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1
Faini, Marie Annette. "Transcriptional Control during Quorum Sensing by LuxR and LuxR Homologues." Thesis, Virginia Tech, 2003. http://hdl.handle.net/10919/31994.
Full textAbstract:
Quorum sensing is a mechanism used by many proteobacteria to regulate expression of target genes in a population-dependent manner. The quorum sensing system of Vibrio fischeri activates the luminescence (lux) operon when the autoinducer signaling molecule (N-3-oxohexanoyl homoserine lactone) is recognized and bound by the activator protein LuxR. LuxR subsequently binds to the lux box centered at à 42.5 bp upstream of the transcription initiation site and activates transcription from the lux operon promoter, resulting in the emission of light at high cell densities. LuxR consists of 250 amino acids arranged into an N-terminal (regulatory) domain and a C-terminal (activation) domain, and is thought to function as an ambidextrous activator capable of making multiple contacts with the alpha and sigma subunits of RNA polymerase (RNAP). Published work describing the results of alanine scanning mutagenesis performed on the C-terminal domain of LuxR (residues 190-250) has identified residues (K198, W201 and I206) that appear to play a role in positive control of transcription initiation. Additional mutagenesis of residues 180-189 has been undertaken via a three-primer or four-primer PCR-based method in this study. Variants of LuxR were screened for their ability to activate luciferase production and to repress transcription from an artificial promoter, and production of full-length LuxR was measured, in an attempt to identify additional positive control variants. No additional positive control variants were found in this study. Work has also been undertaken to identify intergenic suppressors between positive control variants of LuxR and the RNAP alpha subunit, RpoA. Starting with a recombinant Escherichia coli strain encoding the lux operon and LuxR variant I206E, a random chemical mutagenesis was performed on a vector encoding RpoA. Following transformation of the mutated plasmids encoding RpoA, high throughput luminescence assays were used to identify isolates with phenotypes brighter than the control. Isolation of an intergenic suppressor will confirm the existence of protein-protein interactions between LuxR and RpoA within the transcription initiation complex. The ability of other LuxR family members to establish productive protein-protein interactions with RNAP necessary for transcription initiation was also examined. LuxR homologues EsaR of Pantoea stewarti ssp. stewartii, a repressor of known function, and ExpR of Erwinia carotovora subsp. carotovora were also analyzed for their ability to activate the lux operon, as well as to repress transcription from an artificial promoter containing the lux box.Master of Science
2
Collins, Cynthia Hollie Parker Carl Stevens. "Directed evolution of the transcriptional activator LuxR /." Diss., Pasadena, Calif. : Caltech, 2006. http://resolver.caltech.edu/CaltechETD:etd-12272005-160649.
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Brameyer, Sophie. "Cell-cell communication via LuxR solos in Photorhabdus species." Diss., Ludwig-Maximilians-Universität München, 2015. http://nbn-resolving.de/urn:nbn:de:bvb:19-181139.
Full textAbstract:
Bacteria constantly need to monitor their environment and adapt the bacterial group-coordinated behaviour to changing habitats like nutrition alterations or host variations. Commonly cell-cell communication via acyl homoserine lactones (AHLs)is used to synchronise the behaviour of a bacterial population dependent on cell size. This process is referred to as quorum sensing (QS) and predominantly occurs in
Gram-negative bacteria. The typical QS system consists of a LuxI-synthase that synthesises AHLs, and a LuxR-type receptor, which then responds to these AHLs. Upon AHL-binding, the LuxR-type receptor regulates the expression of different target genes and thus influences several processes, like biofilm formation, virulence, antibiotic production or cell-cell interaction. Interestingly, many proteobacteria
possess additional LuxR homologs, but lack a cognate LuxI-type synthase. Those LuxR-type receptors are referred to as LuxR orphans or LuxR solos and can expand the regulatory QS network. Photorhabdus species are insect pathogenic bacteria,
living in symbiosis with entomopathogenic nematodes. They all possess an
exceptionally high number of LuxR solos, but lack LuxI homologs and therefore do
not produce AHLs. The function of these LuxR solos, their role in cell-cell
communication and the identification of their cognate signalling molecules in
Photorhabdus species is the main focus of this work.
In this thesis a novel signalling molecule used for QS could be identified for the
first time in P. luminescens. This novel QS molecule is an α-pyrone named
photopyrone (PPY) and produced endogenously by the photopyrone synthase (PpyS).
The PPYs are specifically recognized by the LuxR solo regulator PluR, which then
activates expression of the pcf (Photorhabdus clumping factor) operon leading to cell
clumping of P. luminescens cells. Moreover, the PpyS/PluR quorum sensing system
and its induced cell clumping contribute to the overall toxicity of P. luminescens.
Furthermore, a second novel signalling molecule sensed by a LuxR solo of
Photorhabdus species could be identified besides PPYs. The insect and human
pathogenic bacteria P. asymbiotica lacks a PpyS homolog as well as a LuxI homolog,
but harbours a pcf operon and a homologue to PluR, which is named PauR. The
signalling molecule sensed by the LuxR-type receptor PauR could be identified,
which is neither an AHL nor a PPY. PauR recognises a 2,5-dialkylresorcinol (DAR)
produced by the DarABC pathway. Upon binding of the cognate signalling molecule,
Summary
XII
PauR activates expression of the pcf operon. This also leads to cell clumping in
P. asymbiotica. Furthermore, the DarABC/PauR QS system also contributes to the
overall pathogenicity of P. asymbiotica against Galleria mellonella insect larvae.
A bioinformatics approach revealed a high number of LuxR solos present in
P. temperata and P. asymbiotica like in P. luminescens. Thereby, several conserved
motives of amino acids could be identified, which are potentially important for signalbinding
and -specificity. Variations in these amino acid motifs are assumed to reflect
the overall variety of signals that can be sensed by LuxR solos.
Furthermore, the specificity of the two LuxR solos PluR and PauR towards their
cognate signalling molecules, PPYs and DARs, respectively, was analysed. Thereby,
it could be shown that the previously identified conserved amino acid motives in the
signal-binding domain (SBD), the TYDQCS-motif of PluR and the TYDQYI-motif of
PauR, are essential but not sufficient for ligand-binding.
Similar as for AHLs, it was unclear how the signalling molecules PPYs and DARs
can cross the bacterial cell membrane. In the last part of this thesis the import
mechanism for the Photorhabdus-specific signalling compounds PPYs and DARs
were identified. Initial evidence could be provided that the membrane-integrated
transporter FadL is mainly involved in the import of these hydrophobic compounds,
and that they are not transported via simple diffusion across the cell membrane, which
is assumed for AHLs.
In conclusion, the data that is compiled presents two LuxR solos of Photorhabdus
species adapted to sense and respond to novel non-AHL signalling molecules used for
QS. Therefore, this thesis reveals that cell-cell communication via LuxR-type
receptors goes far beyond AHL-signalling in nature.
4
Trott, Amy Elizabeth. "Amino Acid Residues in LuxR Critical for its Mechanism of Transcriptional Activation during Quorum Sensing." Thesis, Virginia Tech, 2000. http://hdl.handle.net/10919/34070.
Full textAbstract:
Vibrio fischeri, a symbiotic bioluminescent bacterium, serves as one of the best understood model systems for a mechanism of cell-density dependent bacterial gene regulation known as quorum sensing. During quorum sensing in V. fischeri, an acylated homoserine chemical signal (autoinducer) is synthesized by the bacteria and used to sense their own species in a given environment. As the autoinducer levels rise, complexes form between the autoinducer and the N-terminal domain of a regulatory protein, LuxR. In response to autoinducer binding, LuxR is believed to undergo a conformational change that allows the C-terminal domain to activate transcription of the luminescence or lux operon. To further understand the mechanism of LuxR-dependent transcriptional activation of the lux operon, PCR-based site-directed mutagenesis procedures have been used to generate alanine-substitution mutants in the C-terminal forty-one amino acid residues of LuxR, a region that has been hypothesized to play a critical role in the activation process. An in vivo luminescence assay was first used to test the effects of the mutations on LuxR-dependent activation of the lux operon in recombinant Escherichia coli. Luciferase levels present in cell extracts obtained from these strains were also quantified and found to correlate with the luminescence results. Eight strains encoding altered forms of LuxR exhibited a "dark" phenotype with luminescence output less than 50% and luciferase levels less than 50% of the wildtype control strain. Western immunoblotting analysis with cell extracts from the luminescence and luciferase assays verified that the altered forms of LuxR were expressed at levels approximately equal to wildtype. Therefor, Low luminescence and luciferase levels could be the result of a mutation that either affects the ability of LuxR to recognize and bind its DNA target (the lux box) or to establish associations with RNA polymerase (RNAP) at the lux operon promoter necessary for transcriptional initiation. A third in vivo assay was used to test the ability of the altered forms of LuxR to bind to the lux box (DNA binding assay/repression). All of the LuxR variants exhibiting the "dark" phenotype in the luminescence and luciferase assay were also found to be unable to bind to the lux box in the DNA binding assay. Therefore, it can be concluded that the alanine substitutions made at these positions affect the ability of LuxR to bind to the lux box in the presence and absence of RNA polymerase. Another class of mutants exhibited wildtype phenotypes in the luminescence and luciferase assays but were unable to bind to the lux box in the DNA binding assay. The alanine substitutions made at these amino acid residues may be making contacts with RNAP that are important for maintaining the stability of the DNA binding region of LuxR. Alanine substitutions made at these positions have a defect in DNA binding at the promoter of the lux operon only in the absence of RNAP. None of the alanine substitutions made in the C-terminal forty-one amino acids of LuxR were found to affect activation of transcription of the lux operon without also affecting DNA binding. Taken together, these results support the conclusion that the C-terminal forty-one amino acids of LuxR are important for DNA recognition and binding of the lux box rather than positive control of the process of transcription initiation.Master of Science
5
Poulter, Simon. "The LuxR-family quorum sensing transcriptional regulator CarR in Erwinia and Serratia." Thesis, University of Cambridge, 2011. http://ethos.bl.uk/OrderDetails.do?uin=uk.bl.ethos.609398.
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Brameyer, Sophie [Verfasser], and Ralf [Akademischer Betreuer] Heermann. "Cell-cell communication via LuxR solos in Photorhabdus species / Sophie Brameyer. Betreuer: Ralf Heermann." München : Universitätsbibliothek der Ludwig-Maximilians-Universität, 2015. http://d-nb.info/1069491438/34.
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Qin, Nan. "Analysis of the Regulons Controlled by Transcriptional Regulators LuxR and LitR in Vibrio fischeri." Diss., Virginia Tech, 2008. http://hdl.handle.net/10919/28433.
Full textAbstract:
Quorum sensing is a bacterial signaling system that controls gene expression in a population density-dependent manner. In Gram-negative proteobacteria, the cell density control of luminescence was first observed in the symbiotic marine bacterium Vibrio fischeri and this system is one of the best studied quorum sensing systems. Two-dimensional sodium dodecyl sulfate-polyacrylamide (2D-SDS) gel electrophoresis analysis previously identified several non-Lux proteins in V. fischeri MJ-100 whose expression was dependent on LuxR and 3-oxo-hexanoyl-L-homoserine lactone (3-oxo-C6-HSL). A lacZ reporter was used to show that the promoters for qsrP, acfA, and ribB were directly activated via LuxR-3-oxo-C6-HSL in recombinant Escherichia coli. The sites of transcription initiation were established via primer extension analysis. Based on the position of the lux box-binding site near position â 40, all three promoters appear to have a class II-type promoter structure. Real-time reverse transcription-PCR was used to study the temporal expression of qsrP, acfA, and ribB during the exponential and stationary phases of growth, and electrophoretic mobility shift assays were used to compare the binding affinities of LuxR to the promoters under investigation.
