À partir des ovocytes de la palourde Spisula solidissima, un ADNc codant un récepteur nommé Spi-OAR a été cloné et séquencé. Une analyse de la séquence en acides aminés a indiqué que ce nouveau récepteur possède une forte similarité avec les récepteurs β-adrénergiques et les récepteurs à octopamine. En effet, il est étroitement lié à la classe des récepteurs à octopamine « β-adrénergique-like » couplés à une protéine Gs. L’ADNc de Spi-OAR a été introduit dans un vecteur d'expression (pCEP4) et un épitope reconnaissable par un anticorps commercial a été ajouté au segment N-terminal. Cette construction a été transfectée dans des cellules hôtes (HEK 293) et des études d’immunofluorescence ont montré une expression efficace du récepteur au niveau membranaire. Également, des mesures d'AMPc pour les cellules exprimant Spi-OAR ont révélé une augmentation de ce messager secondaire lors de l'ajout de l'octopamine, et dans une moindre mesure, la tyramine, tandis que la dopamine, la sérotonine et l'histamine n’ont engendré aucun effet. Une légère activité constitutive de ce récepteur dans les cellules hôtes a été observée. De plus, une analyse RT-PCR avec des oligonucléotides spécifiques a révélé l'ARNm de Spi-OAR non seulement dans les ovocytes, mais aussi dans les gonades, le cœur, les muscles adducteurs, les branchies et les ganglions suggérant que ce récepteur soit exprimé de façon ubiquitaire dans divers tissus et dans différents stades embryonnaires chez la palourde. En outre, des études avec des ovocytes isolés n'ont montré aucun effet de l’octopamine sur la réactivation méiotique. Des études éventuelles pourront finalement confirmer le rôle fonctionnel de Spi-OAR. / A cDNA encoding for an octopamine receptor named Spi-OAR was cloned and sequenced from the surf clam Spisula solidissima oocytes. An analysis of its predicted amino acid sequence showed a high degree of similarity with β-adrenergic and octopamine receptors. This receptor qualifies as a novel receptor closely related to the proposed class of insect octopamine « β-adrenergic–like » receptors coupled to Gs protein. This cDNA was introduced into an expression vector (pCEP4), with an added N-terminal FLAG tag sequence, and transfected in host cells (HEK 293). Immunofluorescence studies showed expression of the receptor with a proper localization to the plasma membrane. Measurements of cAMP in transfected cells revealed that addition of octopamine, and to a lower extent, tyramine induced a rise in cAMP while dopamine, serotonine and histamine had no effect. Overexpression of Spi-OAR in mammalian cells induced slight constitutive increase of cAMP. An RT-PCR analysis with specific oligonucleotides revealed the presence of the receptor mRNA not only in oocytes but also in whole gonads, heart, adductor muscle, gills and ganglia suggesting that this receptor is likely ubiquitously expressed. Expression of Spi-OAR was also detected at different embryonic stages. Despite the demonstrated expression of Spi-OAR in oocytes, octopamine had no effect on meiotic reinitiation. Further studies will examine the function of Spi-OAR.

Reward-related learning occurs when previously neutral stimuli acquires an enhanced ability to elicit approach and other responses. Studies in the past have shown that dopamine receptor-mediated 3’,5’-cyclic adenosine monophosphate (cAMP)-dependent intracellular signalling is important for reward-related learning. Until recently, cAMP-dependent protein kinase (PKA) was the only known signalling molecule that was activated by cAMP. However, it has been discovered that another enzyme, exchange protein directly activated by cAMP (Epac), is also activated by cAMP. Thus, it is possible that cAMP mediates reward-related learning by an Epac-dependent signalling pathway. The present study used a conditioned place preference (CPP) paradigm to investigate whether Epac is involved in the acquisition of reward-related learning. Bilateral injections of amphetamine (20 µg/0.5μl/side) into the nucleus accumbens (NAc) have been shown in previous studies to reliably produce a CPP. Thus, amphetamine (20 µg) and Sp-adenosine 3’,5’-cyclic monophosphorothioate triethylamanine (Sp-cAMPS) (0.1, 1.0, 10, 15, 20 µg), an agent that activates both PKA and Epac, or amphetamine (20 µg) and 8-(4-chlorophenylthio)-2’-O-methyladenosine-3’,5’-cyclic monophosphate (8-pCPT) (0.73, 1.27, 1.45, 2.89, 5.78, 11.56 µg), an agent that selectively activates Epac, were co-injected into NAc to determine their effects on the acquisition of CPP. Results showed that 8-pCPT (1.45 µg), but not lower or higher doses, inhibited CPP. Sp-cAMPS (0.1, 15, 20 µg) also inhibited CPP, replicating the results of previous studies. The results implicate Epac in the acquisition of reward-related learning. / Thesis (Master, Neuroscience Studies) -- Queen's University, 2008-04-25 13:29:37.857

