987 resultados para neutral red retention assay
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Assessments of the Atlantic red drum for the northern (North Carolina and north) and southern (South Carolina through east coast of Florida) regions along the U. S. Atlantic coast were recently completed. The joint Red Drum Technical Committee (SAFMC/ASMFC) selected the most appropriate catch matrix (incorporating an assumption on size of recreationally-released fish), selectivity of age 3 relative to age 2, and virtual population analysis (FADAPT). Given gear- and age-specific estimates of fishing mortality (F) for the 1992-1998 period, analyses were made of potential gains in escapement through age 4 and static spawning potential ratio (SPR) from further reductions in fishing mortality due to changes in slot and bag limits. Savings from bag limits were calculated given a particular slot size for the recreational fishery, with no savings for the commercial fisheries in the northern region due to their being managed primarily through a quota. Relative changes in catch-at-age estimates were used to adjust age-specific F and hence calculated escapement through age 4 and static SPR. Adjustment was made with the recreational savings to account for release mortality (10%, as in the stock assessment). Alternate runs for the northern region commercial fishery considered 25% release mortality for lengths outside the slot (instead of 0% for the base run), and 0% vs. 10% gain or loss across legal sizes in F. These results are summarized for ranges of bag limits with increasing minimum size limit (for fixed maximum size), and with decreasing maximum size limit (for fixed minimum size limit). For the southern region, a bag limit of one-fish per angler trip would be required to attain the stated target of 40% static SPR if the current slot limit were not changed. However, for the northern region, a bag limit of one-fish per angler trip appears to be insufficient to attain the stated target of 40% static SPR while maintaining the current slot limit. (PDF contains 41 pages)
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Red hind (Epinephelus guttatus) have been overfished in the Caribbean and were included with seven other regional grouper species deemed vulnerable to risk of extinction. The Puerto Rico Department of Natural and Environmental Resources desired to map spawning red hind aggregations within commonwealth waters as part of their resource management program for the species. Mobile hydroacoustic surveys were conducted over 3-day periods in 2002 and 2003, indexed to the full moon phase in February or March when red hind were known to aggregate. Four vessels concurrently sampled the southwest, south, and southeast coasts of Puerto Rico in 2002. In 2003, three vessels conducted complementary surveys of the northwest, north, and northeast coasts of the island, completing a circuit of the coastal shelf-spawning habitat. These surveys indicated that red hind spawning aggregations were prevalent along the south and west coasts, and sparse along the north coast during the survey periods. Highest spawning red hind concentrations were observed in three areas offshore of the west coast of Puerto Rico, around Mona and Desecheo islands (20,443 and 10,559 fish/km2, respectively) and in the Bajo de Cico seasonal closed area (4,544 fish/km2). Following both 2002 and 2003 surveys, a series of controlled acoustic measurements of known local fish species in net pens were conducted to assess the mean target strength (acoustic backscatter) of each group. Ten species of fish were measured, including red hind (E. guttatus), coney (E. fulvus), white grunt (Haemulon plumieri), pluma (Calamus pennatula), blue tang (Acanthurus coeruleus), squirrel fish (Holocentrus spp.), black durgeon (Melichtyhs niger), ocean file fish (Canthidermis sufflamen), ocean surgeon fish (Acanthurus bahianus), and butter grouper (Mycteroperca spp.). In general, the mean target strength results from the caged fish experiments were in agreement with published target strength length relationships, with the exception of white grunt and pluma.
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Background: Lynch syndrome (LS) is an autosomal dominant inherited cancer syndrome characterized by early onset cancers of the colorectum, endometrium and other tumours. A significant proportion of DNA variants in LS patients are unclassified. Reports on the pathogenicity of the c.1852_1853AA>GC (p.Lys618Ala) variant of the MLH1 gene are conflicting. In this study, we provide new evidence indicating that this variant has no significant implications for LS. Methods: The following approach was used to assess the clinical significance of the p.Lys618Ala variant: frequency in a control population, case-control comparison, co-occurrence of the p.Lys618Ala variant with a pathogenic mutation, co-segregation with the disease and microsatellite instability in tumours from carriers of the variant. We genotyped p.Lys618Ala in 1034 individuals (373 sporadic colorectal cancer [CRC] patients, 250 index subjects from families suspected of having LS [revised Bethesda guidelines] and 411 controls). Three well-characterized LS families that fulfilled the Amsterdam II Criteria and consisted of members with the p.Lys618Ala variant were included to assess co-occurrence and co-segregation. A subset of colorectal tumour DNA samples from 17 patients carrying the p.Lys618Ala variant was screened for microsatellite instability using five mononucleotide markers. Results: Twenty-seven individuals were heterozygous for the p.Lys618Ala variant; nine had sporadic CRC (2.41%), seven were suspected of having hereditary CRC (2.8%) and 11 were controls (2.68%). There were no significant associations in the case-control and case-case studies. The p.Lys618Ala variant was co-existent with pathogenic mutations in two unrelated LS families. In one family, the allele distribution of the pathogenic and unclassified variant was in trans, in the other family the pathogenic variant was detected in the MSH6 gene and only the deleterious variant co-segregated with the disease in both families. Only two positive cases of microsatellite instability (2/17, 11.8%) were detected in tumours from p.Lys618Ala carriers, indicating that this variant does not play a role in functional inactivation of MLH1 in CRC patients. Conclusions: The p.Lys618Ala variant should be considered a neutral variant for LS. These findings have implications for the clinical management of CRC probands and their relatives.
