No / We describe a series of experiments designed to investigate the conformational transition that highly-branched polymers with ligands undergo when interacting with bacteria, a process that may provide a new sensing mechanism for bacterial detection. Fluorescent highly-branched poly(N-isopropyl acrylamide)s (HB-PNIPAM) were prepared by sequential self-condensing radical copolymerizations, using anthrylmethyl methacrylate (AMMA) and fluorescein-O-acrylate (FA) as fluorescent comonomers and 4-vinylbenzyl pyrrole carbodithioate as a branch forming monomer. Differences in reactivity necessitated to first copolymerize AMMA then react with FA in a separate sequential monomer feed step. Modifications of the chain ends produced vancomycin-functional derivatives (HB-PNIPAM-Van). The AMMA and FA labels allow probing of the conformational behaviour of the polymers in solution via Forster resonance energy transfer experiments. It was shown that interaction of this polymer's end groups with Staphylococcus aureus induced a macromolecular collapse. The data thus provide conclusive evidence for a conformational transition that is driven by binding to a bacterium.

Studiens syfte är att utforska hur Aftonbladet och Expressen gestaltade bostadsministern Mehmet Kaplan från den 14 april 2016 fram till 18 april 2016 då han avgick som minister. Syftet är också att se hur förändring av gestaltningen under dessa dagar ter sig. Slutligen kommer studien att jämföra de två tidningarna för att se om och hur de skiljer sig åt i sin gestaltning. De frågeställningar som studien utifrån detta syfte ämnar behandla är följande. -       Hur gestaltas Mehmet Kaplan mellan den 14- 18 april? -       Hur förändras gestaltningen under dessa dagar? -       Hur skiljer sig de olika tidningarnas gestaltning åt?   Metoden som tillämpas är en kvalitativ textanalys, som i praktiken innebär att ställa frågor till texten. Frågorna har byggts upp utifrån Strömbäcks och Entmans teori och definition av gestaltningens fyra grundläggande egenskaper. Vidare har resonemanget om den journalistiska dramaturgins grundläggande element och språkliga redskap inom sensationsjournalistik, använts som stöd i utformandet av analysfrågorna.   Analysmaterialet är hämtat från Retriever, begränsat på redaktionellt digitalt publicerade artiklar mellan datumen 14 - 18 april. Urvalet resulterade i 23 aktiva analysenheter för studien varav är 11 från Aftonbladet och 12 från Expressen.   Studiens resultat är att Mehmet Kaplan gestaltades från Aftonbladet och Expressens håll som otillgänglig, antisemitisk och islamistisk. Samtidigt vidhöll hans politiska kollegor utåt att Kaplan var humanist, demokrat och antirasist. Rapporteringen under de fyra aktuella dagarna var i olika skeenden. En upptrappning, följas av tystnad och sedan explodera i en enorm rapportering, fram tills att Kaplan avgått. Rapporteringen mellan de tidningarna skiljde sig åt, det förklaras i studien på grund av dess olika politiska ideologier. / The study aims to explore how Aftonbladet and Expressen portrayed Housing Minister Mehmet Kaplan from April 142016 until April 18, 2016 when he resigned as minister. The aim is also to see how the change in depiction during these days seems. Finally, the study will compare the two newspapers to see if and how they differ in their interpretation. The issues that the study on the basis that purpose intend to treat is the following. -How portrayed Mehmet Kaplan from 14-18 April? -How to change the design on these days? -How do the various newspaper design at? The method used is a qualitative text analysis, whichin practice means to put questions to the text. The questions have been built up on the basis Strömbäcks and Entmans theory and definition of Gestalt Association four basic characteristics. Furthermore, the reasoning of the journalistic dramaturgy basic elements and linguistic tools in sensational journalism, used to support the design of analytical questions. The analysis material is taken from the Retriever, bounded on digital editorial articles published between the dates of April 14 to 18. The selection resulted in 23 active units of analysis for the study of which 11 are from Aftonbladet and 12 from Expressen. The study's resultis that Mehmet Kaplan figure was from Aftonbladet and Expressen hold as unavailable, anti-Semitic and Islamist. At the sametime maintained his political colleagues out that Kaplan was a humanist, democrat and anti-racist. Reporting during the four days in question were in various stages. An escalation,followed by silence and then explode in a huge reporting, until the Kaplanresigned. The reporting of the newspapers differed, it is explained in the study because of its different political ideologies.

