The glial cells of the brain and the peripheral nervous system retain the capacity to divide and proliferate throughout the lifespan of an individual and thereby have the propensity to give rise to the most adult neurological tumours. Among them, the tumours which arise from different kinds of glial cells are referred to as gliomas. Of the various types of gliomas, astrocytomas are the most common central nervous system neoplasms which make upto 60% of all the primary brain tumours. Being the most prevalent type, the WHO classifies them into grades ranging from I to IV based on their intensity of malignancy. Grade IV astrocytoma or Glioblastoma (GBM) is considered to be the most malignant form with a median survival of 14.6 months, in spite of all therapeutic modalities. GBM is further classified as primary and secondary GBM. Primary GBM manifests de novo without any early history of pre-malignant lesions, on the other hand secondary GBM arises progressively from lower grades over a period of 5-10 years. Like other malignancies, GBM also arises from various genetic and epigenetic variations. Epigenetic variations include all such mitotically and meiotically heritable traits that do not involve changes in DNA sequence. There are three major areas of epigenetics - DNA methylation, histone modifications and non-coding RNAs which are known to have profound effects on gene expression. A lot being known about the genetic derailments in GBM, in this study we looked into the epigenetic aspects of GBM. In our lab, we have carried out various high throughput studies, which unveiled the distorted landscape of DNA methylation and miRNA expression in GBM. This indicates that, in addition to the genetic mechanisms of gene alterations like mutations, copy number aberrations, protein coding genes are also affected by changes in methylation as well as by miRNA misregulation. The study has been divided into two parts. Part one of the study deals with the identification of chromobox homolog 7 (Cbx7), as a hypermethylated and downregulated gene in GBM. More importantly, Cbx7 is a member of the polycomb repressive complex and brings about its function through chromatin modifications. Here we have investigated the role of Cbx7 in gliomagenesis, and why it has to be silenced by methylation for tumorigenesis to ensue. In part two, we elucidated two unique ways of miRNA regulation in GBM. In the first section, we identified miR-326 as a PI3 kinase regulated miRNA and demonstrated its tumour suppressive role in GBM. In the other section, we analysed the copy number aberration data from TCGA and identified miR- 4484 as a miRNA subjected to deletion in GBM. We further went ahead to demonstrate its growth suppressive role in GBM. Part 1: Epigenetic regulation of the chromatin modifier Cbx7; chromobox homolog 7 DNA methylation is involved in the normal cellular control of expression and thereby plays a crucial role in maintaining the homeostasis of the cell. The phenomenon of DNA methylation keeps the various loci of the genome such as the germline specific genes and the repetitive transposable elements silenced, whereas the tumour suppressors and other growth modulator genes are spared from the methylation induced gene repression. One of the important steps that promote tumorigenesis is aberrant hypermethylation, which leads to the silencing of tumour-suppressor genes. Another important epigenetic phenomenon that affects the transcriptibility of the genome is histone modifications, which control the accessibility of the chromatin to the transcriptional machinery. In this section, we identified Cbx7, which happens to be an essential component of the chromatin modifying machinery, as an epigenetically regulated gene in GBM. We observed from the methylation array carried out in our lab, that Cbx7 was one of the highly methylated genes. We also validated that Cbx7 is downregulated in GBM and the same observation was further corroborated from other data sets. The hypermethylated state of Cbx7 was confirmed by DNA bisulphite sequencing and the expression levels of Cbx7 also got alleviated after 5-Aza-2′-deoxycytidine treatment, which is a DNA methylation inhibitor. This indicated that the down regulation of Cbx7 could be attributed to the methylation of its promoter region. In order to figure out the role of Cbx7 in GBM, we carried out transcriptome analysis of Cbx7 overexpressing cells compared to vector control condition by RNA sequencing. Gene ontology analysis revealed a significant enrichment of pathways involved in cell cycle, migration and invasion like processes. In fact, the exogenous overexpression of Cbx7 leads to cell death, reduced colony formation, retarded migration and invasion of cells. In order to explain the above phenotypes brought about by the exogenous expression of Cbx7, we further examined the RNA sequencing data and observed that many of the top most downregulated genes in Cbx7 overexpression state belonged to the Hippo signaling pathway. The effectors of the Hippo pathway, YAP and TAZ which essentially antagonize the pathway activity, are well known for their role in proliferation, migration and invasion in cancer. So we carried out a Gene Set Enrichment Analysis (GSEA) and found that there was a significant negative enrichment of YAP/TAZ targets in the Cbx7 regulated gene set. We validated some of these