Effect of cardiopulmonary bypass on activated partial thromboplastin time waveform analysis, serum procalcitonin and C-reactive protein concentrations

Abstract Introduction Systemic inflammatory response syndrome (SIRS) is a frequent condition after cardiopulmonary bypass (CPB) and makes conventional biological tests fail to detect postoperative sepsis. Biphasic waveform (BPW) analysis is a new biological test derived from activated partial thromboplastin time that has recently been proposed for sepsis diagnosis. The aim of this study was to investigate the accuracy of BPW to detect sepsis after cardiac surgery under CPB. Methods We conducted a prospective study in American Society of Anesthesiologists' (ASA) physical status III and IV patients referred for cardiac surgery under CPB. Procalcitonin (PCT) and BPW were recorded before surgery and every day during the first week following surgery. Patients were then divided into three groups: patients presenting no SIRS, patients presenting with non-septic SIRS and patients presenting with sepsis. Results Thirty two patients were included. SIRS occurred in 16 patients (50%) including 5 sepsis (16%) and 11 (34%) non-septic SIRS. PCT and BPW were significantly increased in SIRS patients compared to no SIRS patients (0.9 [0.5-2.2] vs. 8.1 [2.0-21.3] ng/l for PCT and 0.10 [0.09-0.14] vs. 0.29 [0.16-0.56] %T/s for BPW; P < 0.05 for both). We observed no difference in peak PCT value between the sepsis group and the non-septic SIRS group (8.4 [7.5-32.2] vs. 7.8 [1.9-17.5] ng/l; P = 0.67). On the other hand, we found that BPW was significantly higher in the sepsis group compared to the non-septic SIRS group (0.57 [0.54-0.78] vs. 0.19 [0.14-0.29] %T/s; P < 0.01). We found that a BPW threshold value of 0.465%T/s was able to discriminate between sepsis and non-septic SIRS groups with a sensitivity of 100% and a specificity of 93% (area under the curve: 0.948 +/- 0.039; P < 0.01). Applying the previously published threshold of 0.25%T/s, we found a sensitivity of 100% and a specificity of 72% to discriminate between these two groups. Neither C-reactive protein (CRP) nor PCT had significant predictive value (area under the curve for CRP was 0.659 +/- 0.142; P = 0.26 and area under the curve for PCT was 0.704 +/- 0.133; P = 0.15). Conclusions BPW has potential clinical applications for sepsis diagnosis in the postoperative period following cardiac surgery under CPB

Vers une observation inter-disciplinaire des phénomènes naturels sur les bassins versants de montagne (Hydrogéologie à coût limité du bassin du Vorz (Massif de Belledonne, Isère))

