Introduction: Early growth response 1 (EGR1) is a zinc-finger transcription factor involved in the regulation of cell growth. It can act as either a tumour suppressor or a tumour promoter with a role in the induction of apoptosis in cancer cells by various pathways and is likely to play a role in colorectal cancer (CRC). EGR1 also appears to play a significant role in inflammatory pathways, therefore a possible role in Inflammatory Bowel Disease (IBD) is hypothesised. Patients with IBD have a greater risk of developing CRC, which is increased with duration of symptoms and severity of inflammation and dysplasia. The aim of this study is to determine whether EGR1 is differentially expressed in diseased colon tissue and to investigate novel EGR1-protein interactions in CRC cell lines. Methods: The relative EGR1 expression in CRC cell lines and in normal mucosa and tumours of colorectal cancer patients was determined by qRT-PCR. IBD patient samples were also examined for differential EGR1 expression levels by qRT-PCR, before and after stimulation with inflammatory mediators. Statistical analysis of the data was performed using ‘R’ statistical package, with the mixed-model ANOVA. Statistical significance was set at < 0.05. The genotype of three EGR1 variants was determined in the samples using PCR and sequencing, and the methylation status of regions of the EGR1 promoter was determined using bisulfite sequencing. A yeasttwo hybrid screen was conducted with EGR1 as bait, and screened against a SW480 CRC cell line library. Interesting novel interactions were investigated using immunocytochemistry and immunoprecipitation, as was the novel interaction between EGR1 and NOD2 and between EGR1 and components of the cytoskeleton. Results: Investigation into the relative EGR1 mRNA expression in CRC has shown that there is differential expression of EGR1 between matched normal mucosa and tumour. EGR1 expression is decreased in IBD patients compared with healthy controls. Induction of EGR1 by inflammatory stimuli also appears to be aberrant in these patients. The differential expression of EGR1 was not associated with aberrant methylation of a large region of the EGR1 promoter in either the CRC or IBD patients or with the genotype of EGR1 variants. EGR1 localises to both the cytoplasm and the nucleus in CRC cell lines and this study demonstrate interactions with the IBD susceptibility protein NOD2 and with components of the cyotskeleton. A yeast-two hybrid screen conducted with EGR1 as bait using a CRC cell line library has identified several other novel protein interactions of EGR1 in CRC cell lines. Conclusion: EGR1 is differentially expressed in both CRC and IBD, and in the case of IBD shows aberrant activity, suggesting that EGR1 may play a role in both colorectal diseases. EGR1 interacts with the IBD protein NOD2, and components of the cytoskeleton in CRC cells. Several novel protein interactions with EGR1 have been identified and warrant further study.

Heart failure represents an important cause of death in Western Countries. The pathophysiology of heart failure is mainly associated with abnormalities in intracellular calcium control. We previously showed that Egr-1 negatively regulates expression of sodium-calcium exchanger (NCX) in vivo and in vitro. Here we tested the hypothesis that Egr-1 regulates expression of calcium storage proteins in the sarco-endoplasmic reticulum (SER), calsequestrin (CSQ) and/or ER, calreticulin (CRT) directly or indirectly via Egr-1:NFAT (nuclear factor of activated T-cells) formation. Secondarily, we hypothesized that this will reduce calcium mobilization. We found that undifferentiated 1293F cells, overexpressing Egr-1, have reduced CSQ compared to control H9c2 cells. We demonstrated that Egr-1 negatively regulates CSQ but not CRT expression. The Egr-1 mediated decrease in CSQ is linked to decreased calcium availability. Repression is by a novel NAB-independent (NGFI-A binding protein) activity localized to a.a. region 1-307. We conclude that Egr-1-mediated reductions in calcium storage protein expression alter calcium availability for cardiac contraction/relaxation.

Thesis (Ph. D.)--West Virginia University, 2007. / Title from document title page. Document formatted into pages; contains xv, 156 p. : ill. (some col.). Vita. Includes abstract. Includes bibliographical references (p. 112-153).