In order to fully characterize the LuxR regulon in V. fischeri ES114, microarray analysis was performed in the Greenberg lab (University of Washington) and 18 LuxR-3-oxo-C6-HSL regulated promoters were found including 2 genes (qsrP and acfA) identified previously in MJ-100 in addition to the well-studied lux operon. In collaboration with them, full-length purified LuxR protein was used to show direct interaction between the LuxR protein and 7 genes/operons newly identified out of 13 genes/operons examined. The binding affinity between LuxR proteins and those genes was also measured.
Based on the sequence of the lux boxes of the known genes regulated by LuxR and LitR, a position specific weight matrix (PSWM) was created and used to search through the intergenic regions of the V. fischeri ES114 genome. Some potential LuxR and LitR-regulated genes with high score were tested experimently to confirm direct activation. For the LuxR regulon, these possible LuxR-regulated promoters were cloned into a lacZ reporter and tested for their LuxR dependence. Beyond the genes found in microarray, the promoter of the intergenic region VFA0658-0659 was found to be activated by LuxR and 3-oxo-C6-HSL. For the LitR regulon, two LitR-regulated genes found in the microarray were also identified using PSWM and confirmed by real-time PCR to be dependent on LitR for expression. EMSA experiments showed that LitR can specifically bind to the litR boxes of LitR-regulated genes, litR and VF0170 which confirmed that the regulation is direct.Ph. D.
8
Williams, Joshua W. "Multi-tiered Regulation of luxR Provides Precise Timing and Maintenance of the Quorum Sensing Response of Vibrio fischeri." Diss., Virginia Tech, 2009. http://hdl.handle.net/10919/38580.
Full textAbstract:
The quorum-sensing response of Vibrio fischeri involves a complex network of genes (encoding regulatory proteins as well as sRNAs), that govern host-association and production of bioluminescence. A key regulator of this system is LuxR, which is the transcriptional activator of the lux operon as well as several other genes in. LuxR also autoregulates its own transcription, which we have shown causes bistability and hysteresis in the quorum-sensing response. This behavior allows the system to maintain a stable and robust response in the face of environmental fluctuation or decreases in external autoinducer concentration caused by other sources. There are many factors that are known to regulate luxR expression, including the ArcA redox-responsive regulator, the cAMP-CRP secondary metabolism regulator, and components of the quorum-sensing pathway like LitR. Because of this, LuxR levels are critical in both the timing of quorum-sensing induction, as well as the maintenance of the response over time. This makes it a potential target for multiple levels of regulation in response to factors such as environmental and metabolic conditions, as well as other components of the quorum-sensing network.
Another important global regulatory protein in V. fischeri (and most other species of Gram-negative proteobacteria) is the post-transcriptional regulator CsrA. CsrA controls processes involved in carbon storage and utilization, as well as the transition from exponential to stationary phase growth. We have demonstrated that CsrA is regulated by two sRNAs (CsrB1 and CsrB2) in V. fischeri. Because CsrA regulates changes in cell behavior and is an important metabolic regulator, there is a good possibility that it has some interactions with the quorum-sensing regulon, whose endproduct, bioluminescence, creates a large metabolic demand from the cell.
In an effort to determine at which point in the quorum-sensing regulatory network CsrA regulation is important, epistasis experiments were designed using factorial design, which is a subset of statistical analysis of variance (ANOVA). This method was used to generate a high degree of confidence in the data, so that even minor interactions in the regulatory networks could be established. By altering the levels of CsrA expression in various mutant strains of V. fischeri, we have demonstrated that CsrA acts by an unknown mechanism to increase the transcription of luxR when the quorum-sensing regulator LitR is absent. Our results also demonstrated that CsrA mediates this effect through repression of ArcA activity, which is known to act directly on the luxR and luxI intergenic region as a repressor. This indicates that CsrA may bypass the upstream parts of the quorum-sensing regulatory cascade that lead to litR activation, so that LitR and LuxR may be regulated differently in response to certain conditions.
This work has shown that the interactions between global regulons can coordinately control the amount of quorum-sensing induction by affecting the level of LuxR in the cell. The balance of these regulatory networks allows the cell to tightly regulate the quorum-sensing response. Thus, LuxR serves as a critical regulatory hub in the cell, at which multiple signals can be integrated in order to generate the appropriate cellular response.Ph. D.
9
Callahan, Sean M. (Sean Michael) 1966. "The quorum-sensing regulation of Vibrio fischeri : novel components of the autoinduce/LuxR regulatory circuit." Thesis, Massachusetts Institute of Technology, 1999. http://hdl.handle.net/1721.1/85290.
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Dyszel, Jessica L. "Phenotypes of the LuxR Homolog, SdiA, in Salmonella and E. coli." The Ohio State University, 2009. http://rave.ohiolink.edu/etdc/view?acc_num=osu1243943752.
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Michael, Bindhu. "Detection of Heterologous bacterial pheromones by SdiA, A LuxR Homolog in Salmonella enterica serovar Typhimurium." The Ohio State University, 2001. http://rave.ohiolink.edu/etdc/view?acc_num=osu1407404158.
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Sellick, Katelyn. "A Possible luxR Solo Type Regulator of an Antibiotic-Like Compound from the Soil Bacterium Rhodococcus." Digital Commons @ East Tennessee State University, 2019. https://dc.etsu.edu/etd/3649.
Full textAbstract:
Rhodococcus, a species of bacteria commonly found in the soil, is an under-explored producer of small bioactive compounds including siderophores, pigments and antibiotics. MTM3W5.2 is a strain of Rhodococcus that was previously discovered to produce an antibiotic-like compound that has inhibitory effects on other Rhodococcus strains, including the veterinary pathogen, R. equi. The biosynthetic gene cluster responsible for production of the antibiotic has been identified, and a small gene, BTZ20_3964 at the start of the operon is believed to be a luxR solo regulator of the gene cluster. The goal of this project was to determine this gene’s status as a regulator for the gene cluster. Merodiploids were constructed using the deletion construct, pEX18Km3964AD to obtain a double crossover recombination event to replace the functional gene with the deletion construct. However, evidence indicates that an illegitimate recombination event occurred to produce a merodiploid strain.
13
Swearingen, Matt Charles. "Phenotypes of Salmonella SdiA in Mice and Pigs." The Ohio State University, 2013. http://rave.ohiolink.edu/etdc/view?acc_num=osu1374076511.
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Pennerman, Kayla Kara. "Development of Methods for Structural Characterization of Pantoea stewartii Quorum-Sensing Regulator EsaR." Thesis, Virginia Tech, 2014. http://hdl.handle.net/10919/25297.
Full textAbstract:
The LuxR family of proteins serves as quorum-sensing transcriptional regulators in proteobacteria. At high population densities, a small acyl-homoserine lactone (AHL) molecule, produced by a LuxI homologue, accumulates in the environment. The LuxR proteins bind to their respective AHL when the ligand accumulates to sufficient levels. Once bound to AHL, the holoproteins usually become functional as transcriptional activators. However, there is a subset of LuxR homologues, the EsaR subfamily, which is active without the AHL ligand and becomes inactivated once bound to it. EsaR is the best understood member of this subfamily. It controls virulence in the corn pathogen Pantoea stewartii ssp. stewartii.
Solubility issues have previously limited structural studies of LuxR homologues as the proteins could not be purified without the AHL ligand. A soluble recombinant EsaR protein, HMGE, is biologically active and can be purified in the absence and presence of AHL, unlike most other LuxR homologues. Using HMGE, amino acid substitutions and Förster resonance energy transfer (FRET), experimental methods were designed for determining the dimerization interface of EsaR and for testing the hypothesis that EsaR undergoes a conformational shift when presented with the AHL ligand.
To identify residues of the dimerization interface, heterodimerization assays were designed, involving either coexpression or coincubation of wild-type EsaR and variant HMGE proteins. In this assay, the inability of the proteins to copurify by nickel affinity chromatography would indicate that the modified residue(s) are important for dimerization of EsaR. To determine the conformational change that EsaR undergoes when bound to the AHL ligand, a FRET assay was developed to estimate the distances between amino acid residues in the absence and presence of AHL. Future work will have to include a few modifications to the methods and/or control experiments. This study provides the basis upon which the present methods can be further developed and later used for structural studies of EsaR.Master of Science
15
Chatterjee, Jaidip. "The luminescence induction point of Vibrio harveyi is an integration of multiple regulatory controls : LuxR, MetR, and CRP." Thesis, McGill University, 2000. http://digitool.Library.McGill.CA:80/R/?func=dbin-jump-full&object_id=36889.
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Luminescence is a secondary metabolic process that can be detected and assayed in a non-disruptive, in vivo, and real-time manner. As such the Vibrio harveyi luminescence (lux) system constitutes a model system that can be used to delineate the relatively complex transcriptional control mechanisms that are inherent to secondary metabolic processes. Molecular insights into the control of luminescence induction in V. harveyi is limited to the two-component quorum-sensing system and LuxR. While the quorum-sensing system is thought to facilitate luminescence induction through a mechanism of derepression, LuxR has been shown to be an essential activator of the process by virtue of the absence of luminescence in a luxR null mutant. Additionally, it has been suggested that LuxR activates its own expression. Data presented here demonstrates that LuxR is involved in autorepression. Moreover, it is argued that LuxR may not be a typical activator in that it may mediate activation of lux gene expression through a mechanism of repression. An implication of this proposal is that a second unidentified activator is required for luminescence induction to occur. The search of candidates to fill this proposed activation role focused on two highly conserved and extensively studied activators, MetR and CRP. Both are involved in nutrient dependent pathways as MetR is an activator of methionine biosynthesis genes and CRP is implicated in the regulation of metabolic pathways in response to the presence of preferred carbohydrates. In the specific case of the V. harveyi lux system, MetR is shown to act as a repressor, while CRP is shown to be a critical activator of luminescence induction. The evolutionary and ecological implications of these findings are discussed.
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Geissinger, Jared Scott. "Structure-Function Analysis of the EsaR N-terminal Domain." Thesis, Virginia Tech, 2011. http://hdl.handle.net/10919/46190.
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The LuxR protein family is a class of quorum-sensing regulated bacterial transcription factors that alter gene expression as a function of ligand detection. This coincides with a high population density and/or a low rate of signal ligand diffusion. The majority of LuxR proteins are activated only in the presence of the signal ligand, an acyl-homoserine lactone (AHL). EsaR, from the corn pathogen Pantoea stewartii, represents a subset of LuxR homologues that are active in the absence of AHL and deactivated by its presence. The mechanism by which EsaR responds to AHL in a manner opposite to that of the majority of LuxR homologues remains elusive. Unlike the majority of LuxR homologues, which require AHL for purification, EsaR can be purified and biochemically investigated in the absence and presence of AHL. This work sought to answer questions regarding the structure-function relationship of the LuxR homologue, EsaR.