Les molécules classiques du CMH de classe II présentent des peptides antigéniques aux lymphocytes T CD4+. Cette présentation est régulée par deux molécules non classiques : HLA-DM catalyse la relâche de CLIP et le chargement de peptides et HLA-DO module l’activité de DM. Une expression insuffisante en cellules d’insectes empêche les expériences de cristallisation de DO, probablement en raison de sa conformation, rendant DO instable et inapte à sortir du réticulum endoplasmique (RE). DM corrige la conformation de DO et permet sa sortie du RE. Aussi, par ses ponts disulfures uniques, DM adopte une conformation stable et peut sortir du RE sans lier d’autre molécule. Nous avons tenté de corriger la conformation de DO en introduisant des cystéines pour établir des ponts homologues à ceux de DM. La conformation de DO ne fut pas corrigée. Par ailleurs, nous avons augmenté l’expression de DO en introduisant une séquence partielle de Kozak. Nous avons aussi étudié l’effet de DM sur l’expression de DO. DM a favorisé l’expression de DO, probablement en diminuant sa dégradation. Chaque chaîne du dimère DMαβ est impliquée dans l’oxydation de sa chaîne partenaire. La conformation non-optimale de DO pourrait traduire une incapacité des chaînes α ou β à favoriser l’oxydation de sa partenaire; DM corrigerait ce problème. Notre analyse d’immunobuvardage de type Western a toutefois démontré que DM ne modifie pas l’état d’oxydation de DOα et DOβ. Finalement, nous avons étudié l’interaction DO-DM. L’acide aminé DOαE41 est impliqué dans cette liaison. Certains des acides aminés entre α80 et α84 pourraient être impliqués. Nous avons muté des acides aminés de cette région de DOα. Les résidus testés ne semblent pas impliqués dans la liaison DO-DM. L’obtention de la structure tridimensionnelle de DO et la caractérisation de son état oxydatif et de sa liaison à DM permettront de mieux comprendre son rôle. / Classical MHC class II molecules present antigenic peptides to CD4+ T cells. This presentation is regulated by two non-classical molecules: HLA-DM catalyzes CLIP release and peptide loading and HLA-DO mediates the DM activity. An insufficient expression in insect cells did not allow DO crystal production experiments, probably because of its conformation, rendering DO unstable and unable to leave the endoplasmic reticulum (ER). DM corrects the conformation of DO and allows its egress from the ER. Also, because of its unique disulfide bonds, DM has a stable conformation and can egress from the ER without binding another molecule. We tried to correct the conformation of DO by introducing cysteines to create disulfide bonds homologous to those of DM. However, its conformation was not corrected. Also, we increased DO expression by inserting a partial Kozak sequence. We also studied the effect of DM on DO expression. DM favoured DO expression, probably by reducing its degradation. Each chain of the DMαβ dimer plays a role in the oxidation of its partner chain. The non-optimal conformation of DO might result from an incapacity of its α and β chains to direct each other’s oxidation; DM would correct this problem. Our Western blot analysis showed, however, that DM does not modify the oxidation state of DOα and DOβ. Finally, we studied the DO-DM interaction. The DOαE41 amino acid is involved in this interaction, as some of the α80 to α84 might be. We mutated amino acids in this region of DO. Tested amino acids did not seem involved in DO-DM binding. The tridimensional structure of DO and the characterization of its oxidative state and its DM binding will allow a better understanding of its function.

