Pluripotent cells have the dual abilities to self-renewal and to differentiate into all three germ layers. Pluripotent cells can be isolated from two different stages of mouse embryogenesis. Embryonic stem cells (ESCs) are isolated from the inner cell mass (ICM) of the pre-implantation embryo and are considered to be in a naïve state. On the other hand, cells isolated from epiblast of the post-implantation embryo are referred as epiblast stem cells (EpiSC) and are representative of primed pluripotency. ESCs and EpiSCs are distinct from each other in terms of the morphology, the gene regulatory network and the signalling pathways regulating self-renewal. Under certain conditions, ESCs and EpiSCs can be transitioned into each other. However, the mechanism that regulates this transition from naïve to primed pluripotent state remains to be solved. Nanog, Oct4 and Sox2 form the core gene regulatory network of pluripotency. Additionally, the Id protein family is also important in the maintenance of pluripotency in ESCs. Id proteins function by inhibiting the activity of pro-differentiation factors. Tcf15 is identified as one of the targets of Id inhibition in ESCs. Moreover, Tcf15 has been identified as a repression target of Nanog. In this study, to understand the function of Tcf15, the expression of Tcf15 was characterized in differentiating ESCs. The transient upregulation of Tcf15 mRNA and protein was detected at early stages of differentiation before lineage commitment. Furthermore, Tcf15 protein was heterogeneously expressed in differentiating cells. Mutually exclusive expression of Nanog and Tcf15 proteins were demonstrated in both self-renewing and differentiating ESCs. Further characterization of the effect of Nanog on Tcf15 transcription showed that Tcf15 pre-mRNA was downregulated within 20 minute of Nanog induction. A Nanog binding site was identified at +32kb relative to the Tcf15 transcription start site (TSS). Initially, Nanog binding at this region was confirmed by performing ChIP-PCR experiments. Then, this Nanog binding region was further analysed for its enhancer activity related to the Tcf15 gene. Deletion of the Nanog binding region using CRISPR-Cas9 confirmed that this region acts as Tcf15 enhancer; it was shown that this region was required for the activation of Tcf15 transcription during differentiation. Tcf15 induction experiments were performed in order to the check whether Tcf15 affects Nanog transcription. The results indicate that Nanog is not a direct target of Tcf15, but Tcf15 contributes indirectly to the repression of Nanog. Additional analysis with the Tcf15 enhancer deletion cells showed that Tcf15 is not required for efficient downregulation of naïve markers and the upregulation of primed markers. However, the genes related to the regulation of adhesion properties of cells such as Zyc, Itga3 were induced with lower efficiency in the absence of Tcf15 compared to the wild type cells. In summary, I investigated the reciprocal regulation of Tcf15 and Nanog and the role of Tcf15 for the differentiation. My results suggest that Tcf15 is expressed in the cells that have initiated differentiation but are not lineage-committed. Additionally, Tcf15 can contribute to the regulation of adhesion related genes in order to help the epithelisation of the cells required during the differentiation from naïve to the primed pluripotent state. As a conclusion, Nanog is proposed to help to prevent certain aspects of ESCs differentiation by repressing Tcf15.

It is well documented in the literature that low-birth-weight (LBW) and prematurity are associated with a variety of developmental disabilities. Within this population of LBW children it is estimated that at birth, up to 45% of them experience intraventricular hemorrhage (IVH). Only recently has pediatric research begun to look at the potentially unique effects of IVH, and attempt to separate these out form the effects of LBW in general. The purpose of this study was to investigate the neuropsychological differences that may occur in children with a history of mild or sever IVH, who are now approaching school age. The main objective was to determine whether children, ages 4 and 5, who were diagnosed with a mild IVH at birth would perform differently on a neuropsychological screening from children who were diagnosed with a severe IVH. Twenty-nine 4- and 5-year-olds born at the University of Utah Medical Center and Primary Children's Medical Center constituted the sample for this study. Potential children were identified through the medical records, where documentation of incident and severity of IVH was obtained. Descriptive medical data and documentation of other common sequelae of LBW was also obtained from the medical records. Parents of potential subjects were contacted from the respective medical centers, and interested parents were then contacted by the research team and included in the study. The children were tested on a variety of neuropsychological functions by trained examiners from the Early Intervention Research Institute at Utah State University and from the Neuropsychological Consultation Services in Salt Lake City, Utah. Analysis of this data was used in determining whether or not there were residual differences in the performance of preschool-age children who have a history of IVH at birth. The results did not indicate significant difference between mild and severe IVH groups in performance on the neuropsychological assessment. Discriminant analysis showed no significant results which did not indicate that group membership could be predicted based upon test performance. Individual subtest analyses also did not indicate a significant difference in performance. Further analysis indicated significant relationships between the presence of other common sequelae of LBW/IVH such as seizure disorder and birth asphyxia, and the neuropsychological test results. Further research is needed to determine the reliability of these findings.