targets that were downregulated by Cbx7 overexpression. One of the most downregulated genes that we validated was Connective Tissue Growth Factor (CTGF), which also happens to be a bonafide target of YAP/TAZ. Independent downregulation of CTGF also resulted in reduced migration, thereby phenocopying the effects as were produced by Cbx7 overexpression. Moreover, we also observed that SAPK/JNK was the only kinase whose activity was abolished upon Cbx7 overexpression. Since CTGF is known to activate SAPK/JNK, we assessed the SAPK/JNK activity upon CTGF silencing. We found that levels of phospho-SAPK/JNK were significantly reduced in CTGF silenced condition. In addition to that, the inhibition of the SAPK/JNK by synthetic inhibitor also hampered the migration ability of the cells. We were also able to rescue the loss of migratory potential of glioma cells by the exogenous overexpression of CTGF in Cbx7 stable background. A similar rescue was also achieved by the overexpression of a constitutively active form of SAPK/JNK. This indicates that Cbx7 activates Hippo pathway to inhibit YAP/TAZ dependent transcription, resulting in the downregulation of CTGF, thereby inhibiting CTGF mediated activation of SAPK and thus resulting in the inhibition of glioma cell migration. PART 2: ROLE OF MIRNAS IN GLIOMA DEVELOPMENT AND PROGRESSION miRNAs are a class of small non-coding RNAs that are not translated into functional proteins but still contribute to numerous cellular processes, thereby adding yet another realm of regulation and control. miRNAs bring about gene regulation at the post-transcriptional level, either by degrading the mRNA or by translational repression and in this manner fine tune the expression of protein coding genes. miRNAs are often located in the most fragile sites of the genome which exposes them to grave genetic alterations, thus providing a circumstantial evidence of their etiological role in tumorigenesis. In a malignant state, miRNAs have been found to play pivotal roles in cellular transformation by altering various cellular phenotypes. Owing to their participation in diverse cellular functions, miRNAs have gained a strong foothold in gene regulation. Though a lot has been deciphered about the functional aspect of miRNAs, not much is known about the precise mechanisms which lead to their misregulation and therefore demands in-depth study. The expression of miRNAs can be modulated by a variety of genetic and epigenetic mechanisms. Section I: Role of miR-326 – a PI3 kinase regulated miRNA, in gliomagenesis The TCGA group in the year 2008 identified three major pathways which go disarray in GBM. These include the pro-tumorigenic receptor tyrosine kinase (RTK) pathway, and the p53 and the pRB tumour-suppressive pathways. The RTK signalling includes the PI3 kinase pathway, which is pivotal in gliomagenesis and many other cancers. This directed us to elucidate the set of miRNAs which are controlled by the aberrant functioning of the PI3 kinase pathway. We used synthetic inhibitor LY294002 to abrogate the PI3 kinase signalling and examined the miRNA profile in two glioma cell lines U87 and U251, which have an activated PI3 kinase pathway. Indeed the abrogation of the PI3 kinase pathway resulted in the modulation of a wide array of miRNAs. We validated miR-326 as one of the miRNAs that was upregulated upon PI3 kinase pathway abrogation. Furthermore, we observed that miR-326 was a down regulated miRNA in GBM and different glioma cell lines, as well as in many other publicly available data sets. We also observed that miR-326 is an intragenic miRNA and its host gene Arrestin β1 (ARRB1) also exhibited similar upregulation upon PI3K pathway inhibition. Over-expression of miR-326 resulted in various anti-tumorigenic affects like reduced proliferation, reduced migration and colony suppression. In order to find the targets of miR-326, we analysed the transcriptome by RNA sequencing upon pre-miR-326 transfection. We shortlisted and validated some of the genes which were getting regulated through miRNA over-expression and also explain the functional role of miR-326. Section II: Role of miR-4484 – a copy number deleted miRNA, in gliomagenesis In the TCGA study mentioned above, it was also unfurled that there are many genes in the RTK, p53 and pRB signalling pathways which are made dysfunctional through gene deletions and amplifications. We envisaged whether it is only the protein coding genes which are subjected to such regulations or the non-coding genes like miRNAs as well. In this pursuit, we identified miR-4484 as one of the miRNAs located in the deleted region of uroporphyrinogen III synthase (UROS) gene in the chromosome 11 of the GBM genome. As conceived, miR-4484 was observed to be a downregulated miRNA in association with its host gene UROS. We further elucidated that the downregulation was due to the co-deletion of a locus harbouring both the protein coding gene and the miRNA. In addition, upon over-expression of miR-4484, we observed reduced migration and colony formation, indicating its role as a tumour–suppressor. For seeking the targets of miR-4484, we extracted RNA from miR-4484 over-expression condition and subjected it to RNA sequencing. We shortlisted and validated some of the genes which were getting regulated through miRNA over-expression and possibly explain the functional role of miR-4484.