Le récepteur A des peptides natriurétiques (NPRA) fait partie de la famille des guanylates cyclases membranaires. L’activation du NPRA par ses agonistes naturels, ANP et BNP, induit une production de GMPc qui est responsable de leur rôle dans l’homéostasie cardiovasculaire, l’inhibition de l’hypertrophie et de la fibrose cardiaques et la régulation de la lipolyse. Le NPRA est un homodimère non covalent composé d’un domaine extracellulaire de liaison du ligand (ECD), d’un unique domaine transmembranaire (TM), d’un domaine d’homologie aux kinases et d’un domaine guanylate cyclase. Bien que le NPRA ait un rôle physiologique important, les mécanismes moléculaires régissant son processus d’activation restent inconnus. Nous avons donc analysé les premières étapes du processus d’activation du NPRA. Nous avons d'abord étudié le rôle de la dimérisation des ECD dans l’activation du récepteur. Nous avons utilisé les techniques de liaison de radioligand, de FRET et de modélisation moléculaire, pour caractériser la liaison à l’ECD des agonistes naturels, d’un superagoniste et d’un antagoniste. L’ANP se lie à un dimère d’ECD préformé et la dimérisation spontanée est l’étape limitante du processus de liaison. De plus, comme le démontrent nos études de FRET, tous les peptides, incluant l’antagoniste, stabilisent le récepteur sous sa forme dimérique. Cependant, l’antagoniste A71915 stabilise le dimère d’ECD dans une conformation différente de celle induite par l’ANP. La dimérisation du NPRA semble donc nécessaire, mais non suffisante à l’activation du récepteur. L’état d’activation du NPRA dépend plutôt de l’orientation des sous unités dans le dimère. Nous avons ensuite étudié le mécanisme moléculaire de transduction du signal à travers la membrane. Plusieurs études ont suggéré que l’activation du NPRA implique un changement de conformation du domaine juxtamembranaire (JM). Cependant, les études de cristallographie de l’ECD soluble de NPRA n’ont pas permis de documenter la structure du JM et le changement de conformation impliqué dans la transduction du signal reste inconnu. Pour analyser ce changement de conformation, nous avons d’abord séquentiellement substitué les neuf acides aminés du JM par une cystéine. En étudiant la capacité des mutants à former des dimères covalents de façon constitutive ou induite par l’ANP, nous avons pu évaluer la proximité relative des résidus du JM, avant et après activation du NPRA. Ces résultats ont démontré la proximité élevée de certains résidus spécifiques et sont en contradiction avec les données cristallographiques. Nous avons également démontré que le domaine intracellulaire impose une contrainte conformationnelle au JM à l’état de base, qui est levée après liaison de l’ANP. En introduisant de 1 à 5 alanines dans l’hélice-α transmembranaire, nous avons montré qu’une rotation des TM de 40° induit une activation constitutive du NPRA. Le signal d’activation pourrait donc être transmis à travers la membrane par un mécanisme de rotation des TM. En utilisant nos données expérimentales, nous avons généré le premier modèle moléculaire illustrant la conformation active du NPRA, où les domaines JM et TM sont représentés. Dans son ensemble, cette étude apporte une meilleure compréhension des mécanismes moléculaires régissant les premières étapes du processus complexe d’activation du NPRA. / Natriuretic peptide receptor-A (NPRA) is a member of the particulate guanylate cyclase family. NPRA activation by natural agonists, ANP and BNP, leads to cGMP production, which is responsible for their role in cardiovascular homeostasis, cardiac hypertrophy and fibrosis inhibition and lipolysis regulation. NPRA is a non covalent dimer composed of an extracellular domain (ECD) with a ligand binding site, a single transmembrane region (TM), a kinase homology domain, and a guanylyl cyclase domain. Although NPRA plays an important physiologic role, molecular mecanisms driving its activation process are yet unknown. We thus analysed the first steps of NPRA’s activation process. First, we studied the role of ECD dimerization in receptor activation and determined the sequential steps of this dimerization process. We used radioligand binding, FRET and molecular modeling to characterize the interaction of ECD with natural agonists, a superagonist and an antagonist. ANP binds to preformed ECD dimers and spontaneous