Les voies de signalisation sont des moyens de communication intercellulaires très conservées au cours de l'évolution qui contrôlent tous les aspects du développement des organismes multicellulaires. La signalisation par les récepteurs Notch oriente les destins cellulaires au cours du développement embryonnaire et régule l'homéostasie des tissus. Dans les cellules normales, l'activation de la voie Notch est soumise à une régulation très stricte, ayant pour enjeu de maintenir un équilibre au sein des tissues entre prolifération et différenciation. Cependant des disfonctionnement de la voie Notch sont à l'origine de l'augmentation de la prolifération cellulaire, perturbant ainsi cet équilibre et aboutissant à la transformation maligne des cellules. Le rôle central de NOTCH1 dans l'oncogenèse humaine est soutenu par la présence de mutations activatrices de la voie dans plus de 50% des leucémies aigues lymphoblastiques T (LAL-T), ainsi qu'un large éventail de cancers hématopoïétiques et de tumeurs solides. Au cours des dernières décennies, d'importants progrès ont été réalisés dans la compréhension des mécanismes de transduction du signal Notch et l'identification des processus biologiques qui sont influencés par la voie Notch. Les fonctions physiologiques et pathologiques de NOTCH1 requièrent sa translocation dans le noyau, mais ses partenaires nucléaires et leurs rôles dans la tumorigénèse restent peu connus. De même, les mécanismes moléculaires par lesquelles Notch active la transcription des gènes cibles de la voie restent à élucider.Afin de comprendre les fonctions nucléaires de Notch dans un contexte tumoral, nous avons, par une approche protéomique, caractérisé les complexes protéiques associés à la forme oncogénique de NOTCH1 à partir d'extraits nucléaires de cellules leucémiques T humaines. Nos travaux, associant des approches biochimiques et fonctionnelles, mettent en évidence le rôle central de plusieurs enzymes et facteurs nucléaires dans le contrôle de l'activité transcriptionnelle et les fonctions oncogéniques de Notch.La perturbation de l'expression de plusieurs facteurs nouvellement identifiées induit un arrêt du cycle cellulaire et altère la prolifération in vitro des cellules cancéreuses portant des mutations de la voie Notch. De façon remarquable, l'inhibition des partenaires de NOTCH1 est capable de supprimer la croissance tumorale de cellules leucémiques humaines dans un modèle de xénogreffe murin. Ces travaux auront un impact important dans le développement des nouvelles stratégies capables d'interférer avec les fonctions oncogéniques de Notch et pourraient s'avérer utile dans le traitement des cancers humains. / Activating mutations in NOTCH1, an essential regulator of T-cell development, are frequently found in human T-cell acute lymphoblastic leukemia (T-ALL). However, the precise mechanisms by which Notch causes T-ALL are not fully understood. While several target genes regulated by Notch have been identified in thymocytes and T-ALLs, little is known about the identity of NOTCH1 interacting partners that are required for its oncogenic activity. Using an improved tandem affinity chromatography method, followed by mass spectrometry, we have purified the intracellular active form of NOTCH1 (ICN1) and its nuclear cofactors in human T-ALL cells. We identified and validated a large set of proteins associated with ICN1, including transcriptional activators, repressors and protein-modifying enzymes. Moreover, we found that NOTCH1 interacts with lineage-specifying transcription factors and components of other signaling pathways that could cooperate to confer a specific program of gene expression. This work represents the first comprehensive analysis of Notch partners in the nucleus and provides a framework to elucidate Notch functions and regulation.We