Thesis deals with automatic guided mobile robot focused on obstacle avoidance during ride on planned route. There are summaries of usually used obstacle detecting sensors and algorithms used for path finding. Based on this, own solution is designed. It uses waypoints changes to pass obstacle. MATLAB simulation is created for tests of new designed method. This method is implemented to real robot for real world testing. Reached goals and upgrade possibilities are summarized in bottom of thesis.

The doctoral thesis discusses current options for the navigation of unmanned ground vehicles with a focus on achieving high absolute compliance of the required motion trajectory and the obtained one. The current possibilities of key self-localization methods, such as global satellite navigation systems, inertial navigation systems, and odometry, are analyzed. The description of the navigation method, which allows achieving a centimeter-level accuracy of the required trajectory tracking with the above mentioned self-localization methods, forms the core of the thesis. The new navigation method was designed with regard to its very simple parameterization, respecting the limitations of the used robot drive configuration. Thus, after an appropriate parametrization of the navigation method, it can be applied to any drive configuration. The concept of the navigation method allows integrating and using more self-localization systems and external navigation methods simultaneously. This increases the overall robustness of the whole process of the mobile robot navigation. The thesis also deals with the solution of cooperative convoying heterogeneous mobile robots. The proposed algorithms were validated under real outdoor conditions in three different experiments.

No description available.

La signalisation par l’estrogène a longtemps été considérée comme jouant un rôle critique dans le développement et la progression des cancers hormono-dépendants tel que le cancer du sein. Deux tiers des cancers du sein expriment le récepteur des estrogènes (ER) qui constitue un élément indiscutable dans cette pathologie. L’acquisition d’une résistance endocrinienne est cependant un obstacle majeur au traitement de cette forme de cancer. L’émergence de cancers hormono-indépendants peut est produite par l’activation de ER en absence d’estrogène, l’hypersensibilité du récepteur aux faibles concentrations plasmique d’estrogène ainsi que l’activation de ER par des modulateurs sélectifs. L’activité du ER est fortement influencée par l’environnement cellulaire tel que l’activation de voie de signalisation des facteurs de croissances, la disponibilité de protéines co-régulatrices et des séquences promotrices ciblées. Présentement, les études ont principalement considérées le rôle de ERα, cependant avec la découverte de ERβ, notre compréhension de la diversité des mécanismes potentiels impliquant des réponses ER-dépendantes s’est améliorée. L’activation des voies des kinases par les facteurs de croissance entraîne le développement d’un phénotype tumoral résistant aux traitements actuels. Nos connaissances des voies impliquées dans l’activation de ER sont restreintes. ERα est considéré comme le sous-type dominant et corrèle avec la plupart des facteurs de pronostic dans le cancer du sein. Le rôle de ERβ reste imprécis. Les résultats présentés dans cette thèse ont pour objectif de mieux comprendre l’implication de ERβ dans la prolifération cellulaire par l’étude du comportement de ERβ et ERα suite à l’activation des voies de signalisation par les facteurs de croissance. Nous démontrons que l’activation des récepteurs de surfaces de la famille ErbB, spécifiquement ErbB2/ErbB3, inhibe l’activité transcriptionnelle de ERβ, malgré la présence du coactivateur CBP, tout en activant ERα. De plus, l’inhibition de ERβ est attribuée à un résidu sérine (Ser-255) situé dans la région charnière, absente dans ERα. Des études supplémentaires de ErbB2/ErbB3 ont révélé qu’ils activent la voie PI3K/Akt ciblant à son tour la Ser-255. En effet, cette phosphorylation de ERβ par PI3K/Akt induit une augmentation de l’ubiquitination du récepteur qui promeut sa dégradation par le système ubiquitine-protéasome. Cette dégradation est spécifique pour ERβ. De façon intéressante, la dégradation par le protéasome requiert la présence du coactivateur CBP normalement requis pour l’activité transcriptionnelle des récepteurs nucléaires. Malgré le fait que l’activation de la voie PI3K/Akt corrèle avec une diminution