Depuis quelques années, la plupart des constructeurs automobiles innovent en faisant appel aux techniques de la réalité virtuelle (RV). Cette approche possède un fort potentiel en termes de gain de temps et de réduction des coûts. Elle permet également d'évaluer de nouvelles approches liées au processus de conception lui-même. Cependant, un certain nombre de verrous technologiques et méthodologiques subsistent. Ils concernent en particulier (i) la simplification et la physicalisation des maquettes numériques issues des logiciels de CAO, (ii) le développement de configurations visuo-haptiques adaptées aux différentes tâches impliquées par le prototypage virtuel, et (iii) l'identification des retours sensoriels les plus pertinents, permettant à l'opérateur d'effectuer ces tâches efficacement. Un ensemble de problématiques soulevées par le service R&D de Valeo Lighting Systems (Angers) nous a conduit à traiter les trois aspects évoqués ci-dessus. Les tâches étudiées sont l'accessibilité, l'extraction et la manipulation de lampes sur prototype virtuel à l'échelle 1:1. Dans le manuscrit, nous commençons par aborder les limites du prototypage réel et les apports liés au prototypage virtuel. Ces apports sont examinés en particulier pour les phases de création et de validation des prototypes. Puis, nous présentons un état de l'art exhaustif des dispositifs d'affichage et des interfaces à retour d'effort, que nous avons classées en fonction de leur architecture mécanique et de leur support de référence. Ensuite, nous traitons la simplification et l'intégration de maquettes virtuelles ainsi que l'intégration du modèle biomécanique de l'opérateur humain. Différentes simulations permettant de valider la méthodologie d'intégration proposée sont décrites. Celles-ci sont basées sur l'utilisation de notre plate-forme de réalité virtuelle. Enfin, nous décrivons une série cohérente et progressive d'expérimentations permettant d'évaluer la pertinence et l'influence de différentes modalités sensorielles (visuelle, sonore, vibro-tactile, et kinesthésique) sur la performance humaine. L'objectif est d'identifier les avantages et inconvénients de ces retours d'information dans différentes configurations matérielles. Les résultats sont analysés via différents indicateurs de performance (temps de réalisation des tâches, précision de placement). Des données subjectives sont également recueillies via l'observation des sujets pendant l'exécution des tâches et à partir de questionnaires.