Les protéagineux pourraient contribuer à réduire le déficit en protéines de l’Europe mais ils sont peu cultivés. L’association à une céréale permettrait de contourner leur variabilité de rendement et leur faible compétitivité vis-à-vis des adventices. Cette stratégie serait particulièrement pertinente pour le lupin blanc d’hiver, qui présente un fort potentiel de production de protéines mais un fort risque d’enherbement. Ce travail de thèse étudie en dynamique les interactions entre lupin, céréale et adventices afin de mettre en évidence les traits des espèces permettant d’accroitre leur complémentarité et la compétitivité vis-à-vis des adventices tout en sécurisant la productivité. L’étude d’un réseau de parcelles démontre que l’ajout de triticale permet d’augmenter la compétitivité vis-à-vis des adventices par rapport au lupin pur, d’obtenir un rendement supérieur et plus stable et de maintenir la production de protéines. Un essai en micro parcelles démontre le rôle majeur de la capture précoce d’azote (N) de la céréale pour accroitre la compétitivité vis-à-vis des adventices. Nous montrons que les rapports de compétition varient au cours du cycle et en fonction des espèces ou variétés ainsi que des conditions climatiques et influencent ainsi l’élaboration du rendement. De forts contrastes entre lupin et céréale pour des traits liés à l’acquisition précoce d’N sont décrits ainsi que leurs conséquences sur la mise en place de la complémentarité. Ce travail met en évidence que le choix d’espèces et de variétés aux traits complémentaires permet d’orienter les interactions entre lupin, céréale et adventices et ouvre des perspectives pour l’optimisation des performances. / Pea, faba bean and lupin could help to reduce the European protein deficit but they are not widely grown. Intercropping with a cereal can be seen as a way to circumvent their high yield variability and their low competitive ability against weeds. This strategy could be particularly relevant for winter white lupin, a crop that can produce high amounts of proteins but that is prone to high weed infestations. This doctoral thesis studies the dynamic interactions between lupin, cereal and weeds to highlight species traits that allow increasing their complementarity and their competitive ability against weeds while securing productivity. Results of a multisite study in agricultural fields demonstrate that adding triticale in lupin allows increasing competitive ability against weeds as well as yield and yield stability compared with sole-cropped lupin while maintaining protein production. A plot experiment highlights the key role of cereal early Nitrogen (N) uptake in the increase of competitive ability against weeds. Analysis of resource sharing shows that competitive relationships vary throughout the cropping cycle, depend on species or cultivars as well as climatic conditions and that they influence yield formation. Strong contrasts between lupin and a cereal for traits related to early soil N uptake and their consequences on complementarity are highlighted by the results of rhizotron experiments. This work emphasizes the role of species and cultivar choice to manage interactions between lupin, cereal and weeds and opens perspectives for the optimization of intercrop performances.

Abstract Swedish start-ups seem to efficiently develop new products and services but less successful when it comes to taking them to the market and launching them globally. This research aims to address this gap through investigating Swedish start-ups. In light of this problem, the author argues that there is a need for increasing knowledge regarding the financial success factors and challenges facing Swedish start-ups in the go-to-market phase. The research question is thus: What are the significant challenges and success factors affecting the financing of Swedish start-ups during the go-to-market phase? Four major challenges facing start-ups during the go-to-market phase are identified. These are lack ofsufficient capital,lack of support from the banking sector, lack of support from the regional public leveland, finally, regulations and legal issues. On the other hand, four success factors have been identified, which are support from private investors, shared financial private/public risk, efficient internal operationsand non-traditional financing methods. In order to facilitate the go-to-market financing, it is recommended that start-ups be established in a business incubator environment in order to have access to investor networks and other financial support. Besides, having investors with financial experience in the start-up boards increases the chances of success in the go-to-market phase. The banking sector in Sweden should also play a bigger role in the strategic issues in order to accelerate the start-up’s growth. For further studies, more knowledge regarding the underlying motivations of private capitalists, public funders and loan lenders is desirable. Furthermore, the business incubator’s role in facilitating the financing of start-ups and understanding how start-ups in different sectors should act to increase the rate of success are two important areas for future studies.  Keywords: start-ups, go-to-market, financing, growth, early growth, success factors

