Sampling strategies for characterization of neuropeptides using mass spectrometry and functional implications into cell-cell signaling

In this dissertation, there are developed different analytical strategies to discover and characterize mammalian brain peptides using small amount of tissues. The magnocellular neurons of rat supraoptic nucleus in tissue and cell culture served as the main model to study neuropeptides, in addition to hippocampal neurons and mouse embryonic pituitaries. The neuropeptidomcis studies described here use different extraction methods on tissue or cell culture combined with mass spectrometry (MS) techniques, matrix-assisted laser desorption/ionization (MALDI) and electrospray ionization (ESI). These strategies lead to the identification of multiple peptides from the rat/mouse brain in tissue and cell cultures, including novel compounds One of the goals in this dissertation was to optimize sample preparations on samples isolated from well-defined brain regions for mass spectrometric analysis. Here, the neuropeptidomics study of the SON resulted in the identification of 85 peptides, including 20 unique peptides from known prohormones. This study includes mass spectrometric analysis even from individually isolated magnocellular neuroendocrine cells, where vasopressin and several other peptides are detected. At the same time, it was shown that the same approach could be applied to analyze peptides isolated from a similar hypothalamic region, the suprachiasmatic nucleus (SCN). Although there were some overlaps regarding the detection of the peptides in the two brain nuclei, different peptides were detected specific to each nucleus. Among other peptides, provasopressin fragments were specifically detected in the SON while angiotensin I, somatostatin-14, neurokinin B, galanin, and vasoactive-intestinal peptide (VIP) were detected in the SCN only. Lists of peptides were generated from both brain regions for comparison of the peptidome of SON and SCN nuclei. Moving from analysis of magnocellular neurons in tissue to cell culture, the direct peptidomics of the magnocellular and hippocampal neurons led to the detection of 10 peaks that were assigned to previously characterized peptides and 17 peaks that remain unassigned. Peptides from the vasopressin prohormone and secretogranin-2 are attributed to magnocellular neurons, whereas neurokinin A, peptide J, and neurokinin B are attributed to cultured hippocampal neurons. This approach enabled the elucidation of cell-specific prohormone processing and the discovery of cell-cell signaling peptides. The peptides with roles in the development of the pituitary were analyzed using transgenic mice. Hes1 KO is a genetically modified mouse that lives only e18.5 (embryonic days). Anterior pituitaries of Hes1 null mice exhibit hypoplasia due to increased cell death and reduced proliferation and in the intermediate lobe, the cells differentiate abnormally into somatotropes instead of melanotropes. These previous findings demonstrate that Hes1 has multiple roles in pituitary development, cell differentiation, and cell fate. AVP was detected in all samples. Interestingly, somatostatin [92-100] and provasopressin [151-168] were detected in the mutant but not in the wild type or heterozygous pituitaries while somatostatin-14 was detected only in the heterozygous pituitary. In addition, the putative peptide corresponding to m/z 1330.2 and POMC [205-222] are detected in the mutant and heterozygous pituitaries, but not in the wild type. These results indicate that Hes1 influences the processing of different prohormones having possible roles during development and opens new directions for further developmental studies. This research demonstrates the robust capabilities of MS, which ensures the unbiased direct analysis of peptides extracted from complex biological systems and allows addressing important questions to understand cell-cell signaling in the brain.

Sexuality in the media and emotional well-being among lesbian, gay, & bisexual adolescents