Le 22 Août 2005 une crue intense s'est produite sur le bassin versant du Vorz, détruisant partiellement le hameau de la Gorge. Cet évènement a mis en évidence les difficultés à anticiper les conditions hydrométéorologiques en montagne où elles sont extrêmement variables spatialement et temporellement, et souvent faiblement instrumentées. De ce constat est né le projet de mettre en place un réseau d'instrumentation hydrométéorologique original sur le bassin versant du Vorz, afin d'y observer les phénomènes naturels et hydrologiques s'y produisant, de mieux les appréhender, et de construire les outils et méthodes nécessaires à leur modélisation. Après deux saisons de mesures, les premiers résultats ont montré que le réseau mis en place permet d'obtenir des informations à haute résolution spatiale et temporelle sur les processus hydrométéorologiques. Malgré son installation dans le milieu difficile de la montagne (accessibilité, froid, énergie,...), une très bonne fiabilité a pu être mise en avant, ainsi que des perspectives de transposition à d'autres bassins versants, et ce, pour un faible coût financier. L'originalité du réseau est de réaliser un multi-échantillonnage de nombreux paramètres hydrométéorologiques (pluviométrie, température, neige, insolation,...), avec des résolutions spatiales (10 à 50 mètres) et temporelles (horaire à moins) permettant d'envisager une modélisation hydrologique à différentes échelles, aussi bien pour la gestion des ressources en eau (long terme) que pour la prévention des crues (court terme). Les capteurs mis en place constituent un ensemble complémentaire et indissociable de divers instruments de mesure: iButtons (air et sol), totalisateurs, pluviomètres, appareils photographiques. La mise au point d'un capteur de mesure innovant de cartographie automatique de la couverture neigeuse (SnoDEC), à partir d'images photographiques classiques, prises à pas de temps régulier (5 à 7 images par jours) a été réalisée au cours de ce travail. Il permet de quantifier l'hérogénéité spatiale et temporelle des phénomènes d'enneigement sur le versant, prépondérants sur son hydrologie, au vue de la persistance nivale (5 à 10 mois). L'ensemble de ce dispositif permet de disposer d'une importante base de données, et de mettre en oeuvre différentes techniques d'interpolations des variables hydrométéorologiques sur l'ensemble du bassin versant. Ainsi, des cartographies précises du champ de température et de pluviométrie seront disponibles au pas de temps journalier. En outre, le capteur SnoDEC permettra d'analyser et quantifier l'hétérogénétité spatio-temporelle (altitude, exposition, vitesse de fonte,...) de la couverture nivale. A partir de ces données, on pourra mieux appréhender les mécanismes hydrologiques en jeu sur le site et dessiner les contours des modélisations futures. Dans le même temps, les données disponibles pourront être combinées afin de mettre en évidence des phénomènes difficilement mesurables (limite pluie/neige, inversion thermiques,...), qui serviront à l'avenir à contraindre de manière précise les modèles nivologiques et hydrologiques. Au travers des différents paramètres instrumentés, et grâce à l'utilisation de l'imagerie, ce réseau est capable de mesurer des variables relevant de nombreux champs disciplinaires (dynamique glaciaire, cyle végétatif,...). Il s'inscrit ainsi, par son approche interdisciplinaire, dans une volonté de mise en place d'un réseau de mesure à coût limité, destiné à l'ensemble des acteurs de l'étude et la recherche des milieux de la montagne.In August 2005, a intense flashflood occurs on the Vorz catchment affecting the village of Saint-Agnès. This event highlighed the difficulties to forecast the hydrometeorological conditions in mountain areas where they are extremely variable in space and time (spatially and temporally) and frequently poored monitored. From this observation a project was funded to implement an original meteorological monitoring system on the catchment, in order to observe the natural and hydrologic phenomena to better understand them and to build methods and tools for their modeling. After two years of monitoring, the first results showed that the network implement allows to obtain informations on hydrometeorological process at high spatial and temporal resolution. In spite of the installation in a harsh mountain environment (access, cold, energy,...) a very good reliability, and a lot of perspectives of transpositon on other catchments have been point up for low investment costs.The originality of the network is to achieved a multi-sampling on a lot of hydrometeorological parameters (rain, temprature, snow, insulation,...), with spatial (10 to 50 meters) and temporal (hourly or less) resolution to performed a hydrological modeling at different scale both for the water ressource management (long term) or flashflood prevention (short term). The Sensors use in the network constitute a complementary and indivisible set of monitoring system: iButtion (air and soil temperature), rain gauge, totalizer, cameras. The development of an innovative sensor for automatic cartography of the snow cover (SnoDEC) from terrestrail photographies was achived during this work. This sensor allows to quantify spatial and temporal heterogeneity of snow cover evolution on the catchment, with images taken at regular time steps (5 to 7 frames per day). This heterogenity is essential for understand and modelling the hydrology considering the strong snow persistence (5 to 10 months). The dense network set up on the catchment enable us to collect a large database and implement different interpolation techniques on hydrometeorological process on the catchment. Thus, accurate maps of temperatures and rain are created with a daily or hourly timestep. Furthermore, the SnoDEC sensor will permit to analyse and quantify the spatial and temporal heterogeneity (elevation, aspect, velocity of melting,...) of the snow cover. From the database, we will better understand the hydrological mechanisms occuring on the site, and we will build the first ideas and method for the future modelisation. In the same time, the available data will be combinate in order to highlight phenomena very difficult to measure (rain/snow limit, thermical inversion,...) and that will be use in the future to constraint accurately the snow and hydrologic models. Because of the different parameters monitored and the use of imagery, the network is able to measure variables from many field of study (glacier dynamic, vegetative cycle,...). Thereby, with its interdisciplinary approach the network think to implement a monitoring system at low cost in destination of the actors of study and research in mountain.SAVOIE-SCD - Bib.électronique (730659901) / SudocGRENOBLE1/INP-Bib.électronique (384210012) / SudocGRENOBLE2/3-Bib.électronique (384219901) / SudocSudocFranceF

