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Etude biochimique comparative des "Actin Depolymerizing Factors"(ADFs) d'Arabidopsis : activité inattendue de pontage des filaments d'actine pour les ADFs appartenant à la sous-classe III / Comparative biochemical analysis of Arabidopsis Actin-Depolymerizing Factors (ADFs) : unexpected actin-crosslinking activity for subclass III ADFsTholl, Stéphane 02 March 2012 (has links)
L'organisation et la dynamique du cytosquelette d'actine sont finement régulées par une multitude de "actin-binding proteins" (ABPs). Parmi ces dernières, les ADFs (actin-depolymerizing factors) jouent un rôle majeur dans le turnover des filaments d'actine en induisant leur découpage et en facilitant leur dépolymérisation. Arabidopsis thaliana possède 11 protéines ADFs fonctionnelles qui peuvent être classées en 4 sous-classes sur la base de leur profil d'expression et liens phylogénétiques. Nous démontrons que l’ADF5 et l’ADF9 de la sous-classe III sont des ADFs atypiques puisqu’elles n’induisent pas la dépolymérisation des filaments d’actine. Au contraire, elles montrent une forte capacité à stabiliser et ponter les filaments d’actine en longs câbles in vitro ainsi que in vivo. Nous décrivons la caractérisation d’un nouveau mutant knockout d’Arabidopsis. Les données suggèrent un rôle d’ADF9 dans l’élongation cellulaire. Ainsi, l’hypocotyle est significativement plus long dans les mutants adf9 que dans les plantules sauvages, et ce phénotype est amplifié par des conditions de croissance à l’obscurité dans lesquelles le gène ADF9 est normalement préférentiellement exprimé. L’analyse des cellules épidermiques d’hypocotyle indique que ce phénotype est essentiellement dut à une augmentation de l’élongation cellulaire. De manière surprenante, les plantules mutantes adf9 présentent également des racines plus courtes que les contrôles, suggérant un lien complexe entre l’organisation du cytosquelette d’actine et l’élongation cellulaire. Finalement, la capacité réduite du cal issue des plantules adf9 à proliférer suggère également un rôle d’ADF9 dans la division cellulaire. / Actin cytoskeleton organization and dynamics are tightly regulated by many actin-binding proteins (ABPs). Among ABPs, the actin-depolymerizing factors (ADFs) play a major role in actin filament turnover by promoting actin filament severing and facilitating pointed end depolymerization. Arabidopsis thaliana has 11 functional proteins that can be classified into four subclasses according to their expression profile and phylogenetic relationships. We provide evidence that subclass III ADF5 and ADF9 are unconventional ADFs since they do not display typical actin filament depolymerizing activities. Instead, they exhibit opposite activities with a surprisingly high ability to stabilize and crosslink actin filaments into long and thick actin bundles both in vitro and in live cells. Competition experiments with ADF1 support that ADF9 antagonizes the depolymerizing activity of conventional ADFs. We report the characterization of a not yet described knockout Arabidopsis mutant. Data strongly suggests a role for ADF9 in cell elongation. Indeed, hypocotyls are significantly longer in adf9 mutant than in wild- type seedlings, and this phenotype is enhanced in dark growth conditions in which the ADF9 gene is normally preferentially expressed. The analysis of hypocotyl epidermal cells indicates that this phenotype is essentially due to an increase of cell expansion. Surprisingly, adf9 seedlings exhibit shorter roots than control plants, suggesting a complex link between actin cytoskeleton organization and cell elongation. Finally, the reduced ability of adf9- derived calli to proliferate supports a role for ADF9 in cell division as well.
