CO80 89. Estudio multicéntrico español de la capacidad predictiva de las escalas de riesgo CHADS2 y CHA2DS2vasc en el accidente cerebrovascular tras cirugía coronaria aislada

ObjetivosValidar las escalas de riesgo CHADS2 y CHA2DS-2VASC como modelos predictivos de desarrollo de accidente cerebrovascular (ACV) en cirugía coronaria aislada (CCA).Material y métodosPacientes consecutivos sometidos a CCA en 16 hospitales españoles. Excluidos casos con igual o más de una variable/s incompleta/s. Puntuaciones CHADS2 y CHA2DS2VASC computadas para todos los pacientes, considerándose variable de resultado la aparición de ACV (ataque isquémico transitorio [AIT]/ictus) perioperatorio precoz (primer mes postoperatorio y/o alta hospitalaria). Análisis uni y multivariante. La capacidad discriminativa fue cuantificada por el cálculo del área bajo la curva ROC (AUC).ResultadosVeinte mil novecientos ochenta pacientes incluidos, 282 desarrollaron ACV postoperatorio (1,34%). La incidencia de ACV fue superior en pacientes con insuficiencia cardíaca congestiva (ICC) y/o fracción de eyección inferior al 40% (4,10 vs 0,83%), diabéticos (1,70 vs 1,11%), hipertensos (1,60 vs 0,98%), ACV previo (2,72 vs 1,26%) y a enfermedad arterial periférica (EAP) (3,04 vs 1,04%; p < 0,05). En el análisis multivariante, ICCC (odds ratio [OR]: 4,06), ACV previo (OR: 1,48), EAP (OR: 1,49) constituyeron factores de riesgo independientes para el desarrollo de ACV postoperatorio (p < 0,05). El AUC para CHADS2 fue 0,666, y para CHA2DS2VASc 0,655 (p < 0,0001). La distribución de las tasas de ACV postoperatorio según las puntuaciones de las anteriores escalas se recoge en la figura 1 (p < 0,0001). Figura 1Tasas de ACV postoperatorio en pacientes sometidos a CCA según puntuaciones de las escalas CHADS2 (gris) y CHA2DS2VASC (negro).ConclusionesLas escalas de riesgo CHADS2 y CHA2DS-2VASC pueden resultar útiles en la práctica clínica para estratificar el riesgo de desarrollo de ACV postoperatorio en pacientes sometidos a CCA

Single-step isolation of extracellular vesicles by size-exclusion chromatography

Background: Isolation of extracellular vesicles from plasma is a challenge due to the presence of proteins and lipoproteins. Isolation of vesicles using differential centrifugation or density-gradient ultracentrifugation results in co-isolation of contaminants such as protein aggregates and incomplete separation of vesicles from lipoproteins, respectively. Aim: To develop a single-step protocol to isolate vesicles from human body fluids. Methods: Platelet-free supernatant, derived from platelet concentrates, was loaded on a sepharose CL-2B column to perform size-exclusion chromatography (SEC; n=3). Fractions were collected and analysed by nanoparticle tracking analysis, resistive pulse sensing, flow cytometry and transmission electron microscopy. The concentrations of high-density lipoprotein cholesterol (HDL) and protein were measured in each fraction. Results: Fractions 9–12 contained the highest concentrations of particles larger than 70 nm and platelet-derived vesicles (46%±6 and 61%±2 of totals present in all collected fractions, respectively), but less than 5% of HDL and less than 1% of protein (4.8%±1 and 0.65%±0.3, respectively). HDL was present mainly in fractions 18–20 (32%±2 of total), and protein in fractions 19–21 (36%±2 of total). Compared to the starting material, recovery of platelet-derived vesicles was 43%±23 in fractions 9–12, with an 8-fold and 70-fold enrichment compared to HDL and protein. Conclusions: SEC efficiently isolates extracellular vesicles with a diameter larger than 70 nm from platelet-free supernatant of platelet concentrates. Application SEC will improve studies on the dimensional, structural and functional properties of extracellular vesicles

Improvement in the regulation of the vitamin K antagonist acenocoumarol after a standard initial dose regimen: prospective validation of a prescription model