Molecular cloning, characterization, genomic organization and promoter analysis of the α1,6-fucosyltransferase gene (fut8) expressed in the rat hybridoma cell line YB2/0

<p>Abstract</p> <p>Background</p> <p>The rat hybridoma cell line YB2/0 appears a good candidate for the large-scale production of low fucose recombinant mAbs due to its lower expression of <it>fut8 </it>gene than other commonly used rodent cell lines. However, important variations of the fucose content of recombinant mAbs are observed in production culture conditions. To improve our knowledge on the YB2/0 fucosylation capacity, we have cloned and characterized the rat <it>fut8 </it>gene.</p> <p>Results</p> <p>The cDNAs encoding the rat α1,6-fucosyltransferase (FucT VIII) were cloned from YB2/0 cells by polymerase chain reaction-based and 5' RNA-Ligase-Mediated RACE methods. The cDNAs contain an open reading frame of 1728 bp encoding a 575 amino acid sequence showing 94% and 88% identity to human and pig orthologs, respectively. The recombinant protein expressed in COS-7 cells exhibits a α1,6-fucosyltransferase activity toward human asialo-agalacto-apotransferrin. The rat <it>fut8 </it>gene is located on chromosome 6 q and spans over 140 kbp. It contains 9 coding exons and four 5'-untranslated exons. FISH analysis shows a heterogeneous copy number of <it>fut8 </it>in YB2/0 nuclei with 2.8 ± 1.4 mean copy number. The YB2/0 <it>fut8 </it>gene is expressed as two main transcripts that differ in the first untranslated exon by the usage of distinct promoters and alternative splicing. Luciferase assays allow defining the minimal promoting regions governing the initiation of the two transcripts, which are differentially expressed in YB2/0 as shown by duplex Taqman QPCR analysis. Bioinformatics analysis of the minimal promoter regions upstream exons E-2 and E-3, governing the transcription of T1 and T2 transcripts, respectively, evidenced several consensus sequences for potential transcriptional repressors. Transient transfections of Rat2 cells with transcription factor expression vectors allowed identifying KLF15 as a putative repressor of T1 transcript in Rat2 cells.</p> <p>Conclusion</p> <p>Altogether, these data contribute to a better knowledge of <it>fut8 </it>expression in YB2/0 that will be useful to better control the fucosylation of recombinant mAbs produced in these cells.</p

Genetic Diversity and Pathogenic Potential of Attaching and Effacing Escherichia coli O26:H11 Strains Recovered from Bovine Feces in the United States

Escherichia coli O26 has been identified as the most common non-O157 Shiga toxin-producing E. coli (STEC) serogroup to cause human illnesses in the United States and has been implicated in outbreaks around the world. E. coli has high genomic plasticity, which facilitates the loss or acquisition of virulence genes. Attaching and effacing E. coli (AEEC) O26 strains have frequently been isolated from bovine feces, and there is a need to better characterize the relatedness of these strains to defined molecular pathotypes and to describe the extent of their genetic diversity. High-throughput real-time PCR was used to screen 178 E. coli O26 isolates from a single U.S. cattle feedlot, collected from May to July 2011, for the presence or absence of 25 O26 serogroup-specific and virulence-associated markers. The selected markers were capable of distinguishing these strains into molecularly defined groups (yielding 18 unique marker combinations). Analysis of the clustered regularly interspaced short palindromic repeat 1 (CRISPR1) and CRISPR2a loci further discriminated isolates into 24 CRISPR types. The combination of molecular markers and CRISPR typing provided 20.8% diversity. The recent CRISPR PCR target SP_O26-E, which was previously identified only in stx 2-positive O26:H11 human clinical strains, was identified in 96.4% (161/167 [95% confidence interval, 99.2 to 93.6%]) of the stx-negative AEEC O26:H11 bovine fecal strains. This supports that these stx-negative strains may have previously contained a prophage carrying stx or could acquire this prophage, thus possibly giving them the potential to become pathogenic to humans. These results show that investigation of specific genetic markers may further elucidate our understanding of the genetic diversity of AEEC O26 strains in bovine feces

