La régulation transcriptionnelle de l'expression génique dans le fruit de tomate : caractérisation fonctionnelle de promoteurs fruit-spécifiques et d'un co-facteur de la transcription de type MBF1

Les travaux présentés dans ce manuscrit s'intéressent à la régulation transcriptionnelle de l'expression des gènes au cours du développement du fruit de tomate à travers l'étude des cis-régulations et des trans-régulations. Dans un premier temps, une tentative de caractérisation de promoteur dirigeant l'expression génique spécifiquement dans le fruit a été menée. Pour cela, nous nous sommes focalisés sur l'étude de régions promotrices d'un gène codant une expansine et d'un autre codant une alcool acyl transférase. Ces études ont été menées en évaluant la capacité de ces régions promotrices à diriger l'expression du gène rapporteur codant pour la β-glucuronidase dans des plants de tomate et d'Arabidopsis. Néanmoins, ces approches fonctionnelles n'ont pas permis de dégager un motif consensus capable à lui seul de conférer une expression fruit-spécifique. C'est pourquoi, les travaux de thèse se sont orientés vers la caractérisation d'un co-activateur de transcription de type MBF1 (Multiprotein Bridging Factor1), SlER24 de tomate. Nous avons montré qu'une fusion de SlER24 à un motif répresseur EAR (Ethylene-responsive element-binding associated Amphiphilic Repression) est capable, dans un système cellulaire, de réduire l'expression du gène rapporteur GFP dirigée par un promoteur synthétique de réponse à l'éthylène. Cette validation fonctionnelle par une méthode transitoire, nous a conduit à surexprimer cette construction, de façon stable, dans la tomate MicroTom. Cette surexpression de la construction chimérique SlER24-EAR, induit un retard de germination mais n'a pas d'effet notable sur le développement de la plante. Une fusion similaire de son orthologue AtMBF1c au motif EAR entraîne chez Arabidopsis thaliana une réduction du pourcentage de germination et un nanisme de la plante. Au-delà de l'information sur le rôle des gènes MBF1 dans le développement des plantes, cette étude démontre que la stratégie utilisant le domaine répresseur EAR n'est pas seulement efficace pour l'étude des facteurs de transcription comme cela a été démontré jusque là, mais également pour les co-activateurs ne se liant pas directement à l'ADN. ABSTRACT : The work presented in this manuscript is focused on the study in tomato fruit of gene expression regulation at the transcriptional level and comprises two parts. The first part concerns the isolation and characterization of promoters capable of specifically driving gene expression in tomato fruit. For this purpose, the capacity of native promoter regions and various deletions from two tomato genes, one encoding an expansin and a second encoding an alcohol acyl transferase to drive the expression of the GUS reporter gene, has been studied after stable transformation. This approach does not allow us to define a consensus motif able to drive the activity of a minimal specifically to the fruit. This is why the second part of the thesis was redirected to the characterization of a transcriptional co-activator of the MBF1 family, SlER24, in tomato. It is shown here that a chimerical fusion of SlER24 with the EAR (Ethylene-responsive element-binding associated Amphiphilic Repression) motif is capable of reducing the expression of the GFP reporter gene, driven by a synthetic ethylene-responsive promoter, in a transient cell system. This functional validation led us to express stably the construct in tomato plant using MicroTom for practical reasons. The overexpression of the SlER24/EAR motif fusion caused a germination delay but it had no significant effect on the tomato plant growth. A similar fusion using AtMBF1c from Arabidopsis, caused a reduction in the percentage of seed germination and dwarfism of the plant. Considering the role of the MBF1 genes in plant development, this study demonstrates that the EAR strategy is efficient not only for transcription factors, as demonstrated so far, but also in the case of co-activators known to not bind directly to DN

The Patient's View of Nursing Care after Hip Fracture.

Background: The pathway for patients with a hip fracture described in this study is a fast track. Many studies have focused on prevention of various complications but, so far, the patient's view of nursing care has not been highlighted. Aim: The aim of the study is to illuminate the patient's view on nursing care when treated for a hip fracture. Method: Ten patients were interviewed. A content analysis design was conducted. Findings. From the analysis, four main categories emerged: waiting times; pain/pain relief and mobilisation; attitude/information and sense of security; complications. Conclusion: Patients generally felt satisfied with the nursing provided. The staff created a feeling of security and showed interest and empathy for the patient. However, patients experienced a stressful waiting for surgery, and patients who developed confusion waited more than 24 hours for surgery. Therefore, waiting time must be decreased. Furthermore, patients' descriptions of a variety of pain problem show, for example, that good collaboration between the nurse and physiotherapist is critical for achieving good pain relief before mobilisation. Nursing staff need to be attentive and should elicit the patient's feelings through patient-focused communication in order to relieve anxiety about going home