DNA is subject to attack by DNA damaging agents from both environmental and endogenous sources. In response to DNA damage, living organisms enhance expression of many related genes to facilitate DNA repair and survival. The SOS response is a well-understood prokaryotic regulatory cascade that controls the expression of more than 30 genes in response to DNA damage. However, in eukaryotic organisms from simple budding yeast to human, such a regulatory network has not been reported.<p>Previous research in our laboratory found that among DNA repair mutants of <i>Saccharomyces cerevisiae</i>, only rad6 and rad18 defective in the post-replication repair pathway significantly affected DNA damage induction of several genes examined. Rad6 and Rad18 form a ubiquitin conjugation-ligase complex and are required for the cellular tolerance to damaged DNA. Since the Rad6-Rad18 complex binds to single-stranded DNA, it may act as a DNA damage sensor required for the activation of DNA damage-induced transcription. We performed microarray analysis and found that the induction of up to 379 genes, including those involved in DNA repair, control of replication and transcription, regulation of the cell cycle and cell metabolism, are compromised in the rad6 and rad18 mutants. Although Rad6/Rad18 monoubiquitinates proliferating cell nuclear antigen (PCNA) following DNA damage to initiate a damage tolerance response, PCNA ubiquitination is not required for DNA damage induction. In budding yeast, cell-cycle checkpoints are involved in the control of DNA damage induction of gene expression through phosphorylation of a protein kinase Rad53 by two pathways represented by Rad24 and Sgs1. The Rad6-Rad18 complex appears to function in the Rad24 pathway and parallel to Sgs1. We further demonstrated that the Rad17 subunit of the 9-1-1 complex is subject to Rad6/Rad18- and DNA damage-dependent mono-ubiquitination and that the Rad17-Lys197 residue with flanking sequences homologous to Lys164 of PCNA is absolutely required for the DNA damage induction by Rad6-Rad18. Hence, by ubiquitinating two DNA clamps, PCNA and 9-1-1, the Rad6-Rad18 complex plays a central role in the cellular response to DNA damage by coordinating translesion synthesis, error-free bypass, homologous recombination, as well as transcriptional regulation, reminiscent of roles of RecA in <i>E. coli</i> cells.<p>Several individual genes have also been examined in this study to elucidate the regulatory mechanisms acting on specific DNA damage-inducible genes. In the microarray analysis, DDI2 and DDI3, two identical genes located in duplicated chromosomal regions, were identified due to the highest induction ratio (122-fold) after MMS treatment. Interestingly, DDI2/DDI3 can only be highly induced by SN2-type alkylating agents. Promoter deletion analysis mapped the putative upstream acting sequence (UASDDI2) responsible for 40% of basal expression and 90% of induced expression by MMS.<p>The CRT10 gene was identified through screening of the yeast deletion library for hydroxyurea (HU) resistance. CRT10 encodes a putative 957 amino acid, 110 kDa protein with a leucine repeat and a WD40 repeat near the N-terminus. Deletion of CRT10 resulted in an enhanced resistance to HU reminiscent of the inactivation of two other ribonucleotide reductase (Rnr) suppressors, CRT1 and SML1, which regulate Rnr activity at transcriptional and translational levels, respectively. Epistasis analysis indicates that CRT10 belongs to the CRT1 pathway but not the SML1 pathway. Indeed, deletion of CRT10 enhanced the survival of the mec1 null mutant and increased basal level and DNA damage-induced expression of RNR2 and RNR3, suggesting that Crt10 regulates RNR genes at the transcriptional level. Furthermore, the dun1 mutation is epistatic to crt10 with respect to both HU sensitivity and RNR gene expression. Interestingly, the expression of CRT10 itself is induced by DNA damaging agents and this induction requires DUN1, suggesting that CRT10 plays a role in cellular response to DNA damage and replication blocks. The CRT10 function appears to be achieved by positive regulation of the CRT1 transcript level, indicating that CRT10 is a component of the regulatory circuit.

Cell-cycle checkpoint proteins help maintain genomic integrity by sensing damaged DNA and initiating DNA repair or apoptosis. Checkpoint protein activation to cell-cycle damaging agents can involve post-translational modifications and these alterations provide a means to determine whether DNA in a cell is damaged or not. Steinmoeller et al. (2009) showed that checkpoint proteins are suitable biomarkers for detecting genotoxins in Oncorhynchus mykiss (rainbow trout). In this project, two evolutionarily conserved checkpoint proteins, Rad1 and Hus1, have been cloned from rainbow trout and antibodies against these proteins were developed. This is the first time that either Rad1 or Hus1 has been characterized in rainbow trout. For rtRad1, it was determined that the open-reading frame was 840bp, which encodes 279aa with a predicted protein size of 31kDa. The rtRad1 amino-acid sequence is highly conserved and contains conserved exonuclease and leucine zipper domains. RT-PCR was used to identify alternatively spliced variants of rtRad1 and it appears that these variants encode different sized Rad1 proteins that are tissue and cell-line specific. A Rad1 splice variant that encodes an 18kDa protein appears to be abundant only in heart tissue and in the RTgill-W1 and RTbrain-W1 cell-lines. A genotoxicity study was completed where RTgill-W1 and RTbrain-W1 cells were treated with bleomycin, which induces double-stranded DNA breaks. In RTgill-W1, levels of an 18kDa Rad1 protein increased in a dose-dependent manner while in RTbrain-W1 the Rad1 levels remained the same. It appears that this 18kDa Rad1 protein may be directly involved in maintaining genomic integrity and shows potential to be used as a genotoxicity biomarker. This is the first time that an isoform of Rad1 has shown to be modified in the presence of a damaging agent. Both Rad1 and Hus1 need to be further characterized to determine their usefulness as genotoxicity biomarkers.