This project engages theories of settler colonialism, biopower, and the state of exception to analyze the operations of rights-based narratives of citizenship in relation to political dissent in Canada. I argue that a normalized state of exception founds the white supremacist, settler colonial state, bringing Canadian citizenship into being as a (white) racialized, (cis)gendered, and (hetero)sexualized construct. By examining “resistance narratives” about the Toronto G20 that emerged in the post-G20 climate, my work argues that, in treating the policing practices employed during the G20 as exceptional and in (re)producing the exaltation of white heterosexual cis-masculine citizens, these narratives normalize and reinforce the daily operations of the exception, which targets Indigenous, racialized, and other “Others” in Canada. Finally, my work critically engages with the space of the Eastern Detention Centre (EDC) as a temporary camp set up to detain G20 arrestees, and with the narrative of “Torontonamo” that emerged to describe and explain the EDC. Reading the EDC in the context of other spatial organizations of the exception in Canada, I argue that the “Torontonamo” narrative reasserts race thinking in relation to the normalized operations of the exception. In so doing, it (re)produces white citizen-subjects as the proper recipients of national and international human rights, while abandoning racialized populations to the space of the camp. Ultimately, my work writes against the hegemonic view of the Toronto G20 as an exceptional event in Canadian history. I contend that G20 policing practices were only a hyper-visible example of the normalized operations of the exception within settler colonialism. / Thesis (Master, Gender Studies) -- Queen's University, 2012-09-29 21:16:51.694

This thesis examines, empirically, the securitization of aid delivery at the Dadaab refugee camps in Kenya. Through a series of semi-structured interviews with aid workers, it documents their security concerns, organizatinonal responses to security risks, and discusses the impacts of these concerns and responses on the delivery of aid to the camps. Armed with a biopolitical conceptualization of sovereignty, articulated in the human security paradigm, the humanitarian aid industry has increasingly reached beyond national borders to touch ‘bare life.’ By now, it is widely recognized that humanitarian principles such as neutrality have often failed to protect aid workers from violent attack as they increasingly venture into the world inhabited by “surplus populations.” Drawing on existing research, this study demonstrates how humanitarian aid delivery in high-risk environments, like refugee camps, is essential to the broader task of using aid to securitize and contain high-risk populations and political instability. Paradoxically, without the securitization of aid at the operational level, humanitarian workers are left exposed to the same enduring elements of insecurity that persistently threaten the lives of those they endeavor to help.

The care of planet Earth is the responsibility of all that inhabit it. Our environment sustains us, and choices we make can compromise the Earth's capacity to sustain life.This project explores how landscape architecture can be a tool that can aid in the process of sustaining certain systems of the Earth, while providing form and function for humans at the same time. Coupled with environmental education programming, the site design becomes the framework in which the programming is based.To sustain life, the Earth must give of its resources. It is well understood that the resources on the planet are limited. Altering systems of life support on the planet must embrace a holistic view in that all systems must be maintained, yet at the same time allow us to derive our means from them.Environmental education is a magical arena that allows for nurturing of human developmental needs by providing outlets for their social, emotional, physical, and cognitive growth. At the same time environmental education provides answers to questions that children and adults have about the environment. Landscape Design in the arena of environmental education can provide a direct link to issues of sustaining life support systems and how people learn about maintaining those systems wisely. / Department of Landscape Architecture

During this creative project rationale, developed from scientific fact, community needs and the values inherent to the student was utilized as the foundation for decisions associated with the development of a campground facility for Prairie Creek Reservoir. Comprehensive and site specific problem-solving methods were used to form a broad overview of the park and recreation planning and design procedures available to landscape architects.At the comprehensive scale, a survey was conducted to assess the community need for the camping experience. An inventory of existing camping opportunities available to the residents of Delaware County was used, in conjunction with the survey results and recreation standards, in a supply/demand equation to determine the number and the type of camping facilities needed to meet present and future demand for the camping experience. The concept of camping at Prairie Creek Reservoir Park was viewed in this light.Site scale investigations began with the establishment of user and resource criteria. Natural, cultural and economic data were collected from the landscape contained within, and immediately surrounding, the property leased by the Muncie Park Board at the Prairie Creek Reservoir site. This information was analyzed in accordance with the design criteria to produce a series of computer generated maps. The student identified alternative campground sites within the site with the help of these visual aids. The campground site, best meeting the user needs and resource capabilities, was selected and a series of development proposals for the campground facility was prepared. / Department of Landscape Architecture

