735 resultados para panel de FISH
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Especially in panel surveys, respondent attrition, respondent learning, and interviewer experience effects play a crucial role with respect to data quality. We examine three interview survey quality indicators in the same survey in a cross sectional as well as in a longitudinal way. In the cross sectional analysis we compare data quality in the mature original sample with that in a refreshment sample, surveyed in the same wave. Because in the same wave an interviewer survey was conducted, collecting attitudes on their socio demography, survey attitudes and burden measures, we are able to consider interviewer fixed effects as well. The longitudinal analysis gives more insight in the respondent learning effects with respect to the quality indicators considered by considering the very same respondents across waves. The Swiss Household Panel, a CATI survey representative of the Swiss residential population, forms an ideal modelling database: the interviewer - respondent assignment is random, both within and across waves. This design avoids possible confusion with other effects stemming from a non-random assignment of interviewers, e.g. area effects or effects from assigning the best interviewers to the hard cases. In order to separate interviewer, respondent and wave effects, we build cross-classified multilevel models.
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Reference collections of multiple Drosophila lines with accumulating collections of "omics" data have proven especially valuable for the study of population genetics and complex trait genetics. Here we present a description of a resource collection of 84 strains of Drosophila melanogaster whose genome sequences were obtained after 12 generations of full-sib inbreeding. The initial rationale for this resource was to foster development of a systems biology platform for modeling metabolic regulation by the use of natural polymorphisms as perturbations. As reference lines, they are amenable to repeated phenotypic measurements, and already a large collection of metabolic traits have been assayed. Another key feature of these strains is their widespread geographic origin, coming from Beijing, Ithaca, Netherlands, Tasmania, and Zimbabwe. After obtaining 12.5× coverage of paired-end Illumina sequence reads, SNP and indel calls were made with the GATK platform. Thorough quality control was enabled by deep sequencing one line to >100×, and single-nucleotide polymorphisms and indels were validated using ddRAD-sequencing as an orthogonal platform. In addition, a series of preliminary population genetic tests were performed with these single-nucleotide polymorphism data for assessment of data quality. We found 83 segregating inversions among the lines, and as expected these were especially abundant in the African sample. We anticipate that this will make a useful addition to the set of reference D. melanogaster strains, thanks to its geographic structuring and unusually high level of genetic diversity.
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Colorectal cancer (CRC) is the second leading cause of cancer-related death in developed countries. Early detection of CRC leads to decreased CRC mortality. A blood-based CRC screening test is highly desirable due to limited invasiveness and high acceptance rate among patients compared to currently used fecal occult blood testing and colonoscopy. Here we describe the discovery and validation of a 29-gene panel in peripheral blood mononuclear cells (PBMC) for the detection of CRC and adenomatous polyps (AP). Blood samples were prospectively collected from a multicenter, case-control clinical study. First, we profiled 93 samples with 667 candidate and 3 reference genes by high throughput real-time PCR (OpenArray system). After analysis, 160 genes were retained and tested again on 51 additional samples. Low expressed and unstable genes were discarded resulting in a final dataset of 144 samples profiled with 140 genes. To define which genes, alone or in combinations had the highest potential to discriminate AP and/or CRC from controls, data were analyzed by a combination of univariate and multivariate methods. A list of 29 potentially discriminant genes was compiled and evaluated for its predictive accuracy by penalized logistic regression and bootstrap. This method discriminated AP >1cm and CRC from controls with a sensitivity of 59% and 75%, respectively, with 91% specificity. The behavior of the 29-gene panel was validated with a LightCycler 480 real-time PCR platform, commonly adopted by clinical laboratories. In this work we identified a 29-gene panel expressed in PBMC that can be used for developing a novel minimally-invasive test for accurate detection of AP and CRC using a standard real-time PCR platform.