de l’expression des gènes sous le contrôle de ERβ, on observe une augmentation de la prolifération des cellules cancéreuses. L’inhibition de la dégradation de ERβ réduit cette prolifération excessive causée par le traitement avec Hrgβ1, un ligand de ErbB3. Un nombre croissant d’évidences indique que les voies de signalisations des facteurs de croissance peuvent sélectivement réguler l’activité transcriptionnelle de sous-types de ER. De plus, le ratio ERα/ERβ dans les cancers du sein devient un outil de diagnostique populaire afin de déterminer la sévérité d’une tumeur. En conclusion, la caractérisation moléculaire du couplage entre la signalisation des facteurs de croissance et la fonction des ERs permettra le développement de nouveaux traitements afin de limiter l’apparition de cellules tumorales résistantes aux thérapies endocriniennes actuelles. / It has long been appreciated that estrogenic signaling plays a critical role in the development of hormone-dependent cancers such as breast cancer. Two-thirds of breast cancers express estrogen receptor (ER) which has been demonstrated to play an irrefutable role in tumour development and progression. However the acquisition of endocrine resistance has become a major obstacle in the treatment of hormone-dependent cancers that have acquired a hormone-independent state. Hormone-independent cancers emerge from an array of pathways involving ER activation in the absence of estrogen, hypersensitivity of ER to low serum levels of estrogen and activation by estrogen antagonists. The activity of ER is critically influenced by the cellular environment such as growth factor signaling pathways, availability of coregulatory proteins and the promoter sequence of target genes. The mechanisms studied have mostly considered the role of ERα, however with the discovery of the second subtype, ERβ, the understanding on the diversity of potential mechanisms involving ER-dependent responses have improved. Hormonal-independent activation of ER can occur in estrogen-dependent breast tumours, with concomitant rise in kinase signaling pathways, resulting in the acquisition of a therapeutic resistant phenotype in treated women. Our knowledge is relatively limited on which pathways trigger ER signaling and how these phosphorylation-coupled events affect ER activity. ERα is considered the dominant subtype and correlates with most of the prognostic factors in breast cancers. Conversely the role of ERβ remains unclear. The results presented in this thesis were carried out with the objective of gaining a better understanding of ERβ’s role in cellular proliferation by examining the behavior of ERβ and ERα during the activation of growth factor signaling pathways by cell-surface receptor-tyrosine kinases. We demonstrate here that the activation of cell surface receptors of the ErbB family, specifically ErbB2/ErbB3, inhibits the transcriptional activity of ERβ despite the presence of the coactivator CBP, yet activated ERα. Furthermore the inhibition of ERβ was attributed to a specific serine residue located within the hinge region, not present in ERα. Additional studies of ErbB2/ErbB3-initiated signaling revealed that it triggered the activation of the PI3K/Akt pathway which targeted the serine residue within the hinge region of ERβ. In fact, phosphorylation of ERβ by the PI3K/Akt pathway led to an increase in receptor ubiquitination which promoted its degradation by the ubiquitin-proteasome system which was subtype specific. Interestingly, proteasomal degradation required the presence of the coactivator CBP, which is normally involved in assisting nuclear receptor transcriptional activity. Although the activation of the PI3K/Akt pathway correlated with a decrease in the expression of ERβ target genes it led to an increase in the proliferation of breast cancer cells. Inhibiting the degradation of ERβ reduced the enhanced proliferation of breast cancer cells brought about by the treatment of ErbB3’s ligand, Hrgβ1. Increasing evidence indicates that growth factor signaling pathways can selectively regulate the transcriptional activity of ER subtypes, and the ratio of ERα/ERβ expression in breast tumours is becoming a popular prognostic factor to evaluate the severity of the tumour. Therefore the molecular characterization of the coupling between growth factor signaling and ER function should provide improved therapeutical approaches to overcome or delay the onset of resistance to endocrine therapy in hormone-dependent cancers.

