Characterizing a monotopic membrane enzyme. Biochemical, enzymatic and crystallization studies on Aquifex aeolicus sulfide:quinone oxidoreductase

Monotopic membrane proteins are membrane proteins that interact with only one leaflet of the lipid bilayer and do not possess transmembrane spanning segments. They are endowed with important physiological functions but until now only few of them have been studied. Here we present a detailed biochemical, enzymatic and crystallographic characterization of the monotopic membrane protein sulfide:quinone oxidoreductase. Sulfide:quinone oxidoreductase is a ubiquitous enzyme involved in sulfide detoxification, in sulfide-dependent respiration and photosynthesis, and in heavy metal tolerance. It may also play a crucial role in mammals, including humans, because sulfide acts as a neurotransmitter in these organisms. We isolated and purified sulfide:quinone oxidoreductase from the native membranes of the hyperthermophilic bacterium Aquifex aeolicus. We studied the pure and solubilized enzyme by denaturing and non-denaturing polyacrylamide electrophoresis, size-exclusion chromatography, cross-linking, analytical ultracentrifugation, visible and ultraviolet spectroscopy, mass spectrometry and electron microscopy. Additionally, we report the characterization of its enzymatic activity before and after crystallization. Finally, we discuss the crystallization of sulfide:quinone oxidoreductase in respect to its membrane topology and we propose a classification of monotopic membrane protein crystal lattices. Our data support and complement an earlier description of the three-dimensional structure of A. aeolicus sulfide:quinone oxidoreductase (M. Marcia, U. Ermler, G. Peng, H. Michel, Proc Natl Acad Sci USA, 106 (2009) 9625-9630) and may serve as a reference for further studies on monotopic membrane proteins. (C) 2010 Elsevier B.V. All rights reserved.

74 Front Street (for solo cello)

74 Front Street is intended as a musical response to the work of American sculptor Fred Sandback (1943-2003), in particular his string sculptures which are intentionally uncomplicated, minimal and strikingly beautiful. The relationship between these string works and music is rather distinct; indeed, before formally studying sculpture Sandback made a number of string instruments. He has said that his string works evolved from a desire to rid art of excessive decoration and, while there are instances of activity in 74 Front Street, much of the piece aims to reflect the simplicity of Sandback’s work.

Medieval Women's Writing

Watt, P., Medieval Women's Writing (Cambridge: Polity Press, 2007) RAE2008

Competitive relationships in a fertile grassland community - does size matter?

John Warren, Fred Wilson & Anita Diaz (2002). Competitive relationships in a fertile grassland community - does size matter? Oecologia, 132 (1) pp.125-130 Sponsorship: SEERAD / Leverhulme Trust RAE2008

Garenganze West and East, a review of twenty-one years' pioneer work in the heart of Africa.

http://www.archive.org/details/garenganze00arnouoft/

An Institutional Approach to Donor Control: From Dyadic Ties to a Field‐Level Analysis

Literature on the nonprofit sector focuses on charities and their interactions with clients or governmental agencies; donors are studied less often. Studies on philanthropy do examine donors but tend to focus on microlevel factors to explain their behavior. This study, in contrast, draws on institutional theory to show that macrolevel factors affect donor behavior. It also extends the institutional framework by examining the field‐level configurations in which donors and fundraisers are embedded. Employing the case of workplace charity, this new model highlights how the composition of the organizational field structures fundraisers and donors alike, shaping fundraisers’ strategies of solicitation and, therefore, the extent of donor control.

Living with Jesus: Practical Christologies in Two Boston American Baptist Churches

This dissertation, an exercise in practical theology, undertakes two tasks. First, it examines how the story of Jesus is appropriated and embodied in the corporate practices of worship and mission (congregational christology) and in the daily lives of ordinary believers (everyday christology) at First Baptist Church in Jamaica Plain and Ruggles Baptist Church in Boston, Massachusetts. Second, it places these practical christologies in dialogue with the academic christology of James McClendon to see what creative and critical insights emerge. A key assumption of the study is that doctrinal reflection is precipitated when the story of Jesus interacts with human stories in both autobiographical and public domains. "Living with Jesus" contends that the understandings of Jesus present in the everyday lives of believers and in a congregation's worship and mission merit the attention of scholars in the disciplines of sociology and theology. This dissertation demonstrates that scholarly research on the visible church, everyday religion, and Christian doctrine pays limited attention to the theologies operative in the everyday lives of believers and congregational practices. A gap exists in scholarly knowledge, which "Living with Jesus" attempts to redress. The empirical results of qualitative research fieldwork are set in the context of historical overviews and contemporary snapshots of First Baptist Church in Jamaica Plain and Ruggles Baptist Church. "Living with Jesus" identifies three types of practical christology operative across the two congregations in corporate practices and the everyday lives of individuals: evangelical christology; exemplarist christology; and prophetic christology. The empirical research shows that for a significant minority of people in the sample, the prevailing understanding of Jesus can best be described as a hybrid christology. By paying attention to McClendon's treatment of the Jesus story and placing the three identified practical christologies in dialogue with his christology, it becomes apparent that each practical christology is simultaneously liberating and limiting. This dissertation argues that evaluating a particular practical christology in relation to the Gospel requires an intentional and disciplined effort on the part of congregations, ordinary believers, and theologians. Questions are proposed to assist further christological reflection on worship, mission, pastoral care, and Christian education.