Accroître la compétitivité des cultures vis-à-vis des adventices est un levier essentiel pour concevoir des systèmes de culture moins dépendants des herbicides. Les légumineuses sont des espèces majeures pour diversifier les systèmes mais elles sont réputées peu compétitives vis-à-vis des adventices. Cependant la variabilité entre espèces en particulier pendant la phase précoce a été peu explorée. Ce travail vise à i) étudier, via deux expérimentations en rhizotrons en serre, les traits impliqués dans la croissance et l’acquisition d’azote (N) en début de cycle chez une gamme d’espèces de légumineuse et les conséquences sur la compétition légumineuse-adventice et à ii) quantifier au champ l’intérêt d’associations légumineuse-non légumineuse (colzalégumineuses, maïs-légumineuses pérennes, la légumineuse étant plante de service) sur la croissance et la composition de la flore adventice. Nous montrons qu’il existe une variabilité entre espèces de capture d’N en début de cycle en lien avec les réserves azotées de la semence et l’exploration racinaire. La croissance et l’acquisition d’N de l’adventice sont impactées différemment selon l’espèce de légumineuse pendant la phase précoce. Les essais au champ montrent l’intérêt de combiner des espèces aux traits complémentaires pour l’utilisation des ressources conduisant à un meilleur contrôle des adventices tout en maintenant la productivité de la culture. Cette étude montre aussi que la composition de la flore est modifiée en fonction de la légumineuse insérée dans le système en lien à la fois avec les traits des légumineuses et les traits de réponse des adventices. / Enhancing crop competitiveness against weeds is a relevant way in order to design cropping system less dependent on herbicides. Legumes are key species in diversified cropping systems but they are known to be low competitive against weeds. However, inter-specific variability especially in the early growth had been little studied. Our work aims to i) study, via two greenhouse experiments in rhizotrons, the traits involved in the growth and nitrogen (N) acquisition for a range of legume species and the impacts on weed-legume competition at the beginning of crop cycle and to ii) quantify under field experiments the interest of legume-non legume intercropping (oilseed rapelegumes, maize-perennial legumes, where legume species were grown as service plant) on weed growth and weed species composition. Our results highlighted the inter-specific variability for soil N uptake in the early growth in relation with seed reserves and rooting exploration. Weed growth and N acquisition are influenced differently as a function of legume species in the early growth. Field experiments show the advantage of species combinations with complementary traits for using resources, leading to better weed control while maintaining crop productivity. This study shows also that weed species composition was modified as a function of legume species in relation with both legume traits and weed traits response.