subsequently used Notch nuclear interactome as resource to expand our understanding of how ICN1 orchestrates target gene activation. Biochemical and functional experiments demonstrated that Notch activation in T-ALL cells leads to the assembly of a large multisubunit complex in the nucleus containing ICN-CSL-MAML1 as a core subunit and several classes of transcriptional regulators that act at different steps of the transcriptional activation process. These include the transcriptional activator AF4p12, the histone demethylases LSD1 and PHF8, and the SWI/SNF nucleosome remodeling complex PBAF, all of which are required for expression of Notch-responsive genes. We found that PBAF recruitment to Notch-target enhancers facilitates transcription in a chromatin context, probably through the remodeling of nearby nucleosomal structures, while LSD1 and PHF8 act through their demethylase activity to promote local epigenetic modifications. In the presence of Notch, PHF8 demethylates the repressive dimethyl H3 lysine-27 (H3K27me2) mark and LSD1 removes the repressive H3K9me2 mark from Notch-responsive enhancers to promote gene expression. However, LSD1 is also associated with CSL corepressor complex in the absence of Notch and contributes to CSL-mediated repression of Notch targets by removing the activating H3K4me2 mark, suggesting that LSD1 plays a dual role in Notch signaling regulation. AF4p12 is a poorly-characterized protein that was first identified as one of the MLL translocation partners in leukemias. While its precise role awaits further investigation, our results indicate that AF4p12 is required for RNAPII recruitment at several Notch-target genes, therefore acting as a transcriptional coactivator of ICN1. Importantly, we found that Notch cofactors are recruited to Notch-responsive genes in primary developing T cells and are required for Notch-mediated tumor growth in a xenograft T-ALL model. Thus, this newly characterized Notch-activation complex controls both Notch developmental and oncogenic functions in immature T-cells.In addition, our preliminary data provide insights into regulation of ICN1 stability and identify several protein-modifying enzymes as potential regulators of Notch signaling. Moreover, we propose that non-canonical Notch activities, as well as extensive crosstalk with lineage-specifying transcription factors, might shape the repertoire of Notch regulated genes, therefore contribute to Notch oncogenic functions in T-ALL cells.The identification of Notch-associated proteins in T-ALL uncovered novel aspects of Notch signaling and provided a powerful tool for dissecting mechanisms of Notch function and regulation that could be translated into new therapeutic approaches.

Extracellular stimuli may influence the M1/M2 phenotypic polarization of macrophages. We examined M1/M2 biomarkers, phagocytic activity, and tumoricidal activity in RAW 264.7 mouse macrophages. Macrophages were treated with conditioned media (CM) from 4T1 breast cancer cells, curcumin, 22-oxacalcitriol, LPS, or a combination of the previously listed. Arginase activity, a M2 phenotypic biomarker, was upregulated by the treatment of macrophages with conditioned media. Curcumin, 22- oxacalcitriol, and LPS partially inhibited RAW 264.7 arginase activity in the presence of 4T1 breast cancer media. 22-oxacalcitriol increased the phagocytic ability of RAW 264.7 macrophages in the presence of M2 polarizing substances produced by the 4T1 breast cancer cells. Also, LPS increased RAW 264.7 phagocytic ability in the presence of 4T1 breast cancer CM. This study looked at the potential substances that would possibly reverse the M2 tumor promoting macrophage phenotype seen in the breast cancer tumor environment. / Includes bibliography. / Thesis (M.S.)--Florida Atlantic University, 2017. / FAU Electronic Theses and Dissertations Collection