Functional metagenomic analysis of the human gut microbiome to identify novel salt tolerance genes

The ability to adapt to and respond to increases in external osmolarity is an important characteristic that enables bacteria to survive and proliferate in different environmental niches. When challenged with increased osmolarity, due to sodium chloride (NaCl) for example, bacteria elicit a phased response; firstly via uptake of potassium (K+), which is known as the primary response. This primary response is followed by the secondary response which is characterised by the synthesis or uptake of compatible solutes (osmoprotectants). The overall osmotic stress response is much broader however, involving many diverse cellular systems and processes. These ancillary mechanisms are arguably more interesting and give a more complete view of the osmotic stress response. The aim of this thesis was to identify novel genetic loci from the human gut microbiota that confer increased tolerance to osmotic stress using a functional metagenomic approach. Functional metagenomics is a powerful tool that enables the identification of novel genes from as yet uncultured bacteria from diverse environments through cloning, heterologous expression and phenotypic identification of a desired trait. Functional metagenomics does not rely on any previous sequence information to known genes and can therefore enable the discovery of completely novel genes and assign functions to new or known genes. Using a functional metagenomic approach, we have assigned a novel function to previously annotated genes; murB, mazG and galE, as well as a putative brp/blh family beta-carotene 15,15’-monooxygenase. Finally, we report the identification of a completely novel salt tolerance determinant with no current known homologues in the databases. Overall the genes identified originate from diverse taxonomic and phylogenetic groups commonly found in the human gastrointestinal (GI) tract, such as Collinsella and Eggerthella, Akkermansia and Bacteroides from the phyla Actinobacteria, Verrucomicrobia and Bacteroidetes, respectively. In addition, a number of the genes appear to have been acquired via lateral gene transfer and/or encoded on a prophage. To our knowledge, this thesis represents the first investigation to identify novel genes from the human gut microbiota involved in the bacterial osmotic stress response.

Practice-based personas: integrated user and practice models

Practices are routinised behaviours with social and material components and complex relationships over space and time. Practice-based design goes beyond interaction design to consider how these components and their relationships impact on the formation and enactment of a practice, where technology is just one part of the practice. Though situated user-centred design methods such as participatory design are employed for the design of practice, demand exists for additional methods and tools in this area. This paper introduces practice-based personas as an extension of the persona approach popular in interaction design, and demonstrates how a set of practice-based personas was developed for a given domain – academic practice. The three practice-based personas developed here are linked to a catalogue of forty practices, offering designers both a user perspective and a practice perspective when designing for the domain.

Selected Solo Music for Saxophone by United States Composers: 1975-2005

This dissertation project identifies important works for solo saxophone by United States composers between 1975 and 2005. The quality, variety, expressiveness, and difficulty of the solo saxophone repertoire during these thirty years is remarkable and remedies, to some extent, the fact that the saxophone had been a largely neglected instrument in the realm of classical music. In twentieth-century music, including Jazz, the saxophone developed, nevertheless, a unique and significant voice as is evident in the saxophone repertoire that expands immensely in many instrumental settings, including the orchestra, solo works, and a wide variety of chamber ensembles. Historically, the saxophone in the United States first found its niche in Vaudeville, military bands, and jazz ensembles, while in Europe composers such as Debussy, D'Indy, Schmitt, Ibert, Glazounov, Heiden, and Desenclos recognized the potential of the instrument and wrote for it. The saxophone is well suited to the intimacy and unique timbral explorations of the solo literature, but only by the middle twentieth century did the repertoire allow the instrument to flourish into a virtuosic and expressive voice presented by successive generations of performers – Marcel Mule, Sigurd Rascher, Cecil Leeson, Jean-Marie Londeix, Fred Hemke, Eugene Rousseau, and Donald Sinta. The very high artistic level of theses soloists was inspiring and dozens of new compositions were commissioned. Through the 1960’s American composers such as Paul Creston, Leslie Bassett, Henry Cowell, Alec Wilder, and others produced eminent works for the saxophone, to be followed by an enormous output of quality compositions between 1975 and 2005. The works chosen for performance were selected from thousands of compositions between 1975 and 2005 researched for this project. The three recital dates were: April 6, 2005, in Gildenhorn Recital Hall, December 4, 2005, in Ulrich Recital Hall, and April 15, 2006, in Gildenhorn Recital Hall. Recordings of these recitals may be obtained in person or online from the Michelle Smith Performing Arts Library of the University of Maryland, College Park.

Exercise elicits phase shifts and acute alterations of melatonin that vary with circadian phase

To examine the immediate phase-shifting effects of high-intensity exercise of a practical duration (1 h) on human circadian phase, five groups of healthy men 20-30 yr of age participated in studies involving no exercise or exposure to morning, afternoon, evening, or nocturnal exercise. Except during scheduled sleep/dark and exercise periods, subjects remained under modified constant routine conditions allowing a sleep period and including constant posture, knowledge of clock time, and exposure to dim light intensities averaging (±SD) 42 ± 19 lx. The nocturnal onset of plasma melatonin secretion was used as a marker of circadian phase. A phase response curve was used to summarize the phase-shifting effects of exercise as a function of the timing of exercise. A significant effect of time of day on circadian phase shifts was observed (P < 0.004). Over the interval from the melatonin onset before exercise to the first onset after exercise, circadian phase was significantly advanced in the evening exercise group by 30 ± 15 min (SE) compared with the phase delays observed in the no-exercise group (-25 ± 14 min, P < 0.05). Phase shifts in response to evening exercise exposure were attenuated on the second day after exercise exposure and no longer significantly different from phase shifts observed in the absence of exercise. Unanticipated transient elevations of melatonin levels were observed in response to nocturnal exercise and in some evening exercise subjects. Taken together with the results from previous studies in humans and diurnal rodents, the current results suggest that 1) a longer duration of exercise exposure and/or repeated daily exposure to exercise may be necessary for reliable phase-shifting of the human circadian system and that 2) early evening exercise of high intensity may induce phase advances relevant for nonphotic entrainment of the human circadian system.