Vid GNSS-positionering i samband med fastighetsbildningsåtgärder används vanligtvis den traditionella RTK-mätningen (Real-Time Kinematic) via SWEPOS nätverks-RTK-tjänst. Denna tjänst kräver mobiltelefontäckning eller motsvarande tvåvägskommunikation, vilket kan vara problematiskt inom områden med bristfällig mobiltelefontäckning. Under dessa förhållanden kan istället PPP-mätning (Precise Point Positioning) vara användbart vid fastighetsbildningsåtgärder då dessa tjänster tar emot korrektionsdata i realtid från satelliter. PPP kräver inte någon mobiltelefontäckning, däremot krävs en kommunikationslänk, en RTX-tjänst för att erhålla korrektioner externt från en RTX-satellit. Syftet med studien är att undersöka möjligheten till att nyttja PPP i realtid vid fastighetsbildningsåtgärder som ett alternativ till traditionell GNSS-mätning med nätverks-RTK. För att PPP ska vara ett alternativ till traditionell GNSS-mätning i realtid krävs det att mätosäkerhetskraven inom fastighetsbildning uppfylls. Mätosäkerheten undersöktes genom att utgå ifrån redan kända koordinater (RIX 95-punkter). Mätningarna har genomförts på fem olika platser i Sverige, Göteborg, Vänersborg, Karlstad, Torsby och Malung-Sälen. Mätdata som erhölls från undersökningsplatserna har analyserats samt jämförts med fastighetsbildningskraven. Resultatet av studien erhölls i form av analyserad mätdata med jämförelser mot redan kända (RIX 95) punkter. Avikelsen från känd RIX 95-punkt redovisas i resultatet utifrån tidsaspekten, den systematiska avvikelsen av translativ art, förändringar i avvikelsen från söder till norr samt utifrån två beräkningsmodeller, varav en translation och en transformation. För att få den erhållna mätdatan från RTX-tjänsten att överensstämma bättre med referenspunkten (RIX 95-punkten) togs beräkningsmodellerna fram för att möjliggöra modellering av systematiska avvikelser som uppkommit och därmed uppfylla kraven inom fasighetsbildningsåtgärder. Genom att ha analyserat och granskat olika samband har det framkommit att efter ca 20 minuters mätning, börjar precisionen för mätningarna att bli stabila. Utifrån resultatet är slutsatsen att PPP inte fungerar vid fastighetsbildningsåtgärder för områden inom stomnät, däremot fungerar metoden för skogs- och jordbruksfastigheter utanför stomnät. Förutsatt att en modellering genom translation alternativt transformation som är framtagen i denna studie används för att justera koordinaterna så fungerar PPP-mätning inom samtliga fastighetsbildningsåtgärder. Detta kräver då att mätdata erhålls efter 20 minuters mätning eller mer. / GNSS positioning in conjunction with the real property is usually used the traditional RTK measuring (Real-Time Kinematic) by SWEPOS network RTK service. This service requires mobile phone coverage or equivalent two-way communication, which can be problematic in areas with poor mobile phone coverage. Under these circumstances, PPP (Point Positioning Precise) could be more useful in real property measures when such services receives the correction data in real time from the satellites. PPP does not require any cell phone coverage, however it requires a communication link, a RTX service to obtain corrections externally from a RTX satellite. The purpose of the study is to examine the possibility of using PPP in real time at the real property as an alternative to traditional GNSS measurements with network RTK. The measurement uncertainty was investigated by starting out from already known coordinates (RIX 95 points). The measurements were performed out at five different locations in Sweden, Gothenburg, Vanersborg, Karlstad, Torsby and Malung-Salen. Measurement data obtained from the observations have been analyzed and compared with real property requirements. The results of the study were obtained in the form of data analyzed by comparison of the known (RIX 95) points. The deviation is known from RIX 95 point recognized in income based on the time factor, the bias of the translative case species, changes in deviation from south to north and from two calculation models, a translation and a transformation. To correct the measured values from the RTX service for a better match to the RIX 95 points calculation models were developed to facilitate the modeling of systematic deviations incurred and meet the demands of real property. Analyzing and examining various relationships have shown that after about 20 minutes of measuring, the precision of the measurements starts to become more stable. Based on the results, the conclusion is that the PPP does not work in real property areas within the core network, however, the method works for forestry and agricultural properties outside the core network. Assuming a modelling through translational alternative transformation, developed in this study is used to adjust the coordinates, the PPP measurement is working in all real property registration measures. This requires that the measurement data is obtained after 20 minutes of measurement or more.

