1,721,023 research outputs found
<i>PIGA</i> gene targeting in human somatic cell lines.
No full text<p>(A) A diagram of the promoter-trap <i>PIGA</i> targeting vector. Gray boxes indicate <i>PIGA</i> exon 6, while thick lines indicate <i>PIGA</i> intron 5 and intergenic genomic region. Thin dotted lines indicate homology between the endogenous gene locus and the targeting vector. F1 to F3 and R1 to R3 indicate PCR primers used in the experiment shown in Fig. 2A and B. A filled rectangle indicates the location of the probe used in Southern blot analysis shown in Fig. 2C. XbaI restriction enzyme sites are marked in the diagram. An arrow at the bottom shows the distance between two XbaI sites at the targeted <i>PIGA</i> gene locus. Triangles: loxP; Stop: stop codon (TAA); IRES: internal ribosomal entry site; Neo<sup>R</sup>: neomycin resistance gene; pA: polyadenylation site. (B and C) Human somatic cell lines were infected with the promoter-trap <i>PIGA</i> targeting vector or a vector control (VC), and processed for FLAER staining and then flow cytometric analyses. Shown are the representative dot plots (B) and a graphic representation of the entire results (mean ± s.e.m.; <i>n</i> = 3) (C). In (B), the FLAER-negative ratio for each experiment is noted in the dot plot. TV: promoter-trap <i>PIGA</i> targeting vector.</p
Monitoring of the gene targeting efficiencies in various experimental conditions using the <i>PIGA</i>-based reporter system.
No full text<p>(A) HCT116 and DLD-1 cells were transfected (as a plasmid) or infected (as an AAV) with the promoter-trap <i>PIGA</i> targeting vector, and the ratios of FLAER-negative cell populations were determined in duplicate. In X-axis labels, VC, IRES (plasmid) and IRES indicate a vector control, and the plasmid and AAV versions of promoter-trap <i>PIGA</i> targeting vector, respectively. (B) HCT116 and DLD-1 cells were infected with <i>PIGA</i> targeting vectors in which the Neo<sup>R</sup> gene was driven by the cytomegalovirus (CMV) promoter or the promoter of human phosphoglycerate kinase 1 (<i>PGK1</i>) gene, and the ratios of FLAER-negative cell populations were determined in duplicate. (C) HCT116 was infected with the indicated multiplicity of infection of the AAV-based promoter-trap <i>PIGA</i> targeting vector, and the ratios of FLAER-negative cell populations were determined (mean ± s.e.m.; <i>n</i> = 3).</p
Growth indices of <i>PIGA</i>-targeted and non-targeted cell populations.
No full text<p>Cells derived from HCT116 (left) and DLD-1 (right) were propagated in 96-well tissue culture plates, and cell numbers assayed at Day 3, Day 5, and Day 7 were shown relative to those at Day 1. In each cluster of bar graphs, open bars represent a FLAER-negative bulk cell population and <i>PIGA</i>-targeted (confirmed by Southern blotting) single cell clones #1, #2 and #3 (from left to right); gray bars represent a FLAER-positive bulk cell population and <i>PIGA</i>-non-targeted (determined by analytical PCR) single cell clones #1, #2 and #3 (from left to right); a filled bar represents the parental cell line (mean ± s.d.; <i>n</i> = 6).</p
The journey towards clinical adoption of MALDI-MS-based imaging proteomics: from current challenges to future expectations
Full text linkAmong the spatial omics techniques available, mass spectrometry imaging (MSI) represents one of the most promising owing to its capability to map the distribution of hundreds of peptides and proteins, as well as other classes of biomolecules, within a complex sample background in a multiplexed and relatively high-throughput manner. In particular, matrix-assisted laser desorption/ionisation (MALDI-MSI) has come to the fore and established itself as the most widely used technique in clinical research. However, the march of this technique towards clinical utility has been hindered by issues related to method reproducibility, appropriate biocomputational tools, and data storage. Notwithstanding these challenges, significant progress has been achieved in recent years regarding multiple facets of the technology and has rendered it more suitable for a possible clinical role. As such, there is now more robust and extensive evidence to suggest that the technology has the potential to support clinical decision-making processes under appropriate circumstances. In this review, we will discuss some of the recent developments that have facilitated this progress and outline some of the more promising clinical proteomics applications which have been developed with a clear goal towards implementation in mind
Going Beyond Counting First Authors in Author Co-citation Analysis