Kindergarten as an arena for learning

Bacheloroppgave i barnehagelærerutdanning, Høgskolen i Innlandet, 2018.Norsk: I min bachelor oppgave har jeg valgt å se nærmere på det læringstrykket barnehagen i dag står ovenfor, og hvordan dette læringstrykket kan utfordre barnehagelærerens praksis og forståelse av læring. Jeg har med det formulert problemstillingen slik: «Hvordan utfordrer et stadig økende læringstrykk i barnehagen mitt syn på læring?» For å belyse min problemstilling har jeg valgt å gjøre en diskursanalyse av to utvalgte årsplaner og analysert hvilke læringsdiskurser som råder i de ulike barnehagene. Mine analysefunn vil ligge til grunn for den videre analyse opp mot relevant litteratur. Målet med denne oppgaven har vært å synliggjøre ulike læringssyn som kan eksistere innenfor helhetsmodellen som norske barnehager bygger på.English: In my bachelor thesis I have chosen to take a closer look at the pressure to learn which kindergartens today face and how this pressure can challenge how the kindergarten personnel teach and their understanding of learning. I formulate my way of looking at the problem in this way: “How does an ever increasing pressure to learn challenge my view on learning?” To clarify this problem I have chosen to make a discursive analysis of two different kindergartens and analyzed which learning discourses dominate in those kindergartens. While looking into relevant literature, a further analysis will be based on my previous finds. This paper strives to uncover the different learning views which can exist within the comprehensive model on which Norwegian kindergartens are based

Over-expression of a chimeric gene of the transcriptional co-activator MBF1 fused to the EAR repressor motif causes developmental alteration in Arabidopsis and tomato

Transcriptional co-activators of the Multiprotein Bridging Factor1 (MBF1) type belong to a multigenic family that encode key components of the machinery controlling gene expression by communicating between transcription factors and the basal transcription machinery. Knocking-down the expression of one member of the family has proved difficult probably due to functional redundancy. We show here that a fusion of SlER24, an MBF1 type gene of tomato, to the Ethylene-responsive element-binding associated Amphiphilic Repression (EAR) motif is capable of slowing down significantly the expression of the GFP protein driven by a synthetic ethylene-responsive GCC-rich promoter in a single cell transient expression system. A fusion of AtMBF1c of Arabidopsis to EAR, driven by the 35S promoter, caused a reduction of the percentage of seed germination and dwarfism of the plant. Similar fusion with the SlER24 of tomato in the MicroTom cultivar induced a delay of seed germination and no obvious effect on plant growth. Besides giving information on the role of the MBF1 genes in plant development, this study demonstrates that the EAR strategy is efficient not only for regular transcription factors as demonstrated so far, but also in the case of co-activators known to not bind directly to DNA

Hantavirus Pulmonary Syndrome, French Guiana

A systematic serological survey of patients suffering from symptoms suggestive of Hantavirus pulmonary syndrome allowed us to identify a native case in French Guiana. Partial molecular characterization of the implicated hantavirus revealed its close relationship with the Bolivian Rio Mamore virus. We tentatively named it Maripa virus

Validation of the Body Concealment Scale for Scleroderma (BCSS): Replication in the Scleroderma Patient-centered Intervention Network (SPIN) Cohort

© 2016 Elsevier Ltd Body concealment is an important component of appearance distress for individuals with disfiguring conditions, including scleroderma. The objective was to replicate the validation study of the Body Concealment Scale for Scleroderma (BCSS) among 897 scleroderma patients. The factor structure of the BCSS was evaluated using confirmatory factor analysis and the Multiple-Indicator Multiple-Cause model examined differential item functioning of SWAP items for sex and age. Internal consistency reliability was assessed via Cronbach's alpha. Construct validity was assessed by comparing the BCSS with a measure of body image distress and measures of mental health and pain intensity. Results replicated the original validation study, where a bifactor model provided the best fit. The BCSS demonstrated strong internal consistency reliability and construct validity. Findings further support the BCSS as a valid measure of body concealment in scleroderma and provide new evidence that scores can be compared and combined across sexes and ages

INVESTIGATION OF BENZENE METABOLISM BY LIQUID CHROMATOGRAPHY/ELECTROCHEMISTRY (THIOLS, MICROSOMES, PHENOL)

Long term exposure to benzene has been linked to leukemia and aplastic anemia as well as a number of other blood disorders. The mechanism by which benzene elicits this carcinogenic response is not yet been fully understood. In order to identify the reactive intermediate in benzene metabolism, a sensitive analytical method is needed. Liquid chromatography/electrochemistry offers a distinct advantage over other methods of analysis for the detection and quantitation of benzene metabolites. Most of the metabolites are electroactive and are easily detectable in the picomole range. Because of the specificity of the detector few, if any electroactive compounds interfere in the analysis. All of the metabolites of interest can be identified with a high degree of certainty based on retention time and electrochemical characteristics. The primary metabolite of benzene; phenol, was identified in both mouse liver and porcine bone marrow microsomal incubations. Hydroquinone was detected in the presence in microsomal incubations of benzene and ascorbic acid. The detection of the glutathione conjugate of p-benzoquinone in microsomal incubations by liquid chromatography/electrochemistry provided strong evidence that p-benzoquinone was being produced in these incubations. The role of p-benzoquinone as a reactive intermediate in benzene metabolism was studied. Mechanisms of formation and reduction of the quinone by microsomal enzymes were investigated. The role of glutathione in the detoxification of the quinone intermediate was also investigated. This involved the use of a novel form of LCEC which allows for the simultaneous determination of thiols and disulfides