京都大学 / 0048 / 新制・課程博士 / 博士(農学) / 甲第19036号 / 農博第2114号 / 新制||農||1031(附属図書館) / 学位論文||H27||N4918(農学部図書室) / 31987 / 京都大学大学院農学研究科森林科学専攻 / (主査)教授 髙部 圭司, 教授 髙野 俊幸, 教授 矢﨑 一史 / 学位規則第4条第1項該当 / Doctor of Agricultural Science / Kyoto University / DGAM

Within a Southwestern school system, deficits in early literacy skills exist as is illustrated by kindergarten students not meeting the requirements on the Phonological Awareness and Phonics Inventory (PAPI). To address this deficit in early literacy skills, the school system instituted the use of the Guided Reading Approach (GR); however, it was unknown how the kindergarten teachers were implementing GR. The purpose of this study was to investigate the experiences and challenges of kindergarten teachers who implement GR. Vygotsky's sociocultural theory of learning and constructivist theory provided the conceptual framework. Research questions explored the thoughts, attitudes, and beliefs teachers hold about GR as well as the challenges teachers face when incorporating GR. A case study methodology was used to investigate 6 kindergarten teachers' experiences with implementing GR through the use of interviews and document analysis. Analysis of data revealed that teachers believed that GR was a necessary component of teaching and increased student success. However, teachers did not have enough training, collaboration, or time to invest in GR. A professional training was developed for teachers as a result. The 3-day training will provide teachers with an overview of GR, opportunities for the participants to collaborate with colleagues, and time for the development of GR lesson plans that can immediately be transferred to the classroom. Positive social change may result by helping teachers better understand GR (components and implementation), which may result in an improved reading program, higher student performance, and information to influence others to improve reading programs.