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Muscle LIM protein and Nesprin-1 in Mechanotransduction / Muscle LIM protéine et Nesprin-1 dans MechanotransductionSchwartz, Christine 29 September 2016 (has links)
J’ai étudié trois protéines qui participent à deux vois différentes de méchano-transduction qui est la conversion des stimuli physiques en un signal biochimique.Dans une culture cellulaire en 2D, lorsque les cardiomyocytes sont étirés, MLP est transloqué vers le noyau. Sans translocation, les cellules ne parviennent pas à répondre à la stimulation. Les patients porteurs de mutations dans MLP développent une cardiomyopathie comme les souris MLP knock-out (MLP-/-). Mon objectif a été d’élucider le rôle de MLP dans ces cardiomyopathies en surexprimant des mutations de MLP dans les cardiomyocytes isolés des souris MLP-/- néonataux. Dans les cultures 2D mais pas 3D, MLP n’était pas transloqué vers le noyau après l’étirement des cellules. Bien que je n’aie pas pu résoudre ce problème, j’ai mis au point les expériences nécessaires à la poursuite de ce projet.Nesprins s’intègrent dans un complexe transmembranaire de l’enveloppe nucléaire (EN), le LINC complexe, qui connecte le cytosquelette à l’intérieur du noyau. Les myoblastes isolés des patients porteurs des mutations de Nesprin ou de Lamin, qui est associé au LINC complexe, ont présenté des noyaux déformés ainsi que des anomalies de réponses méchanosensibles : Si cultivées sur supports mous, les cellules affichaient un niveau élevé de fibres musculaires stressées et d’adhésions focales. Le knock-down de FHOD, une cible en aval de ROCK et SRC, qui également étaient actives dans ces myoblastes, a réduit ce phénotype. Bien que l’on ait émis l’hypothèse que les mutations dans Nesprins et Lamins conduisent à une instabilité mécanique de l’EN, ces résultats indiquent que les voies de signalisation par l’EN sont perturbées aussi. / I studied three striated muscle proteins that are participating in two different pathways of mechanotransduction, which is the translation of a physical stimulus into a biochemical signal.When isolated cardiomyocytes are stretched in 2D, MLP shuttles to the nucleus. Without shuttling MLP, these cells fail to respond to the stretch stimulus. Human patients with MLP-mutations develop cardiomyopathies, as well as mice with a knock-out of MLP (MLP-/-). By expressing mutated MLP in neonatal cardiomyocytes of MLP-/- mice, I wanted to elucidate the role of mutant MLP. Surprisingly, MLP did shuttle after stretching of 2D but not 3D cell cultures. Although I could not solve this issue, I prepared the setup for subsequent experiments.Nesprins are part of the nuclear envelope (NE) spanning LINC complex, which connects the cytoskeleton with the nucleus. Myoblasts from patients with mutations in Nesprins or LINC-associated Lamins displayed deformed nuclei and had defects in mechanosensitive responses with an elevated level of stress fibers and focal adhesions on soft surfaces. This phenotype could be rescued by knock-down of formin FHOD1, a downstream target of ROCK and SRC, which also were highly active in the mutant cells. While mutations in Nesprins and Lamins are thought to lead to mechanical instability of the NE, these results indicate that signaling pathways through the NE are disturbed as well.
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The Role of LMO4 in the Regulation of SLK Localization & Activation within Migrating Cells and in Murine Mammary TumorigenesisBaron, Kyla Doreen January 2016 (has links)
The Ste20-like kinase SLK plays a pivotal role in cell migration and focal adhesion turnover. SLK activity is regulated by the LIM domain-binding proteins Ldb1/2. In addition to playing role in tumor initiation and progression, these proteins have been demonstrated to interact with LMO4. Therefore, this project assessed the ability of LMO4 to interact and regulate SLK activity. Results show that LMO4 can directly bind to SLK and activate its kinase activity. LMO4 can be co-precipitated with SLK following the induction of cell migration by scratch wounding. Cre deletion of LMO4 inhibits cell migration and SLK activation, and impairs Ldb1 and SLK recruitment to the leading edge of migrating cells. Src/Yes/Fyn-deficient cells (SYF) express very low levels of LMO4 and do not recruit SLK to the leading edge. Src-family kinase inhibition impairs SLK recruitment to the leading edge, suggesting that both expression of LMO4 and the recruitment of SLK to the leading edge require c-Src activity. In conclusion, cell migration and activation of SLK requires its recruitment to the leading edge by LMO4 in a Src-dependent manner. This study also investigated whether LMO4 deletion through MMTV-Cre-driven excision would impair mammary tumorigenesis in a PyMT mouse model of breast cancer. No difference in Overall Survival was observed between animals with and without LMO4 expression. Western blot analysis and IHC showed that tumors expressed LMO4 protein in animals genotyped as Cre-positive. This result suggests that expression of LMO4 is required for tumor initiation in the PyMT model of murine mammary carcinoma. This project has established a novel cytosolic role for the transcriptional co-activator LMO4 and validated it’s involvement in the regulation of SLK and cell migration. This pathway may provide a novel therapeutic strategy as LMO4 appears to be critical to the initiation and progression of breast cancer.