Fluorescence anisotropy was used to determine the relative DNA-binding affinity of wild type EsaR and three AHL-independent EsaR variants in the presence and absence of AHL. This enabled for quantitative analysis of the relative binding affinities of these AHL-independent variants for the EsaR binding site, the esa box. The results demonstrate that one AHL-independent EsaR variant has a slightly higher affinity for the esa box in the presence, rather than the absence of AHL. The affinity of the other two for the DNA is not impacted by AHL, potentially due to an inability to transduce the signal of ligand detection to the DNA binding domain.
Constructs containing only the EsaR N-terminal domain (NTD) were also developed. These constructs circumvented solubility issues associated with the full-length protein, allowing for additional biochemical analysis. It was determined that the EsaR NTD alone is sufficient for multimerization and ligand binding. Additionally, preliminary X-ray crystallography efforts have established some of the early parameters required to solve the crystal structure of the EsaR ligand binding domain in both the presence and absence of AHL. If pursued, these structures would be the first solved of a LuxR homologue ligand binding domain in both the presence and absence of the native AHL, potentially demonstrating the conformational change that occurs as a result of ligand binding. Collectively, these findings have established some of the groundwork required to resolve the question of what sort of conformational changes occur in EsaR as a result of ligand binding.Master of Science
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Smith, Jenee N. "Characterization of the LuxR Homolog, SdiA, a transcriptional regulator activated by N-acylhomoserine lactone produced by other bacterial species." The Ohio State University, 2007. http://rave.ohiolink.edu/etdc/view?acc_num=osu1180456493.
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Johnson, Deborah Cumaraswamy. "Role of region 4 of the sigma 70 subunit of RNA polymerase in transcriptional activation of the lux operon during quorum sensing." Thesis, Virginia Tech, 2002. http://hdl.handle.net/10919/31680.
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The mechanism of gene regulation used by Gram-negative bacteria during quorum sensing is well understood in the bioluminescent marine bacterium Vibrio fischeri. The cell-density dependent activation of the luminescence (lux) genes of V. fischeri relies on the formation of a complex between the autoinducer molecule, N-(3-oxohexanoyl) homoserine lactone, and the autoinducer-dependent transcriptional activator LuxR. LuxR, a 250 amino acid polypeptide, binds to a site known as the lux box centered at position -42.5 relative to the luxI transcriptional start site. During transcriptional activation of the lux operon, LuxR is thought to function as an ambidextrous activator capable of making multiple contacts with RNA polymerase (RNAP). The specific role of region 4 of the Escherichia coli sigma 70 subunit of RNAP in LuxR-dependent transcriptional activation of the luxI promoter has been investigated. Rich in basic amino acids, this conserved portion of sigma 70 is likely to be surface-exposed and available to interact with transcription factors bound near the -35 element. The effect of 16 single and 2 triple alanine substitution variants of sigma 70 between amino acid residues 590 and 613, was determined in vivo by measuring the rate of transcription from a luxI-lacZ translational fusion via b-galactosidase assays in recombinant E. coli. In vitro work was performed with LuxRDN, the autoinducer-independent C-terminal domain (amino acids 157 to 250) of LuxR because purified, full length LuxR is unavailable. Single-round transcription assays were performed in the presence of LuxRDN and 19 variant RNAPs, one of which contained a C-terminally truncated sigma 70 subunit devoid of region 4. Results indicate that region 4 is essential for LuxRDN-dependent luxI transcription with two specific amino acid residues, E591 and K597, having negative effects on the rate of LuxRDN-dependent luxI transcription in vivo and in vitro. None of the residues tested were identified as having any effect on LuxR-dependent luxI transcription in vivo. These findings suggest that region 4.2 is most likely to be in close proximity to LuxR when bound to the luxI promoter. However, unlike the situation found for other ambidextrous activators, no single residue within region 4.2 of sigma 70 may be critical by itself for LuxR-dependent during transcriptional activation.Master of Science
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McDougald, S. Diane School of Microbiology & Immunology UNSW. "Regulation of starvation and nonculturability in the marine pathogen, Vibrio vulnificus." Awarded by:University of New South Wales. School of Microbiology and Immunology, 2000. http://handle.unsw.edu.au/1959.4/19118.
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Vibrio vulnificus is a model environmental organism exhibiting a classical starvation response during nutrient limitation as well as a non-culturable state when exposed to low temperatures. In addition to these classic global responses, this organism is an opportunistic pathogen that exhibits numerous virulence factors. This organism was chosen as the model organism for the identification of regulators of the viable but nonculturable response (VBNC) and the starvation-induced maintenance of culturability (SIMC) that occurs when cells are starved prior to low temperature incubation. In order to accomplish this, three indirect approaches were used; proteomics, investigation of intercellular signalling pathways and genetic analysis of regulators involved in these responses. Two-dimensional gel electrophoresis was used to identify proteins expressed under conditions that induced SIMC. It was determined that carbon and long-term phosphorus starvation were important in the SIMC response. V. vulnificus was shown to possess genes, luxS and smcR, that are homologues of genes involved in signalling system system 2 in Vibrio harveyi. Signal molecules were produced upon starvation and the entry to stationary phase in V. vulnificus. Furthermore, a null mutation in smcR, a transcriptional regulator was shown to have pleiotropic effects in V. vulnificus, including up-regulation of numerous virulence factors and a defect in starvation survival and development of the SIMC response. We propose that V. vulnificus possesses a signalling system analogous to that of system 2 in V. harveyi, and that this system is involved in the regulation of stationary phase and starvation adaptation in this organism.
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Finney, Angela H. "Role of the C-terminal domain of the a subunit of RNA polymerase in transcriptional activation of the lux operon during quorum sensing." Thesis, Virginia Tech, 2000. http://hdl.handle.net/10919/36285.
Full textAbstract:
Quorum sensing in Gram-negative bacteria is best understood in the bioluminescent marine microorganism, Vibrio fischeri. In V. fischeri, the luminescence or lux genes are regulated in a cell density-dependent manner by the activator LuxR in the presence of an acylated homoserine lactone autoinducer molecule (3-oxo-hexanoyl homoserine lactone). LuxR, which binds to the lux operon promoter at position -42.5, is thought to function as an ambidextrous activator making multiple contacts with RNA polymerase (RNAP). The specific role of the aCTD of RNAP in LuxR-dependent transcriptional activation of the lux operon promoter has been investigated. The effect of seventy alanine substitution variants of the a subunit was determined in vivo by measuring the rate of transcription of the lux operon via luciferase assays in recombinant Escherichia coli. The mutant RNAPs from strains exhibiting at least two fold increased or decreased activity in comparison to the wild-type were further examined by in vitro assays. Since full-length LuxR has not been purified to date, an autoinducer-independent N-terminal truncated form of LuxR, LuxRDN, was used for in vitro studies. Single-round transcription assays were performed using reconstituted mutant RNAPs in the presence of LuxRDN, and fourteen residues in the aCTD were identified as having negative effects on the rate of transcription from the lux operon promoter. Five of these fourteen residues were also involved in the mechanism of both LuxR and LuxRDN-dependent activation in vivo and were chosen for further analysis by DNA mobility shift assays. Results from these assays indicate that while the wild-type aCTD is capable of interacting with the lux DNA fragment tested, all five of the variant forms of the aCTD tested appear to be deficient in their ability to recognize and bind the DNA. These findings suggest that aCTD-DNA interactions may play a role in LuxR-dependent transcriptional activation of the lux operon during quorum sensing.Master of Science
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Pan, Tianyuan [Verfasser], Edmund [Akademischer Betreuer] Maser, and Sabine [Gutachter] Fuchs. "Isolation and identification of two repressors, TetR AND LuxR, FOR 3,17β-hsd and 3α-hsd/cr gene regulation in Comamonas testosteroni / Tianyuan Pan ; Gutachter: Sabine Fuchs ; Betreuer: Edmund Maser." Kiel : Universitätsbibliothek Kiel, 2019. http://d-nb.info/1202630332/34.
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Odenbach, Tina. "Charakterisierung der Hybridsensorkinase LuxN und des Antwortregulators LuxO des Quorum sensing-Systems in Vibrio harveyi." Diss., lmu, 2009. http://nbn-resolving.de/urn:nbn:de:bvb:19-100461.
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Stambrau, Nina. "Der LuxP/AI-2- und LuxQ-abhängige Signaltransduktionsweg des Quorum sensing-Systems von V. harveyi." Diss., lmu, 2008. http://nbn-resolving.de/urn:nbn:de:bvb:19-103233.
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Teplitski, Max I. "Quorum sensing in Sinorhizobium meliloti and effect of plant signals on bacterial quorum sensing." The Ohio State University, 2002. http://rave.ohiolink.edu/etdc/view?acc_num=osu1029777185.
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Damasceno, Alhen Rubens Silveira. "O luxo e o mainstream: a circulação intersígnica das marcas." Universidade de São Paulo, 2017. http://www.teses.usp.br/teses/disponiveis/27/27153/tde-30052017-110852/.
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A sociedade contemporânea é marcada pelo consumo, pelas estratégias mercadológicas que, muitas vezes, são efêmeras e só tentam satisfazer uma pequena parte da nossa constante procura pela completude. A publicidade está inserida nesse contexto ao expressar os valores vigentes na sociedade e também em mediar as possibilidades de inserção dos consumidores nos mais diversos nichos e estratos sociais. As marcas procuram expressar valores, identidades e imagens dos produtos/serviços por meio de ações e manifestações. Tais ações e manifestações podem vir de diversas maneiras, ou seja, em anúncios de mídia impressa, ou na forma de audiovisual; ações de merchandising como as vitrines, as gôndolas, dentre outras e que têm como intuito a geração de vínculos positivos, de associações prazerosas e levar o potencial consumidor ao ato e usufruto do bem ou serviço. O presente trabalho resulta de uma pesquisa entre as marcas de luxo em comparação das marcas mainstream, um aprofundamento intersígnico procurando identificar os efeitos de sentido de cada, suas semelhanças e distinções. O trabalho tem como objetivo compreender a circulação intersígnica das marcas de luxo e das marcas mainstream no contexto do varejo de moda. Os resultados que aqui compartilharemos é proveniente de uma pesquisa que conta com o intercruzamento dos campos da comunicação com o a antropologia visual. A comunicação, mas precisamente, a publicitária, entrará como sua retórica de consumo por meio dos signos que se relacionam para uma produção de sentido eficaz. Já a antropologia visual ficará por conta da pesquisa fotoetnográfica que nos auxiliará no mapeamento e narrativa do consumo de luxo e do mainstream. Como método de análise nos apoiaremos na Semiótica desenvolvida por Charles Sanders Peirce. O registro fotoetnográfico foi realizado nas cidades de: São Paulo, Fortaleza, Brasília e Florianópolis e buscamos fotografar as vitrines das marcas e luxo e das marcas mainstream que nos possibilitem um estudo sobre a circulação intersígnica entre as mais variadas vitrines. Este trabalho não terá como pretensão a análise de recepção por parte dos consumidores, ficaremos restritos só aos possíveis efeitos de sentido que as marcas se utilizam para ampliar seu escopo imagético e identitário.ephemeral and only try to satisfy a small part of our constant search for completeness. Publicity is inserted in this context when expressing the values in force in society and also in mediating the possibilities of insertion of the consumers in the most diverse niches and social strata. Trademarks seek to express values, identities and images of products / services through actions and manifestations. Such actions and manifestations may come in a variety of ways, that is, in print media advertisements, or in the form of audiovisuals; Merchandising actions such as shop windows, gondolas, among others and whose purpose is to generate positive bonds, pleasant associations and lead the potential consumer to the act and usufruct of the good or service. The present work results from a research between the luxury brands in comparison of the mainstream brands, a deepening intersígnico looking for to identify the effects of sense of each, its similarities and distinctions. The work aims to understand the intersection of luxury brands and mainstream brands in the context of fashion retail. The results we share here come from a research that relies on the cross-linking of the fields of communication with visual anthropology. Communication, but precisely the advertising, will enter as its rhetoric of consumption through the signs that relate to a production of effective meaning. The visual anthropology will be based on the photoetnographic research that will help us in the mapping and narrative of the consumption of luxury and the mainstream. As a method of analysis we will rely on the Semiotics developed by Charles Sanders Peirce. Photoetnographic registration was carried out in the cities of: São Paulo, Fortaleza, Brasília and Florianópolis, and we sought to photograph the showcases of the brands and luxury and of the mainstream brands that allow us to study the intersection circulation among the most varied shop windows. This work will not have as a pretension the analysis of reception by consumers, we will be restricted only to the possible effects of meaning that the brands are used to expand their image and identity scope.