Full text linkThe present study examines one of the fundamental aspects of author co-citation analysis (ACA) - the way co-citation
counts are defined. Co-citation counting provides the data on which all subsequent statistical analyses and mappings
are based, and we compare ACA results based on two different types of co-citation counting - the traditional type that
only counts the first one among a cited work's authors on the one hand and a non-traditional type that takes into
account the first 5 authors of a cited work on the other hand. Results indicate that the picture produced through this non-traditional author co-citation counting contains more coherent author groups and is therefore considerably clearer. However, this picture represents fewer specialties in the research field being studied than that produced through the traditional first-author co-citation counting when the same number of top-ranked authors is selected and analyzed. Reasons for these effects are discussed
Variations on the Author
Full text link“Variations on the Author” discusses two of Eduardo Coutinho’s recent films (Um Dia na Vida, from 2010, and Últimas Conversas, posthumously released in 2015) and their contribution to the general question of documentary authorship. The director’s filmography is characterized by a consistent yet self-effacing form of authorial self-inscription: Coutinho often features as an interviewer that rather than express opinions propels discourses; an interviewer that is good at listening. This mode of self-inscription characterizes him as an author who is not expressive but who is nonetheless markedly present on the screen. In Um Dia na Vida, however, Coutinho is completely absent form the image, while Últimas Conversas, on the contrary, includes a confessional prologue that moves the director from the margins to the center of his films. This article examines the ways in which these works stand out in the filmography of a director who offers new insights into the notion of cinematic authorship
Appropriate Similarity Measures for Author Cocitation Analysis
Full text linkWe provide a number of new insights into the methodological discussion about author cocitation analysis. We first argue that the use of the Pearson correlation for measuring the similarity between authors’ cocitation profiles is not very satisfactory. We then discuss what kind of similarity measures may be used as an alternative to the Pearson correlation. We consider three similarity measures in particular. One is the well-known cosine. The other two similarity measures have not been used before in the bibliometric literature. Finally, we show by means of an example that our findings have a high practical relevance.information science;Pearson correlation;cosine;similarity measure;author cocitation analysis
Confirmatory assays examining the <i>PIGA</i> gene locus in FLAER-negative and -positive cell clones.
No full text<p>(A and B) Representative results of PCR examining FLAER-negative and -positive single cell clones derived from HCT116 (A) and DLD-1 (B) infected with the promoter-trap <i>PIGA</i> targeting vector. PCR was conducted with primer pairs F1-R1, F2-R2 and F3-R3 that yielded PCR products of 1,140 bp, 1,184 bp, and 1,031 bp in size, respectively. For the location of primers, see Fig. 1A. Control: parental cell lines. (C) Southern blot analysis of FLAER-negative and -positive single cell clones. Genomic DNA was digested with XbaI, separated on a gel, and hybridized with a probe shown in Fig. 1A. The positions of the DNA size standards are shown to the right in kilobase. P: parental cell lines.</p
Dispelling the Myths Behind First-author Citation Counts
Full text linkWe conducted a full-scale evaluative citation analysis study of scholars in the XML research field to explore just how different from each other author rankings resulting from different citation counting methods actually are, and to demonstrate the capability of emerging data and tools on the Web in supporting more realistic citation counting methods. Our results contest some common arguments for the continued
use of first-author citation counts in the evaluation of scholars, such as high correlations between author rankings by first-author citation counts and other citation
counting methods, and high costs of using more realistic citation counting methods that are not well-supported by the ISI databases. It is argued that increasingly available digital full text research papers make it possible for citation analysis studies to go beyond what the ISI databases have directly supported and to employ more
sophisticated methods
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