Background In a retrospective study we have developed a model which determines the dose of acenocoumarol based on the age of the patient and on the first INR obtained after a standard initial loading dose. The group of patients of this study was used as the control group of the present study. Aim The aim of this study was to prospectively validate the model and to assess whether the use of this model improves the quality of the treatment in the 0-2 months study period. Patients and methods In 197 patients the model was evaluated by (1) in the initial phase: comparison of INRs with the control group, after assessing the dose according to the model, and (2) in the 0-2 months period: calculation of the percentage of time spent in the therapeutic target range compared to the control group. Furthermore, the eventual dose was compared to the dose of the model when the INRs were within the therapeutic target range for the first time and on two successive occasions. Results (1) When dosed according to the model, 50% of INRs in the total group were within the therapeutic target range compared to 45% in the control group, and (2) the percentage time spent within this range was 68 in the total group compared to 63 in the control group (P = 0.0013). When the INRs were within the range for the first time and successively twice, the eventual doses were similar to the model in 59 and 50%, respectively. About 20% of the patients did not achieve two successive INRs within the range. Conclusions Using the model the quality of treatment improved. We advice to use a standardized individualized dose regimen at the initiation of vitamin K antagonist treatmen

Active caspase-3 is removed from cells by release of caspase-3-enriched vesicles

AbstractCleavage of Rho associated Coiled Coil kinase I (ROCK I) by caspase-3 contributes to membrane blebbing. Whether caspase-3 and ROCK I also play a role in the release of membrane vesicles is unknown. Therefore, we transfected a human breast cancer cell line (MCF-7) that is caspase-3 deficient, lacks membrane blebbing, and does not release membrane vesicles, with caspase-3. Cells expressing caspase-3 demonstrate both ROCK I-mediated membrane blebbing, and release of small (400–600nm) membrane vesicles in a ROCK I-independent manner. These membrane vesicles contain caspase-3, and are enriched in caspase-3 activity compared to the releasing cells. Caspase-3-containing vesicles are taken up by untransfected cells but the cells do not show any sign of apoptosis. In conclusion, we show that the release of caspase-3-enriched membrane vesicles and membrane blebbing are two differentially regulated processes. Furthermore, we hypothesize that packaging of caspase-3 into membrane vesicles contributes to cellular homeostasis by the removal of caspase-3, and concurrently, protects the cells' environment from direct exposure to caspase-3 activity

A high-throughput Sanger strategy for human mitochondrial genome sequencing

A population reference database of complete human mitochondrial genome (mtGenome) sequences is needed to enable the use of mitochondrial DNA (mtDNA) coding region data in forensic casework applications. However, the development of entire mtGenome haplotypes to forensic data quality standards is difficult and laborious. A Sanger-based amplification and sequencing strategy that is designed for automated processing, yet routinely produces high quality sequences, is needed to facilitate high-volume production of these mtGenome data sets. We developed a robust 8-amplicon Sanger sequencing strategy that regularly produces complete, forensic-quality mtGenome haplotypes in the first pass of data generation. The protocol works equally well on samples representing diverse mtDNA haplogroups and DNA input quantities ranging from 50 pg to 1 ng, and can be applied to specimens of varying DNA quality. The complete workflow was specifically designed for implementation on robotic instrumentation, which increases throughput and reduces both the opportunities for error inherent to manual processing and the cost of generating full mtGenome sequences. The described strategy will assist efforts to generate complete mtGenome haplotypes which meet the highest data quality expectations for forensic genetic and other applications. Additionally, high-quality data produced using this protocol can be used to assess mtDNA data developed using newer technologies and chemistries. Further, the amplification strategy can be used to enrich for mtDNA as a first step in sample preparation for targeted next-generation sequencing.https://doi.org/10.1186/1471-2164-14-88

Relationship between gastro-intestinal complaints and endotoxaemia, cytokine release and acute phase reaction during and after a long-distance triathlon in highly trained men