dimerization is the rate-limiting step of the ligand binding process. Furthermore, like demonstrated with fluorescence homoquenching, all the studied peptides, including A71915 antagonist, stabilize a dimeric form of the receptor. However, A71915 stabilizes the ECD dimer in a conformation distinct from those induced by ANP. Thus, ECD dimerization is necessary but not sufficient for NPRA activation. The activation state of NPRA seems to depend on the orientation of the receptor subunits within the dimer. Then, we tried to identify the molecular mechanism of signal transduction through the plasma membrane. Previous studies have shown that activation of NPRA involves a conformational change of the juxtamembrane domain (JM). However, crystallographic study of the soluble ECD of NPRA has failed to document JM structure, and the conformational change involved in transmembrane signal transduction is still unknown. To analyse this conformational change, we first sequentially substituted nine amino acids of JM by a cysteine residue. By studying the mutant’s capacity to form ANP-induced or constitutive covalent disulfide dimers, we evaluated the relative proximity of JM residues, before and after NPRA activation. These results demonstrate a high proximity of specific JM residues and are in disagreement with crystallography data. We also demonstrated that intracellular domain imposes a conformational constraint on JM at basal state, which becomes relaxed upon ANP binding. We finally confirmed, with a full-length receptor, that A71915 stabilizes NPRA in a dimeric form where JM are in a conformation distinct from the basal state. By introducing 1 to 5 alanine residues in the transmembrane α-helix, we showed that a TM rotation of 40° leads to constitutive NPRA activation. Activation signal could thus be transmitted through the membrane by a TM rotation mechanism. We finally studied the role of the TM in NPRA dimerization. By using the ToxR system, we demonstrated that the last JM residues are required to stabilize the TM dimer. Using these experimental data, we generated the first molecular model illustrating the active conformation of NPRA, where JM and TM are depicted. In summary, this study allows a better understanding of molecular mecanisms driving the first steps of NPRA’s complex activation process.

Herpesviruses can enter host cells by pH-dependent endocytic pathways in a cell-specific manner. The role of pH in herpesvirus endocytosis is unclear. Herpes simplex virus (HSV) is a paradigm for virus membrane fusion via a complex of glycoproteins. HSV glycoproteins B, D and the heterodimer H-L are necessary and sufficient for membrane fusion. This work analyzes the structure and function of HSV glycoproteins B, D, and H-L at neutral pH, and at the physiological low-pH encountered during endocytic entry. It is demonstrated that mildly acidic low pH triggers specific conformational changes in HSV gB at a pH of 5.7 to 6.0. The antigenic structure of gB functional region I that is critical for fusion is specifically altered by mildly acidic pH both in vitro and during entry into host cells. Point mutations within gB functional region 1 that block membrane fusion still allow conformational changes in region 1. This suggests that specific hydrophobic residues are essential for fusion domain insertion into the host cell membrane but not conformational change. The detected conformational changes were reversible, similar to other class III fusion glycoproteins. Exposure to mildly acidic pH directly triggered the fusion function of HSV glycoproteins and caused gB, but not other glycoproteins, to become more hydrophobic. The oligomeric conformation of gB is altered at a similar pH range. In addition, several approaches were used to monitor gB throughout glycoprotein synthesis and maturation. It is shown that gB may cotranslationally fold and oligomerize as it is synthesized on the ribosome. As gB matures it then alters conformation and/or binding partner to form antigenically distinct populations of gB within the cell and virion. I conclude that intracellular low pH induces changes in gB conformation that, together with additional triggers such as receptor-binding, are essential for virion-cell fusion during herpesviral entry by endocytosis.

Thèse numérisée par la Direction des bibliothèques de l'Université de Montréal.