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The quasi two-day wave: the results of numerical simulation with the COMMA - LIM ModelFröhlich, Kristina, Jacobi, Christoph, Lange, Martin, Pogoreltsev, Alexander 05 January 2017 (has links)
The quasi two-day wave (QTDW), a prominent feature of the mesosphere mainly around solstices, is simulated with the COMMA-LIM Model (Cologne Model of the Middle Atmosphere - Leipzig Institute for Meteorology). The calculations are made approximately one month after the summer solstice in the Northern Hemisphere when the QTDW reaches its maximum in the mesosphere and lower thermosphere. The results show that the QTDW produces a moderate westward forcing of the zonally averaged flow and a poleward driving of the residual mean meridional circulation. / Die Quasi Zwei-Tage Welle (QTDW), eine deutliche Erscheinung in der Mesosphäre kurz nach dem Sommer Solstitium, wird mit dem COMMA-LIM Modell (Cologne Model of the Middle Atmosphere - Leipzig Institute for Meteorology) simuliert. Die Zwei-Tage Welle wurde unter Juli-Bedingungen an der unteren Modellgrenze angeregt, zu der Zeit,
zu der sie ihr Maximum in der Mesosphäre und unteren Thermosphäre erreicht. Die Ergebnisse zeigen eine sich westwärts ausbreitende Welle, die auf den Grundstrom eine moderate Beschleunigung nach Westen ausübt. Die residuelle mittlere Meridional Zirkulation erfährt dadurch eine zum Pol gerichtete Triebkraft.
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Characterization of FHL2 gene and its role in human hepatocellular carcinoma. / CUHK electronic theses & dissertations collectionJanuary 2011 (has links)
Ng, Chor Fung. / Thesis (Ph.D.)--Chinese University of Hong Kong, 2011. / Includes bibliographical references (leaves 156-169). / Electronic reproduction. Hong Kong : Chinese University of Hong Kong, [2012] System requirements: Adobe Acrobat Reader. Available via World Wide Web. / Abstract also in Chinese.
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Functional characterization of FHL2 by microarray analysis and promoter study. / CUHK electronic theses & dissertations collectionJanuary 2013 (has links)
Xu, Jiaying. / Thesis (M.Phil.)--Chinese University of Hong Kong, 2013. / Includes bibliographical references (leaves 98-107). / Electronic reproduction. Hong Kong : Chinese University of Hong Kong, [2012] System requirements: Adobe Acrobat Reader. Available via World Wide Web. / Abstracts also in Chinese.
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Functional study of LIM-homeodomain proteins Lhx1 and Lhx5 in the maintenance of cerebellar Purkinje neurons in the postnatal and adult mouse. / CUHK electronic theses & dissertations collectionJanuary 2012 (has links)
蒲金氏細胞(Purkinje cell)是小腦中的一種主要神經元,其主要作用在於協調身體活動及平衡。蒲金氏細胞之早期分化需要兩個密切相關的LIM同源盒結構域基因Lhx1及Lhx5。在胚胎小腦發育期間,這兩個基因的失活化會導致蒲金氏細胞數量大量減少。但有趣的是,就算在蒲金氏細胞完成分化之後,Lhx1/5之表達依然維持在高水平。這顯示Lhx1/5在產後小腦發育過程中可能有更多作用。為了研究這些可能作用,我把條件性Lhx1/5雙基因剔除小鼠和Pcp2-IRES-Cre轉基因小鼠交配,從而令Lhx1/5在產後第二天的蒲金氏細胞失活化。結果顯示Lhx1/5雙突變體老鼠在出生後兩星期即有顯著但程度不太大的運動失調。但在八星期,牠們出現嚴重的運動協調及身體平衡能力缺失。可是,擁有一個正常的Lhx1或Lhx5等位基因的控制小鼠並沒有這些不正常行為出現。在出生後的三個星期內,缺乏Lhx1/5會導致蒲金氏細胞樹突不正常發展,但小腦的整體細胞結構和分層卻維持正常。另外,這兩個基因對維持蒲金氏細胞已發展的樹突並不起作用,而且在六個月大的成年突變小鼠並沒有蒲金氏細胞退化。利用微陣列及逆轉錄聚合酶鏈式反應,我們在成年突變小鼠的小腦中確定了數個參與在麩胺酸及鈣訊息的突觸基因表達量下降。而這些突觸基因也在其他運動失調小鼠有下降的表達量。研究結果說明了Lhx1及Lhx5對蒲金氏細胞樹突發展有著重要、但功能重疊的作用。 / 在探究Lhx1/5如何控制蒲金氏細胞樹突發展時,我們發現Lhx1/5與Foxp4有蛋白質交互作用。Foxp4屬forkhead家族成員轉錄因子,它表達在小腦原基、遷移中及成熟的蒲金氏細胞。為了初步瞭解Foxp4在蒲金氏細胞發展中的作用,我在產後第十天小腦薄片組織培養中,利用siRNA降低Foxp4基因的表達量。結果發現蒲金氏細胞樹突及關聯的伯格曼膠質細胞支架出現結構性受損。這顯示Foxp4對維持蒲金氏細胞樹突有重要作用。 / 為了進一步研究Foxp4在活體蒲金氏細胞及小腦發育的作用,我把條件性Foxp4基因剔除小鼠和不同的Cre轉基因小鼠交配,從而令Foxp4在不同的發育過程階段中失活化。但是有趣地,我只能在同質結合突變小鼠 (Foxp4Δ/Δ),即Foxp4在生殖細胞時期已經被剔除的情況下,觀察到小腦發育遲緩。當Foxp4在其他發育過程階段中失活化,我並沒有觀察到任何缺陷表型。這個結果顯示了在活體中發生了功能性的彌補,但在小腦薄片組織培養中卻沒有發生。