Le myélome multiple (MM) est une hémopathie maligne, caractérisée par la prolifération au niveau de la moelle osseuse de plasmocytes tumoraux sécrétant le plus souvent une immunoglobuline monoclonale. Malgré de récents progrès thérapeutiques, cette maladie reste aujourd’hui incurable avec une survie médiane d’environ 6 ans. Cependant, il existe une grande hétérogénéité pronostique parmi les patients qui est associée à différentes anomalies génétiques. Parmi celles-ci, la translocation t(4;14) (p16;q32), retrouvée dans 15% des MM, définit l’une des formes de MM actuellement la plus grave. Malgré le rôle potentiel des deux oncogènes FGFR3 et MMSET qu’elle dérégule, les mécanismes moléculaires permettant d’expliquer la gravité des MM avec t(4;14) ne sont pas clairement élucidés. En mettant en œuvre des approches de biologie moléculaire, de séquençage à haut débit et de tests fonctionnels sur une très large série de MM t(4;14), mes travaux ont cherché à identifier les événements génétiques associés au mauvais pronostic de ce sous-groupe de malades. Les résultats montrent l’implication du point de cassure au sein du gène MMSET dans le pronostic des patients. Ils ont également permis de définir le paysage particulier des mutations génétiques affectant les MM t(4;14) en mettent en évidence les fréquences relativement élevées des altérations dans les gènes ATM/ATR et PRKD2. Enfin, ils ont identifiés PKD2 comme une cible thérapeutique et démontré qu’un inhibiteur des PKDs, kb NB 142-70, bloquait la prolifération des cellules de MM in vitro. L’ensemble de ces résultats jette un éclairage nouveau sur la physiopathologie des MM t(4;14) et ouvre la voie vers de nouvelles approches thérapeutiques. / Multiple myeloma (MM) is a hematological malignancy characterized by the proliferationof plasma cells in the bone marrow usually secreting a monoclonal immunoglobulin. Despite recenttherapeutic advances, the disease remains incurable, with a median survival of approximately 6years. However, there is considerable clinical heterogeneity between patients, with differentoutcome primarily associated with discrete genetic abnormalities. Among these, the t (4;14) (p16;q32), which is found in 15% of MM, defines one of the most serious subgroups of MM. Despite thepotential role of two deregulated oncogenes (FGFR3 and MMSET), the molecular mechanisms thatexplain the severity of MM with t (4;14) have not been clearly elucidated. By implementing theapproaches of molecular biology, high throughput sequencing and functional tests on a wide rangeof MM t (4;14), my project sought to identify genetic events associated with the poor prognosis ofthis subgroup of patients. The results showed that the site of the translocation breakpoints withinthe MMSET gene affect patient outcome. They also profiled the landscape of specific geneticmutations affecting t (4;14) MM, and revealed a relatively high frequency of alterations in the ATM/ ATR genes and PRKD2. Finally, this project has identified PKD2 as a therapeutic target anddemonstrated that an inhibitor of PKDs (kb NB 142-70) blocks the proliferation of MM cells in vitro.All these results throw new light on the pathophysiology of MM t (4;14) and opens the way to newtherapeutic approaches.

No description available.

Membrane proteins comprise around 20-30% of most proteomes. They play important roles in most biochemical pathways. All receptors and ion channels are membrane proteins, which make them attractive targets for drug design. Membrane proteins insert and fold co-translationally into the endoplasmic reticular membrane of eukaryotic cells. The protein-conducting channel that inserts the protein into the membrane is called Sec61 translocon, which is a hetero-oligomeric channel that allows transmembrane segments to insert laterally into the lipid bilayer. The focus of this thesis is how the translocon recognizes the transmembrane helices and integrates them into the membrane. We have investigated the sequence requirements for the translocon-mediated integration of a transmembrane α-helix into the ER by challenging the Sec61 translocon with designed polypeptide segments in an in vitro expression system that allows a quantitative assessment of membrane insertion efficiency. Our studies suggest that helices might interact with each other already during the membrane-insertion step, possibly forming helical hairpins that partition into the membrane as a single unit. Further, the insertion efficiency for Nin-Cout vs. Nout-Cin transmembrane helices and the integration efficiency of Alzheimer’s Aβ-peptide fragments has been investigated. Finally, detailed topology mapping was performed on two biologically interesting proteins with unknown topology, the human seipin protein and Drosophila melanogaster odorant receptor OR83b.

