271 resultados para Agrobacterium-mediated transformation


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1974 was the year when the Swedish pop group ABBA won the Eurovision Song Contest in Brighton and when Blue Swede reached number one on the Billboard Hot 100 in the US. Although Swedish pop music gained some international success even prior to 1974, this year is often considered as the beginning of an era in which Swedish pop music had great success around the world. With brands such as ABBA, Europe, Roxette, The Cardigans, Ace of Base, In Flames, Robyn, Avicii, Swedish House Mafia and music producers Stig Andersson, Ola Håkansson, Dag Volle, Max Martin, Andreas Carlsson, Jorgen Elofsson and several others have the myth of the Swedish music miracle kept alive for nearly more than four decades. Swedish music looks to continue reap success around the world, but since the millennium, Sweden's relationship with music has been more focused on relatively controversial Internet-based services for music distribution developed by Swedish entrepreneurs and engineers rather than on successful musicians and composers. This chapter focusses on the music industry in Sweden. The chapter will discuss the development of the Internet services mentioned above and their impact on the production, distribution and consumption of recorded music. Ample space will be given in particular to Spotify, the music service that quickly has fundamentally changed the music industry in Sweden. The chapter will also present how the music industry's three sectors - recorded music, music licensing and live music - interact and evolve in Sweden.

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Flows of cultural heritage in textual practices are vital to sustaining Indigenous communities. Indigenous heritage, whether passed on by oral tradition or ubiquitous social media, can be seen as a “conversation between the past and the future” (Fairclough, 2012, xv). Indigenous heritage involves appropriating memories within a cultural flow to pass on a spiritual legacy. This presentation reports ethnographic research of social media practices in a small independent Aboriginal school in Southeast Queensland, Australia that is resided over by the Yugambeh elders and an Aboriginal principal. The purpose was to rupture existing notions of white literacies in schools, and to deterritorialize the uses of digital media by dominant cultures in the public sphere. Examples of learning experiences included the following: i. Integrating Indigenous language and knowledge into media text production; ii. Using conversations with Indigenous elders and material artifacts as an entry point for storytelling; iii. Dadirri – spiritual listening in the yarning circle to develop storytelling (Ungunmerr-Baumann, 2002); and iv. Writing and publicly sharing oral histories through digital scrapbooking shared via social media. The program aligned with the Australian National Curriculum English (ACARA, 2012), which mandates the teaching of multimodal text creation. Data sources included a class set of digital scrapbooks collaboratively created in a multi-age primary classroom. The digital scrapbooks combined digitally encoded words, images of material artifacts, and digital music files. A key feature of the writing and digital design task was to retell and digitally display and archive a cultural narrative of significance to the Indigenous Australian community and its memories and material traces of the past for the future. Data analysis of the students’ digital stories involved the application of key themes of negotiated, material, and digitally mediated forms of heritage practice. It drew on Australian Indigenous research by Keddie et al. (2013) to guard against the homogenizing of culture that can arise from a focus on a static view of culture. The interpretation of findings located Indigenous appropriation of social media within broader racialized politics that enables Indigenous literacy to be understood as a dynamic, negotiated, and transgenerational flows of practice. The findings demonstrate that Indigenous children’s use of media production reflects “shifting and negotiated identities” in response to changing media environments that can function to sustain Indigenous cultural heritages (Appadurai, 1696, xv). It demonstrated how the children’s experiences of culture are layered over time, as successive generations inherit, interweave, and hear others’ cultural stories or maps. It also demonstrated how the children’s production of narratives through multimedia can provide a platform for the flow and reconstruction of performative collective memories and “lived traces of a common past” (Giaccardi, 2012). It disrupts notions of cultural reductionism and racial incommensurability that fix and homogenize Indigenous practices within and against a dominant White norm. Recommendations are provided for an approach to appropriating social media in schools that explicitly attends to the dynamic nature of Indigenous practices, negotiated through intercultural constructions and flows, and opening space for a critical anti-racist approach to multimodal text production.

