1,720,963 research outputs found
Enhanced three-dimensional particle detection in microcirculation experiments with defocus particle tracking and ghost red blood cells
Full text linkExperimental investigations on the motion of rigid particles in microcirculation environments are still scarce owing to the three-dimensional (3D) motion of the particles and to the particle image masking due to the presence of the red blood cells (RBCs). Despite the recent progress on the 3D tracking of rigid particles in RBC flows with defocus particle tracking (DPT) methods, the problem of particle image masking remains to be solved. Here, we propose, test, and evaluate the use hemoglobin-free RBCs, also known as ghost RBCs, as a replacement for normal RBCs in experiments with rigid particles in microcirculation environments. We performed DPT measurements of a pressure-driven flow of normal and ghost RBC suspensions seeded with rigid particles at three different flow rates. We show that the quasi-transparent appearance of ghost RBCs, as a result of the lack of hemoglobin, eliminates the RBC-induced masking of the defocused particle images and allows to achieve the particle matching standards found in cell-free experiments. In fact, ghost RBC suspensions enable the tracking of the rigid particles across the entire height of the microchannel, which was not possible in normal RBC flows. On a fluid dynamic level, we show that ghost RBC suspensions provide similar conditions to normal RBCs in terms of the velocity of the rigid particles and the rigid particles exhibit similar lateral dynamics in both types of cell suspensions. In essence, the findings from this work demonstrate that ghost RBCs are a well-suited replacement for normal RBCs in experiments aiming at deciphering the motion of rigid particles in microcirculation environments
Engineering of Escherichia coli strains for plasmid biopharmaceutical production: Scale-up challenges
No full textPlasmid-based vaccines and therapeutics have been making their way into the clinic in the last years. The existence of cost-effective manufacturing processes capable of delivering high amounts of high-quality plasmid DNA (pDNA) is essential to generate enough material for trials and support future commercialization. However, the development of pDNA manufacturing processes is often hampered by difficulties in predicting process scale performance of Escherichia coli cultivation on the basis of results obtained at lab scale. This paper reports on the differences observed in pDNA production when using shake flask and bench-scale bioreactor cultivation of E. coli strains MG1655ΔendAΔrecA and DH5α in complex media with 20 g/L of glucose. MG1655ΔendAΔrecA produced 5-fold more pDNA (9.8 mg/g DCW) in bioreactor than in shake flask (1.9 mg/g DCW) and DH5α produced 4-fold more pDNA (8 mg/g DCW) in bioreactor than in shake flask (2 mg/g DCW). Accumulation of acetate was also significant in shake flasks but not in bioreactors, a fact that was attributed to a lack of control of pH.MIT-Portugal ProgramFundacao para a Ciencia e a Tecnologia (Project PTDC/EBB-EBI/113650/2009)Fundacao para a Ciencia e a Tecnologia (PhD Grant SFRH/BD/33786/2009
On the dual effect of glucose during production of pBAD/AraC-based minicircles
No full textMinicircles are promising vectors for DNA vaccination, gene or cell therapies due to their increased transfection efficacy and transgene expression. The in vivo production of these novel vectors involves the arabinose inducible excision of a parental molecule into a minicircircle and a miniplasmid bacterial backbone. Tight control of recombination is crucial to maximize minicircle yields and purity. In this work, a minicircle production system was constructed that relies on the enzymatic activity of ParA resolvase, a recombinase that is expressed under the transcription control of the arabinose inducible expression system pBAD/AraC, and on Escherichia coli BWAA, a strain improved for arabinose uptake. Undesired recombination already after 4 h of incubation in Luria-Bertani broth at 37 °C was observed due to the leaky expression from pBAD/AraC. While addition of glucose to the growth media repressed this leaky expression, it triggered a pH drop to 4.5 during exponential phase in shake flasks, which suppressed growth and plasmid production. The quantitative PCR analysis confirmed only few copies of high-copy number plasmid inside of the E. coli cells. To ensure the stability of minicircle-producing system, seed cultures should be grown at 30 °C with glucose overnight whereas cells for minicircle production should be grown in shake flasks at 37 °C without glucose up to early stationary phase when the recombination is induced by addition of arabinose.MIT-Portugal ProgramFundacao para a Ciencia e a Tecnologia (PhD Grant SFRH/BD/33786/2009)Fundacao para a Ciencia e a Tecnologia (Project PTDC/EBB-EBI/113650/2009
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
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
koamabayili/VECTRON-author-checklist: VECTRON author checklist
No full textWe have done our best to complete the author checklist relating to the use of animals in the hut study. Note that the objective for the hut study was to evaluate the IRS treatment applications for residual efficacy against Anopheles mosquitoes, including the local An. coluzzii mosquito population. Cows were only used to attract mosquitoes into the huts and no tests were carried out directly on the cows. The author checklist is intended for use with studies where experiments are carried out on animals, which is why we have had such difficulty in completing this for the hut study, as many of the questions do not relate to how the cows were used
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