LGB teens’ feelings, desires, and physical attractions run contrary to the heteronormative standards of American society. As such, LGB youth often experience feelings of sadness and dejection that can lead to depression and suicidal tendencies (Russell & Joyner, 2001). Evaluating the factors that could possibly influence the emotional well-being of LGB youth would be an important undertaking given the hindrances LGB adolescents face during sexual socialization. The purpose of this dissertation was to study the portrayal of sexuality in media popular with LGB adolescents and to assess the relationship between media exposure and emotional well-being among LGB teens. In particular, this dissertation distinguished between mainstream media and gay- and lesbian-oriented (GLO) media. GLO media were defined as any media outlet specifically designed, produced, and marketed for gay and lesbian audiences. Two studies were conducted to serve as the initial investigation in a program of research that will be designed to better understand the role of media in the lives of LGB individuals. The first study of this dissertation was a content analysis of the television programs, films, songs, and magazines most popular with LGB teens as determined by self-reports of media consumption in a survey of media use. A total of 96 media vehicles composed the content analysis sample, including 48 television programs, 22 films, 25 musical artists, and 6 magazines. Using a coding scheme that was adapted from previous media sex research, Study 1 measured the frequency of sexual instances as well as the type, nature, and source characteristics for each sexual instance. Results of the content analysis suggest that heterosexuality reigns supreme in mainstream media. When LGB sexuality is depicted in mainstream media, it is often sanitized. LGB sexual talk is rarely sexual; rather it is primarily about the social or cultural components of being lesbian, gay, or bisexual. LGB sexual behavior is also rare in mainstream media, which tend to depict LGB individuals as non-sexually as possible. LGB sexuality in mainstream media exists, but is more about proclaiming LGB identity than actually living it. GLO media depicted LGB sexuality more frequently than mainstream media did. GLO media often depict LGB sexuality in a more realistic manner. LGB sexual talk is about LGB identity, as well as the relational and sexual aspects of being a sexual minority. LGB sexual behavior is commonplace in GLO media, depicting LGB individuals as sexual beings. LGB sexuality in GLO media is prevalent and relatively authentic. The second study was a survey that assessed the relationship between media exposure (both mainstream media and GLO media) and LGB teens’ emotional well-being, considering self-discrepancy as an important mediating variable in that relationship. Study 2 also considered age, sex, and sexual identity commitment as possible moderating variables in the relationship between media exposure and emotional well-being. In Study 2, emotional well-being was defined as lower levels of dejection-related emotions. LGB adolescents (N = 573) completed a questionnaire that was used to investigate the relationships between media exposure and emotional well-being. Results of the survey indicated that mainstream media exposure was not significantly associated with dejection-related emotions. In contrast, GLO media exposure was negatively related to feelings of dejection even when controlling for age, sex, race, perceived social support, school climate, religiosity, geographical location, sexuality of peers, and motivation for viewing LGB inclusive media content. Neither age nor sex moderated the relationships between media exposure variables and dejection, but sexual identity commitment did act as a moderator in the relationship between GLO media exposure and dejection. The negative relationship between GLO media exposure and dejection was stronger for participants lower in sexual identity commitment than for participants higher in sexual identity commitment. In addition, the magnitude of discrepancies between the actual self and the ideal self mediated the relationship between GLO media exposure and dejection for LGB adolescents low in sexual identity commitment. However, self-discrepancy did not mediate the relationship between GLO media exposure and dejection for LGB teens highly committed to their sexual identities. Results of both the content analysis and the survey are discussed in terms of implications for theory and method. Practical implications of this dissertation’s findings are also discussed, as well as directions for future research.

Radiogenic isotopes in the Mediterranean Sea: water mass exchange and precessional variability during the Messinian