另外,條件性Lhx1/5雙基因剔除小鼠和條件性Foxp4基因剔除小鼠的不同表型意味著在控制蒲金氏細胞及小腦發育過程中,有其他蛋白質可能參與在Lhx1/5及Foxp4的轉錄複合子中。我們需要更多的研究去明白Foxp和 LIM同源盒結構域蛋白質在功能上的聯系及它們在中樞神經系統發育中的作用。 / Purkinje cells (PCs) are one of the principal neurons in the cerebellum that is essential for the coordination of fine-tuning body movement and balancing. Early differentiation of PCs requires two closely related LIM-homeodomain genes Lhx1 and Lhx5, as inactivation of both genes results in significant reduction of PC number in embryonic cerebellum. Interestingly, high levels of Lhx1/5 expressions persist even after PC differentiation in the postnatal cerebellum. Hence, there may be additional roles for these two genes during postnatal PC development. To address this question, conditional inactivation approach was used to inactivate both Lhx1/5 in postnatal PCs specifically beginning at postnatal day 2 (P2). Lhx1/5 double conditional knockout (DKO) mutants were generated by crossing Lhx1/5 conditional null mutant mice with Pcp2-IRES-Cre mice. The mutants initially showed modest but noticeable ataxic locomotion at around two weeks after birth. However at 8 weeks old, the mutants displayed severe deficits in motor coordination and body balance. The control animals with one functional copy of either Lhx1 or Lhx5 did not show any abnormality. Deficiencies of both genes could lead to abnormal PC dendritogenesis during the first three weeks of life although the general cytoarchitectural lamination of cerebellar cortex was maintained. However, the two genes were dispensable for the maintenance of developed dendrites in adult mouse and no PC degeneration was observed in the 6 month-old double mutant mouse. Further microarray and semi-quantitative RT-PCR analysis identified down-regulation of several synaptic genes that involved in glutamate and/or calcium signaling in our Lhx1/5 DKO mutant and such disturbance had also been found in other ataxic mouse models. Overall, our findings suggest that Lhx1/5 are required but functionally redundant in dendritogenesis of PCs. / During investigation on how Lhx1/5 control the dendritogenesis of PCs, Lhx1/5 proteins were found to physically interact with Foxp4. Foxp4 belongs to the forkhead transcription factor family that is expressed in developing cerebellum primordium, migrating and mature PCs. To initially examine the function of Foxp4 in PC development, Foxp4 was knocked down by siRNA in organotypic cerebellar slice culture at P10. Impaired organization of PC dendritic arbors and associated Bergmann glial scaffold were resulted, suggesting that Foxp4 is essential for the maintenance of PC dendritic arborization. / To further investigate the function of Foxp4 during the cerebellum and PCs development in vivo, a Foxp4 CKO mouse line was generated and crossed with different lines of Cre-deleter mice, including Zp3-Cre, Pax2-Cre, En1-Cre and Pcp2-IRES-Cre, to inactivate Foxp4 at different developmental stages. Intriguingly, although developmental delay of cerebellum was found in germline deletion of Foxp4 homozygous recombined null mutant, no defective phenotype was observed when Foxp4 was inactivated at other stages. Hence, functional compensation might take place in vivo but not in the cerebellar slice culture. The phenotypic difference between Lhx1/5 DKO and Foxp4 CKO mice imply potential involvement of other proteins in the transcription complex between Lhx1/5 and Foxp4 in regulating the cerebellum and/or PCs development. Thus further investigation is required to understand the functional association between Foxp and LIM-homeodomain protein families during the development of central nervous system. / Detailed summary in vernacular field only. / Detailed summary in vernacular field only. / Detailed summary in vernacular field only. / Tam, Wing Yip. / Thesis (Ph.D.)