Characterization of TRAF6 mediated ubiquitination of presenilins and γ-secretase substrates

Post-translational modification of the γ-secretase protease complexes and their substrates has an important role in controlling receptor-initiated signalling events, which are critically important in the pathogenesis of cancer, inflammatory and Alzheimer’s disease. Our lab has previously characterised an interaction between TRAF6 and presenilin-1, which lead to the identification of interleukin-1 (IL-1) receptor type 1 (IL-1R1) and Toll-like receptor-4 (TLR4) as novel γ-secretase substrates. Subsequently our group showed that TRAF6 promoted ubiquitination and γ-secretase cleavage of IL-1R1. The aim of this project is to study the association between TRAF6 and the presenilins, the critical γ-secretase complex components, and to determine the functional importance of TRAF6-mediated ubiquitination of γ-secretase substrates. Firstly, we show that the full-length presenilins are novel substrates of TRAF6-mediated Lysine-63-linked polyubiquitination. Secondly, we show that co-expression of TRAF6 and the presenilins increases the stability and alters the turnover of the presenilins. Thirdly, we reveal that TRAF6-mediated ubiquitination of presenilin does not affect γ-secretase enzyme activity, but may regulate the full-length presenilin functions such as ER Ca2+ signalling. Previously, we have reported IL-1R1 as a novel substrate of TRAF6-mediated ubiquitination. In this study, we identified five lysine residues in the IL-1R1 intracellular domain targeted by TRAF6-mediated polyubiquitination. Furthermore, mutagenesis of these five lysine residues led to decreased IL-1R1 cell surface expression, precluded the ectodomain shedding and attenuated the responsiveness to IL-1β stimulation, demonstrating the critical role of TRAF6 in IL-1R1 trafficking.

Synthesis of iminophosphine and phosphinoiminol cyclometallated Pt (II) and Pt (IV) chloro complexes and studies into their biological and photophysical properties

This thesis focuses on the synthesis and analysis of novel chloride based platinum complexes derived from iminophosphine and phosphinoamide ligands, along with studies on their reactivity towards substitution and oxidation reactions. Also explored here are the potential applications of these complexes for biological and luminescent purposes. Chapter one provides an extensive overview of platinum coordination chemistry with examples of various mixed donor ligands along with the history of platinum anticancer therapy. It also looks at metals in medicine, both for biological functions as well as for therapeutic purposes and gives a background to some other applications for platinum complexes. Chapter two outlines the design and synthetic strategies employed for the development of novel platinum (II) chloride complexes from iminophosphine and phosphinoamide ligands. Also reported is the cyclometallation of these complexes to form stable tridentate mixed donor platinum (II) compounds. In Chapter three the development of a direct method for displacing a chloride from a platinum metal centre with a desired phosphine is reported. Numerous methods for successful oxidation of the platinum (II) complexes will also be explored, leading to novel platinum (IV) complexes being reported here also. The importance of stabilisation of the displaced anion, chloride, by the solvent system will also be discussed in this chapter. Chapter four investigates the reactivity of the platinum (II) complexes towards two different biomolecules to form novel platinum bio-adducts. The potential application of the platinum (II) cyclometallates as chemotherapeutics will also be explored here using in-vitro cancer cell testing. Finally, luminescence studies are also reported here for the ligands and platinum complexes reported in chapter two and three to investigate potential applications in this field also. Chapter five provides a final conclusion and an overall summary of the entire project as well as identifying key areas for future work.