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Barbosa, Joaquim Carlos da Silva. "A estética e o design nos sapatos da marca Christian Louboutin." Universidade de São Paulo, 2016. http://www.teses.usp.br/teses/disponiveis/27/27152/tde-15052017-152423/.
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Christian Louboutin é um designer da moda, que trata da estética sob a ótica do processo criativo, um produtor de artigos de luxo, que atua em um nicho de mercado com acessórios da moda de luxo, para homens e mulheres com alto poder aquisitivo. Faremos um estudo de caso único, delimitado ao assunto da estética e o design nos sapatos femininos da marca Christian Louboutin, a fim de buscarmos respostas através de alguns questionamentos para as seguintes questões de pesquisa: 1) Como a estética e o design nos sapatos da marca Christian Louboutin desempenham um papel importante no despertar da imagem, de poder, da elegância, da sensualidade e do fetiche nas clientes Louboutin? 2) Como os elementos resultantes dos sentimentos de prazeres e das experiências sensoriais subliminares estimuladas por atributos estéticos, dão origem a diversas sensações e emoções que servem de incentivo no processo de compra? O objetivo deste estudo será o de encontrar os principais atributos estéticos que expressam determinado nível de beleza nos sapatos Christian Louboutin, aqueles que resultam em experiências emocionais sensoriais subliminares que influem nos gostos e nos gastos das clientes, quando compram os sapatos de luxo da marca. A metodologia deste trabalho consistirá de um estudo de caso único que utilizará os métodos de coleta de dados: observação e entrevista. As entrevistas serão realizadas com seis clientes no ponto de venda, associada à determinada fundamentação teórica. Os resultados obtidos demonstraram que não ocorreram divergências significativas nas respostas dadas pelas entrevistadas se contrapomos aos objetivos deste trabalho. Os resultados das entrevistas se alinham ao conceito de estética e do design dado pela fundamentação teórica, e por serem considerados os elementos mais admirados pelas clientes nos sapatos de luxo Christian Louboutin. A conclusão deste estudo de caso é que os elementos estéticos encantam e fascinam as clientes, despertam sensações de poder, paixões e fetiche, além de dar a sensação de ficarem mais lindas, sensuais e atraentes. A estética age como um elemento imprescindível à inovação e a diferenciação nos sapatos, garante a autenticidade da marca, servindo de conexão como o imaginário de suas clientes.Christian Louboutin is a fashion designer who deals with the aesthetics from the perspective of the creative process, a producer of luxury goods, which operates in a niche market with luxury fashion accessories for men and women with high purchasing power. We\'ll make a single case study, defined the subject of aesthetics and design in women\'s shoes brand Christian Louboutin, to seek answers through some questions to the following research questions: 1) How the aesthetics and design in shoes brand Christian Louboutin play an important role in the image of awakening, power, elegance, sensuality and fetish in Louboutin customers? 2) How the resulting elements of the feelings of pleasure and subthreshold sensory experiences stimulated by aesthetic attributes, give rise to different sensations and emotions that serve to encourage the purchase process? The aim of this study is to find the main aesthetic attributes that express certain level of beauty in Christian Louboutin shoes, those that result in subliminal sensory emotional experiences that influence the tastes and spending of customers when they buy luxury brand shoes. The methodology of this work will consist of a single case study that will use the data collection methods: observation and interview. The interviews will be conducted with six customers at point of sale, associated with a particular theoretical framework. The results showed that no significant differences in the answers given by the respondents to counterpose the objectives of this work. The results of the interviews are aligned to the concept of aesthetics and design given by the theoretical basis, and consider the most admired elements by customers in luxury shoes Christian Louboutin. The conclusion of this case study is that the aesthetic elements delight and fascinate customers, arouse feelings of power, passion and fetish, and give the feeling of being the most beautiful, sexy and attractive. The aesthetic acts as an essential element for innovation and differentiation in shoes, guarantees the authenticity of the brand, serving as a connection to the imagination of its customers. Google Tradutor para empresas:Google Toolkit de tradução para appsTradutor de sitesGlobal Market Finder
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Östman, Ludwig. "Rolls-Royce Luxe Nomad." Thesis, Umeå universitet, Designhögskolan vid Umeå universitet, 2018. http://urn.kb.se/resolve?urn=urn:nbn:se:umu:diva-153224.
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The topic of the project has been about the future of luxury mobility and how the brand Rolls-Royce could evolve alongthe technological advancements until the year 2038. As luxury is changing from being about possession to the experience of luxury, the goal was to design a vehicle that could support both new internal and external experiences about the vehicle. The identified obstacle that could limit the scope of new experiences in an autonomous vehicle is motion sickness. As cities grow and technology develops the future cities are crowded so personal space, and privacy is being limited.During the process, the tools used varied from form exploration through sketching, concept principle testing and finalization in 3d software as well as visualization tools to communicate the proposed solutions. Working out of the Rolls-Royce designstudio provided brand insight but also expertise in the field of color & materials as well as advice on engineering solutions from the design team.The result Rolls-Royce Luxe Nomad is a vehicle for exploring the world outside the urban areas. Going far and beyond in search of new and unique experiences but also making the trip there as enjoyable as possible. The concept provides a solution inspired by high-speed trains that could limit those effects. Through understanding the future customers need of privacy a method of controlling the amount of insight was developed to enhance the user experience and giving them an increased sense of control of their space.
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Ihle, Karolina. "Quorum sensing Rezeptorprotein LuxP – gentechnisches Design von LuxP-Derivaten zur Anwendung in der Biosensorik." Doctoral thesis, Saechsische Landesbibliothek- Staats- und Universitaetsbibliothek Dresden, 2011. http://nbn-resolving.de/urn:nbn:de:bsz:14-qucosa-64378.
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Als Quorum sensing (QS) wird die Kommunikation zwischen Bakterien bezeichnet. Diese basiert auf kleinen Signalmolekülen, die Autoinducer (AI) genannt werden. Durch QS werden von Bakterien Verhaltensweisen wie Fähigkeit zur Symbiose, Virulenz, Produktion von Antibiotika und Bildung von Biofilmen reguliert. Die Kommunikation kann innerhalb einer Spezies (Intraspezies-Kommunikation) oder mehreren Spezies (Interspezies-Kommunikation) erfolgen. Gram-negative Bakterien kommunizieren über acetylierte Homoserinlaktone (AHL), Gram-positive Bakterien dagegen benutzen modifizierte Oligopeptide als Autoinducer.
Für die Interspezies-Kommunikation dient der Autoinducer-2 (AI-2). AI-2 entsteht auf dem Weg der spontanen Zyklisierung von 4,5-Dihydroxy-2,3-Pentadion (DPD), der von LuxS synthetisiert wird. Die Universalität des AI-2 als Signalmoleküls basiert auf dessen chemischen Eigenschaften. Als biologisch aktive Formen von DPD gelten S-THMF-Borat (bei marinen Bakterien wie Vibrio harveyi) und R-THMF (z.B. bei Enterobakterien wie Escherichia coli oder Salmonella enterica Serovar Typhimurium). AI-2 wird bei allen Bakterien von einem periplasmatischen Rezeptor gebunden. S-THMF-Borat bindet spezifisch an den Rezeptor LuxP, R-THMF dagegen an den Rezeptor LsrR. Durch die Anbindung des AI-2 verändert sich die Konformation des Rezeptors, was als Signal über weitere Proteine in die Zelle weitergeleitet wird.
In E. coli ist die Expression des Operons lsrACDBFGE von AI-2 abhängig. Der lsr-Promotor wird von dem Repressor LuxR, Phospo-AI-2 sowie dem cAMP-CRP-Komplex reguliert.
In dieser Arbeit wurden die molekularbiologische Grundlagen zur Entwicklung eines AI-2-Biosensors gelegt. Es wurden mehrere Fusionskonstrukte des V. harveyi AI-2 Rezeptors LuxP sowie dessen Derivate mit veränderter Affinität zur AI-2 kreiert, in E. coli exprimiert und aufgereinigt. Auch Rezeptorproteine von Vibrio fischeri sowie E. coli konnten erfolgreich exprimiert werden. Die Expression der Proteine erfolgte in E. coli luxS- Deletionsstämmen, die hierfür konstruiert worden sind. Die AI-2-Rezeptorproteine werden in E. coli vorwiegend in Form von inclusion bodys exprimiert. Nur ein Teil des Proteins ist löslich und kann für die Aufreinigung unter nativen Bedingungen verwendet werden.
Auf der Basis von E. coli luxS- Deletionsstämmen wurde ein Bioassay entwickelt, der für die Detektion von AI-2 verwendet werden kann. Hierfür wurden mehrere Reporterplasmide konstruiert, in denen das rot fluoreszierende Protein DsRed unter die Kontrolle des lsr-Promotors von E. coli kloniert wurden. Dabei konnte unter Verwendung einer dieser Reporterplasmide (pBRDsRed) sowie des luxS-Deletionsstammes KIB1 Bioassay-Bedingungen etabliert werden, die einen Nachweis von AI-2 ermöglichen. Die für den Assay benötigten AI-2-Moleküle wurden in vitro mithilfe der Enzyme Pfs und LuxS und S-Adenosyl-Homocystein (SAH) als Substrat hergestellt.
Der entwickelte AI-2-Bioassay wurde für die Bestimmung der Bindeaktivität der V. harveyi LuxP-Derivate verwendet. Die resultierenden Ergebnisse wiesen eine hohe Reproduzierbarkeit (1,2 bis 11,3 % Standartabweichung) auf.
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Rezaee, Vessal Saeedeh. "Les heuristiques dans le luxe : la psychologie de l'utilisation des signaux liés au luxe." Thesis, Université Grenoble Alpes (ComUE), 2018. http://www.theses.fr/2018GREAG003.