--Chinese University of Hong Kong, 2012. / Includes bibliographical references (leaves 187-203). / Electronic reproduction. Hong Kong : Chinese University of Hong Kong, [2012] System requirements: Adobe Acrobat Reader. Available via World Wide Web. / Abstract also in Chinese. / Abstract --- p.1 / 摘要 --- p.4 / Acknowledgements --- p.6 / Abbreviations --- p.8 / Figure list --- p.12 / Table list --- p.15 / Chapter Chapter 1 --- General Introduction --- p.16 / Chapter 1.1 --- An overview of cerebellum functions and anatomy --- p.16 / Chapter 1.2 --- Purkinje cell development in the mouse --- p.19 / Chapter 1.2.1 --- Embryonic development of mouse cerebellum --- p.19 / Chapter 1.2.2 --- Postnatal development of mouse cerebellum --- p.21 / Chapter 1.3 --- Degeneration of Purkinje cell leads to spinocerebellar ataxia --- p.22 / Chapter 1.4 --- LIM-homeodomain genes Lhx1 and Lhx5 --- p.24 / Chapter 1.4.1 --- LIM-homeodomain --- p.24 / Chapter 1.4.2 --- Lhx1 and Lhx5 are crucial to Purkinje cell differentiation --- p.25 / Chapter 1.5 --- Hypothesis, aim and strategy of the study --- p.26 / Chapter Chapter 2 --- Generation of Lhx5 conditional knockout allele in the mouse --- p.31 / Chapter 2.1 --- Chapter summary --- p.31 / Chapter 2.2 --- Introduction --- p.32 / Chapter 2.3 --- Materials and methods --- p.35 / Chapter 2.3.1 --- Materials --- p.35 / Chapter 2.3.2 --- Construction of Lhx5-conditional targeting vector by recombineering --- p.39 / Chapter 2.3.3 --- Gene targeting in mouse embryonic stem cells --- p.53 / Chapter 2.3.4 --- Generation of Lhx5 CKO mouse --- p.62 / Chapter 2.3.5 --- Histological examination of Lhx5 CKO mouse brain --- p.63 / Chapter 2.4 --- Results --- p.64 / Chapter 2.4.1 --- Generation of Lhx5 conditional targeting construct --- p.64 / Chapter 2.4.2 --- Screening of targeted ES cell clones --- p.64 / Chapter 2.4.3 --- Karyotyping --- p.65 / Chapter 2.4.4 --- Generation of chimeric mice and maintenance of Lhx5 CKO mice --- p.66 / Chapter 2.4.5 --- Histological examination of Lhx5 recombined null mutant mouse --- p.67 / Chapter 2.4.6 --- Gross anatomical examination of Lhx5 recombined null mutant mouse --- p.69 / Chapter 2.5 --- Discussion --- p.71 / Chapter Chapter 3 --- Generation and characterization of Pcp2-CreER[superscript T]² transgenic mouse --- p.75 / Chapter 3.1 --- Chapter summary --- p.75 / Chapter 3.2 --- Introduction --- p.76 / Chapter 3.3 --- Materials and methods --- p.79 / Chapter 3.3.1 --- Materials --- p.79 / Chapter 3.3.2 --- Construction of pPcp2-IRES-CreER[superscript T]²-FRT-Kan-FRT --- p.81 / Chapter 3.3.3 --- Generation of BAC-Pcp2-IRES-CreER[superscript T]² transgene --- p.85 / Chapter 3.3.4 --- Generation of Pcp2-CreER[superscript T]² transgenic mice --- p.90 / Chapter 3.3.5 --- Characterization of Pcp2-CreER[superscript T]² transgenic mice --- p.91 / Chapter 3.4 --- Results --- p.93 / Chapter 3.4.1 --- Construction of BAC-Pcp2-IRES-CreER[superscript T]² --- p.93 / Chapter 3.4.2 --- Production of Pcp2-CreER[superscript T]² transgenic mice --- p.93 / Chapter 3.4.3 --- Expression of Cre recombinase in Pcp2-CreER[superscript T]² transgenic mice --- p.93 / Chapter 3.4.4 --- Histological examination of Pcp2-CreER[superscript T]² transgenic mice --- p.97 / Chapter 3.4.5 --- Behavioral test of Pcp2-CreER[superscript T]² transgenic mice by rotarod --- p.98 / Chapter 3.5 --- Discussion --- p.100 / Chapter Chapter 4 --- Characterization of Lhx1/5 double conditional knockout mouse --- p.103 / Chapter 4.1 --- Chapter summary --- p.103 / Chapter 4.2 --- Introduction --- p.104 / Chapter 4.3 --- Materials and methods --- p.106 / Chapter 4.3.1 --- Mouse strain --- p.106 / Chapter 4.3.2 --- Behavioral tests --- p.106 / Chapter 4.3.3 --- Histological examination of cerebellum --- p.107 / Chapter 4.3.4 --- CreER[superscript T]² induction by tamoxifen --- p.108 / Chapter 