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Nedd4-2, a HECT (homologous with E6-associated protein C-terminus)-type ubiquitin protein ligase, has been implicated in regulating several ion channels, including Navs (voltage-gated sodium channels). In Xenopus oocytes Nedd4-2 strongly inhibits the activity of multiple Navs. However, the conditions under which Nedd4-2 mediates native Nav regulation remain uncharacterized. Using Nedd4-2-deficient mice, we demonstrate in the present study that in foetal cortical neurons Nedd4-2 regulates Navs specifically in response to elevated intracellular Na(+), but does not affect steady-state Nav activity. In dorsal root ganglia neurons from the same mice, however, Nedd4-2 does not control Nav activities. The results of the present study provide the first physiological evidence for an essential function of Nedd4-2 in regulating Navs in the central nervous system.

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Background and purpose Phosphodiesterases PDE3 and/or PDE4 control ventricular effects of catecholamines in several species but their relative effects in failing human ventricle are unknown. We investigated whether the PDE3-selective inhibitor cilostamide (0.3-1μM) or PDE4 inhibitor rolipram (1-10μM) modified the positive inotropic and lusitropic effects of catecholamines in human failing myocardium. Experimental approach Right and left ventricular trabeculae from freshly explanted hearts of 5 non-β-blocker-treated and 15 metoprolol-treated patients with terminal heart failure were paced to contract at 1Hz. The effects of (-)-noradrenaline, mediated through β1-adrenoceptors (β2-adrenoceptors blocked with ICI118551), and (-)-adrenaline, mediated through β2-adrenoceptors (β1-adrenoceptors blocked with CGP20712A), were assessed in the absence and presence of PDE inhibitors. Catecholamine potencies were estimated from –logEC50s. Key results Cilostamide did not significantly potentiate the inotropic effects of the catecholamines in non-β-blocker-treated patients. Cilostamide caused greater potentiation (P=0.037) of the positive inotropic effects of (-)-adrenaline (0.78±0.12 log units) than (-)-noradrenaline (0.47±0.12 log units) in metoprolol-treated patients. Lusitropic effects of the catecholamines were also potentiated by cilostamide. Rolipram did not affect the inotropic and lusitropic potencies of (-)-noradrenaline or (-)-adrenaline on right and left ventricular trabeculae from metoprolol-treated patients. Conclusions and implications Metoprolol induces a control by PDE3 of ventricular effects mediated through both β1- and β2-adrenoceptors, thereby further reducing sympathetic cardiostimulation in patients with terminal heart failure. Concurrent therapy with a PDE3 blocker and metoprolol could conceivably facilitate cardiostimulation evoked by adrenaline through β2-adrenoceptors. PDE4 does not appear to reduce inotropic and lusitropic effects of catecholamines in failing human ventricle.

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Gas-phase transformation of synthetic phosphatidylcholine (PC) monocations to structurally informative anions is demonstrated via ion/ion reactions with doubly deprotonated 1,4-phenylenedipropionic acid (PDPA). Two synthetic PC isomers, 1-palmitoyl-2-oleoyl-sn-glycero-3-phosphocholine (PC16:0/18:1) and 1-oleoyl-2-palmitoyl-sn-glycero-3-phosphocholine (PC18:1/16:0), were subjected to this ion/ion chemistry. The product of the ion/ion reaction is a negatively charged complex, \[PC + PDPA - H](-). Collisional activation of the long-lived complex causes transfer of a proton and methyl cation to PDPA, generating \[PC - CH3](-). Subsequent collisional activation of the demethylated PC anions produces abundant fatty acid carboxylate anions and low-abundance acyl neutral losses as free acids and ketenes. Product ion spectra of \[PC - CH3](-) suggest favorable cleavage at the sn-2 position over the sn-1 due to distinct differences in the relative abundances. In contrast, collisional activation of PC cations is absent of abundant fatty acid chain-related product ions and typically indicates only the lipid class via formation of the phosphocholine cation. A solution phase method to produce the gas-phase adducted PC anion is also demonstrated. Product ion spectra derived from the solution phase method are similar to the results generated via ion/ion chemistry. This work demonstrates a gas-phase means to increase structural characterization of phosphatidylcholines via ion/ion chemistry. Grant Number ARC/CE0561607, ARC/DP120102922

