Fabrication of Nanometer and Micrometer Scale Protein Structures by Site-Specific Immobilization of Histidine-Tagged Proteins to Aminosiloxane Films with Photoremovable Protein-Resistant Protecting Groups

The site-specific immobilization of histidine-tagged proteins to patterns formed by far-field and near-field exposure of films of aminosilanes with protein-resistant photolabile protecting groups is demonstrated. After deprotection of the aminosilane, either through a mask or using a scanning near-field optical microscope, the amine terminal groups are derivatized first with glutaraldehyde and then with N-(5-amino-1-carboxypentyl)iminodiacetic acid to yield a nitrilo triacetic acid (NTA) terminated surface. After complexation with Ni2+, this surface binds histidine-tagged GFP and CpcA-PEB in a site-specific fashion. The chemistry is simple and reliable, and leads to extensive surface functionalization. Bright fluorescence is observed in fluorescence microscopy images of micrometer- and nanometer-scale patterns. X-ray photoelectron spectroscopy is used to study quantitatively the efficiency of photodeprotection and the reactivity of the modified surfaces. The efficiency of the protein binding process is investigated quantitatively by ellipsometry and by fluorescence microscopy. We find that regions of the surface not exposed to UV light bind negligible amounts of His-tagged proteins, indicating that the oligo(ethylene glycol) adduct on the nitrophenyl protecting group confers excellent protein resistance; in contrast, exposed regions bind His-GFP very effectively, yielding strong fluorescence that is almost completely removed on treatment of the surface with imidazole, confirming a degree of site-specific binding in excess of 90%. This simple strategy offers a versatile generic route to the spatially selective site-specific immobilization of proteins at surfaces

WAIT, WHAT JUST HAPPENED? DESUBJECTIVATION, QUEERNESS AND REACHING FOR THE REAL IN CONTEMPORARY PORN CONSUMPTION

While browsing for pornography viewers often appear to choose a clip that might initially seem to offer a prurient thrill. This thesis will ask the question of what the implications are if this choice results in an unfamiliar or unanticipated arousal. It will argue that these patterns of viewing may signal an ‘emergent sexual sensibility’, a way of engaging with pornography that operates apart from the individual’s understandings of their own identitarian frameworks of desire. It will define this pattern of porn viewing not as use, which suggests agency and intentional choice, or consumption, which suggests taking something into the body, but as a form of ‘radical exposure’ due to the unintentional nature of the affects that the pornographic materials have on the individual. Many of these viewers then articulate this arousal in what they write in the comments sections below the clip, which often express shock, surprise, anxiety or humour. This thesis will analyse this unintended arousal by examining these comments from the perspective of queer theory and philosophies of desubjectivation, Lacanian understandings of desire, post-truth political discourse and porn and sexuality studies. It is interested in the possibility that this ‘radical exposure’ is leading to ‘desubjectivation’, a term brought into use by Michel Foucault, and which signals a breaking down of an individual’s identitarian understandings of their own desire, which suggests a form of ‘primary queerness’. With regard to theories of desire, sexuality and pornography, the contribution that this thesis will make is in the way it argues that some viewers of pornography, many of whom seem to have a relatively normative understanding of their own frameworks of desire, appear to be exhibiting desubjectivising, or, as will be explained, queer manifestations of desire

Feminism på köpet - En uppsats om identitetsskapande i ett konsumtionssamhälle

Denna uppsats ämnar förklara fenomenet feministiskt konnoterade budskap, förekomna i kommersiella marknadsföringssyften, mer specifikt videoreklam. Detta görs genom att analysera innehållet och tillhörande diskurs i tre olika reklamfilmer innehållande feministiska budskap i form av uppmaning till kvinnlig egenmakt, förändring och/eller kroppspositivism. För att besvara frågeställningen görs en indelning i två delar: identitetsskapande och konsumtionssamhället. Inom ramen för identitetsskapandet kommer vi utgå från Judith Butlers teori om performativitet. För konsumtionssamhällets förståelse kommer vi utgå från marxistiska teorier om varufetischism och reifikation, med fokus på Karl Marx och Georg Lukács. Vidare analyseras konsumtionssamhället utifrån Jean Baudrillard såväl marxistiska som postmoderna teorier om teckenvärdet. Resultatet vi når är att feminism i marknadsföring förekommer för att konsumenten avser upprätthålla en identitet som både feminin, feminist och konsument. Detta jämförs vidare med beskrivningar av postfeminismen, där en slutsats således blir att detta möjliggörs på grund av den riktning feminismen tagit bland moderna konsumenter