La signalisation par l’estrogène a longtemps été considérée comme jouant un rôle critique dans le développement et la progression des cancers hormono-dépendants tel que le cancer du sein. Deux tiers des cancers du sein expriment le récepteur des estrogènes (ER) qui constitue un élément indiscutable dans cette pathologie. L’acquisition d’une résistance endocrinienne est cependant un obstacle majeur au traitement de cette forme de cancer. L’émergence de cancers hormono-indépendants peut est produite par l’activation de ER en absence d’estrogène, l’hypersensibilité du récepteur aux faibles concentrations plasmique d’estrogène ainsi que l’activation de ER par des modulateurs sélectifs. L’activité du ER est fortement influencée par l’environnement cellulaire tel que l’activation de voie de signalisation des facteurs de croissances, la disponibilité de protéines co-régulatrices et des séquences promotrices ciblées. Présentement, les études ont principalement considérées le rôle de ERα, cependant avec la découverte de ERβ, notre compréhension de la diversité des mécanismes potentiels impliquant des réponses ER-dépendantes s’est améliorée. L’activation des voies des kinases par les facteurs de croissance entraîne le développement d’un phénotype tumoral résistant aux traitements actuels. Nos connaissances des voies impliquées dans l’activation de ER sont restreintes. ERα est considéré comme le sous-type dominant et corrèle avec la plupart des facteurs de pronostic dans le cancer du sein. Le rôle de ERβ reste imprécis. Les résultats présentés dans cette thèse ont pour objectif de mieux comprendre l’implication de ERβ dans la prolifération cellulaire par l’étude du comportement de ERβ et ERα suite à l’activation des voies de signalisation par les facteurs de croissance. Nous démontrons que l’activation des récepteurs de surfaces de la famille ErbB, spécifiquement ErbB2/ErbB3, inhibe l’activité transcriptionnelle de ERβ, malgré la présence du coactivateur CBP, tout en activant ERα. De plus, l’inhibition de ERβ est attribuée à un résidu sérine (Ser-255) situé dans la région charnière, absente dans ERα. Des études supplémentaires de ErbB2/ErbB3 ont révélé qu’ils activent la voie PI3K/Akt ciblant à son tour la Ser-255. En effet, cette phosphorylation de ERβ par PI3K/Akt induit une augmentation de l’ubiquitination du récepteur qui promeut sa dégradation par le système ubiquitine-protéasome. Cette dégradation est spécifique pour ERβ. De façon intéressante, la dégradation par le protéasome requiert la présence du coactivateur CBP normalement requis pour l’activité transcriptionnelle des récepteurs nucléaires. Malgré le fait que l’activation de la voie PI3K/Akt corrèle avec une diminution de l’expression des gènes sous le contrôle de ERβ, on observe une augmentation de la prolifération des cellules cancéreuses. L’inhibition de la dégradation de ERβ réduit cette prolifération excessive causée par le traitement avec Hrgβ1, un ligand de ErbB3. Un nombre croissant d’évidences indique que les voies de signalisations des facteurs de croissance peuvent sélectivement réguler l’activité transcriptionnelle de sous-types de ER. De plus, le ratio ERα/ERβ dans les cancers du sein devient un outil de diagnostique populaire afin de déterminer la sévérité d’une tumeur. En conclusion, la caractérisation moléculaire du couplage entre la signalisation des facteurs de croissance et la fonction des ERs permettra le développement de nouveaux traitements afin de limiter l’apparition de cellules tumorales résistantes aux thérapies endocriniennes actuelles. / It has long been appreciated that