[pt] O processo de revestimento por rotação é caracterizado pelo uso de cilindros girantes para controlar a espessura e aplicar uma fina camada de líquido em um substrato em movimento. A não ser a baixas velocidades dos cilindros, o escoamento bi-dimensional na região de formação dos filmes sobre cada cilindro é instável e o padrão observado experimentalmente consiste em um escoamento tri-dimensional e periódico na direção transversal ao substrato. Esta instabilidade pode limitar a velocidade máxima do processo se a camada líquida depositada sobre o substrato tem que ser uniforme. Para líquidos Newtonianos, a estabilidade deste escoamento é determinada pela competição de forças viscosas e capilares: a instabilidade ocorre acima de um número de capilaridade máximo. Apesar da maioria dos líquidos utilizados em processos de revestimento serem não Newtonianos, as análises disponíveis deste escoamento se limitam a estudos de líquidos Newtonianos. O comportamento não Newtoniano do líquido pode alterar completamente a natureza do escoamento perto da superfície livre; quando pequenas quantidades de polímeros flexíveis de alto peso molecular estão presentes, a instabilidade na direção transversal ocorre a velocidades muito mais baixas, quando comparado ao caso Newtoniano. Os mecanismos responsáveis pela instabilidade a baixas velocidades ainda não são completamente compreendidos. Este escoamento viscoelástico com superfície livre é analisado neste trabalho através de duas equações constitutivas diferenciais, o modelo de Oldroyld-B e o modelo de FENE-P. As equações de conservação de massa, quantidade de movimentos acopladas com os modelos constitutivos, e as equações não-lineares de mapeamento que transformam o problema de superfície livre em um problema de valor de contorno foram resolvidas pelo método de elementos finitos DEVSS-G/SUPG. O sistema de equações algébricas não linear foi resolvido pelo método de Newton com continuação por pseudo-comprimento de arco. Os resultados mostram como o campo de tensão muda com o aumento do número de Weissenberg (elasticidade do líquido), levando a formação de uma camada limite de tensão elástica na superfície livre e tensões elásticas compressivas na direção transversal, que podem explicar o aparecimento da instabilidade a baixas velocidades. Este trabalho também apresenta a formulação de estabilidade linear para escoamentos viscoelásticos com superfícies livres. O modelo dá origem a um problema de auto-valor generalizado, que foi resolvido pelo método de GMRES (ARPACK). Os auto-valores dominantes da matriz Jacobiana indicam a estabilidade do escoamento. Esta formulação foi testada em três escoamentos distintos: escoamento em uma cavidade de tampa móvel, piscina de líquido estática e um escoamento de Couette (simples de cisalhamento). / [en] Roll coating is distinguished by the use of one or more gaps between rotating cylinders to meter and apply a liquid layer to a substrate. Except at low speed, the film splitting flow that occurs in forward roll coating is three-dimensional and results in more or less regular stripes in the machine direction. This instability can limit the speed of the process if a smooth film is required as a final product. For Newtonian liquids, the stability of the film-split flow is determined by the competition of capillary forces and viscous forces: the onset of meniscus nonuniformity is market by a critical value of the capillary number. Although most of the liquids coated industrially are polymeric solutions and dispersions, that are not Newtonian, most of previous theoretical analyses of film splitting flows dealt only with Newtonian liquids. Non-Newtonian behavior can drastically change the nature of the flow near the free surface; when minute amounts of flexible polymer are present, the onset of the three-dimensional instability occurs at much lower speeds than in the Newtonian case. the mechanisms responsible for the early onset of this flow instability is not well understood. This free surface coating flow is analyzed here with differential constitutive models, the Oldroyld-B and the FENE-P equations. The continuity, momentum equations coupled with the constitutive models, and the non-linear mapping equations that transform the free boundary problem into a fixed boundary problem are solved by Newton s method with pseudo-arc-length continuation. The results show how the stress field changes with Weisenberg number, leading to the formation of an elastic boundary layer near the free surface and compressive elastic stresses in the crss-flow direction that may explain the onset of the ribbing instability at the smaller Capillary numbers when viscoelastic liquids are used. This work also presents the formulation for linear stanility analysis of viscoelastic free surface flows. The model leads to a generalized eigenproblem that is solved here using the Arnoldi s method with the software (ARPACK). The leading eigenvalues of the Jacobian Matrix indicate the stability of the flow. The formulation is tested in three different flows: lid-driven cavity, static liquid pool and a couette flow.