Introduction Des études épidémiologiques ont montré qu'une restriction nutritionnelle pendant la grossesse pouvait conduire à une réponse adaptative du fœtus conduisant à une altération durable du métabolisme. Ainsi, les enfants nés avec un petit poids de naissance sont plus à risque de développer des maladies cardiovasculaires à l'âge adulte. L'exposition à des contaminants obésogènes pourrait également jouer un rôle dans l'augmentation du risque d'obésité. L’alimentation est une des principales voies d'exposition à ces contaminants. Après une description de la qualité de l’alimentation pendant la grossesse, l’objectif était d'étudier l'association entre l'exposition prénatale aux contaminants alimentaires et la croissance des enfants.MéthodesLes données de trois cohortes de naissance ont été utilisées : deux études françaises (EDEN et ELFE), et une étude norvégienne (MoBa). Les femmes de ces cohortes, ont rempli un questionnaire de fréquence alimentaire (QFA) portant sur l’alimentation pendant la grossesse. L'évaluation des contaminants a été réalisée en combinant le QFA et des bases de données de contamination, la deuxième Etude de l’Alimentation Total (EAT2) française pour EDEN et plusieurs bases de données de contamination pour MoBa. Premièrement dans ELFE, nous avons créé un score de qualité de l'alimentation et un score spécifique à la grossesse, puis nous avons étudié les facteurs associés à une meilleure qualité alimentaire. Ensuite, nous avons examiné l’association entre la qualité de l'alimentation et la croissance prénatale. Deuxièmement, nous avons étudié la relation entre un contaminant alimentaire : l'acrylamide (AA), et la croissance prénatale, dans EDEN, et la croissance postnatale, dans MoBa. Troisièmement, nous avons étendu nos analyses à tous les contaminants alimentaires de l’EAT2, en analysant les composés pris individuellement, puis considérés en mélange.RésultatsPremièrement, nous avons montré que les recommandations générales et spécifiques étaient globalement bien suivies par les femmes enceintes. Certaines caractéristiques socioéconomiques ou démographiques étaient associées à la fois au score de qualité de l’alimentation et au score spécifique de la grossesse telles que l’âge à l’accouchement, niveau d’étude et revenu du foyer et le tabagisme. Un score élevé de qualité de l’alimentation était associé à un poids de naissance plus élevé et un risque plus faible d'avoir un enfant petit pour l’âge gestationnel (PAG).Deuxièmement, nous avons montré que plus l'exposition pendant la grossesse à l’AA est importante, plus la taille de naissance était faible et plus le risque de PAG est élevé. Dans MoBa, nous avons constaté que l'exposition prénatale à l'AA était associée à une prévalence accrue d'enfants en surpoids ou obèses et à une plus grande vitesse de croissance du poids durant l’enfance.Dans EDEN, sur les 99 composés chimiques sélectionnés, le poids de naissance était associé négativement à l’exposition à quatre contaminants alimentaires et positivement à l’exposition à quatre autres. L'IMC à 5 ans était associé négativement à un contaminant. Aucune de ces associations étaient statistiquement significative après prise en compte de la multiplicité des tests. Lorsque les composés chimiques étaient considérés en mélanges, un mélange de contaminants était positivement associé au poids de naissance et aucun mélange n’était associé à l'IMC à 5 ans.ConclusionUne qualité de l’alimentation élevée est associée à un poids de naissance plus élevé et une diminution du risque de PAG, alors que l'exposition alimentaire à l'AA est associée à une altération de la croissance fœtale. L’exposition prénatale aux contaminants alimentaires, évaluée à partir des données d’EAT, ne semble pas préoccupante vis-à-vis de la croissance prénatale et postnatale précoce car les effets retrouvés sont de faible amplitude et ne sont plus significatifs après prise en compte des tests multiples. / Introduction Maternal diet is the only prenatal source of nutrients and the major source for non-nutrients and can influence foetal growth and offspring’s long-term health. Chemicals known as “obesogens” might also play a role in increasing obesity risk and one of the main route of exposure to these chemicals is through foods. After a description of diet quality during pregnancy, the aim of this thesis was to study the association between prenatal exposure to food chemicals and prenatal and postnatal growth among children.MethodsWe used the data of three birth cohort studies: two French studies (EDEN and ELFE), and a Norwegian study (MoBa). We first described the compliance to dietary guidelines of French pregnant women and the impact of a better diet quality on birth outcomes using the data of the ELFE study. Secondly, we studied the relationship of a specific food contaminant, acrylamide, on birth size in the EDEN and postnatal growth in MoBa. Finally, we extended analyses to all available food chemicals in the second French Total Diet Study (TDS).ResultsThe nutritional guidelines for pregnant women were rather well followed in ELFE. The diet quality score was associated with higher birth weight and lower risk of having a small for gestational (SGA) baby. We showed that prenatal dietary exposure to acrylamide was associated with reduced birth size in EDEN, and to increased postnatal growth in MoBA. In EDEN, on the 99 selected food chemicals, birth weight was associated with eight chemicals (four negatively and four positively). BMI at 5 years was associated with one food chemical. These results were not significant after correction for multiple testing. When using the mixture approach, one mixture of chemical was positively associated with postnatal growth.ConclusionsThe diet quality score was associated with higher birth size. Whereas dietary exposure to acrylamide was associated with impaired foetal growth, when looking at a larger number of food chemicals, we did not find concerning association on child prenatal or postnatal growth. Exposure to food chemical assessed by TDS did not appear to be of major concern for growth but other windows of susceptibility, such as early childhood, and other outcomes, such as cognitive development, should be considered in future studies.