Probing Rotational and Translational Diffusion of Nanodoublers in Living Cells on Microsecond Time Scales

Nonlinear microscopes have seen an increase in popularity in the life sciences due to their molecular and structural specificity, high resolution, large penetration depth, and volumetric imaging capability. Nonetheless, the inherently weak optical signals demand long exposure times for live cell imaging. Here, by modifying the optical layout and illumination parameters, we can follow the rotation and translation of noncentrosymetric crystalline particles, or nanodoublers, with 50 mu s acquisition times in living cells. The rotational diffusion can be derived from variations in the second harmonic intensity that originates from the rotation of the nanodoubler crystal axis. We envisage that by capitalizing on the biocompatibility, functionalizability, stability, and nondestructive optical response of the nanodoublers, novel insights on cellular dynamics are within reach

Impairment of NO-Dependent Relaxation in Intralobar Pulmonary Arteries: Comparison of Urban Particulate Matter and Manufactured Nanoparticles

International audienceBACKGROUND AND OBJECTIVES: Because pulmonary circulation is the primary vascular target of inhaled particulate matter (PM), and nitric oxide is a major vasculoprotective agent, in this study we investigated the effect of various particles on the NO-cyclic guanosine monophosphate (cGMP) pathway in pulmonary arteries. METHODS: We used intrapulmonary arteries and/or endothelial cells, either exposed in vitro to particles or removed from PM-instilled animals for assessment of vasomotricity, cGMP and reactive oxygen species (ROS) levels, and cytokine/chemokine release. RESULTS: Endothelial NO-dependent relaxation and cGMP accumulation induced by acetylcholine (ACh) were both decreased after 24 hr exposure of rat intrapulmonary arteries to standard reference material 1648 (SRM1648; urban PM). Relaxation due to NO donors was also decreased by SRM1648, whereas responsiveness to cGMP analogue remained unaffected. Unlike SRM1648, ultrafine carbon black and ultrafine and fine titanium dioxide (TiO2) manufactured particles did not impair NO-mediated relaxation. SRM1648-induced decrease in relaxation response to ACh was prevented by dexamethasone (an anti-inflammatory agent) but not by antioxidants. Accordingly, SRM1648 increased the release of proinflammatory mediators (tumor necrosis factor-alpha, interleukin-8) from intrapulmonary arteries or pulmonary artery endothelial cells, but did not elevate ROS levels within intrapulmonary arteries. Decreased relaxation in response to ACh was also evidenced in intrapulmonary arteries removed from rats intratracheally instilled with SRM1648, but not with fine TiO2. CONCLUSION: In contrast to manufactured particles (including nanoparticles), urban PM impairs NO but not cGMP responsiveness in intrapulmonary arteries. We attribute this effect to oxidative-stress-independent inflammatory response, resulting in decreased guanylyl cyclase activation by NO. Such impairment of the NO pathway may contribute to urban-PM-induced cardiovascular dysfunction

Tumour-associated carbohydrate antigens in breast cancer

Glycosylation changes that occur in cancer often lead to the expression of tumour-associated carbohydrate antigens. In breast cancer, these antigens are usually associated with a poor prognosis and a reduced overall survival. Cellular models have shown the implication of these antigens in cell adhesion, migration, proliferation and tumour growth. The present review summarizes our current knowledge of glycosylation changes (structures, biosynthesis and occurrence) in breast cancer cell lines and primary tumours, and the consequences on disease progression and aggressiveness. The therapeutic strategies attempted to target tumour-associated carbohydrate antigens in breast cancer are also discussed