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Cette recherche s'intéresse à la façon dont les modes de pensée influencent les préférences des consommateurs pour les produits de luxe. Dans une série d'expérimentation, nous montrons que lorsque les consommateurs sont plus susceptibles de pensée de manière intuitive, ils utilisent davantage le signal lié au luxe dans leur processus de décision. Afin de montrer l'effet de ce signal comme indicateur heuristique, le mode de pensée des consommateurs a été manipulé. Dans un mode de pensée intuitif, les individus ont tendance à utiliser significativement plus le signal lié au luxe comme un signal heuristique dans la prise de décisionIn this research, we discuss consumers’ preference for status products which is affected by their mode of thinking. In a series of experiments, we show that when consumers are more likely to rely on intuitive system thinking, they use status signal more often as a cue in their decision making process. In order to show the effect of the status signal as a heuristic cue, we manipulate consumers’ mode of thinking. Using an online questionnaire, we show that being in intuitive mode of thinking leads individuals to disproportionately rely on status signals as a heuristic cue in their decision making process
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Bezerra, Diogo Henrique Duarte. "Luar-X: extens?o e generaliza??o do sistema de processamento de modelos luar." PROGRAMA DE P?S-GRADUA??O EM ENGENHARIA EL?TRICA E DE COMPUTA??O, 2016. https://repositorio.ufrn.br/jspui/handle/123456789/21944.
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Coordena??o de Aperfei?oamento de Pessoal de N?vel Superior (CAPES)
O sistema de processamento de modelos Luar, utilizado na autoria de documentos NCL, possui conceitos de desenvolvimento baseados no re?so de descri??es por meio de modelos de documentos. Esses conceitos de desenvolvimento podem ser utilizados na autoria de outros documentos baseados em XML por meio da an?lise, adequa??o, projeto e cria??o de uma nova infraestrutura de re?so baseada no sistema Luar. Esta tese apresenta o sistema de processamento de modelos Luar-X, uma generaliza??o e extens?o do sistema de modelos Luar. O Luar-X mant?m os conceitos de desenvolvimento do sistema Luar e modifica, adequa e estende a arquitetura do sistema Luar para permitir o desenvolvimento com linguagens baseadas em XML. Os estudos de caso com as linguagens HTML, SVG e NCL mostram a viabilidade dessa nova infraestrutura de re?so que torna o desenvolvimento de aplica??es e a descri??o de documentos XML ?gil.
The Luar template processing system used in authoring NCL documents, has development concepts based on reuse of descriptions through document templates. These development concepts can be used in the authorship of other XML-based documents through the analysis, adequacy, design and creation of a new reuse infrastructure based on Luar system. This view shows the processing system of models Luar-X, a generalization and extension Luar model system. The Luar-X keeps the concepts of development of Luar system and he changes, adapts and extends the Luar system architecture to enable development with XML-based languages. The case studies with HTML, SVG and NCL show the feasibility of this new reuse infrastructure, which makes application development and description of agile XML documents.
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Corbel, Céline. "Produits de luxe et commerce électronique." Grenoble 2, 2006. http://www.theses.fr/2006GRE21033.
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Internet est un espace de liberté et offre de nouvelles possibilités commerciales pour toutes les industries. Les maisons de luxe utilisent le Web comme un moyen de communication, puisque chacune d'entre elles y possèdent déjà un site. La question se pose néanmoins de savoir si la vente des produits de luxe est compatible avec la nature d'Internet, étant donné l'aura qui entoure ces produits et qui les distingue nettement des autres objets et si les mécanismes juridiques actuels permettent d'utiliser Internet comme un nouveau canal de distribution. L'industrie du luxe doit être présente sur la toile, comme acteur commercial mais également comme défenseur de ses droits de propriété intellectuelle. En effet, le Net est une source de risques pour cette industrie: le phénomène des ventes parallèles et des contrefaçons, y est largement accru et des infractions propres aux nouvelles technologies ont vu le jour. Cette thèse propose d'analyser les règles juridiques existantes qui permettent à l'industrie du luxe de commercialiser ses produits sur Internet et de lutter contre ses dangersThe Internet provides freedom and offers new commercial possibilities for all industries. Companies that sell luxury products already use the Internet for marketing, since each has a website. Nevertheless, the issue arises of knowing whether using the Internet to sell luxury products is compatible with the nature of Internet, given that the aura of luxury products clearly distinguishes them from other products available on the market and whether current laws allow such companies to use the Internet as a new distribution channel. As such, luxury industry companies use the Internet not only as commercial actors but as defenders of intellectual property rights. Indeed, Internet is also a source of risks for the luxury industry: instances of parallel sales and counterfeiting have increased greatly, and new technologies have given rise to new kinds of infringement. This thesis analyzes the current legal framework and posits that it allows luxury industry companies to market their products on the Internet while adequately countering its threats
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Souza, Anne Tess Guimarães Araújo de. "O estudo do comportamento do consumidor de bens de luxo : os determinantes da atitude face ao luxo." Master's thesis, FEUC, 2013. http://hdl.handle.net/10316/24762.
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Dissertação de mestrado em Gestão (Marketing), apresentada à Faculdade de Economia da Universidade de Coimbra, sob a orientação de Arnaldo Fernandes Matos Coelho.De entre os diversos estudos que influenciam o marketing, uma das ramificações que se encontra em vias de desenvolvimento é o estudo do comportamento dos consumidores de produtos de luxo. Procurar entender este ambiente de luxo, envolve não apenas a compra de produtos caros mas também outros fatores relacionados diretamente com a história e cultura de uma marca. As estratégias empresariais de luxo encontram-se cada vez mais adaptadas ao mercado consumista, isto é, executando novas estratégias para a fidelização de clientes. A presente investigação possui como objetivo o estudo do comportamento do consumidor de bens de luxo, bem como os determinantes da atitude destes face ao mesmo. Pretende-se identificar quais são os impactos da intenção de compra na perceção do luxo e na atitude face ao luxo, assim como os seus antecedentes que possuem significância para este consumidor. Para atingir este objetivo, efetuou-se uma revisão de literatura académica sobre a temática. A presente investigação consistiu num estudo empírico, utilizando metodologia quantitativa e descritiva, não-probabilística. Através da aplicação de 409 inquéritos a consumidores de bens de luxo franceses e brasileiros, o resultado da análise dos questionários revelou a existência de comportamentos diferentes mas, simultaneamente, semelhantes entre as duas culturas distintas, que, por vezes, revelaram-se motivadas pelos mesmos princípios. O presente estudo pretendeu efetuar a sua contribuição académica através do enriquecimento dos conhecimentos teóricos existentes sobre a temática, a partir de uma pesquisa empírica, para encontrar diversos contributos para as empresas de marcas de luxo, bem como para as estratégias empresariais do segmento de consumo de luxo. Os resultados do presente estudo pretendem criar interesse académico, e empresarial, para a elaboração de propostas de marketing pela sua plausível relevância científica.
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Maldonado, Maura Carneiro. "Usando o luxo, consumindo o espaço: uma investigação sobre as relações entre espaço, consumo de moda e luxo." Universidade Federal da Paraíba, 2009. http://tede.biblioteca.ufpb.br:8080/handle/tede/3846.
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Previous issue date: 2009-07-30Coordenação de Aperfeiçoamento de Pessoal de Nível Superior - CAPES
Luxury consumption in Brazil has been growing significantly, this country being considered one of the 10 largest luxury markets in the world.Though it is restricted to just 1% of the population, the sector already sets in movement US$2,5 billion per year. This strong commercial presence stimulates theoretical and academic analysis about the theme. Some studies were developed in the sense of shedding light on the behaviour f luxury consumers and the symbolic aspects associated to it. However, the proposal of this dissertation is to review the relations between the consumption of luxury fashion and the space thereby occupied. The main assumption is that there is an exchange between the meanings of this type of consumption, its potential consumers and the marketplace where these elements complete one another and are realized in the social field. The research was made through an ethnographic qualitative investigation with two main sources of data: in-depth interviews and participant observation and the exam of information was made through discourse analysis. The motivations for luxury consumption were identified as the quality of products in themselves, the beauty and distinction attached to it and the hedonism in the differentiated experience of acquisition. As to space, the analysis of the data demonstrate that high prestige shops follow spatial codes and references consciously or unconsciously developed by high acquisition groups. The placing of this type of establishment must be in harmony with the aspects of luxury products, being also a reflex of those. It is important that there exists an identification between the offered product, the shopping point and the identity of the consumers. The symbolic aspects of space turn to the perception of the product, thereby having the power to highlight its qualities, as well as it may bring about a negative view of the products, as a consequence of the lowering of the values connected to it such as authenticity, beauty, status and prestige amongst others.
O consumo de luxo no Brasil vem crescendo rapidamente, sendo inclusive este país considerado um dos 10 maiores mercados de luxo do mundo. Apesar de estar restrito a apenas 1% da população, o setor já movimenta U$ 2,5 bilhões anuais. Esta forte presença comercial estimula a análise acadêmica e teórica sobre o tema. Alguns estudos foram desenvolvidos no sentido de compreender o comportamento do consumidor de luxo e os aspectos simbólicos a ele associados. No entanto, é proposta desta dissertação analisar as relações entre consumo de moda de luxo e o espaço por ele ocupado. Parte-se da idéia de que existe uma troca entre os significados deste tipo de consumo, os consumidores potenciais e os locais de compra onde estes elementos se completam e se caracterizam no campo social. A pesquisa valeu-se de um método qualitativo de investigação, com duas fontes de dados principais: entrevistas em profundidade e observação participante e o exame destas informações foi feito a partir da análise de conteúdo. Foram identificadas como motivações para o consumo de luxo a qualidade intrínseca ao produto, a beleza e a distinção por ele proporcionado e o hedonismo alcançado pela experiência diferenciada da compra. Sobre o espaço, a análise dos dados indica que é esperado que as lojas de alto prestígio sigam os códigos e referências espaciais estabelecidos consciente ou inconscientemente pelos grupos de alto poder aquisitivo. O posicionamento deste tipo de estabelecimento deve estar harmonizado com os aspectos do produto de luxo, sendo também reflexo destes. É importante que exista uma identificação entre o produto ofertado, o ponto de venda e a identidade dos consumidores. Os aspectos simbólicos do espaço recaem sobre a percepção do produto tendo o poder de elevar as suas qualidades, bem como pode acarretar uma visão pejorativa dos produtos fruto da diminuição dos valores relacionados a ele como autenticidade, beleza, status, prestígio, entre outros.
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Koromyslov, Maxime. "L'impact de la délocalisation du luxe sur les attitudes des consommateurs : le cas des maisons françaises de luxe." Thesis, Nancy 2, 2009. http://www.theses.fr/2009NAN22002/document.