DNA is subject to attack by DNA damaging agents from both environmental and endogenous sources. In response to DNA damage, living organisms enhance expression of many related genes to facilitate DNA repair and survival. The SOS response is a well-understood prokaryotic regulatory cascade that controls the expression of more than 30 genes in response to DNA damage. However, in eukaryotic organisms from simple budding yeast to human, such a regulatory network has not been reported.<p>Previous research in our laboratory found that among DNA repair mutants of <i>Saccharomyces cerevisiae</i>, only rad6 and rad18 defective in the post-replication repair pathway significantly affected DNA damage induction of several genes examined. Rad6 and Rad18 form a ubiquitin conjugation-ligase complex and are required for the cellular tolerance to damaged DNA. Since the Rad6-Rad18 complex binds to single-stranded DNA, it may act as a DNA damage sensor required for the activation of DNA damage-induced transcription. We performed microarray analysis and found that the induction of up to 379 genes, including those involved in DNA repair, control of replication and transcription, regulation of the cell cycle and cell metabolism, are compromised in the rad6 and rad18 mutants. Although Rad6/Rad18 monoubiquitinates proliferating cell nuclear antigen (PCNA) following DNA damage to initiate a damage tolerance response, PCNA ubiquitination is not required for DNA damage induction. In budding yeast, cell-cycle checkpoints are involved in the control of DNA damage induction of gene expression through phosphorylation of a protein kinase Rad53 by two pathways represented by Rad24 and Sgs1. The Rad6-Rad18 complex appears to function in the Rad24 pathway and parallel to Sgs1. We further demonstrated that the Rad17 subunit of the 9-1-1 complex is subject to Rad6/Rad18- and DNA damage-dependent mono-ubiquitination and that the Rad17-Lys197 residue with flanking sequences homologous to Lys164 of PCNA is absolutely required for the DNA damage induction by Rad6-Rad18. Hence, by ubiquitinating two DNA clamps, PCNA and 9-1-1, the Rad6-Rad18 complex plays a central role in the cellular response to DNA damage by coordinating translesion synthesis, error-free bypass, homologous recombination, as well as transcriptional regulation, reminiscent of roles of RecA in <i>E. coli</i> cells.<p>Several individual genes have also been examined in this study to elucidate the regulatory mechanisms acting on specific DNA damage-inducible genes. In the microarray analysis, DDI2 and DDI3, two identical genes located in duplicated chromosomal regions, were identified due to the highest induction ratio (122-fold) after MMS treatment. Interestingly, DDI2/DDI3 can only be highly induced by SN2-type alkylating agents. Promoter deletion analysis mapped the putative upstream acting sequence (UASDDI2) responsible for 40% of basal expression and 90% of induced expression by MMS.<p>The CRT10 gene was identified through screening of the yeast deletion library for hydroxyurea (HU) resistance. CRT10 encodes a putative 957 amino acid, 110 kDa protein with a leucine repeat and a WD40 repeat near the N-terminus. Deletion of CRT10 resulted in an enhanced resistance to HU reminiscent of the inactivation of two other ribonucleotide reductase (Rnr) suppressors, CRT1 and SML1, which regulate Rnr activity at transcriptional and translational levels, respectively. Epistasis analysis indicates that CRT10 belongs to the CRT1 pathway but not the SML1 pathway. Indeed, deletion of CRT10 enhanced the survival of the mec1 null mutant and increased basal level and DNA damage-induced expression of RNR2 and RNR3, suggesting that Crt10 regulates RNR genes at the transcriptional level. Furthermore, the dun1 mutation is epistatic to crt10 with respect to both HU sensitivity and RNR gene expression. Interestingly, the expression of CRT10 itself is induced by DNA damaging agents and this induction requires DUN1, suggesting that CRT10 plays a role in cellular response to DNA damage and replication blocks. The CRT10 function appears to be achieved by positive regulation of the CRT1 transcript level, indicating that CRT10 is a component of the regulatory circuit.