A study of diet and health in the elderly; the gut microbiota as a source of bioactive agents

The global proportion of older persons is increasing rapidly. Diet and the intestinal microbiota independently and jointly contribute to health in the elderly. The habitual dietary patterns and functional microbiota components of elderly subjects were investigated in order to identify specific effector mechanisms. A study of the dietary intake of Irish community-dwelling elderly subjects showed that the consumption of foods high in fat and/or sugar was excessive, while consumption of dairy foods was inadequate. Elderly females typically had a more nutrient- dense diet than males and a considerable proportion of subjects, particularly males, had inadequate intakes of calcium, magnesium, vitamin D, folate, zinc and vitamin C. The association between dietary patterns, glycaemic index and cognitive function was also investigated. Elderly subjects consuming ‘prudent’ dietary patterns had better cognitive function compared to those consuming ‘Western’ dietary patterns. Furthermore, fully-adjusted regression models revealed that a high glycaemic diet was associated with poor cognitive function, demonstrating a new link between nutrition and cognition. An extensive screening study of the elderly faecal-derived microbiota was also undertaken to examine the prevalence of antimicrobial production by intestinal bacteria. A number of previously characterised bacteriocins were isolated (gassericin T, ABP-118, mutacin II, enterocin L-50 and enterocin P) in this study. Interestingly, a Lactobacillus crispatus strain was found to produce a potentially novel antimicrobial compound. Full genome sequencing of this strain revealed the presence of three loci which exhibited varying degrees of homology with the genes responsible for helveticin J production in Lb. helveticus. An additional study comparing the immunomodulatory capacity of ‘viable’ and ‘non-viable’ Bifidobacterium strains found that Bifidobacterium-fermented milks (BFMs) containing ‘non-viable’ cells could stimulate levels of IL-10 and TNF-α in a manner similar to those stimulated by BFMs containing ‘viable’ cells in vitro.

The legitimation of risk and democracy: A case study of Bt cotton in Andhra Pradesh, India

This thesis explores the inter-related attempts to secure the legitimation of risk and democracy with regard to Bt cotton, a genetically modified crop, in the state of Andhra Pradesh in India. The research included nine months of ethnographic fieldwork, extensive library and newspaper research, as well as university attendance in India, undertaken between June, 2010 and March, 2011. This comparative study (involving organic, NPM and Bt cotton cultivation) was conducted in three villages in Telangana, a region which was granted secession from Andhra Pradesh in July, 2013, and in Hyderabad, the state capital. Andhra Pradesh is renowned for its agrarian crisis and farmer suicides, as well as for the conflict which Bt cotton represents. This study adopts the categories of legitimation developed by Van Leeuwen (2007; 2008) in order to explore the theory of risk society (Beck, 1992; 1994; 1999; 2009), and the Habermasian (1996: 356-366) core-periphery model as means of theoretically analysing democratic legitimacy. The legitimation of risk and democracy in relation to Bt cotton refers to normative views on the way in which power should be exercised with regard to risk differentiation, construction and definition. The analysis finds that the more legitimate the exercise of power, the lower the exposure to risk as a concern for the collective. This also has consequences for the way in which resources are distributed, knowledge constructed, and democratic praxis institutionalised as a concern for social and epistemic justice. The thesis argues that the struggle to legitimate risk and democracy has implications not only for the constitution of the new state of Telangana and the region’s development, but also for the emergence of global society and the future development of humanity as a whole.

Fabrication and characterization of doped and porous silicon nanowires as anodes for lithium ion batteries