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Kiristyvä kansainvälinen kilpailu pakottaa automaatiojärjestelmien valmistajat ottamaan käyttöön uusia menetelmiä, joiden avulla järjestelmien suorituskykyä ja joustavuutta saadaan parannettua. Agenttiteknologiaa on esitetty käytettäväksi olemassa olevien automaatiojärjestelmien kanssa vastaamaan automaatiolle asetettaviin uusiin haasteisiin. Agentit ovat itsenäisiä yhteisöllisiä toimijoita, jotka suorittavat niille ennalta määrättyjä tehtäviä. Ne tarjoavat yhtenäisen kehyksen kehittyneiden toimintojen toteutukselle. Agenttiteknologian avulla automaatiojärjestelmä saadaan toimimaan joustavasti ja vikasietoisesti. Tässä työssä selostetaan agenttiteknologian ajatuksia ja käsitteitä. Lisäksi selvitetään sen soveltuvuutta monimutkaisten ohjausjärjestelmien kehittämiseen ja etsitään käyttökohteita sen soveltamiselle levytehtaassa. Työssä käsitellään myös aatteita, jotka ovat johtaneet agenttiteknologian käyttöön automaatiojärjestelmissä, sekä selostetaan agenttiavusteisen esimerkkisovelluksen rakenne ja testitulokset. Tutkimuksen tuloksena löydettiin useita kohteita agenttiteknologian käytölle levytehtaassa. Esimerkkisovellus osoittaa sen sopivan hyvin kehittyneiden toimintojen toteutukseen automaatiojärjestelmissä.
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OBJECTIVES: Immunohistochemistry (IHC) has become a promising method for pre-screening ALK-rearrangements in non-small cell lung carcinomas (NSCLC). Various ALK antibodies, detection systems and automated immunostainers are available. We therefore aimed to compare the performance of the monoclonal 5A4 (Novocastra, Leica) and D5F3 (Cell Signaling, Ventana) antibodies using two different immunostainers. Additionally we analyzed the accuracy of prospective ALK IHC-testing in routine diagnostics. MATERIALS AND METHODS: Seventy-two NSCLC with available ALK FISH results and enriched for FISH-positive carcinomas were retrospectively analyzed. IHC was performed on BenchMarkXT (Ventana) using 5A4 and D5F3, respectively, and additionally with 5A4 on Bond-MAX (Leica). Data from our routine diagnostics on prospective ALK-testing with parallel IHC, using 5A4, and FISH were available from 303 NSCLC. RESULTS: All three IHC protocols showed congruent results. Only 1/25 FISH-positive NSCLC (4%) was false negative by IHC. For all three IHC protocols the sensitivity, specificity, positive (PPV) and negative predictive values (NPV) compared to FISH were 96%, 100%, 100% and 97.8%, respectively. In the prospective cohort 3/32 FISH-positive (9.4%) and 2/271 FISH-negative (0.7%) NSCLC were false negative and false positive by IHC, respectively. In routine diagnostics the sensitivity, specificity, PPV and NPV of IHC compared to FISH were 90.6%, 99.3%, 93.5% and 98.9%, respectively. CONCLUSIONS: 5A4 and D5F3 are equally well suited for detecting ALK-rearranged NSCLC. BenchMark and BOND-MAX immunostainers can be used for IHC with 5A4. True discrepancies between IHC and FISH results do exist and need to be addressed when implementing IHC in an ALK-testing algorithm.
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Background Chronic obstructive pulmonary disease (COPD) is a heterogeneous disease whose assessment and management have traditionally been based on the severity of airflow limitation (forced expiratory volume in 1 s (FEV1)). Yet, it is now clear that FEV1 alone cannot describe the complexity of the disease. In fact, the recently released Global Initiative for Chronic Obstructive Lung Disease (GOLD), 2011 revision has proposed a new combined assessment method using three variables (symptoms, airflow limitation and exacerbations). Methods Here, we go one step further and propose that in the near future physicians will need a"control panel" for the assessment and optimal management of individual patients with complex diseases, including COPD, that provides a path towards personalised medicine. Results We propose that such a"COPD control panel" should include at least three different domains of the disease: severity, activity and impact. Each of these domains presents information on different"elements" of the disease with potential prognostic value and/or with specific therapeutic requirements. All this information can be easily incorporated into an"app" for daily use in clinical practice. Conclusion We recognise that this preliminary proposal needs debate, validation and evolution (eg, including"omics" and molecular imaging information in the future), but we hope that it may stimulate debate and research in the field.