estrogenic signaling plays a critical role in the development of hormone-dependent cancers such as breast cancer. Two-thirds of breast cancers express estrogen receptor (ER) which has been demonstrated to play an irrefutable role in tumour development and progression. However the acquisition of endocrine resistance has become a major obstacle in the treatment of hormone-dependent cancers that have acquired a hormone-independent state. Hormone-independent cancers emerge from an array of pathways involving ER activation in the absence of estrogen, hypersensitivity of ER to low serum levels of estrogen and activation by estrogen antagonists. The activity of ER is critically influenced by the cellular environment such as growth factor signaling pathways, availability of coregulatory proteins and the promoter sequence of target genes. The mechanisms studied have mostly considered the role of ERα, however with the discovery of the second subtype, ERβ, the understanding on the diversity of potential mechanisms involving ER-dependent responses have improved. Hormonal-independent activation of ER can occur in estrogen-dependent breast tumours, with concomitant rise in kinase signaling pathways, resulting in the acquisition of a therapeutic resistant phenotype in treated women. Our knowledge is relatively limited on which pathways trigger ER signaling and how these phosphorylation-coupled events affect ER activity. ERα is considered the dominant subtype and correlates with most of the prognostic factors in breast cancers. Conversely the role of ERβ remains unclear. The results presented in this thesis were carried out with the objective of gaining a better understanding of ERβ’s role in cellular proliferation by examining the behavior of ERβ and ERα during the activation of growth factor signaling pathways by cell-surface receptor-tyrosine kinases. We demonstrate here that the activation of cell surface receptors of the ErbB family, specifically ErbB2/ErbB3, inhibits the transcriptional activity of ERβ despite the presence of the coactivator CBP, yet activated ERα. Furthermore the inhibition of ERβ was attributed to a specific serine residue located within the hinge region, not present in ERα. Additional studies of ErbB2/ErbB3-initiated signaling revealed that it triggered the activation of the PI3K/Akt pathway which targeted the serine residue within the hinge region of ERβ. In fact, phosphorylation of ERβ by the PI3K/Akt pathway led to an increase in receptor ubiquitination which promoted its degradation by the ubiquitin-proteasome system which was subtype specific. Interestingly, proteasomal degradation required the presence of the coactivator CBP, which is normally involved in assisting nuclear receptor transcriptional activity. Although the activation of the PI3K/Akt pathway correlated with a decrease in the expression of ERβ target genes it led to an increase in the proliferation of breast cancer cells. Inhibiting the degradation of ERβ reduced the enhanced proliferation of breast cancer cells brought about by the treatment of ErbB3’s ligand, Hrgβ1. Increasing evidence indicates that growth factor signaling pathways can selectively regulate the transcriptional activity of ER subtypes, and the ratio of ERα/ERβ expression in breast tumours is becoming a popular prognostic factor to evaluate the severity of the tumour. Therefore the molecular characterization of the coupling between growth factor signaling and ER function should provide improved therapeutical approaches to overcome or delay the onset of resistance to endocrine therapy in hormone-dependent cancers.