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In many ways, China’s education system is quite different from systems of education in the West. Rich descriptions of school transformation, however, have revealed that the factors that fuelled transformation in schools in China are also evident in schools in Australia, England, Finland, Wales and the United States. This paper draws on an international project that examined how secondary schools from six countries achieved success by developing and drawing on their resources, referred to as four forms of capital: financial, intellectual, social and spiritual. It describes how five secondary schools in Chongqing, Western China, viewed each form of capital and how the four forms of capital were strengthened and aligned through outstanding governance to support the success of all students. The case is made that, although some aspects of the forms of capital found in schools in China may be viewed differently, the approaches adopted by these schools share a number of common elements with approaches to school transformation identified in Western schools. It is argued that these common elements from a range of international settings constitute a rich evidence base for understanding school transformation and for new insights in governance and leadership.

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The mechanisms involved in the control of embryonic stem (ES) cell differentiation are yet to be fully elucidated. However, it has become clear that the family of fibroblast growth factors (FGFs) are centrally involved. In this study we examined the role of the FGF receptors (FGFRs 1-4) during osteogenesis in murine ES cells. Single cells were obtained after the formation of embryoid bodies, cultured on gelatin-coated plates, and coaxed to differentiate along the osteogenic lineage. Upregulation of genes was analyzed at both the transcript and protein levels using gene array, relative-quantitative PCR (RQ-PCR), and Western blotting. Deposition of a mineralized matrix was evaluated with Alizarin Red staining. An FGFR1-specific antibody was generated and used to block FGFR1 activity in mES cells during osteogenic differentiation. Upon induction of osteogenic differentiation in mES cells, all four FGFRs were clearly upregulated at both the transcript and protein levels with a number of genes known to be involved in osteogenic differentiation including bone morphogenetic proteins (BMPs), collagen I, and Runx2. Cells were also capable of depositing a mineralized matrix, confirming the commitment of these cells to the osteogenic lineage. When FGFR1 activity was blocked, a reduction in cell proliferation and a coincident upregulation of Runx2 with enhanced mineralization of cultures was observed. These results indicate that FGFRs play critical roles in cell recruitment and differentiation during the process of osteogenesis in mES cells. In particular, the data indicate that FGFR1 plays a pivotal role in osteoblast lineage determination.

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With the emergence of personal and ubiquitous computing systems in the last decade, interaction designers have started designing products by employing quality oriented aspects such as user experience, playfulness, enchantment and others. In order to explore novel forms of mediated interactions, designers need to focus beyond the basic user requirements and usability issues. We present a procedure and results of a design exercise that we carried out with students of a master's course on Visual Design. Our intention was to explore new forms of mediated interaction by using a specific design exercise. We provide the details of the resulted design concepts and discuss the usefulness of our design exercise.

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High-energy synchrotron in situ X-ray powder diffraction has been used to elucidate the mechanism of the hydriding phase transformation in a LaNi5 model hydrogen storage intermetallic in real time. The transformation proceeds at 10 °C via the transient growth of an interfacial phase, the γ phase, with lattice parameters intermediate between those of the α (dilute solid solution) and β (concentrated hydride) phases. The γ phase forms to partially accommodate the 24% change in unit cell volume between the α and β phases during hydriding and dehydriding. The α, γ and β phases coexist at the nanoscopic level.

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The shared nature of genetic information presents new challenges for legal understandings of the self. Within traditional legal discourses the individual is conceptualised as separate and autonomous. In contrast, the genetic individual is understood as inherently relational. This paper analyses the transformation of our understandings of the personal. The transformative processes are assessed through discussion of the changing meanings of privacy in the context of genetic information within families; changing views over access to information about biological parentage by children conceived through assisted reproductive technology; preimplantation genetic diagnosis and the changing context of reproductive decisionmaking.

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This project explored the potential for halogen bonds to predictably organise metal-containing molecular building blocks in crystalline materials. A novel method for the halogen bond mediated crystal engineering of metal complexes was discovered, which led to the preparation of new materials with potential applications in molecular switching devices and advanced memory storage systems.