La maladie de Huntington (MH) est une affection neurodégénérative héréditaire dont la mutation conduit à une expansion anormale d’un segment polyglutamine dans la protéine Huntingtine (Htt). La Htt mutée, bien qu’ubiquitaire dans le cerveau, conduit à une neurodégénérescence préférentielle du striatum. Cette atteinte pourrait en partie s’expliquer par la présence de produits de gènes sélectivement exprimés dans le striatum. Le laboratoire étudie depuis plusieurs années l’implication potentielle de marqueurs moléculaires du striatum dans la vulnérabilité des neurones de cette structure cérébrale vis-à-vis de la Htt mutée. Durant ma thèse, j’ai étudié plus spécifiquement trois de ces marqueurs du striatum: l’ARN long intergénique non-codant Abhd11os et les protéines µ-crystalline (CRYM) et doublecortin-like kinase 3 (DCLK3). Une étude préliminaire avait montré l’effet neuroprotecteur de ces marqueurs du striatum contre la toxicité induite par un fragment court de la Htt mutée dans un modèle murin aigu de la MH. J’ai donc étudié plus en détails les caractéristiques de ces "modificateurs" de la MH, ainsi que les mécanismes moléculaires potentiels permettant d’expliquer leur effet neuroprotecteur dans un contexte de la MH. J’ai également mené une expérience de thérapie génique en surexprimant le marqueur striatal DCLK3 dans un modèle transgénique de la MH. Cette étude nous a permis de valider le haut potentiel thérapeutique de cette protéine.L’élucidation précise des mécanismes d’action de ces modificateurs de la MH reste encore à résoudre, mais plusieurs pistes sont maintenant possiblement envisagées par rapport à leurs caractéristiques moléculaires. Outre la découverte de candidats neuroprotecteurs qui pourrait permettre de développer de nouvelles cibles thérapeutiques, cette étude a permis d’envisager de nouvelles hypothèses permettant d’expliquer la vulnérabilité striatale dans la MH et de donner une vue d’ensemble des voies sur lesquelles il serait possible d’agir pour induire des effets neuroprotecteurs dans ce contexte. / Huntington’s disease (HD) is a neurodegenerative disorder caused by the mutation of huntingtin (Htt) gene, which leads to an abnormal polyglutamine expansion in the Htt protein.Whereas mutant Htt (mHtt) is ubiquitously expressed in the brain, it preferentially affects the striatum. Our hypothesis is that genes products selectively expressed in the striatum could be involved in the high vulnerability of the striatum. From this hypothesis, numerous teams studied “markers of the striatum”, that are genes product enriched in the striatum whose expression are up- or down-regulated in HD compared to healthy condition.During my thesis, I studied three of these striatal markers: the long intergenic non-coding RNA Abhd11os, and the two proteins µ-crystallin (CRYM) and doublecortin-like kinase 3 (DCLK3). A preliminary study from the laboratory has shown that these three markers have neuroprotective effects against a toxic fragment of mHtt in vivo. So, the aims of my thesis were to further characterize these three ill-defined disease modifiers and to better understand the putative molecular mechanisms underlying their neuroprotective effects against mHtt.I also conducted a translational study on DCLK3, whose results validate the high therapeutic potential of this protein.The elucidation of the mechanisms underlying the neuroprotective effects of these disease modifiers against mHtt toxicity will require further studies, but new trails can be envisioned, according to their characteristics. My study has enlightened new therapeutic targets and more globally gives an overview of molecular mechanisms to modulate to induce neuroprotective effects in this context, leading to new hypothesis explaining striatal vulnerability in HD.