4.3.5 --- Gene expression profiling using microarray --- p.109 / Chapter 4.3.6 --- Transmission electron microscopy --- p.110 / Chapter 4.3.7 --- Statistical analysis --- p.111 / Chapter 4.4 --- Results --- p.112 / Chapter 4.4.1 --- Early postnatal developmental delay in female DKO mutant --- p.112 / Chapter 4.4.2 --- Lhx1/5 DKO mutants displayed significant motor deficit --- p.114 / Chapter 4.4.3 --- Abnormal Purkinje cell dendritic arborization in the adult Lhx1/5 DKO mutant mouse --- p.117 / Chapter 4.4.4 --- Reduction in the number of synaptic vesicles in the adult Lhx1/5 DKO mutant --- p.119 / Chapter 4.4.5 --- Abnormal Purkinje cell dendrite development in the Lhx1/5 DKO mutant mouse --- p.120 / Chapter 4.4.6 --- Lhx1/5 were not required for the maintenance of developed Purkinje cell dendrite --- p.122 / Chapter 4.4.7 --- Comparison of gene expression profiles in the Lhx1/5 DKO mutant and control --- p.128 / Chapter 4.5 --- Discussion --- p.130 / Chapter Chapter 5 --- Foxp4 - a potential interacting partner of Lhx1/5 --- p.137 / Chapter 5.1 --- Chapter summary --- p.137 / Chapter 5.2 --- Introduction --- p.138 / Chapter 5.3 --- Materials and methods --- p.139 / Chapter 5.3.1 --- Co-immunoprecipitation --- p.139 / Chapter 5.3.2 --- Foxp4 expression pattern and knockdown in cerebellar slice culture --- p.140 / Chapter 5.3.3 --- Generation of Foxp4 CKO mouse --- p.146 / Chapter 5.4 --- Results --- p.148 / Chapter 5.4.1 --- Lhx1 and Lhx5 physically interacted with Foxp4 --- p.148 / Chapter 5.4.2 --- Foxp4 expression during mouse cerebellum development --- p.150 / Chapter 5.4.3 --- Effective gene silencing by siRNA in cerebellar slice culture --- p.151 / Chapter 5.4.4 --- Silencing gene expression of Foxp4 at P5 exerted no observable effect on Purkinje cell survival or differentiation --- p.154 / Chapter 5.4.5 --- Developed Purkinje cell dendritic arbors and associated Bergmann glial fibers were impaired when Foxp4 was knockdown at P10 --- p.157 / Chapter 5.4.6 --- Generation of Foxp4 targeting construct and conditional knockout mouse --- p.159 / Chapter 5.4.7 --- Developmental delay of cerebellum in Foxp4 recombined homozygous mutants --- p.163 / Chapter 5.4.8 --- Normal cerebellum development in adult En1-Cre; Foxp4[superscript fx/fx] and Pax2-Cre; Foxp4[superscript fx/fx] mutants --- p.165 / Chapter 5.4.9 --- Purkinje cell-specific knockout of Foxp4 did not impair Purkinje cell maintenance, motor activity and learning --- p.167 / Chapter 5.5 --- Discussion --- p.171 / Chapter 5.6 --- Acknowledgements --- p.177 / Chapter Chapter 6 --- General discussion, future works and conclusion --- p.179 / Chapter 6.1 --- Evolutionary conserved function of Lhx1 and Lhx5 in neurons --- p.180 / Chapter 6.2 --- LHX1 and LHX5 in human diseases --- p.181 / Chapter 6.3 --- Transcription complex between LIM-homeodomain and forkhead domain proteins may be important in the cerebellum development --- p.182 / Chapter 6.4 --- Future works --- p.183 / Chapter 6.5 --- Conclusion --- p.186 / References --- p.187
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Exploring the Use of Consumer Grade Technology for Kinematic Assessment of the Upper Limb Following a StrokeTran, Johnathan 20 June 2014 (has links)
Upper limb deficits post stroke affect up to 60% of stroke survivors. The assessment of motor deficits post stroke is important for identifying rehabilitation goals and assessing treatment efficacy. Current clinical tools used to assess motor impairment utilize clinical observation to describe the performance of diagnostic motor tasks. However there are some concerns regarding the ability of these scales to fully describe the quality of performance, and detect small but important changes which reflect motor recovery. Kinematic analysis has been increasingly suggested