During the late Miocene, exchange between the Mediterranean Sea and Atlantic Ocean changed dramatically, culminating in the Messinian Salinity Crisis (MSC). Understanding Mediterranean-Atlantic exchange at that time could answer the enigmatic question of how so much salt built up within the Mediterranean, while furthering the development of a framework for future studies attempting to understand how changes may have impacted global thermohaline circulation. Due to their association with specific water masses at different scales, radiogenic Sr, Pb, and Nd isotope records were generated from various archives contained within marine deposits to endeavour to understand better late Miocene Mediterranean-Atlantic exchange. The archives used include foraminiferal calcite (Sr), fish teeth and bone (Nd), dispersed authigenic ferromanganese oxyhydroxides (Nd, Pb), and a ferromanganese crust (Pb). The primary focus is on sediments preserved at one end of the Betic corridor, a gateway that once connected the Mediterranean to the Atlantic through southern Spain, although other locations are investigated. The Betic gateway terminated within several marginal sub-basins before entering the Western Mediterranean; one of these is the Sorbas Basin, a well-studied location whose sediments have been astronomically tuned at high temporal resolution, providing the necessary age control for sub-precessional resolution records. Since the climatic history of the Mediterranean is strongly controlled by precessional changes in regional climate, the aim was to produce records at high (sub-precessional) temporal resolution, to be able to observe clearly any precessional cyclicity driven by regional climate which could be superimposed over longer trends. This goal was achieved for all records except the ferromanganese crust record. The 87Sr/86Sr isotope record (Ch. 3) shows precessional frequency excursions away from the global seawater curve. As precessional frequency oscillations are unexpected for this setting, a numerical box model was used to determine the mechanisms causing the excursions. To enable parameterisation of model variables, regional Sr characteristics, data from general circulation model HadCM3L, and new benthic foraminiferal assemblage data are employed. The model results imply that the Sorbas Basin likely had a positive hydrologic budget in the late Miocene, very different to that of today. Moreover, the model indicates that the mechanism controlling the Sr isotope ratio of Sorbas Basin seawater was not restriction, but a lack of density-driven exchange with the Mediterranean. Beyond improving our understanding of how marginal Mediterranean sub-basins may evolve different isotope signatures, these results have implications for astronomical tuning and stratigraphy in the region, findings which are crucial considering the geological and climatic history of the late Miocene Mediterranean is based entirely on marginal deposits. An improved estimate for the Nd isotope signature of late Miocene Mediterranean Outflow (MO) was determined by comparing Nd isotope signatures preserved in the deeper Alborán Sea at ODP Site 978 with literature data as well as the signature preserved in the Sorbas Basin (Ch. 4; -9.34 to -9.92 ± 0.37 εNd(t)). It was also inferred that it is unlikely that Nd isotopes can be used reliably to track changes in circulation within the shallow settings characteristic of the Mediterranean-Atlantic connections; this is significant in light of a recent publication documenting corridor closure using Nd isotopes. Both conclusions will prove useful for future studies attempting to understand changes in Mediterranean-Atlantic exchange. Excursions to high values, with precessional frequency, are also observed in the radiogenic Pb isotope record for the Sorbas Basin (Ch. 5). Widening the scope to include locations further away from the gateways, records were produced for late Miocene sections on Sicily and Northern Italy, and similar precessional frequency cyclicity was observed in the Pb isotope records for these sites as well. Comparing these records to proxies for Saharan dust and available whole rock data indicates that, while further analysis is necessary to draw strong conclusions, enhanced dust production during insolation minima may be driving the observed signal. These records also have implications for astronomical tuning; peaks in Pb isotope records driven by Saharan dust may be easier to connect directly to the insolation cycle, providing improved astronomical tuning points. Finally, a Pb isotope record derived using in-situ laser ablation performed on ferromanganese crust 3514-6 from the Lion Seamount, located west of Gibraltar within the MO plume, has provided evidence that plume depth shifted during the Pliocene. The record also suggests that Pb isotopes may not be a suitable proxy for changes in late Miocene Mediterranean-Atlantic exchange, since the Pb isotope signatures of regional water masses are too similar. To develop this record, the first published instance of laser ablation derived 230Thexcess measurements are combined with 10Be dating.

Avian flu: the creation of expectations in the interplay between science and the media

This paper examines the emerging cultural patterns and interpretative repertoires in reports of an impending pandemic of avian flu in the UK mass media and scientific journals at the beginning of 2005, paying particular attention to metaphors, pragmatic markers ('risk signals'), symbolic dates and scare statistics used by scientists and the media to create expectations and elicit actions. This study complements other work on the metaphorical framing of infectious disease, such as foot and mouth disease and SARS, tries to link it to developments in the sociology of expectations and applies insights from pragmatics both to the sociology of metaphor and the sociology of expectations.

Marketplace forces and the history of retailing : The cycle of control

This paper traces the history of store (retailer-controlled) and national (manufacture controlled)brands; identifies the key historical characteristics of the past 200 years of marketing history;describes the four main time periods of U.S. retail marketing (1800 - 2000); and comments on the most likely developments within the current phases of brand marketing. Will the future focus on technology and new forms of communications? The Internet exemplifies an unconventional retailing environment, with etailer numbers growing rapidly. The central proposition of this paper is that a "cycle of control" - a pattern of marketing developments within the history of retailing and national marketing communications - Can indicate the success of marketing strategies in the future.