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Background: The presence of EGFR kinase domain mutations in a subset of NSCLC patients correlates with the response to treatment with the EGFR tyrosine kinase inhibitors gefitinib and erlotinib. Although most EGFR mutations detected are short deletions in exon 19 or the L858R point mutation in exon 21, more than 75 different EGFR kinase domain residues have been reported to be altered in NSCLC patients. The phenotypical consequences of different EGFR mutations may vary dramatically, but the majority of uncommon EGFR mutations have never been functionally evaluated. Results: We demonstrate that the relative kinase activity and erlotinib sensitivity of different EGFR mutants can be readily evaluated using transfection of an YFP-tagged fragment of the EGFR intracellular domain (YFP-EGFR-ICD), followed by immunofluorescence microscopy analysis. Using this assay, we show that the exon 20 insertions Ins770SVD and Ins774HV confer increased kinase activity, but no erlotinib sensitivity. We also show that, in contrast to the common L858R mutation, the uncommon exon 21 point mutations P848L and A859T appear to behave like functionally silent polymorphisms. Conclusion: The ability to rapidly obtain functional information on EGFR variants of unknown relevance using the YFP-EGFR-ICD assay might prove important in the future for the management of NSCLC patients bearing uncommon EGFR mutations. In addition, our assay may be used to determine the response of resistant EGFR mutants to novel second-generation TKIs.
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Background: The diagnosis of invasive candidiasis is difficult because there are no specific clinical manifestations of the disease and colonization and infection are difficult to distinguish. In the last decade, much effort has been made to develop reliable tests for rapid diagnosis of invasive candidiasis, but none of them have found widespread clinical use. Results: Antibodies against a recombinant N-terminal fragment of the Candida albicans germ tube-specific antigen hyphal wall protein 1 (Hwp1) generated in Escherichia coli were detected by both immunoblotting and ELISA tests in a group of 36 hematological or Intensive Care Unit patients with invasive candidiasis and in a group of 45 control patients at high risk for the mycosis who did not have clinical or microbiological data to document invasive candidiasis. Results were compared with an immunofluorescence test to detect antibodies to C. albicans germ tubes (CAGT). The sensitivity, specificity, positive and negative predictive values of a diagnostic test based on the detection of antibodies against the N-terminal fragment of Hwp1 by immunoblotting were 27.8 %, 95.6 %, 83.3 % and 62.3 %, respectively. Detection of antibodies to the N-terminal fragment of Hwp1 by ELISA increased the sensitivity (88.9 %) and the negative predictive value (90.2 %) but slightly decreased the specificity (82.6 %) and positive predictive values (80 %). The kinetics of antibody response to the N-terminal fragment of Hwp1 by ELISA was very similar to that observed by detecting antibodies to CAGT. Conclusion: An ELISA test to detect antibodies against a recombinant N-terminal fragment of the C. albicans germ tube cell wall antigen Hwp1 allows the diagnosis of invasive candidiasis with similar results to those obtained by detecting antibodies to CAGT but without the need of treating the sera to adsorb the antibodies against the cell wall surface of the blastospore.
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The alkaline comet assay is a method of detecting DNA strand breaks and alkali labile sites in individual cells. The method was used to detect DNA strand breaks in isolated blood cells (leukocytes) of carp (Cyprius carpio). DNA damage have been induced by exposure of the cells to sediment extract. Therefore comet assay can be applied as in vitro bioassay for investigations on toxicity of marine sediments.
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Micro-fabrication technology has substantial potential for identifying molecular markers expressed on the surfaces of tissue cells and viruses. It has been found in several conceptual prototypes that cells with such markers are able to be captured by their antibodies immobilized on microchannel substrates and unbound cells are flushed out by a driven flow. The feasibility and reliability of such a microfluidic-based assay, however, remains to be further tested. In the current work, we developed a microfluidic-based system consisting of a microfluidic chip, an image grabbing unit, data acquisition and analysis software, as well as a supporting base. Specific binding of CD59-expressed or BSA-coupled human red blood cells (RBCs) to anti-CD59 or anti-BSA antibody-immobilized chip surfaces was quantified by capture efficiency and by the fraction of bound cells. Impacts of respective flow rate, cell concentration, antibody concentration and site density were tested systematically. The measured data indicated that the assay was robust. The robustness was further confirmed by capture efficiencies measured from an independent ELISA-based cell binding assay. These results demonstrated that the system developed provided a new platform to effectively quantify cellular surface markers effectively, which promoted the potential applications in both biological studies and clinical diagnoses.