The tablet is a new IT-tool which have started to get introduced into school, but research is still missing and especially in the pre-school environment. The purpose with this study is to investigate teachers uptake, how they use the tablet in everyday pedagogy work as well as the organizational conditions that that facilitate and hinders. Through my observations and interviews in a Swedish K-5 school I have found that the teachers find tablets more appealing to use in comparison to computers. These findings are partly supported by Davis et al.’s (1989) TAM-theory who considers there to be three types of aspects that affect the way individuals adapt to technology. Firstly the teachers’ motivation increased when they saw the usefulness in the tablet in both educational and administrative possibilities. Secondly, the tablet is perceived as an ease of use-artifact which in it self gives the teachers incentive for usage. Thirdly, the infrastructure problem that exists around computers creates an escape to the more appealing tablets.

Cell fusion is an important technique that is used in the field of medicine and biomedical research. For instance, fusion can be used to create hybridomas and novel types of secretory hybrid cells. It may also be used to engineer cultured insulin-secreting pancreatic B-cell lines for the treatment of diabetes. Historically, the applications listed above have been accomplished by a number of methodologies including dielectrophoresis, centrifugation, polyethylene glycol (PEG) and viral fusion proteins. However, these approaches often fail to produce the desired results due to poor cell viability, lack of 1:1 fusion, and use of non-physiological environments. It is proposed that the application of an electrical field generated by corona charge (corona fields) and subsequent treatment with direct current (DC) pulse technology will overcome these deficiencies. Isolated and pre-labeled neuronally committed human teratocarcinoma (NT2) cells in monoculture or co-culture, were seeded in chambers, constructed in the laboratory, and allowed to adhere to the chamber bottom prior to corona treatment. A corona generator, also constructed in the laboratory, was used to expose cells to positive and negative electrical charges to induce cell-cell contact. The cells were then pulsed with DC voltage to induce fusion. During the experiments, cells were photographed sequentially to record cell movement/contact and fusion. The project was designed to identify optimal corona-based electrofusion parameters for viable, 1:1 cell fusion. Optimal results for cell-cell contact were obtained using a cell density of 2.35 times ten to the fourth power cells per microliter Dulbecco's Modified Eagle Medium (DMEM) in a grounded circular plate corona chamber following at least 3 minutes of settling time. Corona charges from (+) 6.1 kilivolt and (-) 5.5 kilivolt potentials were determined as being most favorable for cell movement and viability. Fusion was best achieved by first exposing either a circular or square ungrounded corona chamber configuration to 3 minutes (+) corona charge followed by 3 minutes (--) corona charge; disturbing the cells in the chamber with mechanical force; and then exposing them to 8-15 sequences of a 2,500 Volts per centimeter DC pulse at 100 microseconds.

Cell-cycle checkpoint proteins help maintain genomic integrity by sensing damaged DNA and initiating DNA repair or apoptosis. Checkpoint protein activation to cell-cycle damaging agents can involve post-translational modifications and these alterations provide a means to determine whether DNA in a cell is damaged or not. Steinmoeller et al. (2009) showed that checkpoint proteins are suitable biomarkers for detecting genotoxins in Oncorhynchus mykiss (rainbow trout). In this project, two evolutionarily conserved checkpoint proteins, Rad1 and Hus1, have been cloned from rainbow trout and antibodies against these proteins were developed. This is the first time that either Rad1 or Hus1 has been characterized in rainbow trout. For rtRad1, it was determined that the open-reading frame was 840bp, which encodes 279aa with a predicted protein size of 31kDa. The rtRad1 amino-acid sequence is highly conserved and contains conserved exonuclease and leucine zipper domains. RT-PCR was used to identify alternatively spliced variants of rtRad1 and it appears that these variants encode different sized Rad1 proteins that are tissue and cell-line specific. A Rad1 splice variant that encodes an 18kDa protein appears to be abundant only in heart tissue and in the RTgill-W1 and RTbrain-W1 cell-lines. A genotoxicity study was completed where RTgill-W1 and RTbrain-W1 cells were treated with bleomycin, which induces double-stranded DNA breaks. In RTgill-W1, levels of an 18kDa Rad1 protein increased in a dose-dependent manner while in RTbrain-W1 the Rad1 levels remained the same. It appears that this 18kDa Rad1 protein may be directly involved in maintaining genomic integrity and shows potential to be used as a genotoxicity biomarker. This is the first time that an isoform of Rad1 has shown to be modified in the presence of a damaging agent. Both Rad1 and Hus1 need to be further characterized to determine their usefulness as genotoxicity biomarkers.