A new educational technology for media and communication studies : mapping media in Australia and Sweden

This chapter reports on Australian and Swedish experiences in the iterative design, development, and ongoing use of interactive educational systems we call ‘Media Maps.’ Like maps in general, Media Maps are usefully understood as complex cultural technologies; that is, they are not only physical objects, tools and artefacts, but also information creation and distribution technologies, the use and development of which are embedded in systems of knowledge and social meaning. Drawing upon Australian and Swedish experiences with one Media Map technology, this paper illustrates this three-layered approach to the development of media mapping. It shows how media mapping is being used to create authentic learning experiences for students preparing for work in the rapidly evolving media and communication industries. We also contextualise media mapping as a response to various challenges for curriculum and learning design in Media and Communication Studies that arise from shifts in tertiary education policy in a global knowledge economy.

Report on relationship management and culture change

Brisbane Water (BW), a commercialised business arm of Brisbane City Council (BCC) entered into an alliance with a number of organisations from the private sector in order to design, construct, commission and undertake upgrades to three existing wastewater treatment plants located at Sandgate, Oxley Creek, and Wacol in Brisbane. The alliance project is called the Brisbane Water Environmental Alliance (BWEA). This report details the efforts of a team of researchers from the School of Management at Queensland University of Technology to investigate this alliance. This is the second report on this project, and is called Stage 2 of the research. At the time that Stage 2 of the research project was conducted, the BWEA project was nearing completion with a further 8 months remaining before project completion. The aim of this report is to explore individuals’ perceptions of the effectiveness and functioning of the BWEA project in the latter stages of the project. The second aim of this report is to analyse the longitudinal findings of this research project by integrating the findings from Stage 1 and Stage 2 of the project. This long-term analysis of the functioning and effectiveness of the alliance is important because at the current time, researchers have little knowledge of the group developmental processes that occur in large-scale alliances over time. Stage 2 of this research project has a number of aims including assessing performance of the BWEA project from the point of view of a range of stakeholders including the alliance board and alliance management team, alliance staff, and key stakeholders from the client organisation (Brisbane Water). Data were collected using semi-structured interviews with 18 individuals including two board members, one external facilitator, and four staff members from the client organisation. Analysis involved coding the interview transcripts in terms of the major issues that were reported by interviewees.

The media and communications in Australia (3rd ed.)

Traditional media are under assault from digital technologies. Online advertising is eroding the financial basis of newspapers and television, demarcations between different forms of media are fading, and audiences are fragmenting. We can podcast our favourite radio show, data accompanies television programs, and we catch up with newspaper stories on our laptops. Yet mainstream media remain enormously powerful. The Media and Communications in Australia offers a systematic introduction to this dynamic field. Fully updated and revised to take account of recent developments, this third edition outlines the key media industries and explains how communications technologies are impacting on them. It provides a thorough overview of the main approaches taken in studying the media, and includes new chapters on social media, gaming, telecommunications, sport and cultural diversity. With contributions from some of Australia's best researchers and teachers in the field, The Media and Communications in Australia is the most comprehensive and reliable introduction to media and communications available. It is an ideal student text, and a reference for teachers of media and anyone interested in this influential industry.

Identity, relationships and culture

The purpose of this paper is to consider contemporary developments in the knowledge area dealing with culture, relationships and individual identity. It has three main objectives: the first is to examme the ways in which knowledge is currently produced within the discipline; the second is to look at an alternative approach to the issues, an approach which stresses more piecemeal and contingent ways of understanding; finally, this paper looks at how this new theoretical knowledge is being applied, or can be applied, to areas which impact directly upon educational practices.

Hypercapitalism : an investigation into the relationship between language, new media, and social perceptions of value

Overall, this thesis purports to make two significant contributions to knowledge. The first is a foundational critique of political economy in the context of an emergent global knowledge economy. The second is a method for analysing evaluations in language. The relationships that give coherence to those two contributions are as follows. The widely-heralded emergence of a knowledge economy indicates that more intimate aspects of human activity have become exposed to commodification on a massive scale, specifically, activities associated with thought and language. Correspondingly, more abstract forms of value have developed as the products of thought and language have become dominant commodity forms. Historical investigation shows that value has moved from an objective category in political economy, pertaining to such substances as precious metals and land, to become situated today predominantly in “expert” expressions of language, or more precisely, their institutional contexts of production. These are now propagated and circulated on a global scale. Legal, political, and technological developments are key in the development of new, more abstract forms of labour and value, although the relationships connecting these are neither simple nor direct. They are, however, inseparably related in the trajectories that this thesis describes. Consequently they are dealt with inseparably throughout.