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La délocalisation du luxe demeure encore aujourd’hui difficilement appréhendable. La complexité relève, d’une part, du concept du luxe qui est difficilement définissable, et de la sensibilité du sujet qui demeure encore tabou dans le secteur du luxe. Pourtant, plusieurs cas de délocalisations dans le luxe sont aujourd’hui connus. Compte tenu de l’imaginaire que représentent le luxe et les marques de luxe, il convient de s’interroger sur l’impact de la délocalisation sur le consommateur, notamment sur ses attitudes envers le produit et la marque. Cette thèse constitue une première recherche sur la question de la délocalisation appliquée aux marques françaises du luxe. Elle mobilise, d’une part, les théories sur le luxe et la marque en tant que champ d’application et, d’autre part, la littérature sur l’effet « pays d’origine » en tant que cadre théorique. Notre recherche étudie l’effet de la délocalisation sur les attitudes des consommateurs envers le produit délocalisé et la marque de luxe. L’influence modératrice de deux nouvelles variables est considérée, à savoir la centralité du pays d’origine au sein de l’image de marque et les attitudes des consommateurs envers la délocalisation du luxe. Sur le plan méthodologique, deux études qualitatives auprès des consommateurs (17) et des professionnels du luxe (34) ont été réalisées, suivie de l’étude quantitative auprès d’un échantillon de 555 consommateurs interrogés en deux temps, selon un plan factoriel complet. Ainsi, notre recherche a mis en lumière l’impact négatif de la délocalisation du luxe sur les attitudes envers le produit délocalisé et la marque de luxe. L’effet est plus prononcé en cas de délocalisation de la fabrication par rapport au design, lors de l’abandon de l’origine française au profit d’un pays de délocalisation inacceptable et se ressent davantage sur la qualité perçue que sur les deux autres variables attitudinales. L’impact des variables modératrices ressort limitéThe relocation of the luxury sector is, even nowadays, still difficult to comprehend. On one hand, this complexity is due to the concept of luxury which is not easy to define, and on the other hand, it is due to the topic’s sensitivity which is still taboo in the luxury sector. Yet there have been several cases of luxury relocations. Considering the imaginative world represented by luxury brands, we should look at the impact of relocation on consumers, notably their attitudes towards the product and the brand. This thesis is a first research study on the question of relocating French luxury brands. It uses both the theories about luxury and brands as areas of application, and the literature on the Country-of-Origin Effect as a theoretical framework. Our research studies the effect of relocation on consumers’ attitudes towards the relocated product and luxury brand. The moderating influence of two new variables has been taken into account, i.e. the centrality of the country of origin at the heart of the brand image and consumers’ attitudes towards luxury relocation. In terms of methodology, two qualitative surveys were carried out: one on consumers (17), the other on luxury professionals (34). These were followed by a quantitative survey using a sample of 555 consumers. The survey was carried out in two parts, based on a complete factorial design. Our research therefore brings out the negative impact of luxury relocation upon attitudes to the relocated product and brand. This effect is even more noticeable in the case of production relocation in comparison with design one or when the French origin is abandoned for an unacceptable country of manufacture. In this case, the effect is felt more on the perceived quality than the two other attitudinal variables. The impact of the moderating variables comes out as a limited one
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Dubois-Pélerin, Éva. "Le luxe privé à Rome et en Italie au Ier siècle après J.-C. /." Naples : Paris : Centre Jean Bérard ; diff. de Boccard, 2008. http://catalogue.bnf.fr/ark:/12148/cb41398609p.
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Selosse, Philippe. "Le régime juridique du produit de luxe." Thesis, Lille 2, 2017. http://www.theses.fr/2017LIL20002/document.
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Le produit de luxe n’est pas un bien comme les autres. Ses qualités matérielles et immatérielles lui confèrent une valeur particulière qui oblige son producteur à le vendre dans un environnement commercial adapté. La règle de droit peut-elle considérer cette particularité économique ? Paradoxalement, la France est leader mondiale du marché des produits de luxe, mais nul n’est en mesure d’affirmer avec précision ce qu’est le luxe. Intégrer une notion aussi insaisissable au sein d’un raisonnement juridique semble difficile. Pourtant, les atteintes subies par les titulaires des droits du produit de luxe ont convaincu le juge européen de mettre en place des règles protectrices spéciales. Le but poursuivi est légitime. Il s’agit de protéger les investissements réalisés pour vendre et promouvoir le produit de luxe. Mais cette démarche légale s’appuie sur une méthode de qualification qui n’est pas satisfaisante. L’«aura», le «prestige» ou la «sensation» de luxe qui émanent du produit marqué, sont des critères trop subjectifs pour assurer l’application systématique et cohérente de règles protectrices. C’est pourquoi, outre la démonstration d’un corpus de règles applicables au produit de luxe, il convient d’analyser les fondements de sa reconnaissance par le droit positif, ainsi que l’instauration d’un régime unifié reposant sur des critères de définition précis, prenant en considération les qualités intrinsèques de ce bien particulierThe luxury product is not a product like any other. Its material and immaterial qualities confer a special value that requires its producer to sell it in a proper business environment. The rule of law can it consider this economic feature ? Paradoxically, France is world's leading luxury goods market, but no one is able to state precisely what is luxury. The law seems unsuited to integrate a concept as elusive as luxury. Yet, violations suffered by the owners of luxury product rights have convinced the European judge to set up special protective rules. The aim is legitimate. This is to protect the investments made to sell and promote luxury products. But this legal approach is based on a method of qualification which is not satisfactory. The "will", "prestige" or the "feel" of luxury emanating frombranded product, are too subjective criteria to ensure systematic and consistent implementation of protective rules. Therefore, in addition to the demonstration of a body of rules applicable to the luxury product, it should analyze the foundations of its recognition by positive law and the establishment of a unified system based on criteria precise definition, taking into account the intrinsic qualities of that particular property
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Carreira, Joana Fernandes. "Motivações de compra de marcas de luxo." Master's thesis, Instituto Superior de Economia e Gestão, 2016. http://hdl.handle.net/10400.5/13009.
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Mestrado em MarketingA noção de luxo tem mudado muito ao longo dos anos, o mercado de luxo está em crescimento em mercados emergentes e que, cada vez mais, as estratégias de marketing deste tipo de marcas tem vindo a mudar. O conceito de luxo tem vindo a ser desenvolvido ao longo dos tempos, se anteriormente o legado e a expertise associada da marca eram duas das principais características que definiam as marcas de luxo atualmente já o conseguimos fazer também sob o ponto de vista do consumidor, ligado às suas motivações, atitudes e interesses. Neste sentido, esta dissertação tem como objectivo investigar quais as motivações de compra dos consumidores das marcas de luxo através de um modelo que reúne características e conceitos relevantes tanto ao nível da marca, como brand equity, tanto como o ponto de vista do consumidor.
The luxury concept has changed a lot over the years. The luxury market is growing in emerging markets and, luxury brands has been changing their marketing strategies. The definition of luxury has been modified over the years, previously the legacy of a brand and their expertise associated, were two of the main characteristics that defined luxury brands. However, we can also define luxury in the consumer's perspective, based on their motivations, attitudes and interests. This work aims to investigate what are the main buying motivations of luxury brands consumer's, through a model that brings together relevant features and concepts of a brand perspective, as brand equity, and of consumer's perspective.
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Sousa, Sandra de Abreu. "Alta-costura e o objeto de luxo." Master's thesis, Universidade de Lisboa, Faculdade de Arquitetura, 2018. http://hdl.handle.net/10400.5/16519.
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Dissertação de Mestrado em Design, com a especialização em Moda apresentada na Faculdade de Arquitetura da Universidade de Lisboa para obtenção do grau de Mestre.Esta dissertação tem como titulo “Alta-Costura e o Objeto de Luxo, Estágio no Ateliê Storytailors”, pretendendo identificar e analisar a adaptação de conceitos de Alta-Costura à realidade de um ateliê português. O trabalho desenvolvido neste estágio levou a uma profunda aprendizagem e colaboração em vários projetos, quer em coleções, quer em encomendas de ateliê, como vestidos de noiva e de cerimonia, desde o seu início colaborando nas diversas fases dos respetivos projetos, onde foram acompanhadas todas as fases da criação e da construção das peças, desde a tirada de medidas do cliente até à criação de moldes, corte, montagem, provas (fitting) e acabamentos finais. Participou-se ainda no desenvolvimento das coleções na área de design, na criação de peças para desfile, sob o conceito de desperdício zero, aplicado no decorrer do estágio. As metodologias utilizadas neste estágio foram tanto de carater intervencionista, tendo a aluna participado nas áreas de design e de produção da empresa Storytailors, como não-intervencionista, no sentido da observação e do registo do trabalho desenvolvido pelos colaboradores da empresa. A aluna pretende demonstrar no exemplo do ateliê Storytailors, como elementos da Alta-Costura podem ser introduzidos no trabalho de um ateliê português. Além disto a aluna pôde verificar a importância da vertente prática desenvolvida no estágio, aplicando conhecimentos teóricos anteriormente adquiridos, em contexto profissional.
ABSTRACT: This dissertation has as title "Haute Couture and the Luxury Object, Internship at Atelier Storytailors", intending to identify and analyze the adaption of concepts of Haute Couture to the reality of a Portuguese atelier. The work developed in this internship led to a profound learning and collaboration in various projects in both collections and atelier orders such as wedding gowns and ceremonial garments, collaborating in several stages of the respective projects, accompanying all stages of the creation and construction of garments, from taking the client measures until pattern making, cutting, assembling, fitting and finishings. The intern participated in the development of collections in the design area, creating garments for fashion shows under the concept of zero waste, applied in the course of the internship. The methodologies used in this stage were both of interventionist character, the student participating in the areas of design and production of the company Storytailors, as non-interventionist, in the sense of the observation and registration of the work developed by the staff of the company. The student intends to demonstrate at the example of the atelier Storytailors, how elements of haute-couture can be introduced into the work of a Portuguese atelier. Also, the student could verify the importance of practice, applying during the internship, the previously acquired theoretical knowledge in a professional context.
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Dereumaux, René Maurice. "Le luxe, application à l'industrie du meuble." Nice, 2006. http://www.theses.fr/2006NICE0043.
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Ce travail de recherche s'intéresse à une industrie essentielle dans la production de richesses des pays développés en général et de la France en particulier, l'industrie du luxe. Ostentatoire et simplement superflu pour l'observateur superficiel, ce secteur est cependant parmi les plus dynamiques de l'activité économique et sociale. Il est non seulement un créateur important de valeur ajoutée, mais également de prestige et de différenciation sociale. S'il constitue un marché en expansion, notamment de part le développement de pays très peuplés comme l'Inde ou la Chine, il reste cependant très méconnu et relativement peu étudié. Cette analyse prend pour cadre la stratégie de diversification des marques de luxe au sein d'un secteur traditionnel, l'industrie du meuble. Cette stratégie s'est récemment illustrée par le développement de filiales consacrées à cette industrie par deux grands noms de maisons de luxe : Giorgio Armani et Ralph Lauren. La recherche s'articule en deux temps. Après un premier temps consacré à " l'analyse sectorielle " où les deux activités sont détaillées et également appréhendées dans un cadre dynamique, le second temps revient sur la stratégie de diversification en elle-même, en expliquant ses raisons et ses chances de réussite. Cette deuxième partie appuie finalement son raisonnement sur le cas d'Armani Casa en reprenant les étapes qui ont mené à la diversification et les qualités que doit posséder une marque de Luxe pour la réussir. Enfin, il s'est dégagé de l'observation du secteur du Luxe que, parmi ses fondamentaux, la relation à l'art et la gestion de l'image de marque comptait particulièrement dans la réussite de l'activité traditionnelle comme dans les démarches de développement que représente une diversificationThis work deals with an essential part of the wealth production of developed countries in general and of France particularly: the luxury industry. Ostentatious and superfluous at the first look, this sector is however one of the most dynamic of the socio-economic activity. It's not only a great producer of both added value and Glamour but also of prestige and social differentiation. It constitutes a market on expansion, considering the boom of the Chinese and Indian economies, but it still stays widely unknown and it experiments a relative lack of studies. This analysis takes place in the diversification strategy of the Luxury brands in a traditionnal sector, the furniture industry. This strategy has been illustrated recently by the development of furniture branch for two important actors of the luxury industry: Giorgio Armani and Ralph Lauren. The research is organised on two steps. After the first part called: “the sector analysis” where the main activities are described and presented in a dynamic perpective, the second deals with the diversification strategy itself, giving the causes of its choice and the chance to reach the objectives of the firms. This second part is particulary illustrated with the Armani Casa case, wich authorizes to parse all the steps from the mother brand to the new branch including the basic qualities needed to success. This work has allowed to understand that the Luxury sector as a whole has a strong link with Art and with the Brand image. This link must be strong both on the fundamental activity and on the new developments that a diversification symbolises
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Godey, Bruno. "La sensibilité au luxe, une variable personnelle centrale pour comprendre les antécédents de la valeur perçue d’une marque de luxe." Caen, 2012. http://www.theses.fr/2012CAEN0697.
