1,721,056 research outputs found
Developing a new efficient CRISPR-Cas9 base Homology Directed Repair oriented gene editing plasmid for Streptomyces
Full text linkThe CRISPR-Cas9 editing mechanism revolutionized genetic engineering, providing a precise
and versatile tool for editing the genomes of virtually any organism. Streptomyces are soil
dwelling bacteria with a complex life cycle and a reputation for producing a wide array of
bioactive compounds, including many clinically important antibiotics. Despite their
significance, genetic manipulation in Streptomyces has traditionally been challenging. Their
complex and large genomes, slow growth, and the presence of multiple, sometimes
redundant, biosynthetic gene clusters made traditional methods of mutagenesis laborious and
time-consuming. The implementation of CRISPR-Cas9 in Streptomyces therefore has a
profound impact on both basic research and industrial applications. In particular, it is important
for the development of new antibiotics at a time when rising antibiotic resistance is growing
into a global health crisis.
This study sought to develop a new efficient CRISPR-Cas9 gene editing plasmid that works
based on the homology directed repair system in Streptomyces. The fundamental goal of the
study is to characterise the phenotype of a deletion mutant in the gene cluster, SCO6357-53,
by using the newly developed CRISPR-Cas9 plasmid. Previous study has shown that
SCO6357-53 is a suspected teicoplanin tolerance/resistance cluster in Streptomyces
coelicolor. A natural product glycopeptide, teicoplanin, is an important alternative front-line
therapy to vancomycin for the treatment of Gram-positive nosocomial infections but, unlike
vancomycin, its mechanism of resistance is not clearly understood. While this study sought to
characterise this cluster, an inability to create the new CRISPR-Cas9 based editing plasmid
prevented any genetic manipulations to be carried out. Extensive plasmid DNA analysis using
restriction enzyme digestion, PCR and sequencing was conducted. An initial inability to
produce the desired plasmid led to a reconfiguring of the DNA assembly, however this second
method also failed to produce the desired resulting plasmid. Whole plasmid DNA sequencing
analysis revealed that a range of different unexpected rearrangements of DNA occurred, with
either the loss of DNA segments which are functionally vital or a gain of DNA due to an
unexpected insertion of external DNA fragments in various loci in the reconfigured plasmid.
Although additional unwanted DNA fragments were present, one plasmid sample seemed to
preserve all functionally vital regions for the HDR oriented targeted gene editing, so further
cloning based on Gibson assembly was undertaken. However, unfortunately this also failed to
obtain a desired product. Thus, an inability to construct the required HDR oriented gene editing
plasmid, pCRISPR-Cas9KM, meant that further investigation into the teicoplanin resistance
cluster could not be carried out. In light of this, further advancements into generating this actinomycete-codon optimised
genome editing tool needs to be carried out. By investigating the knockout of this teicoplanin
resistance-conferring cluster, further insights into the associated resistance mechanisms can
be determined. Gaining insights into these resistance mechanisms will provide a fundamental
keystone in tackling the rising Anti-Microbial Resistance crisis. Through understanding these
mechanisms, current antibiotics can be developed in a way that circumvent the arising
bacterial resistance mechanisms, thus helping to alleviate the increasingly prevalent AMR
crisis
Sport across cultures: Applications of the human capital model in refugee communities
No full textWhen working with the world’s most vulnerable populations there are questions surrounding the salience of physical activity promotion programs given the multitude of basic needs that must first be met. Indeed, physical activity may be a low priority for individuals seeking safety, reunification with loved ones, and food for their families, as a subsistence lifestyle makes excess weight gain, diabetes, and cardiovascular disease irrelevant. Yet, when working with people from a refugee background for whom these challenges all too frequently apply, opportunities for sport and activity have repeatedly surfaced as desirable and needed, yet are utterly deficient. If we conceptualize physical activity purely as a chronic disease prevention tool, its significance within under-resourced communities is most assuredly lost; however, if we harness the power of physical activity to serve as an agent of positive social change, then it instantly becomes more meaningful and necessary
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
Community development and community resilience: An integrative approach
No full textThroughout history, communities and civilizations have sought to enhance the quality of community life and the well-being of its people. However, more recently there has been greater interest in attending to the details of community development by capitalizing on the improved ability to capture community well-being and successes scientifically. That interest invites greater attention to the development of indicators that can quantify those qualities of life that lead to strong and healthy communities. The selection of meaningful indicators is dependent upon several factors including a process that stimulates meaningful involvement of community stakeholders, but the single most important is the identification of an underlying model to guide the work. Indicators do not have meaning in themselves. For these measures to provide a coherent assessment of the community, an integrative approach to understanding what constitutes a healthy and strong community in a dynamic environment is required. A resilience perspective serves that purpose and provides a framework that is broad, neutral, and conceptually strong enough to structure development of significant sets of indicators. Exemplary community indicator processes across the nation, particularly recent efforts in the Phoenix, Arizona region, provide evidence supporting the value of indicator development for\ud
community building
Why It Hurts to Write: An Analysis of Pain in the Writing Process
No full textFrom the Faculty Nominator: Greg Bortnichak’s and Kate Murray’s collaboratively researched and written English 221 paper combines excellent interdisciplinary research to reach conclusions that are important for all serious writers, as well as for writing teachers and Writing Center tutors. I was impressed by the coherent authorial persona from which the paper appears to arise, even though it was produced by two experienced writers with distinctly differing personal styles. They connect the classical and modern philosophy of pain with neurological and social-psychological studies to explain pain as a spiritual, biological, and intellectual event. This contextualizes their thorough survey of recent research on pain as part of the composing process experienced by writers of academic prose, poetry, and fiction. Their description of how pain functions as part of intense intellectual effort suggests ways writers can endure pain or even use it creatively.One of the things that scared me the most about finishing college were the memories I had from the two-month period I had dedicated to researching and writing this paper. It was one of the most excruciating experiences of my life. The actual writing did not commence until after a full month of research had been accumulated, summarized, and organized by my dear friend and co-author, Kate Murray. Kate and I then met nearly every night for another few weeks for free-form stream-of-consciousness discussions about everything the sources brought to mind, and just how on earth we were going to synthesize all of them into a cohesive piece. My best guess as to why we chose the topic of “Pain in the Writing Process” is that we were both hurting from the mere anticipation of writing this particular essay, and we needed help figuring out why. In order to make the narrative voice more consistent, as well as to accommodate personal needs of my co-author, I agreed to the majority of the writing and began a ceaseless two-week battle between the Paper and I. In a way it became my trip to the literary sweat-lodge. I hoped to bring back something that could transcend mere facts and theory. The end result was something I could barely recognize. I had a very hard time revisiting this paper for final edits. Whenever I would try, I was strangely overcome with an eerie feeling that I was traveling back to an old home filled with memories that may best be forgotten. It was a challenging experience, through and through, that reminded me of how brave anyone is who willingly experiences trauma to produce a piece of honest writing. To anyone who struggles with their craft: I know there is someone, somewhere who will find themselves in your words. Jack Kerouac describes the challenge of an artist quite beautifully: “[they] fill empty space with substance of our lives.” Culling that which is everything out of the ether is no small feat. Be proud.Arnold Sandershttp://blogs.goucher.edu/verge/verge3
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
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