內政部土地測量局為擴大RTK有效作業範圍，同時降低主站布設密度，已規劃透過網際網路高速、寬頻之數據傳輸技術，預定於九十五年度前完成建置全國性電子化GPS（ｅ-GPS）衛星定位測量基準網，就其連續定位觀測資料，建構區域性定位誤差內插模式，並配合虛擬基準站（Virtual Base Station，VBS）即時動態定位技術（Real-Time Kinematic, RTK）（ 簡稱VBS-RTK），獲得高精度的定位成果，提供多目標定位服務及加值應用。 本研究利用全國性e-GPS衛星定位基準站即時動態定位系統北區服務網為基準網，探討以VBS-RTK定位技術應用於圖根測量及直接辦理土地複丈之可行性。首先檢核實驗區內控制點與圖根點成果發現，縱坐標（N）較差絕對值之平均值為1.9公分，橫坐標（E）較差平均值為2.0公分。經分析後得知，本實驗區之VRS-RTK測量成果與原坐標成果間有系統誤差存在，透過六參數轉換後，其坐標較差值均能符合圖根點測量規範。其次，以VBS-RTK直接辦理土地複丈，其界址點坐標成果較差亦均符合界址點位置檢查與原坐標值之較差不得超過6公分之規定。 / For expanding the effective range and decreasing the density of base stations, Land Survey Bureau, Ministry of the Interior (LSB) have set up a national e-GPS base stations network in 2006. In order to obtain high precision position result and provide multi-goals positioning service with plus value application, the system will be involved regional position error interpolating mode and Virtual Base Station（VBS）technique. The research area is located within the north service net of the national e-GPS base stations network, feasible discussion of using to Supplementary control survey and direct to transact the land resurvey with the VBS-RTK technique. Firstly, The 4th and the higher order control points and the traverse points located at the research area were examined with the VRS-RTK. Compared with the original data, the results have shown that the average of the absolute value of errors is 1.9 cm at the N-axis, 2.0 cm at the E-axis. After transformations either by affine-6-parameters, the result shows that all of the errors are satisfy the stipulation of rules.Secondly, using the VBS-RTK technique to undertake land survey correction, the all errors are satisfy the stipulation of rules.( less than 6 cm )

The first half of the Masters thesis is the result of a survey made on the behalf of the Swedish company KORDAB International AB. The survey includes an overview of different positioning systems and some wireless communication techniques available on the market today. Positioning systems discussed are GPS, DGPS, AGPS and GSM positioning. Mobile Internet connections through mobile phones and communication through radio modems are mentioned and described as examples of wireless communication techniques. Examples of techniques described are HSCSD, GPRS, UMTS and MOBITEX. KORDAB is in the starting blocks to implement a real time positioning feature into their own technical information system GEOSECMA. This survey will give them a base which will help them to decide on which system to use for this feature. The second half of the thesis includes parts concerning a prototype made to exemplify how KORDAB could implement the real time positioning feature into GEOSECMA. The NMEA 0183 protocol, reference systems and transformation are described to give the necessary background knowledge for the construction and functionality of the prototype. The prototype is a GPS-communicator made as an interface between a GPS-receiver and GEOSECMA and its functionality is also described in this second half of the thesis. Feasible applications are also discussed to show the possibilities real time positioning gives. One application discussed is “Zoom and Auto highlighting”. This application is designed to help user of GEOSECMA to zoom in the map and highlighting the nearest object at current location.

<p>The first half of the Masters thesis is the result of a survey made on the behalf of the Swedish company KORDAB International AB. The survey includes an overview of different positioning systems and some wireless communication techniques available on the market today. Positioning systems discussed are GPS, DGPS, AGPS and GSM positioning. Mobile Internet connections through mobile phones and communication through radio modems are mentioned and described as examples of wireless communication techniques. Examples of techniques described are HSCSD, GPRS, UMTS and MOBITEX. KORDAB is in the starting blocks to implement a real time positioning feature into their own technical information system GEOSECMA. This survey will give them a base which will help them to decide on which system to use for this feature. </p><p>The second half of the thesis includes parts concerning a prototype made to exemplify how KORDAB could implement the real time positioning feature into GEOSECMA. The NMEA 0183 protocol, reference systems and transformation are described to give the necessary background knowledge for the construction and functionality of the prototype. The prototype is a GPS-communicator made as an interface between a GPS-receiver and GEOSECMA and its functionality is also described in this second half of the thesis. Feasible applications are also discussed to show the possibilities real time positioning gives. One application discussed is “Zoom and Auto highlighting”. This application is designed to help user of GEOSECMA to zoom in the map and highlighting the nearest object at current location.</p>