Propelentes sólidos são materiais energéticos que produzem gases em alta pressão por meio de uma reação de combustão. Qualquer propelente sólido inclui dois ou mais dos seguintes componentes: oxidante (nitratos e percloratos); combustível (resinas orgânicas ou polímeros); compostos químicos combinando oxidantes e combustíveis (nitrocelulose ou nitroglicerina); aditivos para facilitar processos de produção ou alterar a taxa de queima e inibidores (fita de etilcelulose), para restringir superfícies de combustão. Pequenas percentagens de aditivos são usadas para modificar diversas propriedades mecânicas, químicas e balísticas dos propelentes sólidos: acelerar ou desacelerar a velocidade de combustão (catalisadores e inibidores de combustão, respectivamente); assegurar a estabilidade química para prevenir a deterioração durante a estocagem; controlar as propriedades de processamento durante a produção de propelente (tempo de cura, fluidez para extrusão ou moldagem, etc.); controlar as propriedades de absorção de radiação no propelente em combustão; aumentar a resistência mecânica e diminuir a deformação elástica; e, finalmente, minimizar a sensibilidade térmica. No caso de propelentes sólidos Base Dupla (mistura de duas bases ativas: a nitrocelulose e a nitroglicerina), é possível alterar sua velocidade de queima principalmente pelo emprego de pequenos teores de modificadores balísticos, em geral sais orgânicos de cobre e chumbo. Neste trabalho, estudou-se a aceleração da velocidade de queima de uma formulação conhecida de propelente Base Dupla - BD, alterando o teor total dos modificadores balísticos cromato de cobre e estearato de chumbo (ou plastabil - nome comercial) na receita original, bem como a proporção entre eles. Estas alterações na formulação original devem, idealmente, preservar os parâmetros de desempenho estabelecidos para as propriedades químicas (estabilidade química) e mecânicas (densidade da massa e ensaios de tração), ao mesmo tempo otimizando o desempenho balístico, pelo aumento da velocidade de queima. Os resultados experimentais mostram que para os parâmetros de qualidade elongação e velocidade de queima a interação entre os fatores, Proporção Sal de Chumbo/Sal de Cobre (Fator A) e Teor de Modificadores Balísticos (Fator B) foram significativos, ou seja, quanto maior os fatores pior o resultado com as propriedades. Com os parâmetros de resistência a tração e densidade da massa, o fator A e B respectivamente influenciam negativamente quando aumentado em sua concentração. Para o parâmetro estabilidade química não houve nenhum sinal de melhora ou influencia dos fatores. No caso da velocidade de queima a interação AB é o que mais influencia. Melhorando significativamente a velocidade de queima. / Solid propellants are energetic materials which produce a considerable amount of high-pressure gases by means of a combustion reaction. Any solid propellant formulation includes at least two of the following items: oxidizer (nitrates and perchlorates); fuel (organic resins or polymers); chemical compounds combining oxidizers and fuels (nitrocellulose or nitroglycerine); additives to easy production operations or to modify the burning rate and inhibitors (tape ethyl-cellulose), to restrict the combustion surfaces. Small amounts of additives are employed to modify the mechanical, chemical and ballistic features of the solid propellants: to accelerate or diminish the burning rate (catalysts and inhibitors of burning, respectively); to assure the chemical stability in order to prevent the deterioration during stocking; to control the processing properties during propellant production (curing time, extrusion or casting rheology); to control the radiation absorption in the burning propellant; to enhance the mechanical resistance and to reduce the strain; and, finally, to get the thermal sensitivity to a minimum level. In the case of Double-Base solid propellants (blend of two energetic bases: nitrocellulose and nitroglycerine), it\'s possible to control its burning rate mainly by the use of small amounts of ballistic modifiers, generally copper and lead organic salts. This work has studied the burning rate acceleration of a known Double-Base propellant formulation, by changing the total amount of the ballistic modifiers copper chromate and lead stearate (commercially known as plastabil) in the original formulation, as well as the proportion between them. These changes at the original recipe should preserve, ideally, the performance levels required for the chemical (chemical stability) and mechanical properties (density and stress-strain evaluation), optimizing, at the same time, the ballistic performance, through the burning rate enhancement. Results show that for the parameters of quality and elongation rate of burning the interaction between factors, Proportion of Lead Salt / Salt Copper (Factor A) and content Ballistic Modifiers (Factor B) were significant, ie, the higher the worst factors result with the properties. With the parameters of tensile strength and mass density, the factor A and B respectively negatively influence increased when its concentration. For the chemical stability parameter there was no sign of improvement or influences of factors. In the case of burning rate AB interaction is what most influences. Significantly improving the speed of burning.