The aim of the present study was to establish whether gastro-intestinal (GI) complaints observed during and after ultra-endurance exercise are related to gut ischaemia-associated leakage of endotoxins [lipopolysaccharide (LPS)] into the circulation and associated cytokine production. Therefore we collected blood samples from 29 athletes before, immediately after, and 1, 2 and 16 h after a long-distance triathlon for measurement of LPS, tumour necrosis factor-a and interleukin-6 (IL-6). As the cytokine response would trigger an acute-phase response, characteristic variables of these responses were also measured, along with creatine kinase (CK) to obtain an indicator of muscle damage. There was a high incidence (93 % of all participants) of GI symptoms; 45 % reported severe complaints and 7 % of the participants abandoned the race because of severe GI distress. Mild endotoxaemia (5-15 pg/ml) was evident in 68 % of the athletes immediately after the race, as also indicated by a reduction in IgG anti-LPS levels. In addition, we observed production of IL-6 (27-fold increase immediately after the race), leading to an acutephase response (20-fold increase in C-reactive protein and 12 % decrease in pre-albumin 16 h after the race). The extent of endotoxaemia was not correlated with the GI complaints or the IL-6 response, but did show a correlation with the elevation in C-reactive protein (r(s) 0.389; P = 0.037). Creatine kinase levels were increased significantly immediately post-race, and increased further in the follow-up period. Creatine kinase levels did not correlate with those of either IL-6 or C-reactive protein. It is therefore concluded that LPS does enter the circulation after ultra-endurance exercise and may, together with muscle damage, be responsible for the increased cytokine response and hence GI complaints in these athletes

Seismic imaging of dyke swarms within the Sorgenfrei–Tornquist Zone (Sweden) and implications for thermal energy storage

There is a great interest and demand for green-type energy storage in Sweden for both short- and long-term (hours, days, weeks and seasons) periods. While there are a number of approaches proposed (e.g., compressed air, geothermal and thermal), only a few have commercially been demonstrated through upscaling projects. Among these, the thermal energy storage (TES) that stores energy (excess heat or cold) in fluids is particularly interesting. The excess energy can be stored underground in excavated caverns and used for large district heating and cooling purposes as well as for balancing and regulating electrical energy in power grids. For an upscaling underground TES project within the Tornquist suture zone of Scania in the southwest of Sweden, three high-resolution seismic profiles, each approximately 1&thinsp;km long, were acquired. Geologically, the site sits within the southern margin of the Romeleåsen fault zone in the Sorgenfrei–Tornquist Zone (STZ), where dolerite dyke swarms of Carboniferous–Permian age are observed striking in the SE–NW direction for hundreds of kilometers both on land and in offshore seismic and magnetic data (from Scania to Midland Valley in the UK). These dykes, 10–50&thinsp;m thick, in the nearby quarries (within both Precambrian gneiss and quartzite) express themselves mostly in a subvertical manner. They can therefore act as a good water/fluid barrier, which can be an important geological factor for any TES site. For the data acquisition, combined cabled and wireless recorders were used to provide continuity on both sides of a major road running in the middle of the study area. Bedrock depressions are clearly depicted in the tomograms, suggesting the possibility of zones of weaknesses, highly fractured and/or weathered, in the bedrock and confirmed in several places by follow-up boreholes. Several steeply dipping (60–65°) reflections were imaged down to 400&thinsp;m depth and interpreted to originate from dolerite dykes. This interpretation is based on their orientations, strong amplitudes, regular occurrences and correlation with downhole logging data. In addition, groundwater flow measurements within the unconsolidated sediments and in bedrock suggest steeply dipping structures are the dominant factor in directing water mainly along a SE–NW trend, which is consistent with the strike of the dyke swarm within the STZ. To provide further insight on the origin of the reflections, even the historical crustal-scale offshore BABEL (Baltic and Bothnian Echoes from the Lithosphere) lines (A-AA-AB) were revisited. Clear multiphase faults and signs of intrusions or melt source in the lower crust are observed, as well as a Moho step across the Tornquist zone. Overall, we favor that the reflections are of dolerite origin and their dip component (i.e., not subvertical) may imply a Precambrian basement (and dykes) tilting, block rotation, towards the NE as a result of the Romeleåsen reverse faulting. In terms of thermal storage, these dykes then may be encountered during the excavation of the site and can complicate underground water flow should they be used as a fluid barrier in case of leakage.</p

Accuracy and Reproducibility in Quantification of Plasma Protein Concentrations by Mass Spectrometry without the Use of Isotopic Standards

Medical Biochemistr