We investigated the gestational expression of androgen receptor (AR) and defined its regulation and that of its co-repressors, PSF and p54nrb, by steroid hormones and myometrial stretch in vivo in pregnant and non-pregnant rats. Our data demonstrate that, 1) myometrial AR expression decreases prior to term; 2) AR expression is up-regulated by MPA treatment and down-regulated by mechanical stretch; (3) myometrial PSF protein expression is down-regulated by estrogen signaling and by mechanical stretch, and up-regulated by androgen signaling; (4) while myometrial PSF mRNA expression is also down-regulated by stretch, the regulation by estrogen and P4 on PSF mRNA appear to be opposite to the effects on PSF protein. We conclude that the decreased androgen signaling in late pregnancy (as a result of decreased AR and PSF expression mediated by hormonal and mechanical signals) may contribute to the mechanisms leading to labor initiation.

We investigated the gestational expression of androgen receptor (AR) and defined its regulation and that of its co-repressors, PSF and p54nrb, by steroid hormones and myometrial stretch in vivo in pregnant and non-pregnant rats. Our data demonstrate that, 1) myometrial AR expression decreases prior to term; 2) AR expression is up-regulated by MPA treatment and down-regulated by mechanical stretch; (3) myometrial PSF protein expression is down-regulated by estrogen signaling and by mechanical stretch, and up-regulated by androgen signaling; (4) while myometrial PSF mRNA expression is also down-regulated by stretch, the regulation by estrogen and P4 on PSF mRNA appear to be opposite to the effects on PSF protein. We conclude that the decreased androgen signaling in late pregnancy (as a result of decreased AR and PSF expression mediated by hormonal and mechanical signals) may contribute to the mechanisms leading to labor initiation.

Thesis (Ph. D.)--University of Washington, 2007. / Vita. Includes bibliographical references (leaves 61-63).

Thesis (Ph. D.)--University of Washington, 1998. / Vita. Includes bibliographical references (leaves [100]-113).

Thesis (Ph. D.) -- University of Texas Southwestern Medical Center at Dallas, 2005. / Vita. Bibliography: 123-135.

Submitted by LIEGE ABDALLAH KAWAI null (liegekawai@gmail.com) on 2017-12-13T13:56:50Z No. of bitstreams: 1 liege kawai - tese final.pdf: 5173025 bytes, checksum: 74d38826e72cc45ba04c0050ac6a409b (MD5) / Approved for entry into archive by Elza Mitiko Sato null (elzasato@ibilce.unesp.br) on 2017-12-13T14:57:31Z (GMT) No. of bitstreams: 1 kawai_la_dr_sjrp.pdf: 5173025 bytes, checksum: 74d38826e72cc45ba04c0050ac6a409b (MD5) / Made available in DSpace on 2017-12-13T14:57:31Z (GMT). No. of bitstreams: 1 kawai_la_dr_sjrp.pdf: 5173025 bytes, checksum: 74d38826e72cc45ba04c0050ac6a409b (MD5) Previous issue date: 2017-11-22 / Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES) / Corynebacterium pseudotuberculosis (C. pseudotuberculosis), é uma bactéria gram positiva anaeróbia facultativa, pleomórfica, que não esporula, não forma cápsula, e que apresenta 2 biotipos ou biovares, sendo o biovar equi capaz de infectar preferencialmente equinos, enquanto o biótipo denominado ovis acomete pequenos ruminantes. Esta bactéria faz parte do grupo CMNR (Corynebacterium, Mycobacterium, Nocardia e Rhodococcus), que demonstra grande importância veterinária e médica, uma vez que estes micro-organismos comumente infectam animais, podendo infectar o homem, causando perdas econômicas pela ineficácia ou alto custo de terapias existentes. Um exemplo seria a linfadenite caseosa (LC) causada por C. pseudotuberculosis, que afeta particularmente a pecuária de caprinos e ovinos, com a condenação da carcaça e redução da produção de lã e de carne. A transmissão da doença e contágio dos animais é direta, muitas vezes através da alimentação e ingestão de água em local contaminado por animais doentes. Essa doença possui incidência na pecuária mundial, principalmente de caprinos e ovinos, havendo registros de ocorrência no Brasil, Europa, Oriente Médio, Austrália, Nova Zelândia, África do Sul, Canadá e Estados Unidos e mesmo com todos os avanços tecnológicos, ainda não há métodos de prevenção totalmente eficazes, como vacinas e medicamentos, tampouco para o tratamento de animais infectados, que geralmente são de custo elevado, por longos períodos e sem a garantia de cura ou de isenção de reincidência da LC. Deste modo, técnicas mais rápidas e fáceis para detecção e diagnóstico da doença, bem como para seu tratamento, se tornam imprescindíveis, evitando não só a disseminação da doença, mas também suas consequentes perdas econômicas. Atualmente, devido ao uso indiscriminado de antibióticos para o tratamento de infecções de origem bacteriana, bem como à constante exposição destes micro-organismos a essas drogas em ambientes hospitalares, observa-se o desenvolvimento de micro-organismos resistentes às terapias disponíveis, sendo um desafio mundial a descoberta e elaboração de tratamentos eficazes para a prevenção, controle e eliminação destes patógenos, como alternativa aos já existentes. Visando terapias alternativas e direcionadas para infecção por C. pseudotuberculosis, as proteínas tioredoxina, tioredoxina redutase e diphtheria toxin repressor foram estudadas no presente trabalho, a fim de melhor compreender este micro-organismo. / Corynebacterium pseudotuberculosis (C. pseudotuberculosis), is a gram-positive, facultative anaerobe, pleomorphic, non-sporulating bacterium with two biotopes or biovars, being the biovar equi capable of infecting horses, whereas the biotype called ovis infects small ruminants. It is part of the CMNR group (Corynebacterium, Mycobacterium, Nocardia and Rhodococcus), which demonstrates great veterinary and medical importance, since these common microorganisms infect animals and can infect humans, causing economic losses due to the inefficiency or high cost of existing therapies. An example is a caseous lymphadenitis (LC) caused by C. pseudotuberculosis, which particularly affects the goats and sheep livestock, with carcass condemnation and reduction of wool and meat production. The transmission of disease and the contagion of animals is direct, often through feeding and drinking water in places contaminated by sick animals. This disease has an incidence in the world livestock, mainly of goats and sheep, with occurrence records in Brazil, Europe, the Middle East, Australia, New Zealand, South Africa, Canada and the United States and even with all technological advances, still there are no totally effective prevention methods, such as vaccines and medications, nor for the treatment of infected animals, which are usually of a high cost, for long periods and without guarantee of cure or exemption from recurrence of LC. In this way, faster and easier techniques for the detection of the diagnosis of this disease as well as for its treatment become essential, avoiding not only a spread of the disease, but also its consequent economic losses. Currently, the use of indiscriminate antibiotics for the treatment of infections of bacterial origin, as well as the constant exposure of these microorganisms to these drugs in hospital environments, shows the development of microorganisms resistant to the available therapies, being one world-wide challenge the elaboration of effective treatments for the prevention, control and elimination of these pathogens, as an alternative to the existing ones. Aiming alternative therapies for infection by C. pseudotuberculosis, proteins such as thioredoxin, thioredoxin reductase, diphtheria toxin repressor, were studied in the present work, for a better comprehension of this microorganism.