Boron (B) is a rare element on Earth crust with a natural abundance of 0.001%. However, boron content of water and soils may be significantly high in the regions with rich boron reserves. In addition, extensive use of agrochemicals in soils as well as various natural processes increases the boron concentration in water. Despite B is an essential element for all living creatures, it may pose risks at high level exposures. World Health Organization (WHO) has recommended a daily intake of 1 to 13 mg B for adults. Turkey has almost 70% of world boron reserves principally in four regions: K&uuml / tahya, Emet / Balikesir, Bigadi&ccedil / Eskisehir, Kirka and Bursa, Kemalpasa. The boron content of water in these regions may go up to significant levels. Therefore, it is important to determine B in drinking water from these regions. Electrothermal atomic absorption spectrometry (ETAAS) is a relatively sensitive technique for determination of boron. However, the technique suffers from formation of molecular boron compounds. Therefore, use of chemical modifiers and pyrolytically coated graphite tubes modified with refractory carbide forming elements (Ta, W, Zr, Pd, Ru, Os) were utilized to develop a reliable and sensitive method. Based on optimization studies, Tantalum (Ta) coated tube and co-injection of 5.0 &micro / L 0.01 mol/L Ca(NO3)2, 5.0 &micro / L 0.05 mol/L citric acid together with 15.0 &micro / L sample solution prepared in 1000 mg/L Mg(NO3)2 have been chosen as optimum conditions. Optimum temperatures for pyrolysis and atomization temperatures were determined as 1100 and 2700 &deg / C, respectively. Under these conditions, a detection limit of 0.088 mg/L and a characteristic mass of 186 pg for 15.0 &micro / L sample volume were obtained. The accuracy of the method was checked by EnviroMAT-Waste Water EU-L-1 CRM and NIST 1573a Tomato Leaves SRM analyses. Drinking water samples were collected from Balikesir, Bigadi&ccedil / and K&uuml / tahya, Emet and analyzed by the developed method. Samples were also analyzed by more sensitive techniques / ICP-OES and ICP-MS for a comparison study. The results are compatible with each other.

<p>Medical information must be available for all people in the world. Therefore it is important to translate medical research articles into foreign languages. The aim of this essay was to translate a German medical research article called “Troponinerhöhung und EKG-Veränderungen bei Schlaganfall und Subarachnoidalblutung” into Swedish and to analyse how problems that appeared during the translation process could be solved. The analysis was based on Vinay and Darbelnets, Kollers and Ingos translation theories and dealt with the question how to translate German noun phrases into Swedish. Special attention was paid to noun phrases consisting of medical terms and noun phrases containing extended modifiers.</p><p>The medical terms in the source text were divided into three categories depending on their origin, Greek and Latin terms, German terms and English terms. The translation of a large number of Greek and Latin terms was based on the translation procedure called borrowing. Calque was represented especially in the translation of terms of German origin, whereas borrowing or paraphrasing was preferred when translating English terms.</p><p>There were 63 noun phrases with extended modifiers in the source text. Two fifths of these corresponded to Swedish noun phrases with extended modifiers, whereas the rest must be translated into Swedish noun phrases with adjective premodifiers or relative clauses or into verbal expressions.</p>

Στην παρούσα εργασία η ενασχόλησή μας αφορούσε τη σύνθεση και τον χαρακτηρισμό βιοενεργών γυαλιών. Οι συνθέσεις που πραγματοποιήθηκαν ήταν καθαρής SiO2, μικτών γυαλιών SiO2–ΜΟ (Μ=Ca, Mg) αλλά επίσης και γυαλιών σύστασης SiO2 –CaΟ–P2O5. Πραγματοποιήθηκαν παρασκευές με διάφορα ποσοστά τροποποιητών (Ca, Mg). Μετά την παρασκευή των υλικών ακολούθησε ο φυσικοχημικός χαρακτηρισμός τους με διάφορες τεχνικές όπως: προσδιορισμός ειδικής επιφάνειας και όγκου πόρων (ΒΕΤ), ηλεκτρονική μικροσκοπία σάρωσης (SEM), θερμοσταθμική ανάλυση (TGA), περίθλαση ακτίνων Χ (XRD) και φασματοσκοπία απορρόφησης υπερύθρου (IR). Μετά ακολούθησε μελέτη της βιοενεργότητας με εμβάπτιση σε διάλυμα SBF. Τα αποτελέσματα έδειξαν ότι τα γυαλιά