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Notre question de recherche résulte du constat de l’existence d’un état de tension caractéristique de la prise de décision du consommateur en situation de choix d’une marque de luxe. De nombreux choix de marques sont le fruit d’une volonté de la part du consommateur de faire coïncider sa perception de soi, sa personnalité avec celles de la marque. Toutefois, dans le domaine spécifique du luxe, il est traditionnellement mis en avant que les motivations qui président à l’achat des marques sont essentiellement de nature ostentatoire. Nous cherchons donc à expliquer dans quelle mesure les motivations du consommateur à nouer une relation particulière d’attachement avec une marque de luxe reposent sur un modèle intégrateur formé à partir de sa perception de l’appartenance d’une marque à la catégorie du luxe et de la congruence d’image avec une marque de luxe. La valeur perçue d’une marque de luxe peut également être influencée par des variables psychologiques individuelles. Si certaines d’entre-elles permettent de distinguer efficacement les consommateurs pour l’ensemble des biens de consommation, il nous semble qu’elles ne prennent pas en compte l’intégralité des particularismes du rapport au luxe des individus. Nous avons donc élaboré le concept de « sensibilité au luxe » afin de nous permettre d’analyser les différences de logique de prise de décision du consommateur en matière de marques de luxe. L’intensité de la sensibilité au luxe peut en effet nous donner la possibilité de comprendre les raisons pour lesquelles certains consommateurs vont rechercher une marque de luxe qui leur ressemble et d’autres, une marque qui pour eux symbolise particulièrement le luxeOur research question resulted from acknowledging the existence of a state of tension, characterizing the consumer decision-making process for luxury brands. Many brand choices are driven by a desire from consumers to correlate their self perception, and personality, with that of the brand. However, in the specific field of luxury, it has been traditionally emphasized that motivations driving brand purchases are essentially ostentatious. We seek to explain how the motivation of consumers to establish an attachment relationship with a particular luxury brand is based on an integrative model, formed from the perception of a brand belonging to the luxury category and from the image congruence with a luxury brand. The perceived value of a luxury brand can also be influenced by individual psychological variables. Whilst some of them effectively distinguish consumers for all consumer goods, it seems they do not take into account all the particularities of the individuals’ relationship to the luxury dimension. We have developed the concept of “luxury sensitivity” to allow us to analyze the differences in consumer decision-making logic for luxury brands. The intensity of luxury sensitivity can provide us with the opportunity to understand the reasons why some consumers will choose a luxury brand that resembles them and others a brand that especially symbolizes luxury
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Neto, Bruno Miguel Custódio Rosa. "Marketing na hotelaria de luxo. Caso: região Alentejo." Master's thesis, Universidade de Évora, 2019. http://hdl.handle.net/10174/26057.
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O setor do turismo é potenciador do desenvolvimento económico do país. Todos
os anos os lucros neste setor não param de crescer, este setor encontra-se numa fase de
crescimento, colocando Portugal em competição com as principais cidades europeias.
Grande parte de receita deste setor tem origem no setor hoteleiro, que ganha cada vez
mais força com o avançar dos anos sendo alvo de investimentos por parte dos
empreendedores. A hotelaria não fornece apenas alojamento para o turista, mas também
serviços de restauração, SPA e eventos. No Alentejo, nos últimos anos tem existido um
elevado investimento no setor hoteleiro.
O presente estudo tem como objetivo geral encontrar as ferramentas e métodos
com que as unidades hoteleiras de cinco estrelas da região Alentejo usam o marketing.
Em termos metodológicos aplicou-se um questionário às unidades hoteleiras com a
classificação de 5 estrelas na região Alentejo por NUTS II.
O objetivo geral passará por analisar o modo como os empreendimentos
hoteleiros de luxo na região Alentejo atuam ao nível do Marketing, de modo que serão
analisadas individualmente as características dos empreendimentos, compreender o tipo
de ocupação dos hóspedes, identificar se recorrem a estudos de mercado, analisar o
planeamento dos empreendimentos; Marketing in luxury hotels. Case: Alentejo region
Abstract:
The tourism sector is an enabler of the country's economic development. Every year
profits in this sector continue to grow, putting this sector is in a phase of growth, placing
Portugal in competition with the main European destinations. Much of this sector's revenue
comes from the hotel sector, which is gaining more and more strength year after year,
being the object of investments by the entrepreneurs. The hotel industry provides not only
tourist accommodation, but also catering services, SPA and events. In the Alentejo region,
in recent years there has been a high investment in the hotel sector.
The present study has as prime objective to find the tools and methods with which
the five-star hotels of the Alentejo region use marketing. In methodological terms, a
questionnaire was applied to the hotels with the classification of 5 stars in the Alentejo
region by NUTS II.
The main goal will be to analyse the way the luxury hotelier companies
use Marketing, by individually examining the characteristics of each hotelier
company, by understanding the type of guests and occupation, by identifying if
these are based of market studies, and by analysing the strategic planning of the
hotelier companies.
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Luz, António Maria Zarco Adrião Alves da. "Atitudes face às imitações de produtos de luxo." Master's thesis, Instituto Superior de Economia e Gestão, 2015. http://hdl.handle.net/10400.5/13150.
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Mestrado em MarketingEsta dissertação tem como propósito compreender, no contexto português, os fatores que influenciam as atitudes dos consumidores em relação às imitações de produtos de luxo. Para tal foi realizada a pesquisa de literatura, desenvolvido um modelo concetual que se foca nos fatores sociais, individuais, sociodemográficos e consumo prévio, e delimitadas as hipóteses objeto do estudo. Finalmente, foi definida a população alvo e concretizado o estudo das características dos consumidores, através da análise dos resultados de um questionário distribuído online. Foram obtidas 810 respostas completas e validadas para a análise dos dados. Conclui-se que o consumo pelo status tem uma influência negativa e significativa nas atitudes dos consumidores face às imitações de produtos de luxo, sendo que os consumidores que já tiveram uma experiência prévia com este tipo de produtos os encaram de modo mais favorável. Como sugerido pela literatura, conclui-se que as atitudes face às imitações de produtos de luxo estão positivamente relacionadas com a intenção de compra. Ou seja, aqueles que têm uma atitude favorável relativamente às imitações de produtos de luxo estarão mais recetivos a adquiri-los no futuro. This dissertation aims to understand, in the Portuguese context, the factors that influence the attitudes of consumers towards counterfeits of luxury goods. To this end, literature was researched, a conceptual model that focuses on social, individual, socio-demographic and prior consumption factors was developed, and hypotheses to be studied were defined. Finally the target population was settled and the study of the characteristics of the consumers was completed through the analysis of the results of a questionnaire distributed online. 810 replies were validated for data analysis. It was concluded that consumption by status has a significant negative influence on the attitudes of consumers with regard to counterfeits of luxury goods. However, consumers who have already had a previous experience with this type of products have a more favorable attitude towards them. As suggested by the literature, it is concluded that the attitudes towards counterfeits of luxury goods are positively related to the intention to purchase. That is, people who have a favorable attitude towards counterfeits of luxury goods will be more likely to acquire them in the future.
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Carvalheiro, João Tiago da Cruz. "Antecedentes do comprometimento afetivo com automóveis de luxo." Master's thesis, Universidade de Aveiro, 2012. http://hdl.handle.net/10773/10357.
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Mestrado em GestãoEsta dissertação propõe estudar a imagem da marca, a qualidade percebida, a satisfação global e a confiança, como antecedentes do comprometimento afetivo de marcas de automóveis de luxo de determinados segmentos. Considera-se ainda a qualidade e a satisfação como mediadores entre a imagem, e a confiança e o comprometimento afetivo. Os resultados mostram que efetivamente a qualidade e a satisfação podem atuar como mediadores. A personalidade da marca, as atitudes positivas face à marca e a satisfação global podem exercer um efeito mais forte na construção do comprometimento afetivo com a marca de carros de luxo dos segmentos considerados, do que a qualidade percebida.
This dissertation aims to study brand image, perceived quality, overall satisfaction and trust as antecedents of affective commitment of luxury car brands in certain segments. Quality and satisfaction act as mediators between image, and trust and affective commitment. The results show that quality and satisfaction can really act as mediators. Brand personality, brand attitude and overall satisfaction may have a stronger effect on affective commitment with the luxury car brand, than perceived quality.
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Zhu, Jinge. "Mechanistic and inhibitory studies of S-ribosylhomocysteinase (LuxS)." Connect to this title online, 2005. http://rave.ohiolink.edu/etdc/view?acc%5Fnum=osu1115841808.
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Thesis (Ph. D.)--Ohio State University, 2005.Title from first page of PDF file. Document formatted into pages; contains xxii, 200 p.; also includes graphics (some col.). Includes bibliographical references (p. 190-200). Available online via OhioLINK's ETD Center.
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Andrade, Renata Paiva de. "Condomínios de luxo na periferia de São Paulo." reponame:Repositório Institucional do FGV, 1993. http://hdl.handle.net/10438/5345.
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Previous issue date: 1993-06-02T00:00:00ZEnfoca a análise dos condomínios de luxo na periferia de São Paulo à partir das motivações que levam as famílias a procur essa alternativa de moradia, das novas relações de sociabilidade que estabelecem entre seus moradores e das formas de segregação presentes
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Strehlau, Suzane. "O luxo falsificado e suas formas de consumo." reponame:Repositório Institucional do FGV, 2004. http://hdl.handle.net/10438/2532.
