolysis, and mislocalisation.

Pro-inflammatory gene expression and neurotoxic effects of activated microglia are attenuated by absence of CCAAT/enhancer binding protein beta

Background. Microglia and astrocytes respond to homeostatic disturbances with profound changes of gene expression. This response, known as glial activation or neuroinflammation, can be detrimental to the surrounding tissue. The transcription factor CCAAT/enhancer binding protein ß (C/EBPß) is an important regulator of gene expression in inflammation but little is known about its involvement in glial activation. To explore the functional role of C/EBPß in glial activation we have analyzed pro-inflammatory gene expression and neurotoxicity in murine wild type and C/EBPß-null glial cultures. Methods. Due to fertility and mortality problems associated with the C/EBPß-null genotype we developed a protocol to prepare mixed glial cultures from cerebral cortex of a single mouse embryo with high yield. Wild-type and C/EBPß-null glial cultures were compared in terms of total cell density by Hoechst-33258 staining; microglial content by CD11b immunocytochemistry; astroglial content by GFAP western blot; gene expression by quantitative real-time PCR, western blot, immunocytochemistry and Griess reaction; and microglial neurotoxicity by estimating MAP2 content in neuronal/microglial cocultures. C/EBPß DNA binding activity was evaluated by electrophoretic mobility shift assay and quantitative chromatin immunoprecipitation. Results. C/EBPß mRNA and protein levels, as well as DNA binding, were increased in glial cultures by treatment with lipopolysaccharide (LPS) or LPS + interferon ¿ (IFN¿). Quantitative chromatin immunoprecipitation showed binding of C/EBPß to pro-inflammatory gene promoters in glial activation in a stimulus- and gene-dependent manner. In agreement with these results, LPS and LPS+IFN¿ induced different transcriptional patterns between pro-inflammatory cytokines and NO synthase-2 genes. Furthermore, the expressions of IL-1ß and NO synthase-2, and consequent NO production, were reduced in the absence of C/EBPß. In addition, neurotoxicity elicited by LPS+IFN¿-treated microglia co-cultured with neurons was completely abolished by the absence of C/EBPß in microglia.