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Background Red colour in kiwifruit results from the presence of anthocyanin pigments. Their expression, however, is complex, and varies among genotypes, species, tissues and environments. An understanding of the biosynthesis, physiology and genetics of the anthocyanins involved, and the control of their expression in different tissues, is required. A complex, the MBW complex, consisting of R2R3-MYB and bHLH transcription factors together with a WD-repeat protein, activates anthocyanin 3-O-galactosyltransferase (F3GT1) to produce anthocyanins. We examined the expression and genetic control of anthocyanins in flowers of Actinidia hybrid families segregating for red and white petal colour. Results Four inter-related backcross families between Actinidia chinensis Planch. var. chinensis and Actinidia eriantha Benth. were identified that segregated 1:1 for red or white petal colour. Flower pigments consisted of five known anthocyanins (two delphinidin-based and three cyanidin-based) and three unknowns. Intensity and hue differed in red petals from pale pink to deep magenta, and while intensity of colour increased with total concentration of anthocyanin, no association was found between any particular anthocyanin data and hue. Real time qPCR demonstrated that an R2R3 MYB, MYB110a, was expressed at significant levels in red-petalled progeny, but not in individuals with white petals. A microsatellite marker was developed that identified alleles that segregated with red petal colour, but not with ovary, stamen filament, or fruit flesh colour in these families. The marker mapped to chromosome 10 in Actinidia. The white petal phenotype was complemented by syringing Agrobacterium tumefaciens carrying Actinidia 35S::MYB110a into the petal tissue. Red pigments developed in white petals both with, and without, co-transformation with Actinidia bHLH partners. MYB110a was shown to directly activate Actinidia F3GT1 in transient assays. Conclusions The transcription factor, MYB110a, regulates anthocyanin production in petals in this hybrid population, but not in other flower tissues or mature fruit. The identification of delphinidin-based anthocyanins in these flowers provides candidates for colour enhancement in novel fruits.

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Awareness within work environments should not be seen limited to important work-related information, activities and relationships. Mediating somewhat casual and engaging encounters related to non-work issues could also lead to meaningful and pleasurable experiences. This paper explores a design approach to support playfully mediated social awareness within an academic environment. Using ethnographic exploration and understanding the current and aspired practices, we provide details of two broad (and some times overlapping) categories of interaction for supporting and enhancing playfully mediated social awareness amongst staff members: 1) Self-Reflections and 2) Casual Encounters. We implement these two categories of interaction in an intelligent, asynchronous, large screen display called Panorama, for the staff room of our computer science department. Panorama attempts to mediate non-critical, non-work related information about the staff-members in an engaging manner to enhance social awareness within the department. We particularly emphasize on the soft design issues like reflections, belonging, care, pleasure and playfulness utilized in our design approach. The result of a two-phase assessment study suggests that our conceptualization of social awareness and the Panorama application has the potential to be easily incorporated into our academic environment.

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Background We describe novel plasmid vectors for transient gene expression using Agrobacterium, infiltrated into Nicotiana benthamiana leaves. We have generated a series of pGreenII cloning vectors that are ideally suited to transient gene expression, by removing elements of conventional binary vectors necessary for stable transformation such as transformation selection genes. Results We give an example of expression of heme-thiolate P450 to demonstrate effectiveness of this system. We have also designed vectors that take advantage of a dual luciferase assay system to analyse promoter sequences or post-transcriptional regulation of gene expression. We have demonstrated their utility by co-expression of putative transcription factors and the promoter sequence of potential target genes and show how orthologous promoter sequences respond to these genes. Finally, we have constructed a vector that has allowed us to investigate design features of hairpin constructs related to their ability to initiate RNA silencing, and have used these tools to study cis-regulatory effect of intron-containing gene constructs. Conclusion In developing a series of vectors ideally suited to transient expression analysis we have provided a resource that further advances the application of this technology. These minimal vectors are ideally suited to conventional cloning methods and we have used them to demonstrate their flexibility to investigate enzyme activity, transcription regulation and post-transcriptional regulatory processes in transient assays.