By using Si(100) with different dopant type (n++-type (As) or p-type (B)), it is shown how metal-assisted chemically (MAC) etched silicon nanowires (Si NWs) can form with rough outer surfaces around a solid NW core for p-type NWs, and a unique, defined mesoporous structure for highly doped n-type NWs. High resolution electron microscopy techniques were used to define the characteristic roughening and mesoporous structure within the NWs and how such structures can form due to a judicious choice of carrier concentration and dopant type. Control of roughness and internal mesoporosity is demonstrated during the formation of Si NWs from highly doped n-type Si(100) during electroless etching through a systematic investigation of etching parameters (etching time, AgNO3 concentration, %HF and temperature). Raman scattering measurements of the transverse optical phonon confirm quantum size effects and phonon scattering in mesoporous wires associated with the etching condition, including quantum confinement effects for the nanocrystallites of Si comprising the internal structure of the mesoporous NWs. Laser power heating of NWs confirms phonon confinement and scattering from internal mesoporosity causing reduced thermal conductivity. The Li+ insertion and extraction characteristics at n-type and p-type Si(100) electrodes with different carrier density and doping type are investigated by cyclic voltammetry and constant current measurements. The insertion and extraction potentials are demonstrated to vary with cycling and the occurrence of an activation effect is shown in n-type electrodes where the charge capacity and voltammetric currents are found to be much higher than p-type electrodes. X-ray photo-electron spectroscopy (XPS) and Raman scattering demonstrate that highly doped n-type Si(100) retains Li as a silicide and converts to an amorphous phase as a two-step phase conversion process. The findings show the succinct dependence of Li insertion and extraction processes for uniformly doped Si(100) single crystals and how the doping type and its effect on the semiconductor-solution interface dominate Li insertion and extraction, composition, crystallinity changes and charge capacity. The effect of dopant, doping density and porosity of MAC etched Si NWs are investigated. The CV response is shown to change in area (current density) with increasing NW length and in profile shape with a changing porosity of the Si NWs. The CV response also changes with scan rate indicative of a transition from intercalation or alloying reactions, to pseudocapactive charge storage at higher scan rates and for p-type NWs. SEM and TEM show a change in structure of the NWs after Li insertion and extraction due to expansion and contraction of the Si NWs. Galvanostatic measurements show the cycling behavior and the Coulombic efficiency of the Si NWs in comparison to their bulk counterparts.

What students can tell about lecturers when they are teaching: manifestations of confident and under-confident lecturers

Delivering a lecture requires confidence, a sound knowledge and well developed teaching skills (Cooper and Simonds, 2007, Quinn and Hughes, 2007). However, practitioners who are new to lecturing large groups in higher education may initially lack the confidence to do so which can manifest itself in their verbal and non-verbal cues and the fluency of their teaching skills. This results in the perception that students can identify the confident and non-confident teacher during a lecture (Street, 2007) and so potentially contributing to a lecturer’s level of anxiety prior to, and during, a lecture. Therefore, in the current educational climate of consumerisation, with the increased evaluation of teaching by students, having the ability to deliver high-quality, informed, and interesting lectures assumes greater significance for both lecturers and universities (Carr, 2007; Higher Education Founding Council 2008, Glass et al., 2006). This paper will present both the quantitative and qualitative data from a two-phase mixed method study with 75 nurse lecturers and 62 nursing students in one university in the United Kingdom. The study investigated the notion that lecturing has similarities to acting (Street, 2007). The findings presented here are concerned with how students perceived lecturers’ level of confidence and how lecturers believed they demonstrated confidence. In phase one a specifically designed questionnaire was distributed to both lecturers and students and a response rate of 91% (n=125) was achieved, while in phase two 12 in-depth semi-structured interviews were conducted with lecturers. Results suggested that students in a lecture could identify if the lecturer was confident or not by the way they performed a lecture. Students identified 57 manifestations of non-confidence and lecturers identified 85, while 57 manifestations of confidence were identified by students and 88 by lecturers. Overall, these fell into 12 main converse categories, ranging from body language to the use of space within the room. Both students and lecturers ranked body language, vocal qualities, delivery skills, involving the students and the ability to share knowledge as the most evident manifestations of confidence. Elements like good eye contact, smiling, speaking clearly and being fluent in the use of media recourses where all seen as manifestations confidence, conversely if these were poorly executed then a presentation of under confidence was evident. Furthermore, if the lecturer appeared enthusiastic it was clearly underpinned by the manifestation of a highly confidence lecturer who was secure in their knowledge base and teaching abilities: Some lecturers do appear enthusiastic but others don’t. I think the ones that do know what they are talking about, you can see it in their voice and in their lively body language. I think they are also good at involving the students even. I think the good ones are able to turn boring subjects into lively and interesting ones. (Student 50) Significantly more lecturers than students felt the lecturer should appear confident when lecturing. The lecturers stated it was particularly important to do so when they did not feel confident, because they were concerned with appearing capable. It seems that these students and lecturers perceived that expressive and apparently confident lecturers can make a positive impact on student groups in terms of involvement in lectures; the data also suggested the reverse, for the under confident lecturer. Findings from phase two indicated that these lecturers assumed a persona when lecturing, particularly, but not exclusively, when they were nervous. These lecturers went through a process of assuming and maintaining this persona before and during a lecture as a way of promoting their internal perceptions of confidence but also their outward manifestation of confidence. Although assuming a convincing persona may have a degree of deception about it, providing the knowledge communicated is accurate, the deception may aid rather than hinder learning, because enhances the delivery of a lecture. Therefore, the deception of acting a little more confidently than one feels might be justified when the lecturer knows the knowledge they are communicating is correct, unlike the Dr Fox Effect where the person delivering a lecture is an actor and does not know the subject in any detail or depth and where the deception to be justified (Naftulin, et al., 1973). In conclusion, these students and lecturers perceive that confident and enthusiastic lecturers communicate their passion for the subject in an interesting and meaningful manner through the use of their voice, body, space and interactions in such a way that shows confidence in their knowledge as well as their teaching abilities. If lecturers, therefore, can take a step back to consider how they deliver lectures in apparently confident ways this may increase their ability to engage their students and not only help them being perceived as good lecturers, but also contribute to the genuine act of education.

