Reaction-Based Probes for Imaging Mobile Zinc in Live Cells and Tissues

Chelatable, or mobile, forms of zinc play critical signaling roles in numerous biological processes. Elucidating the action of mobile Zn(II) in complex biological environments requires sensitive tools for visualizing, tracking, and manipulating Zn(II) ions. A large toolbox of synthetic photoinduced electron transfer (PET)-based fluorescent Zn(II) sensors are available, but the applicability of many of these probes is limited by poor zinc sensitivity and low dynamic ranges owing to proton interference. We present here a general approach for acetylating PET-based probes containing a variety of fluorophores and zinc-binding units. The new sensors provide substantially improved zinc sensitivity and allow for incubation of live cells and tissue slices with nM probe concentrations, a significant improvement compared to the μM concentrations that are typically required for a measurable fluorescence signal. Acetylation effectively reduces or completely quenches background fluorescence in the metal-free sensor. Binding of Zn(II) selectively and quickly mediates hydrolytic cleavage of the acetyl groups, providing a large fluorescence response. An acetylated blue coumarin-based sensor was used to carry out detailed analyses of metal binding and metal-promoted acetyl hydrolysis. Acetylated benzoresorufin-based red-emitting probes with different zinc-binding sites are effective for sensing Zn(II) ions in live cells when applied at low concentrations (∼50–100 nM). We used green diacetylated Zinpyr1 (DA-ZP1) to image endogenous mobile Zn(II) in the molecular layer of mouse dorsal cochlear nucleus (DCN), confirming that acetylation is a suitable approach for preparing sensors that are highly specific and sensitive to mobile zinc in biological systems.National Institutes of Health (U.S.) (NIH grant GM065519)National Institutes of Health (U.S.) (NIH grant R01-DC007905)National Institutes of Health (U.S.) (NIH Fellowship (F32- EB019243))National Institutes of Health (U.S.) (NIH Fellowship (T32-DC011499))National Institutes of Health (U.S.) (NIH Fellowship (F32-DC013734)

Bodily relations and reciprocity in the art of Sonia Khurana

This article explores the significance of the ‘somatic’ and ‘ontological turn’ in locating the radical politics articulated in the contemporary performance, installation, video and digital art practices of New Delhi-based artist, Sonia Khurana (b. 1968). Since the late 1990s Khurana has fashioned a range of artworks that require new sorts of reciprocal and embodied relations with their viewers. While this line of art practice suggests the need for a primarily philosophical mode of inquiry into an art of the body, such affective relations need to be historicised also in relation to a discursive field of ‘difference’ and public expectations about the artist’s ethnic, gendered and national identity. Thus, this intimate, visceral and emotional field of inter- and intra-action is a novel contribution to recent transdisciplinary perspectives on the gendered, social and sentient body, that in turn prompts a wider debate on the ethics of cultural commentary and art historiography

Constellations of identity: place-ma(r)king beyond heritage

This paper will critically consider the different ways in which history and belonging have been treated in artworks situated in the Citadel development in Ayr on the West coast of Scotland. It will focus upon one artwork, Constellation by Stephen Hurrel, as an alternative to the more conventional landscapes of heritage which are adjacent, to examine the relationship between personal history and place history and argue the primacy of participatory process in the creation of place and any artwork therein. Through his artwork, Hurrel has attempted to adopt a material process through which place can be created performatively but, in part due to its non-representational form, proves problematic, aesthetically and longitudinally, in wholly engaging the community. The paper will suggest that through variants of ‘new genre public art’ such as this, personal and place histories can be actively re-created through the redevelopment of contemporary urban landscapes but also highlight the complexities and indeterminacies involved in the relationship between artwork, people and place

Lifeworld Inc. : and what to do about it

Can we detect changes in the way that the world turns up as they turn up? This paper makes such an attempt. The first part of the paper argues that a wide-ranging change is occurring in the ontological preconditions of Euro-American cultures, based in reworking what and how an event is produced. Driven by the security – entertainment complex, the aim is to mass produce phenomenological encounter: Lifeworld Inc as I call it. Swimming in a sea of data, such an aim requires the construction of just enough authenticity over and over again. In the second part of the paper, I go on to argue that this new world requires a different kind of social science, one that is experimental in its orientation—just as Lifeworld Inc is—but with a mission to provoke awareness in untoward ways in order to produce new means of association. Only thus, or so I argue, can social science add to the world we are now beginning to live in