The objective of the field of Synthetic Biology is to implement novel functionalities in a biological context or redesign existing biological systems. To achieve this, it employs tried and tested engineering principles, such as standardisation and the design-build-test cycle. A crucial part of this process is the convergence of modelling and experiment. The aim of this thesis is to improve the design principles employed by Synthetic Biology in the context of Gene Regulatory Networks (GRNs). Small Ribonucleic Acids (sRNAs), in particular, are focussed on as a mechanism for post-transcriptional expression regulation, as they present great potential as a tool to be harnessed in GRNs. Modelling sRNA regulation and its interaction with its associated chaperone Host-Factor of Bacteriophage Qβ (Hfq) is investigated. Inclusion of Hfq is found to be necessary in stochastic models, but not in deterministic models. Secondly, feedback is core to the thesis, as it presents a means to scale-up designed systems. A linear design framework for GRNs is then presented, focussing on Transcription Factor (TF) interactions. Such frameworks are powerful as they facilitate the design of feedback. The framework supplies a block diagram methodology for visualisation and analysis of the designed circuit. In this context, phase lead and lag controllers, well-known in the context of Control Engineering, are presented as genetic motifs. A design example, employing the genetic phase lag controller, is then presented, demonstrating how the developed framework can be used to design a genetic circuit. The framework is then extended to include sRNA regulation. Four GRNs, demonstrating the simplest forms of genetic feedback, are then modelled and implemented. The feedback occurs at three different levels: autoregulation, through an sRNA and through another TF. The models of these GRNs are inspired by the implemented biological topologies, focussing on steady state behaviour and various setups. Both deterministic and stochastic models are studied. Dynamic responses of the circuits are also briefly compared. Data is presented, showing good qualitative agreement between models and experiment. Both culture level data and cell population data is presented. The latter of these is particularly useful as the moments of the distributions can be calculated and compared to results from stochastic simulation. The fit of a deterministic model to data is attempted, which results in a suggested extension of the model. The conclusion summarises the thesis, stating that modelling and experiment are in good qualitative agreement. The required next step is to be able to predict behaviour quantitatively.