to augment clinical assessment; however, current kinematic tools are not well suited to the time and financial constraints of a clinical environment. The objective of this thesis was to investigate the feasibility of utilizing low-cost, depth sensing technology (Kinect sensor) to augment the current upper limb stroke assessment. Study one characterizes the accuracy of the Kinect sensor, and defines optimal markers and conditions for data collection. Results revealed sufficient ability to quantify metrics for the hand, and the trunk. Study two explored the feasibility of clinical use for the Kinect sensor, specifically its ability to distinguish kinematic performance between the affected and less-affected limbs within an individual, and differences in the affected limb between individuals. Results from study 2 indicated that the Kinect is able to identify interlimb differences and correlations with upper limb impairment scores for some kinematic metrics. Findings from this thesis suggest a potential use for the Kinect in a clinical environment for the purposes of upper limb stroke assessment; however, there are many factors and limitations which need to be considered prior to its use.
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The effect of mission trips on mission-mindednessCho, Hyun Chul. January 2005 (has links)
Thesis (D. Min.)--Southwestern Baptist Theological Seminary, 2005. / Added title page in Korean: Sŏnʼgyo yŏhaeng i sŏnʼgyo ŭisik pyŏnhwa e michʻinŭn yŏnghyang. 880-02 Includes bibliographical references (leaves 93-101).
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Desenvolvimento de novos biolubrificantes hidr?ulicos derivados dos ?leos de maracuj? e de moringa in natura e epoxidados / Development of new hydraulic biolubricants derived from passion fruit and moringa oils in natura and epoxidizedSilva, Maria Susana 02 February 2015 (has links)
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Previous issue date: 2015-02-02 / Conselho Nacional de Desenvolvimento Cient?fico e Tecnol?gico - CNPq / O aumento da consci?ncia ambiental, maximizando a biodegradabilidade e
minimizando a ecotoxicidade, ? a for?a motriz principal para os novos desenvolvimentos
tecnol?gicos. Deste modo, os lubrificantes biodegrad?veis utilizados em ?reas
ambientalmente sens?veis podem ser mais explorados. O objetivo desse trabalho foi obter
novos biolubrificantes a base de ?leo de maracuj? (Passiflora edulis Sims f. flavicarpa
Degener) e de moringa (Moringa oleifera Lamarck), in natura e epoxidados e otimizar o uso
de um novo pacote de aditivos atraves da metodologia de planejamento experimental para sua
utiliza??o como fluido hidr?ulico. Na primeira etapa do trabalho foi realizada a otimiza??o do
processo de epoxida??o dos ?leos utilizando o planejamento experimental fracionado 24-1
,
variando a temperatura, tempo de rea??o, raz?o molar de ?cido f?rmico e a raz?o molar de
per?xido de hidrog?nio. Em seguida, foi investigada a seletividade, termodin?mica e a
cin?tica da obten??o dos dois ep?xidos a 30, 50 e 70 ?C. Os resultados dessa primeira etapa
confirmaram que para a epoxida??o do ?leo de maracuj? s?o necess?rias 3 horas de rea??o, 30
?C e uma raz?o H2O2/C=C/HCOOH (1:1:1) e para o ?leo de moringa foram necess?rias 3
horas de rea??o, 30 ?C e uma raz?o de H2O2/C=C/HCOOH (1:1:1,5). Os resultados das
convers?es finais foram iguais a 83,09% (?0,3) para o ep?xido de ?leo de maracuj? e 91,02%
(?0,4) para o ep?xido do ?leo de moringa. Em seguida, foi feito o planejamento experimental
fatorial 23
para avaliar quais as melhores concentra??es do inibidor de corros?o e antidesgaste
(IC), antioxidante (BHA) e extrema press?o (EP). Os biolubrificantes obtidos nessa etapa
foram caracterizados segundo as normas DIN 51524 (Part 2 HLP) e DIN 51517 (Part 3 CLP).