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior / This article discusses the distinction that the National Program for Strengthening Family Farming (PRONAF) promotes among farmers in Cachoeira do Sul / RS, given that we are referring to a very heterogeneous social category, which includes over 80% of farms in Brazil. Which was built to provide the basis for public policy, created in the mid 90's and throughout its history has gone through many changes. Currently PRONAF operates in almost all municipalities in the country, have varying interest rates and financing agricultural credit in accordance with the guidelines of the different economic groups of farmers. But even acting in these conditions, the program has failed to meet the needs of the great heterogeneity of producers embraces the concept of family farming. Thus, this study aimed to: understand what are the differences between those farmers who access and those without access to PRONAF and the reasons for that decision, considered the characteristics of the plant and in the same socioeconomic study site. Thus, we performed a search in Cachoeira do Sul / RS, which was used for a qualitative and quantitative methodology, but with greater attention to qualitative aspects of research, since it had as its goal the understanding and quantification not numbers. We use the techniques of observation, semi-structured open interviews, a field diary, analysis of documents and photographs of the site. Where 35 farmers were interviewed in eleven locations within the municipality, 18 and 17 non-pronafianos pronafianos, and mediators of the organs (EMATER, STR, AF and Branch). The survey results indicate that PRONAF promotes a fairly sharp distinction between the farmers, non-pronafianos generally have lower incomes, live in smaller properties, make use of traditional technologies, have few social relationships and afraid to go into debt with bank branches. These differences are known mediators of the organs, but they eschew the situation and the distance between pronafianos and nonpronafianos in Cachoeira do Sul. / O presente trabalho aborda a diferenciação que o Programa Nacional de Fortalecimento da Agricultura Familiar (PRONAF) promove entre os agricultores familiares no Município de Cachoeira do Sul/RS, tendo em vista que se refere a uma categoria social bastante heterogênea, que incorpora mais de 80% dos estabelecimentos agropecuários no Brasil. A qual foi construída para dar base a essa política pública, criada na metade dos anos 90 e que ao longo de sua história passou por muitas mudanças. Atualmente o PRONAF atua em praticamente todos os municípios do país, possui taxas de juros diferenciadas e financia crédito agrícola de acordo com o enquadramento econômico dos distintos grupos de agricultores familiares. Mas, mesmo atuando nessas condições, o programa não tem conseguido atender aos anseios da grande heterogeneidade de produtores que abarca o conceito de agricultura familiar. Dessa forma, este estudo teve como objetivo principal: compreender quais são as diferenças entre aqueles agricultores que acessam e aqueles que não acessam ao PRONAF e os motivos para essa tomada de decisão, considerado as características da unidade de produção e da condição socioeconômica dos mesmos no local de estudo. Nesse sentido, realizamos uma pesquisa no Município de Cachoeira do Sul/RS, para a qual foi utilizada uma metodologia quali-quantitativa, mais com maior atenção aos aspectos qualitativos da pesquisa, já que a mesma tinha como objetivo a compreensão e não a quantificação em números. Fizemos uso das técnicas de observação, da entrevista aberta semi-estruturada, de um diário de campo, de análise de documentos e fotografias do local. Onde foram entrevistados 35 agricultores familiares em onze localidades do interior do Município, sendo 18 pronafianos e 17 não-pronafianos, além dos órgãos mediadores (EMATER, STR, AF e Agências Bancárias). Os resultados da pesquisa apontam que o PRONAF promove uma diferenciação bastante acentuada entre os agricultores familiares, os não-pronafianos em geral possuem menores rendas, vivem em propriedades menores, fazem uso de tecnologias tradicionais, possuem poucas relações sociais e tem medo de se endividar com as agências bancárias. Essas diferenças são do conhecimento dos órgãos mediadores, mas, os mesmos se abstêm da situação e da distância que existe entre pronafianos e não-pronafianos em Cachoeira do Sul.

Doctor of Philosophy / Department of Diagnostic Medicine/Pathobiology / A. Sally Davis / William Wilson / Rift Valley fever virus (RVFV) is a zoonotic arbovirus that is a significant threat to livestock and humans. It is listed as #3 for most dangerous animal threats and is in the top 10 pathogens needing urgent research in preventative and control measures. Although RVFV has never been reported in the US or Europe, outbreaks outside the African continent have sparked renewed interest in developing diagnostics and vaccines to protect both agriculture and public health. Having specific and versatile diagnostics is critical for vaccine development and application. For example, diagnostic tools that aid in identifying key immunogens and understanding the virus-host interaction directly contribute to developing protective vaccines. Additionally, vaccines that are used prophylactically or in response to an outbreak require diagnostic tests to differentiate infected from vaccinated animals (DIVA). This is critical for assessing the return to ‘disease free’ status after an outbreak. Unfortunately, there are limited RVFV diagnostic tests that are versatile and DIVA compatible with the newest RVFV vaccines. We describe the development of several diagnostic tools that are DIVA compatible for detecting RVFV nucleic acid, antibodies, and antigens. First, we evaluate a fluorescence microsphere immunoassay (FMIA) for the detection of antibodies against a RVFV surface glycoprotein and the nucleocapsid protein. The targets developed in this assay provide the basis for a DIVA-compatible serological assay with a candidate RVFV Gn/Gc subunit vaccine, as well as, offer a multiplexing platform that can simultaneously screen for several ruminant diseases. Second, we describe a novel chromogenic in situ hybridization (ISH) assay to detect RVFV in formalin-fixed, paraffin-embedded (FFPE) tissues. This molecular assay offers a highly sensitive, multiplexing platform that detects RVFV RNA on the cellular level of diagnostic tissue samples. Moreover, we demonstrate the first application of ISH as a DIVA-compatible assay for candidate RVFV gene-deletion vaccines. Third, we provide working protocols for western blot (WB), immunohistochemistry (IHC), and immunofluorescence (IF) that use monoclonal or polyclonal antibodies against key RVFV antigens. These tools can be applied to pathogenesis research and used in the development of vaccine and therapeutic countermeasures against RVFV. The RVFV diagnostic methods developed and evaluated in this dissertation can serve as a model for developing diagnostic strategies for other transboundary animal diseases.