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In the past four decades, women have made major inroads into occupations previously dominated by men. This paper examines whether occupational feminization is accompanied by a decline in wages: Do workers suffer a wage penalty if they remain in, or move into, feminizing occupations? We analyze this question over the 1990s and 2000s in Britain, Germany, and Switzerland, using longitudinal panel data to estimate individual fixed effects for men and women. Moving from an entirely male to an entirely female occupation entails a loss in individual earnings of 13 percent in Britain, 7 percent in Switzerland, and 3 percent in Germany. The impact of occupational feminization on wages is not linear, but sets apart occupations holding more than 60 percent of women. Moving into such female occupations incurs a wage penalty. Contrary to the prevailing idea in economics, differences in productivity-human capital, job-specific skills, and time investment-do not fully explain the wage gap between male and female occupations. The wage penalty associated with working in a female occupation is also much larger where employer discretion is greater-in the private sector-than where wagesetting is guided by formal rules-the public sector. These findings suggest that wage disparities across male and female occupations are due to gender devaluation.
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The purpose of this thesis is to study factors that explain the bilateral fiber trade flows. This is done by analyzing bilateral trade flows during 1990-2006. It will be studied also, whether there are differences between fiber types. This thesis uses a gravity model approach to study the trade flows. Gravity model is mostly used to study the aggregate data between trading countries. In this thesis the gravity model is applied to single fibers. This model is then applied to panel data set. Results from the regression show clearly that there are benefits in studying different fibers in separate. The effects differ considerably from each other. Furthermore, this thesis speaks for the existence of Linder’s effect in certain fiber types.
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AMPK, a master metabolic switch, mediates the observed increase of glucose uptake in locomotory muscle of mammals during exercise. AMPK is activated by changes in the intracellular AMP:ATP ratio when ATP consumption is stimulated by contractile activity but also by AICAR and metformin, compounds that increase glucose transport in mammalian muscle cells. However, the possible role of AMPK in the regulation of glucose metabolism in skeletal muscle has not been investigated in other vertebrates, including fish. In this study, we investigated the effects of AMPK activators on glucose uptake, AMPK activity, cell surface levels of trout GLUT4 and expression of GLUT1 and GLUT4 as well as the expression of enzymes regulating glucose disposal and PGC1α in trout myotubes derived from a primary muscle cell culture. We show that AICAR and metformin significantly stimulated glucose uptake (1.6 and 1.3 fold, respectively) and that Compound C completely abrogated the stimulatory effects of the AMPK activators on glucose uptake. The combination of insulin and AMPK activators did not result in additive nor synergistic effects on glucose uptake. Moreover, exposure of trout myotubes to AICAR and metformin resulted in an increase in AMPK activity (3.8 and 3 fold, respectively). We also provide evidence suggesting that stimulation of glucose uptake by AMPK activators in trout myotubes may take place, at least in part, by increasing the cell surface and mRNA levels of trout GLUT4. Finally, AICAR increased the mRNA levels of genes involved in glucose disposal (hexokinase, 6-phosphofructokinase, pyruvate kinase and citrate synthase) and mitochondrial biogenesis (PGC-1α) and did not affect glycogen content or glycogen synthase mRNA levels in trout myotubes. Therefore, we provide evidence, for the first time in non-mammalian vertebrates, suggesting a potentially important role of AMPK in stimulating glucose uptake and utilization in the skeletal muscle of fish.
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Postprint (published version)
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Postprint (published version)
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Epitheliocystis is an infectious disease affecting gills and skin of various freshwater and marine fishes, associated with high mortality and reduced growth of survivors. Candidatus Piscichlamydia salmonis and Clavochlamydia salmonicola have recently been identified as aetiological agents of epitheliocystis in Atlantic Salmon. In addition, several other members of the Chlamydiales order have been identified in other fish species. To clarify the pathogenicity of Chlamydia-like organisms towards fishes, we investigated the permissivity of two fish cell lines, EPC-175 (Fathead Minnow) and RTG-2 (rainbow trout) to three Chlamydia-related bacteria: Waddlia chondrophila, Parachlamydia acanthamoebae and Estrella lausannensis. Quantitative PCR and immunofluorescence demonstrated that W. chondrophila and, to a lesser extent, E. lausannensis were able to replicate in the two cell lines tested. Waddlia chondrophila multiplied rapidly in its host cell and a strong cytopathic effect was observed. During E. lausannensis infection, we observed a limited replication of the bacteria not followed by host cell lysis. Very limited replication of P. acanthamoebae was observed in both cell lines tested. Given its high infectivity and cytopathic effect towards fish cell lines, W. chondrophila represents the most interesting Chlamydia-related bacteria to be used to develop an in vivo model of epitheliocystis disease in fishes.