O processo de epoxida??o dos ?leos foi capaz de melhorar a estabilidade oxidativa e diminuir
o ?ndice de acidez total quando comparados aos ?leos in natura. Em termos de desempenho
f?sico-qu?mico como lubrificantes, o melhor fluido foi o ep?xido do ?leo de moringa
aditivado (EMO-ADI), seguido do ep?xido do ?leo maracuj? aditivado (EMA-ADI) e, por
?ltimo, o ?leo de maracuj? in natura sem aditivos (OMA). Para finalizar, foi feita a
investiga??o de seus comportamentos tribol?gicos sob condi??es de lubrifica??o lim?trofe. O
desempenho tribol?gico dos lubrificantes desenvolvidos foi analisado em um equipamento
HFRR (High Frequency Reciprocating Rig) e atrav?s da medi??o do coeficiente de atrito, que
ocorre durante o contato e a forma??o do filme lubrificante, durante o ensaio. O desgaste foi
avaliado atrav?s de microscopia ?ptica e microscopia eletr?nica de varredura (MEV).
Observou-se que, em todos os ensaios, foi formado um filme lubrificante e que a adi??o de EP
e IC nas bases vegetais de ?leo de maracuj? e moringa in natura n?o ocasionou uma redu??o
significativa no desgaste. Os biolubrificantes desenvolvidos a partir de ?leo de maracuj? e
moringa modificados via epoxida??o apresentaram propriedades tribol?gicas superiores aos
fluidos comerciais testados, mostrando serem potenciais lubrificantes para substitui??o dos
fluidos comerciais de base mineral. / With the increasing environmental awareness, maximizing biodegradability
and minimizing ecotoxicity is the main driving force for new technological developments.
Thus, can be developed new biodegradable lubricants for use in environmentally sensitive
areas. The aim of this study was to obtain new bio-lubricants from passion fruit (Passiflora
edulis Sims f. flavicarpa Degener) and moringa (Moringa oleifera Lamarck) epoxidized oils
and develop a new additive package using experimental design for their use as a hydraulic
fluid. In the first stage of this work was performed the optimization of the epoxidation process
of the oils using fractional experimental design 24-1
, varying the temperature, reaction time,
ratio of formic acid and hydrogen peroxide. In the second step was investigated the
selectivity, thermodynamics and kinetics of the reaction for obtaining the two epoxides at 30,
50 and 70 ?C. The result of the experimental design confirmed that the epoxidation of passion
fruit oil requires 2 hours of reaction, 50 ?C and a ratio H2O2/C=C/HCOOH (1:1:1). For
moringa oil were required 2 hours reaction, 50 ?C and a ratio of H2O2/C=C/HCOOH
(1:1:1.5). The results of the final conversions were equal to 83.09% (? 0.3) for passion fruit
oil epoxide and 91.02 (?0,4) for moringa oil epoxide. Following was made the 23
factorial
design to evaluate which are the best concentrations of corrosion inhibitor and anti-wear (IC),
antioxidant (BHA) and extreme pressure (EP) additives. The bio-lubricants obtained in this
step were characterized according to DIN 51524 (Part 2 HLP) and DIN 51517 (Part 3 CLP)
standards. The epoxidation process of the oils was able to improve the oxidative stability and
reduce the total acid number, when compared to the in natura oils. Moreover, the epoxidized
oils best solubilized additives, resulting in increased performance as a lubricant. In terms of
physicochemical performance, the best lubricant fluid was the epoxidized moringa oil with
additives (EMO-ADI), followed by the epoxidized passion fruit oil with additives (EPF-ADI)
and, finally, the passion fruit in natura oil without additives (PFO). Lastly, was made the
investigation of the tribological behavior under conditions of boundary lubrication for these
lubricants. The tribological performance of the developed lubricants was analyzed on a HFRR
equipment (High Frequency Reciprocating Rig) and the coefficient of friction, which occurs
during the contact and the formation of the lubricating film, was measured. The wear was
evaluated through optical microscopy and scanning electron microscopy (SEM). The results
showed that the addition of extreme pressure (EP) and anti-wear and corrosion inhibitor (CI)
additives significantly improve the tribological properties of the fluids. In all assays, was
formed a lubricating film that is responsible for reducing the coefficient of metal-to-metal
wear. It was observed that the addition of EP and IC additives in the in natura vegetable oils
of passion fruit and moringa did not favor a significant reduction in wear. The bio-lubricants
developed from passion fruit and moringa oils modified via epoxidation presented satisfactory
tribological properties and shown to be potential lubricants for replacement of commercial
mineral-based fluids.
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