The diploma thesis consists of a theoretical and a practical part. The objective of the theoretical part is to introduce basic terms and ideas related to emotions (with focus on emotional regulation) and to map different views on the possibilities of emotional regulation. Further, the thesis mainly deals with the repressive coping style. Included in the conclusion of the theoretical part is a short chapter dealing with motivation, especially the implicit motives. This whole part of the thesis is based on specialised literature. The practical part contains an empirical research, before the beginning of which a small number of repressors had been found using a screening method. The research itself maps the subjective views of these repressors on the regulation of their own emotions and their implicit motivational tendencies. Conclusions and a summary of results are given at the end of the practical part.

No description available.

Histone Nuclear Factor P (HiNF-P) is a known transcriptional regulator that is critical for the activation of replication dependent histone H4 genes during S phase. HiNF-P is a 65 kDa zinc finger protein that binds to its consensus binding sequence in the Cell Cycle Control Element (Site II) of the proximal promoter region of 11 of the 14 histone H4 genes. HiNF-P is a known co-factor of the global histone gene regulator and cyclinE/CDK2 substrate p220NPAT, however it was not known if this regulatory function reflected a physical interaction. In addition, other HiNF-P interacting proteins have yet to be identified. The work presented in this thesis identifies and characterizes HiNF-P interactions with various proteins within the cell, including p220NPAT. A yeast two-hybrid interaction screen identified candidate interacting proteins of HiNF-P and provided insight into novel cellular functions and transcriptional targets. A candidate yeast two-hybrid approach identified an interaction between HiNF-P and p220NPAT. This direct physical interaction links the cyclin E/CDK2 signaling pathway governing the G1/S phase transition with replication dependent histone gene transcription in S phase. An unbiased yeast two-hybrid screen for HiNF-P interacting proteins revealed an interactome library which suggests roles of HiNF-P in multiple cellular processes. This screen identified 67 candidate HiNF-P interacting proteins that are RNA processing factors, known and putative gene regulators, uncharacterized proteins, proliferation related proteins, as well as metabolic and signaling proteins. Identification of multiple RNA binding and processing factors, including the splicing cofactor, SRm300, links HiNF-P to mRNA processing. HiNF-P is potentially functioning in mRNA processing by interacting with these proteins directly and functioning in complex with them, or more likely, by recruiting these and other splicing factors to sites of transcription. We identified a number of known and putative gene regulators which are candidate HiNF-P interacting proteins. We isolated the atypical C2CH zinc finger protein, THAP7, a known transcriptional repressor. THAP7 interacts with HiNF-P by co-immunoprecipitation and co-immunofluorescence experiments. We show forced expression of THAP7 abrogates HiNF-P/p220 mediated activation of histone H4 gene transcription. THAP7 may represent a novel co-factor of HiNF-P and p220 mediated regulation of histone H4 genes. Identification of interacting proteins of HiNF-P that are involved in transcriptional regulation provides insight into other transcriptional targets of HiNF-P. HiNF-P is localized throughout the nucleus, presumably at multiple gene foci. These interacting proteins may represent novel co-factors of HiNF-P regulation of these other multiple target genes. HiNF-P has been identified as a regulator of cell cycle dependent histone genes, therefore we were interested in identifying other proliferation related proteins with which HiNF-P is interacting. We identified a number of proteins thought to be involved in cellular proliferation, including Ki-67 and an unknown protein XTP2. The functions of these proteins have not been identified. An interaction with HiNF-P might suggest a role for these proteins in histone gene regulation. In addition, Ki-67 has been implicated transcriptional control of ribosomal genes, although no role of HiNF-P in this function has been identified. HiNF-P is a known regulator of histone gene expression via a functional interaction with the global histone gene regulator and cyclin E/CDK2 substrate, p220. This thesis demonstrates HiNF-P directly interacts with the N-terminus of p220. This interaction requires multiple regions within the N-terminus including a LisH-like domain known to function in protein-protein interactions, a region (aa 121-145) known to be required for histone gene transactivation, and another uncharacterized region (209-318). In addition a phylogenically conserved region within the C-terminus of HiNF-P, the HiNF-P Specific Conserved Region (PSCR) is necessary for this interaction. Mutational analysis of these regions abrogates this interaction. HiNF-P and p220 co-localize at specific foci within the cell corresponding to Cajal bodies, which are known sites of histone gene clusters. This work shows that this interaction is necessary for histone gene transcriptional activation and HiNF-P dependent recruitment of p220 to histone H4 gene promoters. In addition HiNF-P as well as p220 interact with the Stem Loop Binding Protein (SLBP) and co-localize in situ. SLBP is a necessary factor for histone pre-mRNA processing events which also occur at Cajal bodies. These interactions provide evidence of the coupling of transcription and processing of histone genes and the involvement of common factors in both processes. This would allow for rapid production of abundant histone proteins which is needed during S phase. This thesis has identified multiple candidate interacting proteins of HiNF-P. These proteins establish HiNF-P as a protein involved in many cellular processes and mechanisms beyond transcriptional control of cell cycle dependent histone genes.