All cells and organisms consume energy for survival. A robust system has evolved in vertebrates to serve as an energy reservoir. In particular, specialized cells, adipocytes, are responsible for the dynamic storage of energy by accumulating and releasing fatty acids. Fluctuating energy demands require adipose tissue to adjust in size, however complications can arise in both extremes giving rise to systemic diseases, such as obesity and diabetes mellitus (T2D). In mammals, leptin production in adipocytes is up-regulated by feeding and insulin to provide long-term post-prandial satiety. Although this regulatory connection is central to all physiological effects of leptin, the molecular mechanism remains unknown for leptin production. Here, we show that the transcription factor Egr1 is rapidly but transiently induced by insulin in adipose cells both in vitro and in vivo in a mTORC1-dependent fashion. Induction of Egr1 was immediately followed by an increase in leptin transcription. Chromatin immunoprecipitation and luciferase assays demonstrate that Egr1 directly binds to and activates the leptin promoter. Interestingly, the lipid droplet protein Fat specific protein 27 (FSP27) may work as a co-factor for Egr1 in regulating leptin expression. By using siRNA-mediated knock out of Egr1 along with its over-expression in adipocytes, we demonstrate that Egr1 is both necessary and sufficient for the stimulatory effect of insulin on leptin transcription. Knockout of the mTORC1-regulated translation repressor 4EBP1/2 increases leptin transcription both in vitro and in vivo. Adipose specific doxycycline-inducible constitutively active Rheb transgenic mouse lines contained higher circulating leptin and transcription of leptin following doxycycline treatment and were able to maintain elevated leptin levels following a 16 hour fast. Thus, insulin and nutrients, such as amino acids and glucose, activate leptin expression via the mTORC1-Egr1 regulatory axis.

No description available.

Angiopoietin-1 (Ang-1), the main ligand for the endothelial cell (EC)-selective Tie-2 receptors, promotes survival, proliferation, migration and differentiation of these cells. Despite its importance in various aspects of vascular biology, the mechanisms of action of the Ang-1/Tie-2 receptor pathway have not been fully explored. / To identify the downstream modulators of Ang-1, we evaluated changes in the transcriptome of human umbilical vein endothelial cells (HUVECs) treated with Ang-1 protein for four hours by employing the oligonucleotide rnicroarray technology. Eighty-six genes were significantly upregulated by this treatment and forty-nine genes were significantly downregulated. These genes are involved in the regulation of cell cycle, proliferation, apoptosis, transcription and differentiation. Furthermore, we found that the Erk1/2, PI3-Kinase and mTOR pathways are implicated in promoting gene expression in HUVECs in response to Ang-1. Analysis of the microarray data employing the Ingenuity Pathways analysis software to place the regulated genes in the context of biological networks revealed several highly connected nodes including the chemokine Interleukin-8 (IL-8) and the transcription factor Early growth response-1 (Egr-1). Due to the importance of these genes in promoting angiogenesis, we decided to evaluate their roles in Ang-1/Tie-2 receptor signaling and biological effects. / Ang-1 induced IL-8 expression in a time- and dose-dependent manner in ECs through both transcriptional and post-transcriptional mechanisms. To study the functional role of Ang-1-induced IL-8, we generated HUVECs that overexpress Ang-1. In these cells, neutralizing IL-8 significantly reduced EC proliferation and migration. IL-8 promoter activity experiments and gel shift assays revealed the involvement of the transcription factor AP-1 in Ang-1-induced IL-8. Ang-1 stimulated the phosphorylation of c-Jun through activation of Erk1/2, JNK and PI-3 kinase pathways. Similarly, Ang-1 provoked the expression and DNA binding of Egr-1 in HUVECs. Employing siRNA and DNAzyme to specifically knock-down Egr-1, we found that Ang-1-induced Egr-1 also promotes EC proliferation and migration. / We conclude that Ang-1 provokes a coordinated response intended to promote EC survival, proliferation, and angiogenesis and to inhibit EC apoptosis. Ang-1 induces EC proliferation and migration in part through the secretion of the soluble mediator Interleukin-8 and through induction of the transcription factor Egr-1.