Os objetivos deste trabalho foram avaliar diferentes modelos de seleção com base nos efeitos aleatórios maternos considerados para características de crescimento e perímetro escrotal, estimar parâmetros genéticos para pesos do nascimento aos 12 meses (pesos ao nascer, a desmama e aos 12 meses de idade); perímetro escrotal aos 12 meses e correlações genéticas entre as características para bovinos compostos Montana Tropical, em análises uni, bicaracterísticas. Estimar parâmetros genéticos para as características categóricas de musculosidade, precocidade e conformação aos 12 meses em modelos uni e bicaracterísticas utilizando diferentes metodologias para análise (modelos mistos, modelos thresholds bayesianos e modelos lineares generalizados mistos) e compará-los. Nas análises feitas para as características de crescimento e perímetro escrotal os efeitos maternos influenciaram os pesos do nascimento aos 12 meses de idade. As estimativas de herdabilidade direta obtidas das análises bicaracterísticas foram superiores àquelas obtidas das análises unicaracterísticas e as estimativas pela análise bicaracterística para as herdabilidades foram 0,27 para peso ao nascer; 0,18 para peso à desmama; 0,20 para peso aos 12 meses; e 0,19 para perímetro escrotal aos 12 meses. As correlações genéticas estimadas entre pesos obtidos em idades jovens com peso ao ano foram moderadas a baixas (< 0,60). A correlação genética obtida entre perímetro escrotal e características de crescimento foram 0,04, e <0,01 com peso a desmama indicando baixa associação entre as características e de 0,38 com peso aos 12 meses o que pode ao longo do tempo gerar animais mais pesados aos 12 meses. A seleção com base em características de crescimento em qualquer idade pode promover ganhos genéticos moderados no peso corporal de animais do composto Montana Tropical. É importante considerar nas análises os pesos prévios à seleção para estimar parâmetros genéticos para pesos após a seleção. Para as características morfológicas as estimativas de herdabilidade foram de baixas a moderadas e houve diferença entre o modelo linear e o de limiar e o modelo linear generalizado misto, na obtenção de estimativas de herdabilidades e correlações genéticas, de características categóricas morfológicas multinomiais. As estimativas dos parâmetros genéticos obtidas por modelo de limiar foram superiores aos demais métodos avaliados, com valores de 0,42; 0,37 e 0,25 para musculosidade, precocidade e conformação aos 12 meses, e as correlações genéticas estimadas em conjunto com peso a desmama para musculosidade, precocidade e conformação aos 12 meses foram 0,89; 0,22 e 0,83 respectivamente. Os escores visuais de conformação, precocidade e musculatura aos 12 meses podem responder rapidamente à seleção individual. / The objectives this study were to evaluate different models of selection based on maternal random effects considered for growth traits and scrotal circumference; to estimate genetic parameters for weights from birth to 12 months (birth weight, weaning and 12 months old), scrotal circumference at 12 months and to estimate genetic correlations between traits for cattle composite Montana Tropical, by univariate and two-trait analysis. To estimate genetic parameters for categorical traits of muscling, precocity and conformation at 12 months in uni-and two-trait models using different methodologies for analysis (mixed models, Bayesian models thresholds and generalized linear mixed models) and compare them. In the analysis made for growth traits and scrotal circumference, maternal effects influenced the weights from birth to 12 months of age. Direct heritability estimates obtained from two-trait analyzes were higher than those obtained using univariate models, and the estimates of heritability for the two traits analysis were 0.27 for birth weight, 0.18 for weaning weight, 0.20 for weight at 12 months , and 0.19 for scrotal circumference at 12 months. The genetic correlation obtained between weights at young ages and yearling weight were moderate to low ( < 0.60). Genetic correlation obtained between scrotal circumference and growth traits were 0.04, and < 0.01 with weaning weight indicating a low correlation between the traits and 0.38 from weight at 12 months which may over time provide heavier animals at 12 months. Selection based on growth traits at any age can provide moderate genetic gains in body weight of animals of the composite Montana Tropical. It is important to consider in the analysis to selecting the prior weights to estimate genetic parameters for weights after selection. For the morphological traits, heritability estimates were low to moderate and there was a difference between the threshold and linear model and generalized linear mixed model, for estimates of heritability and genetic correlations of morphological multinomial categorical traits. Estimates of genetic parameters by the threshold model were higher than the other methods evaluated, with values of 0.42, 0.37 and 0.25 for muscling, precocity, and conformation at 12 months, and the genetic correlations estimated in with weaning weight for muscling, precocity and conformation at 12 months were 0.89, 0.22 and 0.83 respectively. The visual scores of conformation, precocity and muscling at 12 months can respond quickly to individual selection.