Localizado na modalidade de aprendizagem um computador por aluno e no contexto da proposta metodológica de PAs (Projetos de Aprendizagem), este estudo tem por objetivo apresentar uma dinâmica de acompanhamento da conceituação a partir da enunciação sobre as produções escritas dos alunos. Parte-se da concepção de enunciação como apropriação ativa da língua, na qual se expõe a compreensão em muitos casos ausente no registro textual. Ferramentas do laptop em utilização na escola pólo do pré-piloto do Projeto UCA (Um Computador por Aluno) em Porto Alegre e o ambiente virtual de aprendizagem Amadis dão suporte ao trabalho. Sustentam a análise o Método Clínico de Piaget e a teoria de reações compensadoras proposta pelo mesmo autor, aplicáveis à amplitude de temas contemplados nos PAs e aos diferentes percursos empreendidos pelos estudantes em suas pesquisas. / Located in 1:1 (one computer per student) learning modality and in the context of Learning Projects methodology, this study aims to present a dynamics of monitoring the process of conceptualization considering students’ written productions. For the purpose of this study, enunciation is understood as an active process of language ownership, which exposes comprehension often lacking in textual speech. Resources from the laptop used in the trial school of UCA (One Computer per Child) Project pre-pilot in Porto Alegre and the virtual learning environment Amadis support the work. The analysis is based on Jean Piaget Clinical Method and on the Cognitive Compensation Reactions theory, proposed by the same author, applicable to the variety of themes covered in the projects and to the different paths taken by students in their researches.

Localizado na modalidade de aprendizagem um computador por aluno e no contexto da proposta metodológica de PAs (Projetos de Aprendizagem), este estudo tem por objetivo apresentar uma dinâmica de acompanhamento da conceituação a partir da enunciação sobre as produções escritas dos alunos. Parte-se da concepção de enunciação como apropriação ativa da língua, na qual se expõe a compreensão em muitos casos ausente no registro textual. Ferramentas do laptop em utilização na escola pólo do pré-piloto do Projeto UCA (Um Computador por Aluno) em Porto Alegre e o ambiente virtual de aprendizagem Amadis dão suporte ao trabalho. Sustentam a análise o Método Clínico de Piaget e a teoria de reações compensadoras proposta pelo mesmo autor, aplicáveis à amplitude de temas contemplados nos PAs e aos diferentes percursos empreendidos pelos estudantes em suas pesquisas. / Located in 1:1 (one computer per student) learning modality and in the context of Learning Projects methodology, this study aims to present a dynamics of monitoring the process of conceptualization considering students’ written productions. For the purpose of this study, enunciation is understood as an active process of language ownership, which exposes comprehension often lacking in textual speech. Resources from the laptop used in the trial school of UCA (One Computer per Child) Project pre-pilot in Porto Alegre and the virtual learning environment Amadis support the work. The analysis is based on Jean Piaget Clinical Method and on the Cognitive Compensation Reactions theory, proposed by the same author, applicable to the variety of themes covered in the projects and to the different paths taken by students in their researches.

The Master’s thesis is focused on the research influence the dose and type of scattered reinforcement in a reference concrete on physical and mechanical properties of the resulting composite. The investigated properties of composite belongs compressive strength, tensile strength in the flexural and tensile strength in the transverse, static modulus of elasticity in compressive strength, dynamic modulus of elasticity determined using NDT tests. The reference concrete mixture is certified as structural concrete of the class, which is commonly used on construction sites.