Municipal services: organisations, companies and alternatives

A review of the impact of international institutions and multinational companies on municipal services worldwide, and of campaigns for alternatives.

The effects of cadence and power output upon physiological and biomechanical responses to incremental arm-crank ergometry

The aim of this study was to examine the effects of cadence and power output on physiological and biomechanical responses to incremental arm-crank ergometry (ACE). Ten male subjects (mean +/- SD age, 30.4 +/-5.4 y; height, 1.78 +/-0.07 m; mass, 86.1 +/-14.2 kg) undertook 3 incremental ACE protocols to determine peak oxygen uptake (VO2 peak; mean of 3 tests: 3.07 +/- 0.17 L.min-1) at randomly assigned cadences of 50, 70, or 90 r.min-1. Heart rate and expired air were continually monitored. Central (RPE-C) and local (RPE-L) ratings of perceived exertion were recorded at volitional exhaustion. Joint angles and trunk rotation were analysed during each exercise stage. During submaximal power outputs of 50, 70, and 90 W, oxygen consumption (VO2) was lowest for 50 r.min-1 and highest for 90 r.min-1 (p < 0.01). VO2 peak was lowest during 50 r.min-1 (2.79 +/-0.45 L.min-1; p < 0.05) when compared with both 70 r.min-1 and 90 r.min-1 (3.16 +/-0.58, 3.24 +/-0.49 L.min-1, respectively; p > 0.05). The difference between RPE-L and RPE-C at volitional exhaustion was greatest during 50 r.min-1 (2.9 +/- 1.6) when compared with 90 r.min-1 (0.9 +/- 1.9, p < 0.05). At VO2 peak, shoulder range of motion (ROM) and trunk rotation were greater for 50 and 70 r.min-1 when compared with 90 r.min-1 (p < 0.05). During submaximal power outputs, shoulder angle and trunk rotation were greatest at 50 r.min-1 when compared with 90 r.min-1 (p < 0.05). VO2 was inversely related to both trunk rotation and shoulder ROM during submaximal power outputs. The results of this study suggest that the greater forces required at lower cadences to produce a given power output resulted in greater joint angles and range of shoulder and trunk movement. Greater isometric contractions for torso stabilization and increased cost of breathing possibly from respiratory-locomotor coupling may have contributed increased oxygen consumption at higher cadences.