Barnehagen som læringsarena

Norsk: I min bachelor oppgave har jeg valgt å se nærmere på det læringstrykket barnehagen i dag står ovenfor, og hvordan dette læringstrykket kan utfordre barnehagelærerens praksis og forståelse av læring. Jeg har med det formulert problemstillingen slik: «Hvordan utfordrer et stadig økende læringstrykk i barnehagen mitt syn på læring?» For å belyse min problemstilling har jeg valgt å gjøre en diskursanalyse av to utvalgte årsplaner og analysert hvilke læringsdiskurser som råder i de ulike barnehagene. Mine analysefunn vil ligge til grunn for den videre analyse opp mot relevant litteratur. Målet med denne oppgaven har vært å synliggjøre ulike læringssyn som kan eksistere innenfor helhetsmodellen som norske barnehager bygger på

La régulation transcriptionnelle de l'expression génique dans le fruit de tomate (Caractérisation fonctionnelle de promoteurs fruit-spécifiques et d'un co-facteur de la transcription de type MBF1)

The work presented in this manuscript is focused on the study in tomato fruit of gene expression regulation at the transcriptional level and comprises two parts. The first part concerns the isolation and characterization of promoters capable of specifically driving gene expression in tomato fruit. For this purpose, the capacity of native promoter regions and various deletions from two tomato genes, one encoding an expansin and a second encoding an alcohol acyl transferase to drive the expression of the GUS reporter gene, has been studied after stable transformation. This approach does not allow us to define a consensus motif able to drive the activity of a minimal specifically to the fruit. This is why the second part of the thesis was redirected to the characterization of a transcriptional co-activator of the MBF1 family, SlER24, in tomato. It is shown here that a chimerical fusion of SlER24 with the EAR (Ethylene-responsive element-binding associated Amphiphilic Repression) motif is capable of reducing the expression of the GFP reporter gene, driven by a synthetic ethylene-responsive promoter, in a transient cell system. This functional validation led us to express stably the construct in tomato plant using MicroTom for practical reasons. The overexpression of the SlER24/EAR motif fusion caused a germination delay but it had no significant effect on the tomato plant growth. A similar fusion using AtMBF1c from Arabidopsis, caused a reduction in the percentage of seed germination and dwarfism of the plant. Considering the role of the MBF1 genes in plant development, this study demonstrates that the EAR strategy is efficient not only for transcription factors, as demonstrated so far, but also in the case of co-activators known to not bind directly to DNALes travaux présentés dans ce manuscrit s'intéressent à la régulation transcriptionnelle de l'expression des gènes au cours du développement du fruit de tomate à travers l'étude des cis-régulations et des trans-régulations. Dans un premier temps, une tentative de caractérisation de promoteur dirigeant l'expression génique spécifiquement dans le fruit a été menée. Pour cela, nous nous sommes focalisés sur l'étude de régions promotrices d'un gène codant une expansine et d'un autre codant une alcool acyl transférase. Ces études ont été menées en évaluant la capacité de ces régions promotrices à diriger l'expression du gène rapporteur codant pour la b-glucuronidase dans des plants de tomate et d'Arabidopsis. Néanmoins, ces approches fonctionnelles n'ont pas permis de dégager un motif consensus capable à lui seul de conférer une expression fruit-spécifique. C'est pourquoi, les travaux de thèse se sont orientés vers la caractérisation d'un co-activateur de transcription de type MBF1 (Multiprotein Bridging Factor1), SlER24 de tomate. Nous avons montré qu'une fusion de SlER24 à un motif répresseur EAR (Ethylene-responsive element-binding associated Amphiphilic Repression) est capable, dans un système cellulaire, de réduire l'expression du gène rapporteur GFP dirigée par un promoteur synthétique de réponse à l'éthylène. Cette validation fonctionnelle par une méthode transitoire, nous a conduit à surexprimer cette construction, de façon stable, dans la tomate MicroTom. Cette surexpression de la construction chimérique SlER24-EAR, induit un retard de germination mais n'a pas d'effet notable sur le développement de la plante. Une fusion similaire de son orthologue AtMBF1c au motif EAR entraîne chez Arabidopsis thaliana une réduction du pourcentage de germination et un nanisme de la plante. Au-delà de l'information sur le rôle des gènes MBF1 dans le développement des plantes, cette étude démontre que la stratégie utilisant le domaine répresseur EAR n'est pas seulement efficace pour l'étude des facteurs de transcription comme cela a été démontré jusque là, mais également pour les co-activateurs ne se liant pas directement à l'ADNTOULOUSE-ENSAT-Documentation (315552324) / SudocSudocFranceF