Effect of random and hub gene disruptions on environmental and mutational robustness in Escherichia coli

BACKGROUND: Genome-wide profiling has allowed the regulatory interaction networks of many organisms to be visualised and the pattern of connections between genes to be studied. These networks are non-random, following a power-law distribution with a small number of well-connected 'hubs' and many genes with only one or a few connections. Theoretical work predicts that power-law networks display several unique properties. One of the most biologically interesting of these is an intrinsic robustness to disturbance such that removal of a random gene will have little effect on network function. Conversely, targeted removal of a hub gene is expected to have a large effect. RESULTS: We compared the response of Escherichia coli to environmental and mutational stress following disruption of random or hub genes. We found that disruption of random genes had less effect on robustness to environmental stress than did the targeted disruption of hub genes. In contrast, random disruption strains were slightly less robust to the effect of mutational stress than were hub disruption strains. When we compared the effect of each disruption on environmental and mutational stress, we found a negative relationship, such that strains that were more environmentally robust tended to be less robust to mutational stress. CONCLUSION: Our results demonstrate that mutant strains of E. coli respond differently to stress, depending on whether random or hub genes are disrupted. This difference indicates that the power-law distribution of regulatory interactions has biological significance, making random disruptions less deleterious to organisms facing environmental stress. That E. coli can reduce the effect of environmental stress without reducing the phenotypic effect of additional mutations, indicates that robustness and evolvability need not be antagonistic

Reflections on participation: Social or sporting capital?

ConnectSport Research Director, Dr Dan Parnell, of the Business School at Manchester Metropolitan University, and Louise Morby, Senior Lecturer in Sport Development at Leeds Beckett University and Equity Lead at the UK Sport Development and Research Network (UKSDN), provide some reflections on a recent conference focused on participation in sport and physical activity

Los casos de conciencia en la novela pastoril del Siglo de Oro: casuismo y probabilismo en la <em>Arcadia</em> (1598) de Lope de Vega

La <em>Arcadia</em> (1598) de Lope de Vega’s <em>Arcadia</em> (1598) was one of the most successful books in Lope’s already exceptional career, but, in spite of that success, modern critics insist in their criticism of the work, which they see as disorganized and superficial. In order to explain this contrast between these complaints and <em>Arcadia</em>’s success in Early-Modern times, we analyze one of the aspects of the work that critics reject the most: characterization, in particular of the two protagonists, the shepherds Anfriso and Belisarda, as well as some secondary characters. We claim that Golden Age readers must have appreciated the <em>Arcadia</em>’s characters partly because Lope designed them following a way of thinking typical of the time but profoundly strange to ours: moral theology’s casuistry and probabilism. In this context we examine the novel’s «cases» in detail, explaining how they must have been read and enjoyed at the time. In addition, this contextualization allows us to explore the reasons behind Lope’s interest for casuistry, and to relate this way of thinking to his experience as a playwright.<br><br>La <em>Arcadia</em> (1598) de Lope de Vega fue uno de los libros más exitosos de la ya monstruosa carrera del Fénix, pero pese a ello la crítica actual insiste en criticar la obra tachándola de desorganizada y superficial. Para explicar el desfase entre estas críticas y el contrastable éxito de la <em>Arcadia</em> en su tiempo, este trabajo analiza uno de los aspectos más censurados por los estudiosos: la construcción de los personajes del libro, concretamente la de los dos protagonistas, los pastores Anfriso y Belisarda, y la de algunos personajes secundarios. En particular, proponemos que el lector áureo debió de apreciar los personajes de la Arcadia porque estaban diseñados de acuerdo con un hábito mental típico de la época, pero totalmente ajeno a nuestro modo de pensar, el casuismo y probabilismo de la teología moral. En este contexto examinamos en detalle los «casos» de la novela, explicando cómo debió de leerlos y disfrutarlos el público áureo. Además, esta línea de investigación permite explorar la naturaleza del interés del Fénix por la casuística, y las relaciones de este modo de pensar con su oficio de dramaturgo

Preterm neonatal morbidity and mortality by gestational age: a contemporary cohort

Although preterm birth less than 37 weeks gestation is the leading cause of neonatal morbidity and mortality in the United States, the majority of data regarding preterm neonatal outcomes come from older studies, and many reports have been limited to only very preterm neonates. Delineation of neonatal outcomes by delivery gestational age is needed to further clarify the continuum of mortality and morbidity frequencies among preterm neonates