που παρασκευάστηκαν είχαν μεσοπορώδη δομή ενώ όσον αφορά τη βιοενεργότητα τα γυαλιά σύστασης SiO2–CaO and SiO2–CaΟ–P2O5 προκάλεσαν σχηματισμό απατίτη στην επιφάνειά τους. / In the present thesis our work was focused on the study of the synthesis and characterization of bioactive glasses. Pure SiO2 and mixed glasses composed of SiO2–ΜΟ (Μ=Ca, Mg) and SiO2–CaΟ–P2O5 were prepared using different amounts of modifiers (Ca, Mg). Next the prepared materials were characterized by using various techniques such as: BET, SEM, TGA, XRD and IR. Finally their bioactivity was studied in vitro after immersion in SBF solution. It was found that the prepared materials showed a mesoporous structure. Regarding the bioactivity glasses with compositions of SiO2–CaO and SiO2–CaΟ–P2O5 induced the formation of apatite layer on their surface.

It is well established that epigenetic modifications to the genome are crucial for the exquisite control of gene expression required for an organism to develop and differentiate. These modifications are maintained through mitotic rounds of cell division, but must be cleared and reset through meiosis in order for the cells of the early embryo to achieve totipotency. Although we know these mechanisms exist, the rules determining which modifications are established where on the genome and the genes involved in these processes remain poorly characterised. Much of what is known about epigenetic processes has come from studies in non-mammalian organisms, such as Drosophila. However, in our laboratory we have developed a mammalian system for identifying modifiers of epigenetic gene silencing. An ENU mutagenesis screen is being carried out using an inbred mouse line carrying a GFP transgene, with an erythroid-specific promoter, that is particularly sensitive to changes in epigenetic modifications. Currently, 14 mutant lines that display a heritable shift in GFP expression have been recovered. These have been termed Modifiers of Murine Metastable Epialleles (Mommes). When I began my PhD in 2005, we had not identified any of the mutations underlying the phenotypes observed. To confirm the efficacy of the screen, I have tested the effect of heterozygosity for null alleles of two known epigenetic modifiers, Dnmt3a and Dnmt3b, on expression of the GFP transgene. Heterozygosity for the Dnmt3b knockout allele does shift expression while heterozygosity for the Dnmt3a knockout allele does not. This highlights the limitations of the screen. With this particular screen we will only detect modifiers that are expressed during haematopoiesis in the bone marrow. I have also worked on MommeD5. MommeD5 is a semi-dominant, homozygous embryonic lethal mutation that acts as an enhancer of variegation. I have found that the MommeD5 allele carries a 7 bp deletion in the major histone deacetylase, Histone deacetylase 1 (Hdac1), and this significantly alters the C-terminus of the mutant protein. The finding of Hdac1 attests to the screen design. The MommeD5 homozygous mutants die at approximately the same time as the published knockout of Hdac1 and the heterozygous mutants show increased levels of Hdac2 and acetylated histone H3, as reported in Hdac1-deficient embryonic stem cells. In addition, I have studied the effect of heterozygosity for each of the mutations on the phenotype of the mouse. In general, heterozygous Momme mutants are viable and fertile, but show subtle abnormal phenotypes. However, in the case of MommeD5 none were observed and this may relate to the compensatory upregulation of other histone deacetylases. In the case of Dnmt3a and Dnmt3b a sex ratio distortion is seen in the colonies, with less males seen than expected. Also, Dnmt3a heterozygous mutant males that inherited the mutant allele from the dam are smaller and show an increased range of body weights compared to their wild-type male littermates. This may be an example of intangible variation, i.e. phenotypic variation observed in isogenic individuals raised in standardised environments. These results suggest that epigenetic mechanisms have a role in intangible variation, also known as developmental noise. Despite the fact that it is now acknowledged by many that stochastic events occur at the level of the cell, the idea that it can happen at the level of the whole organism is rarely considered.

