1,720,968 research outputs found
Molecular mechanisms controlling bacilysin biosynthesis in plant growth promoting rhizobacterium - Bacillus amyloliquefaciens FZB42
Full text linkBacillus amyloliquefaciens FZB42 ist ein grampositives Bakterium, das in der Rhizosphäre das Pflanzenwachstum fördert (PGPR - Plant Growth Promotion) und pathogene Organismen hemmt. Abgesehen von dieser Fähigkeit produziert es eine Vielzahl von sekundären Metaboliten, die sowohl ribosomale als auch nicht-ribosomale Peptide umfassen. In dieser Arbeit erfolgte die Untersuchung der transkriptionellen Aktivierung und Regulation der Bacilysin- Biosynthese an den Promotoren der bac- und ywfH- Gene. Durch 5´-Deletionsanalysen wurde der Promotor von Bacilysin identifiziert. Die A (Sigmafaktor A) - abhängige Transkription startet über die konservierten Promotorelemente (-10 und -35) von den bac- und ywfH Genen. Die Untersuchungen der Promotoraktivitäten vom Wildtyp und den erzeugten Regulationsmutanten erfolgten über in vivo ß-Galaktosidase-(Reporter)-Assays. Die Ergebnisse der Reporter-Aktivitäten zeigten, dass Transkriptionsregulatoren die Expression der Bacilysin- Gene aktivieren. Mehrere globale Regulatoren wie DegU, ComA, Hpr und AbrB beeinflussen die Genexpression. In dieser Arbeit wurde mithilfe von DNaseI Footprinting-Analysen die DegU-Bindung an die bac- und ywfH- Promotoren bestätigt.Die negative Regulation der Bacilysin-Biosynthese wird durch den Regulator der transienten Phase Hpr bewerkstelligt. Eine direkte Hpr-Bindung an bac Promotor wurde mit DNaseI Footprint-Analysen gezeigt. Der Promotor des monocistronischen Gens ywfH wurde aber durch Hpr nicht beeinflusst. Die anderen Transkriptionsregulatoren, wie ComA und AbrB, regulieren die Genexpression von Bacilysin indirekt über DegQ und Hpr. In dieser Arbeit konnte demonstriert werden, dass der globale Regulator AbrB den Promotor vom hpr-Gen direkt kontrolliert. Zusammenfassend liefert diese Studie neue Informationen über die genetische Regulation der Bacilysin- Biosynthese in B. amyloliquefaciens FZB42.Bacillus amyloliquefaciens FZB42 is a Gram-positive, pathogen-suppressing and plant-growth promoting rhizobacterium. Apart from this ability, it produces a vast array of secondary metabolites, which includes both ribosomal and non-ribosomal peptides. In this work, the transcriptional activation and regulation of bacilysin biosynthesis were studied at the promoters of bac and ywfH genes. The promoter of bacilysin was identified using 5''-deletion analysis. Sigma factor A (σA) was found to start transcription via conserved promoter elements (-10 and -35) of bac and ywfH genes. lacZ reporter fusion studies were performed in wild type and regulatory mutants. The results show the involvement of transcriptional regulators to activate the expression of bacilysin genes. Several global regulators such as DegU, ComA, Hpr and AbrB were identified and found to influence gene expression. In particular, I confirmed DegU binding in bac and ywfH promoters using radioactive DNase I footprinting. Furthermore, Hpr, a transition state regulator was found negatively to control bacilysin biosynthesis. Hpr binding to bac promoter was demonstrated using radioactive DNase I footprinting. Remarkably, Hpr does not influence the promoter of the monocistronic gene, ywfH. The other transcriptional regulators, such as ComA and AbrB, were correlated indirectly to affect the gene expression of bacilysin via DegQ and Hpr, respectively. The gene regulation of hpr was studied in this work. It was demonstrated that AbrB, a global regulator, directly controls the promoter of the hpr gene. However, the consensus sequence for AbrB binding was not identified, since it covers the entire promoter region in the DNA-protein interaction study. To conclude, this study provides new information regarding the genetic regulation of bacilysin biosynthesis in B. amyloliquefaciens FZB42
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
Generation of iPSC-derived human forebrain organoids assembling bilateral eye primordia
No full textInduced pluripotent stem cell-derived brain organoids enable the developmental complexities of the human brain to be deconstructed. During embryogenesis, optic vesicles (OVs), the eye primordium attached to the forebrain, develop from diencephalon. However, most 3D culturing methods generate either brain or retinal organoids individually. Here we describe a protocol to generate organoids with both forebrain entities, which we call OV-containing brain organoids (OVB organoids). In this protocol, we first induce neural differentiation (days 0–5) and collect neurospheres, which we culture in a neurosphere medium to initiate their patterning and further self-assembly (days 5–10). Then, upon transfer to spinner flasks containing OVB medium (days 10–30), neurospheres develop into forebrain organoids with one or two pigmented dots restricted to one pole, displaying forebrain entities of ventral and dorsal cortical progenitors and preoptic areas. Further long-term culture results in photosensitive OVB organoids constituting complementary cell types of OVs, including primitive corneal epithelial and lens-like cells, retinal pigment epithelia, retinal progenitor cells, axon-like projections and electrically active neuronal networks. OVB organoids provide a system to help dissect interorgan interactions between the OVs as sensory organs and the brain as a processing unit, and can help model early eye patterning defects, including congenital retinal dystrophy. To conduct the protocol, experience in sterile cell culture and maintenance of human induced pluripotent stem cells is essential; theoretical knowledge of brain development is advantageous. Furthermore, specialized expertise in 3D organoid culture and imaging for the analysis is needed
Cilium induction triggers differentiation of glioma stem cells
Full text linkGlioblastoma multiforme (GBM) possesses glioma stem cells (GSCs) that promote self-renewal, tumor propagation, and relapse. Understanding the mechanisms of GSCs self-renewal can offer targeted therapeutic interventions. However, insufficient knowledge of GSCs’ fundamental biology is a significant bottleneck hindering these efforts. Here, we show that patient-derived GSCs recruit elevated levels of proteins that ensure the temporal cilium disassembly, leading to suppressed ciliogenesis. Depleting the cilia disassembly complex components is sufficient to induce ciliogenesis in a subset of GSCs via relocating platelet-derived growth factor receptor-alpha (PDGFR-α) to a newly induced cilium. Importantly, restoring ciliogenesis enabled GSCs to switch from self-renewal to differentiation. Finally, using an organoid-based glioma invasion assay and brain xenografts in mice, we establish that ciliogenesis-induced differentiation can prevent the infiltration of GSCs into the brain. Our findings illustrate a role for cilium as a molecular switch in determining GSCs’ fate and suggest cilium induction as an attractive strategy to intervene in GSCs proliferation
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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