Veränderungen in der lokalen Chromatinstruktur beeinflussen die dynamische Regulation von Genen, welche für die Differenzierung notwendig sind. PU.1 ist ein Master-Transkriptionsfaktor in der Hämatopoese und wird streng reguliert, um ein zelllinienspezifisches Expressionsmuster zu erzielen. Hohe Konzentrationen von PU.1 sind für myeloische Differenzierung erforderlich. In B-Zellen wird PU.1 mittelstark exprimiert und muss aktiv runterreguliert werden, um eine Ausdifferenzierung der multipotenten Vorläuferzellen zu T-Zellen zu ermöglichen. Derzeit ist wenig über die Regulierung von PU.1 in T-Zellen bekannt. Darüber hinaus wurde eine abnormale Expression von PU.1 in verschiedenen Leukämieerkrankungen beobachtet. Mittels eines genome-wide Chromatin-Interaktions-Screens konnten wir einen cis-Repressor mit insulierender Kapazität identifizieren, welcher mittels eines Chromatinloops die Promotoraktivität von PU.1 in T-Zellen, jedoch nicht in myeloischen oder B-Zellen blockiert. Sowie Looping als auch Insulation erfordern die Bindung des Chromatin-Regulatorprotein CTCF. Im Gegensatz zu normalen myeloischen Zellen finden wir, dass Krebszellen aus myeloischen Leukämie Patienten diese T-Zell-spezifische repressive Chromatinstruktur aufweisen, was einen räumlichen Kontakt des Insulator mit dem PU.1 Promotor ermöglicht. Die Ergebnisse dieser Arbeit beschrieben das CTCF gesteuerte „long distance looping“ als ein neuer molekularer epigenetischer Mechanismus, um Transkriptionsfaktor PU.1 in T-Zellen runterzuregulieren, und zeigen zum ersten mal, dass Krebszellen die Chromatinstruktur anderer Zelllinien imitieren können, um die Expression von Differenzierungsgenen zu blockieren. / Alterations in the local chromatin structure orchestrate the dynamic regulation of differentiation promoting genes. PU.1 is a master transcription factor in hematopoiesis. PU.1 gene must be tightly regulated to achieve lineage specific expression pattern. High levels of PU.1 are required for myeloid commitment: it is expressed at intermediate level in B-cells and must be actively silenced to permit T cell development from early multipotent progenitors. However, little is known of how PU.1 is regulated in T-cells. Moreover, aberrant PU.1 expressions have been observed in multiple leukemias. Using a genome-wide chromatin interaction screen we identified a cis-repressor with insulating capacity that undergoes long-distant chromatin looping to block PU.1 promoter activity in T cells but not myeloid or B cells. Looping and repression requires binding of the chromatin regulator protein CTCF. In contrast to normal myeloid cells, we found that cancer cells from myeloid leukemia patients adopt the T cell specific repressive chromatin structure bringing the insulator into spatial contact with the PU.1 promoter. These results identify CTCF controlled long-distant insulator looping as a novel mechanism to silence lineage-opposing transcription factor expression, and reveal that cancer cells can mimic the chromatin confirmation of another lineage to block expression of differentiation driving genes.