Tsai Sau-Na. / Thesis submitted in: 2003. / Thesis (M.Phil.)--Chinese University of Hong Kong, 2004. / Includes bibliographical references (leaves 157-170). / Abstracts in English and Chinese. / Thesis committee --- p.i / Statement --- p.ii / Abstract --- p.iii / Acknowledgements --- p.vii / General Abbreviations --- p.ix / Abbreviations of Chemicals --- p.xii / Table of contents --- p.xiv / List of figures --- p.xx / List of tables --- p.xxii / Chapter 1. --- Literature Review --- p.1 / Chapter 1.1 --- Salinization is a global problem --- p.1 / Chapter 1.2 --- Causes of soil salinization in agricultural lands --- p.2 / Chapter 1.3 --- Toxicity of salinity in plants --- p.4 / Chapter 1.3.1. --- Physiological drought --- p.4 / Chapter 1.3.2. --- Nutritional imbalance --- p.5 / Chapter 1.3.3 --- Specific ion toxicity --- p.6 / Chapter 1.4 --- Plant adaptation to salinity --- p.7 / Chapter 1.5 --- Ion transport proteins in plant --- p.10 / Chapter 1.5.1 --- "Pump, channel and carrier" --- p.10 / Chapter 1.5.2 --- Pumps --- p.11 / Chapter 1.5.2.1 --- P-ATPase --- p.11 / Chapter 1.5.2.2 --- V-ATPase --- p.12 / Chapter 1.5.2.3 --- PPiase --- p.12 / Chapter 1.5.3 --- Cation channels --- p.13 / Chapter 1.5.3.1 --- K+ channels --- p.13 / Chapter 1.5.3.1.1 --- Shaker family --- p.14 / Chapter 1.5.3.1.1.1 --- KIRCs --- p.16 / Chapter 1.5.3.1.1.2 --- KORCs --- p.17 / Chapter 1.5.3.1.1.3 --- VICs --- p.18 / Chapter 1.5.3.1.2 --- Kir family --- p.18 / Chapter 1.5.3.1.2 --- KCO family --- p.19 / Chapter 1.5.3.2 --- Ca2+ channels --- p.20 / Chapter 1.5.3.2.1 --- TPC family --- p.20 / Chapter 1.5.3.2.2 --- CNGC family --- p.21 / Chapter 1.5.4 --- Anion Channels --- p.22 / Chapter 1.5.5 --- Carriers --- p.23 / Chapter 1.5.5.1 --- High affinity K+ carriers --- p.23 / Chapter 1.5.5.1.1 --- HKT transporter --- p.24 / Chapter 1.5.5.1.2 --- HAK/KUP transporter --- p.25 / Chapter 1.5.5.2 --- Cation/H+ antiporters --- p.26 / Chapter 1.5.5.2.1 --- Na+/H+ antiporter --- p.27 / Chapter 1.5.5.2.2 --- Ca2+/H+ antiporters --- p.30 / Chapter 1.6 --- Ion homeostasis and salt tolerance --- p.31 / Chapter 1.6.1 --- Ion transporters involved in ion homeostasis during salt stress --- p.31 / Chapter 1.6.2 --- Sodium uptake under salt stress --- p.32 / Chapter 1.6.4 --- Sodium extrusion --- p.36 / Chapter 1.6.5 --- Intracellular compartmentation --- p.37 / Chapter 1.6.6 --- Genetic engineering of ion transporter for improvement of salt tolerance --- p.40 / Chapter 1.7 --- Soybean as a target for studies of salt tolerance --- p.41 / Chapter 1.7.1 --- Economic importance of soybean --- p.41 / Chapter 1.7.2 --- Salt tolerant soybean in China --- p.43 / Chapter 1.7.3 --- Previous studies of Wenfeng7 and Union in our laboratory --- p.43 / Chapter 1.7.4 --- Hypothesis and research strategy of my project --- p.46 / Chapter 2. --- Materials and methods --- p.49 / Chapter 2.1 --- Materials --- p.49 / Chapter 2.1.1. --- Plant materials --- p.49 / Chapter 2.1.2. --- Bacteria strains and plasmid vectors --- p.50 / Chapter 2.1.3. --- Growth media for soybeans and A. thaliana --- p.50 / Chapter 2.1.4. --- Chemicals and reagents used --- p.50 / Chapter 2.1.5. --- Solutions used --- p.51 / Chapter 2.1.6. --- Commercial kits used --- p.51 / Chapter 2.1.7. --- Equipment and facilities used --- p.51 / Chapter 2.1.8. --- Primers used --- p.51 / Chapter 2.2 --- Methods --- p.52 / Chapter 2.2.1 --- Cloning of ion transporters --- p.52 / Chapter 2.2.1.1. --- Sample preparation --- p.52 / Chapter 2.2.1.2 --- Total RNA extraction --- p.52 / Chapter 2.2.1.3 --- Primer design for RACE --- p.53 / Chapter 2.2.1.4 --- 5´ة& 3´ة RACE of ion transporters --- p.54 / Chapter 2.2.1.5 --- Subcloning of RACE cDNA fragments --- p.56 / Chapter 2.2.1.6 --- PCR screening of white colonies --- p.57 / Chapter 2.2.1.8 --- Preparation of recombinant plasmid for sequencing --- p.57 / Chapter 2.2.1.9 --- Sequencing and homology search --- p.58 / Chapter 2.2.1.10 --- Cloning of full length coding regions of ion transporters --- p.58 / Chapter 2.2.1.11 --- "Sequence comparison, analysis and multialignment" --- p.62 / Chapter 2.2.2 --- Gene expression profiles --- p.62 / Chapter 2.2.2.1 --- Sample stepwise treatment with different concentration of NaCl --- p.62 / Chapter 2.2.2.2 --- Sample treatment with different Hoagland's solution supplement with 1.2% NaCl --- p.63 / Chapter 2.2.2.3 --- Preparation of single-stranded DIG-labeled PCR probes --- p.64 / Chapter 2.2.2.4 --- Testing the concentration of DIG-labeled probes --- p.65 / Chapter 2.2.2.5 --- Northern blot technique --- p.66 / Chapter 2.2.2.6 --- RT-PCR (Reverse-transcription polymerase chain reaction) --- p.67 / Chapter 2.2.3 --- Functional test using transgenic plants --- p.68 / Chapter 2.2.3.1 --- Preparation of chimeric gene constructs and recombinant plasmids --- p.68 / Chapter 2.2.3.2 --- "Eletroporation of Agrobacterium, tumefaciens" --- p.69 / Chapter 2.2.3.3 --- Seed sterilization and plant growth --- p.70 / Chapter 2.2.3.4 --- Vacuum infiltration transformation of Arabidopsis thaliana --- p.71 / Chapter 2.2.3.5 --- Selection of hemizygous and homozygous transgenic plants --- p.72 / Chapter 2.2.3.6 --- Genomic DNA extraction and PCR screening --- p.72 / Chapter 2.2.3.7 --- RT-PCR and Northern Blot of transgenic plants --- p.73 / Chapter 2.2.3.8 --- Functional test on MS plate supplemented with NaCl --- p.73 / Chapter 2.2.3.9 --- Functional test on sand supplemented with Hoagland's solution and NaCl --- p.74 / Chapter 3. --- Results --- p.76 / Chapter 3.1 --- "Cloning of Nhx, AKT1 and CLC from Wenfeng7 and Union" --- p.76 / Chapter 3.1.1 --- "Cloning of 5'- & 3'- RACE cDNA fragments of Nhx, AKT1 and CLC" --- p.76 / Chapter 3.1.2 --- "Cloning of full length coding regions of Nhx, AKT1 and CLC from Wenfeng7 and Union" --- p.77 / Chapter 3.1.3 --- "Sequence comparison, analysis and multialignment" --- p.82 / Chapter 3.1.3.1 --- Sequence analysis and multialignment of GmNhx1 and GmNhx2 --- p.82 / Chapter 3.1.3.2 --- Sequence analysis and multialignment of GmAKTl --- p.92 / Chapter 3.1.3.3 --- Sequence analysis and multialignment of GmCLC --- p.101 / Chapter 3.2 --- "Gene expression profiles of GmNhx, GmCLC and GmAKTl" --- p.111 / Chapter 3.2.1 --- Induction of GmNhx and GmCLC gene expression by NaCl in different Hoagland's solution --- p.111 / Chapter 3.2.2 --- RT-PCR using gene specific primers to distinguish the gene expression of GmNhx1 and GmNhx2 --- p.116 / Chapter 3.2.3 --- RT-PCR analysis of the transcripts of GmAKTl in Wenfeng7 and Union --- p.118 / Chapter 3.3 --- Functional analysis of transgenic plants in salt tress --- p.120 / Chapter 3.3.1 --- "Construction of chimeric gene of GmNhx1´ة GmNhx2, GmCLC and GmAKT1 into V7 vector" --- p.120 / Chapter 3.3.2 --- Transformation of chimeric gene constructs into A. tumefaciens --- p.122 / Chapter 3.3.3 --- Vacuum infiltration transformation of Arabidopsis thaliana and selection of transgenic plants --- p.123 / Chapter 3.3.4 --- PCR screening of transgene from transgenic plants --- p.130 / Chapter 3.3.5 --- PT-PCR and Northern blot analysis of the transgene transcripts --- p.133 / Chapter 3.3.6 --- Functional test of transgenic plants under salt stress --- p.135 / Chapter 4. --- Discussion --- p.139 / Chapter 4.1 --- "Isolation of GmNhx, GmAKTl and GmCLC from Wenfeng7 and Union" --- p.139 / Chapter 4.1.1. --- GmNhx1 and GmNhx2 are putative vacuolar Na+/H+ antiporters from Wenfeng7 and Union --- p.139 / Chapter 4.1.2. --- GmAKT1 is an inward-rectifying K+ channel from Wenfeng7 and Union --- p.141 / Chapter 4.1.3 --- GmCLC is a putative vacuolar voltage-dependent chloride channel from Wenfeng7 and Union --- p.144 / Chapter 4.2 --- "Gene expression profiles of GmNhx, GmAKT1 and GmCLC from Wenfeng7 and Union" --- p.146 / Chapter 4.2.1 --- Differential expression between GmNhx1 and GmNhx2 in Wenfeng7 and Union --- p.146 / Chapter 4.2.2 --- Coordinated expression of GmNhx and GmCLC in wenfeng7 and Union --- p.147 / Chapter 4.2.3 --- GmAKT1 is preferentially expressed in roots of wenfeng7 and Union and presented in low abundance --- p.148 / Chapter 4.3 --- Functional tests of transgenic Arabidopsis plants --- p.150 / Chapter 4.3.1 --- Screening of heterozygous and homozygous transgenic plant --- p.150 / Chapter 4.3.2 --- Function tests of heterozygous and homozygous transgenic plants under salt stress --- p.151 / Chapter 4.3.3 --- Gene silencing in transgenic plants --- p.152 / Chapter 5. --- Conclusion and perspectives --- p.155 / References --- p.157 / "Appendix I: Buffer, restriction and modifying enzymes" --- p.171 / Appendix II: Major chemicals and reagents used in this research --- p.171 / Appendix III: Major common solutions used in this research --- p.174 / Appendix IV: Commercial kits used in this research --- p.177 / Appendix V: Major equipment and facilities used --- p.177