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L-selectin plays a crucial role in inflammation cascade by initiating the tethering and rolling of leukocytes on endothelium wall. While many L-selectin molecules are rapidly shed from the cell surface upon activation, the remaining membrane-anchored L-selectin may still play an important role in regulating leukocyte rolling and adhesion with different binding kinetics. Here we developed an in vitro model to activate Jurkat cells via interlukin-8 (IL-8) and quantified the two-dimensional (2D) binding kinetics, using a micropipette aspiration assay, of membrane-anchored L-selectin to P-selectin glycoprotein ligand 1 (PSGL-1) ligand coupled onto human red blood cells (RBCs). The data indicated that L-selectin shedding reduced the amount of membrane-anchored L-selectin and lowered both its reverse and forward rates. These results suggested that the rolling dynamics of activated leukocytes was determined by two opposite impacts: reducing the surface presentation would enhance the rolling but lowering the kinetic rates would decrease the rolling. This finding provides a new insight into understanding how L-selectin shedding regulates leukocyte rolling and adhesion.
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Durante los últimos años hemos venido observando la tendencia a incorporar capacidad de proce- samiento y comunicación a dispositivos que hasta entonces se utilizaban de modo independiente. La evolución de los móviles a smartphones es un claro ejemplo de dicha tendencia, aunque también cabe mencionar otros ejemplos, como es el caso de los denominados hogares inteligentes, en los que elementos del hogar se encuentran interconectados y pueden actuar de modo conjunto. Esta ten- dencia no se limita a sistemas independientes, sino que propone interconectar todos los elementos disponibles para conformar la denominada Internet de los Objetos/Cosas o Internet of Things, IoT. Una de las mayores dificultades que se presenta en estos sistemas es que las características de es- tos nuevos dispositivos inteligentes, en general limitados en términos de cómputo, almacenamiento, autonomía o comunicación, queda a menudo lejos de los equipos informáticos tradicionales. Esta cuestión, junto con la ausencia de estándares para gestionar estos dispositivos, constituye un impor- tante problema a abordar. Considerando este marco, en este proyecto se ha desarrollado una aplicación orientada a este tipo de dispositivos. Más concretamente, la aplicación tiene como soporte una red de sensores inalámbricos, WSN, con el objetivo de realizar seguimiento de individuos. Cabe destacar que el desarrollo de la aplicación se ha realizado utilizando Contiki OS, sistema ope- rativo diseñado especialmente para dispositivos con características limitadas como los presentados anteriormente y firme candidato a convertirse en estándar.
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El seguimiento de distintas especies de animales contribuye en gran medida a su estudio y, por tanto, a su conservación y control. Los avances tecnológicos de los últimos años han facilitado las posibilidades de seguimiento con la creación de distintos dispositivos que permiten conocer los movimientos de la especie que se desea estudiar. Uno de los sistemas más utilizados consiste en la utilización de dispositivos GPS incorporados al espécimen sobre el que se realiza el seguimiento y cuya señal es recogida por satélites que se encargan de almacenar y posteriormente reenviar la información para su almacenamiento y procesamiento en el laboratorio. El principal problema de este sistema es su elevado coste. Existen alternativas que no presentan un coste tal alto, tales como el uso de módulos basados en telefonía móvil. Sin embargo, tienen limitaciones de cobertura, por lo que no es aplicable en todos los ámbitos. Este proyecto forma parte de una propuesta que ofrece realizar seguimiento de ejemplares de una especie de ave, la gaviota Patiamarilla, en Gipuzkoa mediante la utilización de una red de sensores y que tiene varias ventajas frente a las opciones presentadas anteriormente. En este proyecto en concreto se ha diseñado e implementado el módulo que permite recoger la información obtenida por el conjunto de sensores (cada ejemplar lleva incorporado un sensor que permite registrar su posición) y enviarla a un servidor centralizado para su posterior consulta y análisis. Adicionalmente, también se permite consultar el último estado registrado de cada dispositivo de seguimiento, además de contemplar la posibilidad de actualizar su software.
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A search for dielectron decays of heavy neutral resonances has been performed using proton-proton collision data collected at √s = 7 TeV by the Compact Muon Solenoid (CMS) experiment at the Large Hadron Collider (LHC) in 2011. The data sample corresponds to an integrated luminosity of 5 fb−1. The dielectron mass distribution is consistent with Standard Model (SM) predictions. An upper limit on the ratio of the cross section times branching fraction of new bosons, normalized to the cross section times branching fraction of the Z boson, is set at the 95 % confidence level. This result is translated into limits on the mass of new neutral particles at the level of 2120 GeV for the Z′ in the Sequential Standard Model, 1810 GeV for the superstring-inspired Z′ψ resonance, and 1940 (1640) GeV for Kaluza-Klein gravitons with the coupling parameter k/MPl of 0.10 (0.05).