A set S of vertices in a graph G=(V,E) is a total dominating set of G if every vertex of V is adjacent to some vertex in S. In this thesis, we consider total dominating sets of minimum cardinality which have the additional property that distinct vertices of V are totally dominated by distinct subsets of the total dominating set.

A set S of vertices in a graph G = (V,E) is a total dominating set of G if every vertex of V is adjacent to a vertex in S. We consider total dominating sets of minimum cardinality which have the additional property that distinct vertices of V are totally dominated by distinct subsets of the total dominating set.

<p>“Fröknarna von Pahlen”, is a series of novels written by the author Agnes von Krusenstjerna. Especially the fourth and fifth parts, published in 1933, raised questions about sexuality, especially about what was conceived as perverse and provoking descriptions. “Fröknarna von Pahlen” became a part of heated debates about what is acceptable to write about. How can the so called Pahlen feud be understood from an intersectional perspective, and from a focus on values, and by discussing imagined communities? The purpose is to find out what is going on in these debates. Six different values are being pointed out; art and skill, truth, freedom of speech and freedom of the press, the value in the young, the value in female perspectives, and moral values. There is a number of knot points tied to these values, and differentiating processes such as sex, class, age, ethnicity, religion etc. are all intertwined in these debates. From an intersectional understanding, none of these processes are more primal than another. The knot points are both of an emotional nature and thematic. The individual voices that emerge in the feud are named small narratives, and the more intersubjective narratives are called grand narratives. These narratives are being investigated rhetorically; for instance how some stories can appear more as truths than others, and it is analyzed how they separate people in groups and create hierarchies. They are also being seen from an emotional perspective; how individual feelings are a part of emotions, larger contexts and meaning coherences. These feelings are also understood as actions. Throughout the investigation there is a hermeneutic will to make things intelligible, and respect and point out the many different perspectives. This is being made in a cultural relativistic attempt. By focusing on imagined communities, different comradeships and groups in the feud can be pointed out. People can consider themselves parts of these groups, but they can also, more or less involuntarily, be considered as parts of these groups. In the writers opinion, the most important question is how “extreme” sexual descriptions an author is allowed to bring forth.<em></em><em></em></p>

“Fröknarna von Pahlen”, is a series of novels written by the author Agnes von Krusenstjerna. Especially the fourth and fifth parts, published in 1933, raised questions about sexuality, especially about what was conceived as perverse and provoking descriptions. “Fröknarna von Pahlen” became a part of heated debates about what is acceptable to write about. How can the so called Pahlen feud be understood from an intersectional perspective, and from a focus on values, and by discussing imagined communities? The purpose is to find out what is going on in these debates. Six different values are being pointed out; art and skill, truth, freedom of speech and freedom of the press, the value in the young, the value in female perspectives, and moral values. There is a number of knot points tied to these values, and differentiating processes such as sex, class, age, ethnicity, religion etc. are all intertwined in these debates. From an intersectional understanding, none of these processes are more primal than another. The knot points are both of an emotional nature and thematic. The individual voices that emerge in the feud are named small narratives, and the more intersubjective narratives are called grand narratives. These narratives are being investigated rhetorically; for instance how some stories can appear more as truths than others, and it is analyzed how they separate people in groups and create hierarchies. They are also being seen from an emotional perspective; how individual feelings are a part of emotions, larger contexts and meaning coherences. These feelings are also understood as actions. Throughout the investigation there is a hermeneutic will to make things intelligible, and respect and point out the many different perspectives. This is being made in a cultural relativistic attempt. By focusing on imagined communities, different comradeships and groups in the feud can be pointed out. People can consider themselves parts of these groups, but they can also, more or less involuntarily, be considered as parts of these groups. In the writers opinion, the most important question is how “extreme” sexual descriptions an author is allowed to bring forth.