The epigenetic contribution to phenotype is now well established. Studies over the past decade have shown that proteins that are able to establish and propagate epigenetic modifications are essential for mammalian development. Some of the genes involved in these processes have been identified, but the roles of many remain unknown. The mutagenesis screens for modifiers of position effect variegation in Drosophila suggest that there are over 200 genes that are able to modify epigenetic variegation. We emulated this screen in the mouse to identify mammalian modifiers of a variegating transgene. The screen aimed to identify novel genes involved in epigenetic reprogramming, and to generate mouse models to study the impact of disruption to the epigenome. Inbred male mice carrying a variegating GFP transgene expressed in erythrocytes were mutagenised with ENU. Offspring were screened by flow cytometry and in the initial rounds of mutagenesis, 11 dominant mutant lines were identified. These lines were called MommeDs (Modifiers of murine metastable epialleles, dominant). This thesis describes the mapping and phenotypic characterisation of three Momme lines: MommeD7, MommeD8 and MommeD9. The MommeD9 mutation enhances variegation and was mapped to a 3.4 Mb interval on Chromosome 7. A mutation in a 5? splice site was found in the Trim28 gene. Analysis of Trim28 mRNA and protein in heterozygotes showed that the mutant allele was null. Homozygotes die before mid-gestation. Heterozygotes are viable but display variable and complex phenotypes, including infertility, obesity, behavioural abnormalities and premature death. Obese MommeD9 mice have liver steatosis, impaired glucose tolerance and other indicators of metabolic syndrome. This phenotype has not previously been reported for mice haploinsufficient for Trim28. There is considerable variability of phenotypes among inbred MommeD9 heterozygotes, which suggests a role for epigenetics in phenotypic noise or “intangible variation”. MommeD8 is a semi-dominant enhancer of variegation. Some homozygotes are viable but some die around birth. Viable homozygotes weigh less than wildtype littermates and have increased CpG methylation at the GFP transgene enhancer element. The mutation was mapped to a 4 Mb interval on chromosome 4. Extensive candidate gene sequencing failed to find a mutation and so DNA from mutant and wildtype individuals were sequenced across the entire linked interval by 454 Sequencing technology. MommeD8 individuals carry two point mutations, one is intergenic and the other lies in an intron of the Ppie gene. Analysis of Ppie mRNA in heterozygotes and homozygotes shows that mutants have reduced transcript levels, suggesting that a deficiency in Ppie causes the increased silencing of GFP. The Ppie gene has not been reported to be involved in epigenetic reprogramming and little is known about its function. Mice heterozygous for MommeD7 have a marked increase in expression of GFP. Heterozygotes have a range of hematopoietic abnormalities including splenomegaly, anaemia and reticulocytosis. Homozygotes die at birth and appear pale. The increased GFP in the peripheral blood appears to be the consequence of an increase in reticulocytes. The mutation is linked to a 1.5 Mb interval on Chromosome 7. MommeD7 mice appear to have hematopoietic abnormalities that affect the expression of the erythroid-specific GFP reporter transgene. MommeD7 mice serve as a reminder that, as well as discovering bona fide modifiers of epigenetic reprogramming, the ENU screen can also identify hematopoietic mutants.

Thesis (Ph.D.)--University of Alberta, 2009. / A thesis submitted to the Faculty of Graduate Studies and Research in partial fulfillment of the requirements for the degree of Doctor of Philosophy, Department of Biochemistry. Title from pdf file main screen (viewed on August 11, 2009). Includes bibliographical references.