CYP17 is an ACTH/cAMP inducible gene in the human adrenal cortex encoding a cytochrome P450 enzyme with sterol 17α-hydroxylase activity and 17,20 lyase activity essential for biosynthesis of cortisol and androgens. Studies carried out during the past decade have shown that acclerated transcription of inducible eukaryotic genes involves sequential chromatin modifications by cooperative promoter-specific transcription factors and the class of proteins called transcriptional coregulators. In the present work, we aimed to first identify important chromatin modifications and chromatin modifying complexes at the CYP17 transcription start site and nearby steroidogenic factor-1 (SF-1) binding site. Then, we asked what modifications to SF-1 occur during the interaction of this nuclear receptor with the CYP17 promoter, and what their function may be. Finally, we asked how ACTH/cAMP signaling affects SF-1-containing chromatin-modifying complexes during the early phase of transcriptional induction of CYP17. Results from chromatin immunoprecipitation (ChIP) and mammalian two hybrid experiments identified complexes including one comprised of SF-1, steroid receptor coactivator-1 (SRC-1), and the histone acetyltransferase general control nonderepressed 5 (GCN5) as cAMP-inducible, but sensitive to the SF-1 antagonist sphingosine, and able to act in stimulating CYP17 transcription. Moreover, ATPases on the promoter coincided with manipulation of nucleosome histone H2 dimer content. Next, we found that SF-1 phosphorylation by glycogen synthase kinase 3beta (GSK3beta), reciprocal dephosphorylation by phosphatase(s), and acetylation by GCN5 at nearby sites at the ligand binding pocket opening were required for efficient CYP17 transcription. This leads us to propose that ligand binding to SF-1 is controlled by these post-translational modifications. Finally, we determined that the corepressors E1A C-terminal binding proteins (CtBP) 1 and 2 are protein kinase A (PKA) targets and are sensitive to PKA-dependent NADH accumulation. These effects of PKA activation by ACTH/cAMP in adrenal cortex cells enforce CYP17 transcription concomitant with dimerization of CtBP1 and CtBP2.

Adenoviruses have been extensively studied in the field of gene regulation, since their genes are subjected to a tightly controlled temporal expression during the virus lifetime. The early-to-late shift in adenoviral gene expression distinguishes two completely different programs in gene expression. The adenoviral L4-33K protein, which is the subject of this thesis, was previously implicated to be a key player in the transition from the early to the late phase of infection. Here we show that L4-33K activates late gene expression by functioning as a virus-encoded alternative RNA splicing factor activating splicing of transcripts containing weak 3’ splice sites; a feature common to the viral genes expressed at late times of infection. The splicing enhancer activity of L4-33K was mapped to a tiny arginine/serine (RS) repeat in the carboxyl-terminal domain of the protein. Also, the subcellular distribution to the nucleus with enrichment in the nuclear membrane and subnuclear redistribution to viral replication centers during a lytic infection was observed to depend on this motif. RS repeats are common features for the cellular splicing factors serine/arginine-rich (SR) proteins, which in turn are regulated by reversible phosphorylation. We further show that L4-33K is phosphorylated by two cellular protein kinases, the double-stranded DNA-dependent protein kinase (DNA-PK) and protein kinase A (PKA) in vitro. Interestingly, DNA-PK and PKA have opposite effects on the control of the temporally regulated L1 alternative RNA splicing. DNA-PK functions as an inhibitor of the late specific L1-IIIa pre-mRNA splicing whereas PKA functions as an activator of L1-IIIa pre-mRNA splicing. In summary, this thesis describes L4-33K as an SR protein related viral alternative splicing factor. A tiny RS repeat conveys splicing enhancer activity as well as redistribution of L4-33K to replication centers. Finally, DNA-PK and PKA that phosphorylates L4-33K are suggested to be novel regulatory factors controlling adenovirus alternative splicing.

Thesis (Ph. D. in Religion)--Vanderbilt University, May 2009. / Title from title screen. Includes bibliographical references.

Thesis (Ph. D.) -- Georgia State University, 2007. / Includes abstract. Includes bibliographical references (leaves 319-350). Original electric version created as PDF file.