In drei Betrieben mit maschineller Milchgewinnung wurden an 193 Mutterschafen der Rasse Ostfriesisches Milchschaf 17 Euter- und Zitzenmerkmale erfasst und eine Melkbarkeitsprüfung durchgeführt. Bei zwei bis vier Untersuchungen je Tier bis zum fünften Laktationsmonat konnten 9.231 Eutermerkmalsmessungen und 5.204 Eutermerkmalsbeurteilungen in die Auswertung einbezogen werden. Zusätzlich standen die einzeltierbezogenen Gehalte somatischer Zellen in der Milch zur Verfügung. Die festgestellten Beziehungen zwischen Euterform- und Leistungsmerkmalen lassen den Schluss zu, dass mit der Selektion nach Euterformmerkmalen eine direkte züchterische Einflussnahme auf Melkmaschineneignung und Eutergesundheit sowie eine indirekte züchterische Einflussnahme auf die Melkbarkeit möglich ist. Die Milchleistung beeinflusst signifikant die Euterdimension. Für eine Euterbeurteilung eignen sich besonders Eutertiefe, Bodenabstand, Hintereuteraufhängung, Euterband, Zitzenplatzierung und Zitzengröße. Diese Merkmale sind von wirtschaftlicher Relevanz, mit guter Sicherheit erfassbar, durch eine mittlere Erblichkeit gekennzeichnet und weisen zu anderen ebenfalls wirtschaftlich relevanten Merkmalen enge phänotypische Beziehungen auf. Ein Model zur Euterformbeschreibung kann als Grundlage für eine zukünftige, vergleichbare Leistungsprüfung dienen. Die Euterform verändert sich mit zunehmender Anzahl Laktationen, wobei die Unterschiede zwischen erster und zweiter Laktation am größten sind. Der Laktationsmonat hatte im Untersuchungszeitraum nur einen geringen Einfluss. Daraus schlussfolgernd eignet sich der erste Laktationsabschnitt bis zum vierten Laktationsmonat innerhalb der zweiten Laktation für eine frühestmögliche sichere Beurteilung der Euterform. Die Erfassung von Milchleistung und Melkbarkeit ist nur mittels Milchleistungs- und Melkbarkeitsprüfung bzw. Melktest möglich. Die systematischen Faktoren Betrieb und Laktationsnummer zeigen signifikante Einflüsse auf die Ergebnisse und sollten in der Ergebnisberechnung berücksichtigt werden. Die Eutergesundheit hat ihre Bedeutung für Nutzungsdauer und Leistungsfähigkeit der Tiere, aber auch im Hinblick auf die Verarbeitungsfähigkeit und Qualität der Milch und Milchprodukte. Eine Selektion der Tiere mit dauerhaft hohen Zellzahlen trägt zur Gesunderhaltung der Herde und zur Erzeugung von Qualitätsprodukten bei. / 193 lactating Eastfrisian Milk Sheep were used to investigate 17 udder and teat traits and were subjected to a milkability test. In up to 4 test per ewe during the first five month of lactation a total of 9231 udder measurements and 5204 udder assessments were included in this investigation. Repeated information on somatic cell count per individual animal were also available. Correlation between udder morphology and performance traits indicate that a selection for udder morphology traits will have a direct effect on machine milkability and udder health; a indirect effect can be assumed for all milkability traits. Milk yield is positive correlated with udder dimension. Depth of udder, distance to floor, hind udder attachment, udder ligament, position of teats and teat size are all useful traits for udder assessment. All these traits are economically relevant, can be a measured with sufficient accuracy, and do possess a medium range heritability and show positive phenotypic correlations to other economically relevant traits. Description of udder morphology requires a standard model for a comparative performance recording. Udder morphology is systematically affected by number of lactation, with large differences between the first and the second lactation. Months of lactation had only a minor influence. Results of this investigation indicate that udder assessment can best be implemented during the second lactation within the first four month of lactation. Milkability tests should be implemented and incorporated in a breeders strategy to improve udder morphology and udder health. A range of systematic factors is influencing milkability which requires a consequent registration of these influencing factors and correction before traits are being used for breeding value estimation. Standard milk performance recording together with the registration of somatic cells in combination with assessment of udder morphology and teat morphology will provide the bases for a consequent selection to improve milk yield and udder health.