Thesis. 1977. Ph.D.--Massachusetts Institute of Technology. Dept. of Biology. / Microfiche copy available in Archives and Science. / Vita. / Bibliography : leaves 115-120. / Ph.D.

Tillage erosion is a major contributor to the total soil erosion in cultivated topographically complex lands. No study has been carried out on tillage erosion associated with cereal-based production systems in the Canadian Prairies, and there is a need to examine tillage erosivity of secondary tillage and seeding implements and the effect of slope curvature on tillage translocation. With both tillage and water erosion occurring in a cultivated topographically complex landscape, it is valuable to investigate the relative contributions of and the possible linkage and interactions between these two erosion processes. Tillage translocation causes the mixture of subsoil into the till-layer, which may considerably affect soil properties and therefore the related biophysical processes. In this study, using plot tracers, we examined tillage translocation caused by four tillage implements: air-seeder, spring-tooth-harrow, light-cultivator and deep-tiller in southern Manitoba, Canada. We determined that secondary tillage and seeding implements could be as erosive as primary tillage implements in a cereal-based production system. In the majority of cases, tillage translocation could be explained by slope gradient alone; however, slope curvature also significantly affected tillage translocation. In two field sites in the North America Great Plains (NAGP), measured 137Cs inventories were converted into total soil erosion rates. Tillage and water erosion rates were estimated using models. The comparisons of the model estimates to 137Cs estimates showed that both tillage and water erosion significantly contributed to the total soil erosion in undulating slopes while tillage erosion was the predominant erosion process in hummocky hilltops. The contributions of and the linkage and interactions between water and tillage erosion showed predictable patterns in different landform elements, with the knowledge of which, landscape segmentation could be used to assess the potential of soil erosion. Further investigation of tillage translocation was demonstrated with four hypothetic landscapes: plane slope, symmetric hill, asymmetric hill and irregular hill, and is tested against field data. A Visual Basic coded program (TillTM) was developed to simulate the redistribution of soil constituents and soil mass. We determined that the pattern of soil mass redistribution was dependent on topography, while the pattern of soil constituent redistribution was affect by topographic features, tillage patterns and temporal scales. / February 2007

Thèse de doctorat : Sciences du vivant. Aspects moléculaires et cellulaires de la biologie : Strasbourg 1 : 2008. / Titre provenant de l'écran-titre. Bibliogr. p. 173-190.

Reproduction de : Thèse d'université : Génie biologique et médical : Grenoble 1 : 1994. / Titre provenant de l'écran-titre.

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