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Previous issue date: 2004-08-17T00:00:00ZThe concept of a luxury is dynamic and historical, belonging to a society in a particular moment and venue. As its consumption is widespread, it ceases to represent a social distinction. The luxury is not in the object itself, but in the judgment of a person on its value. The knowledge on luxury brands is sophisticated and requires specific qualifications. Not all prestigious products are necessarily a luxury and within the luxury category there is a scale of prestige. Taste, although it has a portion of individual origin, is shaped by the environment, and this will influence the attitudes and the behavior in consumption and socialization. The Habitus theory by Bourdieu offers an explanation based on the space of social positions organized around three dimensions: the total volume of social, cultural and economical capital, its composition and path taken over time. The cultural form of the incorporated capital, the habitus, implies in the assimilation of cultural conditions. The knowledge about false brands and their corresponding genuine items allow the owner to show all of ones habitus, that incorporated knowledge that only time and dedication can generate. The various origins and types of forgeries allow for showing a specific knowledge. The manner of relating oneself with the forged product, the manner of using them, how to cope with the subject in front of others in a social interaction generate an exhibition of habitus. The interviewed consumer of a habitus resulting from the stability in the popular stratum demonstrates little brand knowledge, being restricted to a name and a label. The ancestral nature in the elite stratum assures a tranquility in the consumption of forgeries, since, as tastemakers, they may express their distance from the need and their possibility of imposing the rules for distinction. For the group interviewed showing a recent social mobility, the forged brand may be used as leverage to increase social capital. When the social space occupied is still unstable, there continue to be reflections of social mobility, the brand is used as a crutch for generating connections, demonstrating a social position to those who are not a part of ones most intimate group of relations. The forged product may be a demonstration of distinction, if used suitably, with elements allied with the consumption by the dominant class.
O conceito do luxo é dinâmico e histórico, e pertence a uma sociedade em determinado momento e lugar. Na medida que seu consumo é divulgado, deixa de representar uma distinção social. O luxo não está no objeto, mas no julgamento de um sujeito sobre o seu valor. O conhecimento sobre marcas de luxo é sofisticado e requer competências específicas. Nem todos produtos de prestígio são necessariamente de luxo e dentro da categoria de luxo existe uma gradação de prestígio. O gosto, embora tenha uma parcela de origem individual, é moldado pelo meio, e isso vai influenciar as atitudes e os comportamentos de consumo e socialização. A teoria de Habitus de Bourdieu fornece uma explicação baseada no espaço de posições sociais organizada em torno de três dimensões: o volume total de capital social, cultural e econômico, sua composição e trajetória no tempo. A forma cultural do capital incorporada, o habitus, implica na assimilação de disposições culturais. O conhecimento sobre marcas falsas e seu correspondente genuíno possibilitam o seu proprietário de demonstrar todo seu habitus, aquele conhecimento incorporado que apenas o tempo e a dedicação trazem. As diversas origens e tipos de falsificações permitem exibir um conhecimento específico. A forma de se relacionar com o produto falsificado, o modo de usar, como abordar o assunto com as outras pessoas numa interação social, proporciona uma exibição do habitus. O consumidor entrevistado de habitus oriundo da estabilidade no estrato popular demonstra poucos conhecimentos sobre a marca, esta se restringe a um nome e uma etiqueta. A ancestralidade no estrato de elite assegura uma tranqüilidade no consumo de falsificados, porque, como formadores do gosto, podem expressar sua distância da necessidade e sua possibilidade de impor as regras de distinção. Para o grupo entrevistado que apresenta mobilidade social recente a marca falsa pode servir como alavanca para aumentar o capital social. Quando o espaço social ocupado ainda está instável, ainda há reflexos da mobilidade social, a marca serve como muleta para criar vínculos, demonstrar uma posição social para aqueles que não pertencem ao seu grupo mais íntimo de relações. A marca falsificada de luxo pode ser enquadrada como produto de luxo. O produto falso pode ser uma demonstração de distinção, se usado adequadamente, com elementos aliados ao consumo da classe dominante.
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Kambam, Pavan Kumar Reddy. "Engineering of the LuxI-LuxR quorum sensing systems for applications in industrial biotechnology." 2010. https://scholarworks.umass.edu/dissertations/AAI3397713.
Full textAbstract:
Quorum-sensing systems enable autonomous activation of gene expression and have been widely used in the construction of artificial genetic circuits, but exhibit poor tunability. We anticipate that tunable quorum sensing would facilitate its application in synthetic biology and industrial biotechnology. We engineered LuxI, which synthesizes signal molecule, 3-oxo homoserine lactone (OHHL), using a protein engineering method referred to as directed evolution. An antibiotic-based genetic selection method was developed and two rounds of mutant library screening resulted in two mutants with approximately 80-fold increase in OHHL yields. To demonstrate the application of the genetic selection in engineering LuxI homologs, a similar method was applied to alter the substrate specificity of RhlI, a key enzyme of the RhlI-RhlR quorum-sensing system involved in the pathogenesis of Pseudomonas aeruginosa. In particular, production of its cognate signal molecule, BHL (butanoyl homoserine lactone), has been enhanced more than two-fold, and synthesis of additional signal molecule, HHL (hexanoyl homoserine lactone), has been improved from an undetectable level to a level comparable to BHL, indicating a significant change in substrate specificity. We demonstrated the application of the engineered LuxI-LuxR quorum-sensing systems for regulation of gene expression. Although inducible gene expression systems are commonly used to enhance production of toxic proteins in lab cultures, the high cost of inducers prevents their application at an industrial scale, and therefore an auto-inducible and tunable gene expression system is an attractive alternative. Engineered LuxI-LuxR autoinduction system is highly tunable to a desired cell density for the optimal production of proteins of various toxicities. In particular, expression of Nun toxic protein when expressed under engineered LuxI-LuxR system with delayed autoinduction in the mid-late log phase was detectable on western blotting whereas constitute expression resulted in complete inhibition of cell growth. By coupling with the Cre-loxP site-specific recombination system, cell density-dependent deactivation of gene expression was also accomplished. Taking together, we have created a highly tunable and auto-inducible system for activating or deactivating gene expression. Using LuxI-LuxR and its homologous systems, we designed a synthetic symbiotic two species ecosystem for application to mixed cultures. Dynamic simulation and bifurcation analysis showed that the designed system admits a stable coexistence and the steady-state fraction of each species could be altered by varying signaling molecule synthesis rates. The design also revealed stable periodic solutions over certain domains of parameter space.
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Sayut, Daniel J. "Engineering of artificial cellular circuits based on the LuxI-LuxR quorum-sensing system." 2010. https://scholarworks.umass.edu/dissertations/AAI3427568.
Full textAbstract:
Natural cellular networks are very good at processing diverse inputs, generating complicated responses, and confounding researchers with their complexities. As an alternative to traditional cellular engineering approaches, the field of synthetic biology attempts to avoid the complexities of natural systems by focusing on the bottom-up construction of artificial cellular circuits. By rationally building up circuit complexity, synthetic biologists hope to both create novel systems capable of programming unique cellular responses, and gain insights into the design principles of natural systems. Circuits that allow for the programming of intercellular responses are of particular interest, and researchers have focused on the use of bacterial communication mechanisms (quorum sensing) to construct such circuits. At their most basic, quorum-sensing systems are composed of three main components, making them amenable to genetic manipulation. These components, however, have properties that have been finely tuned through evolution to function in very specific ways, and repurposing them for our own uses requires methods to overcome their naturally evolved properties. This thesis details our work in the construction and engineering of synthetic circuits based on components of the LuxI-LuxR quorum-sensing system. Using these components, we demonstrate methods for altering both the sensitivity and the form of the quorum-sensing response through the creation of three unique systems: an ultrasensitive positive feedback loop, a logical AND gate, and a coupled feedback loop oscillator. Construction and tuning of each circuit’s properties were achieved through a mixture of rational and evolutionary approaches, with particular emphasis on the directed evolution of the LuxR transcriptional activator. Mathematical modeling was also used during the construction of the more complex circuits to predict the properties that were essential to their functionalities. With the construction and characterization of these circuits, we have provided both well-defined modules that can be used in the construction of more complex systems, and developed methods that will allow for the creation and engineering of additional synthetic circuits.
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Sayut, Daniel Jon. "Engineering of Artificial Cellular Circuits Based on the LuxI-LuxR Quorum-Sensing System." 2010. https://scholarworks.umass.edu/open_access_dissertations/306.
Full textAbstract:
Natural cellular networks are very good at processing diverse inputs, generating complicated responses, and confounding researchers with their complexities. As an alternative to traditional cellular engineering approaches, the field of synthetic biology attempts to avoid the complexities of natural systems by focusing on the bottom-up construction of artificial cellular circuits. By rationally building up circuit complexity, synthetic biologists hope to both create novel systems capable of programming unique cellular responses, and gain insights into the design principles of natural systems. Circuits that allow for the programming of intercellular responses are of particular interest, and researchers have focused on the use of bacterial communication mechanisms (quorum sensing) to construct such circuits. At their most basic, quorum-sensing systems are composed of three main components, making them amenable to genetic manipulation. These components, however, have properties that have been finely tuned through evolution to function in very specific ways, and repurposing them for our own uses requires methods to overcome their naturally evolved properties. This thesis details our work in the construction and engineering of synthetic circuits based on components of the LuxI-LuxR quorum-sensing system. Using these components, we demonstrate methods for altering both the sensitivity and the form of the quorum-sensing response through the creation of three unique systems: an ultrasensitive positive feedback loop, a logical AND gate, and a coupled feedback loop oscillator. Construction and tuning of each circuit's properties were achieved through a mixture of rational and evolutionary approaches, with particular emphasis on the directed evolution of the LuxR transcriptional activator. Mathematical modeling was also used during the construction of the more complex circuits to predict the properties that were essential to their functionalities. With the construction and characterization of these circuits, we have provided both well-defined modules that can be used in the construction of more complex systems, and developed methods that will allow for the creation and engineering of additional synthetic circuits.
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Collins, Cynthia Hollie. "Directed Evolution of the Transcriptional Activator LuxR." Thesis, 2006. https://thesis.library.caltech.edu/5146/1/CHC_Thesis.pdf.
Full textAbstract:
LuxR-type proteins are responsible for density-dependent transcriptional regulation in quorum-sensing systems that employ acyl-homoserine lactones (acyl-HSLs) as signal molecules. The Vibrio fischeri lux operon, which includes LuxR, has been well studied and shown to function in E. coli. The components of the lux operon have been used to engineer de novo genetic circuits because they provide a versatile intercellular communications system. We have used directed evolution to engineer LuxR to generate new components for engineering genetic circuits with a wide range of communications functions and to explore the evolution of LuxR specificity and response.
We developed and validated a screening system to identify mutants of LuxR that activate gene expression with non-cognate acyl-HSLs. We screened libraries of luxR mutants for variants exhibiting increased gene activation in response to octanoyl-HSL (C8HSL). Eight LuxR variants were identified that showed a 100-fold increase in sensitivity to C8HSL. These variants displayed increased sensitivities to a broadened range of acyl-HSLs while maintaining a wild-type or greater response to LuxR’s cognate signal, 3-oxo-hexanoyl-homoserine lactone (3OC6HSL).
To generate a LuxR with a new signaling specificity, we used a dual selection system to identify LuxR variants that either activate gene expression (ON) or do not (OFF) under desired sets of conditions. The dual selection system was evaluated prior to its use, and a 490-fold enrichment in functional clones per round of ON/OFF selection was observed. We used the dual selection system to identify a LuxR variant that responds to straight-chain acyl-HSLs, but no longer responds to 3OC6HSL. A single mutation in the N terminal signal-binding domain reduces the response to acyl-HSLs having a carbonyl substituent at the third carbon of the acyl chain.
We used the dual selection system to identify LuxR variants that activate transcription upon binding to a promoter containing a mutated operator site. An amino acid position critical for modulating the DNA-binding specificity of LuxR for the new DNA target was identified. The dual selection system provides a rapid and reliable method for identifying LuxR variants that have the desired response, or lack thereof, to a given set of acyl-HSL signals or DNA targets.