Component Interactions and Electron Transfer in Toluene/o-Xylene Monooxygenase

The multicomponent protein toluene/o-xylene monooxygenase (ToMO) activates molecular oxygen to oxidize aromatic hydrocarbons. Prior to dioxygen activation, two electrons are injected into each of two diiron(III) units of the hydroxylase, a process that involves three redox active proteins: the ToMO hydroxylase (ToMOH), Rieske protein (ToMOC), and an NADH oxidoreductase (ToMOF). In addition to these three proteins, a small regulatory protein is essential for catalysis (ToMOD). Through steady state and pre-steady state kinetics studies, we show that ToMOD attenuates electron transfer from ToMOC to ToMOH in a concentration-dependent manner. At substoichiometric concentrations, ToMOD increases the rate of turnover, which we interpret to be a consequence of opening a pathway for oxygen transport to the catalytic diiron center in ToMOH. Excess ToMOD inhibits steady state catalysis in a manner that depends on ToMOC concentration. Through rapid kinetic assays, we demonstrate that ToMOD attenuates formation of the ToMOC–ToMOH complex. These data, coupled with protein docking studies, support a competitive model in which ToMOD and ToMOC compete for the same binding site on the hydroxylase. These results are discussed in the context of other studies of additional proteins in the superfamily of bacterial multicomponent monooxygenases.National Institute of General Medical Sciences (U.S.) (5-R01-GM032134)United States. National Institutes of Health (T32GM008334

The impact of the arts in social work education: a systematic review

Evidence on the effectiveness of arts-based approaches in professional education has been gathering momentum in the last decade embracing disciplines such as medicine, the allied professions, social work and social care. Key texts have emerged promoting the use of the arts in professional education and there have been some attempts to capture empirical evidence on its value. This paper reports on a systematic review of the current body of knowledge on the impact of the arts in social work education. We introduce the rationale for undertaking a systematic review and the methodology and approach used. We then discuss the three significant themes from our synthesis of the evidence reviewed. These were; positioning social work practice through linking micro and macro thinking; the cultivation of leadership beyond verbal reasoning and art as pedagogy. The findings are discussed in the context of what the arts can offer challenges in social work education

Eco-aesthetic dimensions: Herbert Marcuse, ecollogy and art

In his last book, The Aesthetic Dimension (1978), Marcuse argued that a concern for aesthetics is justified when political change is unlikely. But the relation between aesthetics and politics is oblique: “Art cannot change the world, but it can contribute to changing the consciousness … of the men and women who could change the world.” (p. 33). Marcuse also linked his critique of capitalism to environmentalism in the early 1970s: “the violation of the Earth is a vital aspect of the counterrevolution.” (Ecology and Revolution, in The New Left and the 1960s, Collected Papers 3, 2005, p. 173). This article revisits Marcuse’s ideas on aesthetics and ecology, and reviews two recent art projects which engage their audiences in ecological issues: The Jetty Project (2014) by Wolfgang Weileder—which used recycled material and community participation to construct a temporary monument within a wider conservation project on the Tyne, N-E England—and Fracking Futures by HeHe (Helen Evans and Heiko Hansen)—which turned the interior of the gallery at FACT, Liverpool, into what appeared to be a fracking site. The aim is not to evaluate the projects, nor to test the efficacy of Marcuse’s ideas, more to ask again whether art has a role in a shift of attitude which might contribute to dealing with the political and economic causes of climate change

Increase of Direct C-C Coupling Reaction Yield by Identifying Structural and Electronic Properties of High-Spin Iron Tetra-azamacrocyclic Complexes

Macrocyclic ligands have been explored extensively as scaffolds for transition metal catalysts for oxygen and hydrogen atom transfer reactions. C–C reactions facilitated using earth abundant metals bound to macrocyclic ligands have not been well-understood but could be a green alternative to replacing the current expensive and toxic precious metal systems most commonly used for these processes. Therefore, the yields from direct Suzuki–Miyaura C–C coupling of phenylboronic acid and pyrrole to produce 2-phenylpyrrole facilitated by eight high-spin iron complexes ([Fe3+L1(Cl)2]+, [Fe3+L4(Cl)2]+, [Fe2+L5(Cl)]+, [Fe2+L6(Cl)2], [Fe3+L7(Cl)2]+, [Fe3+L8(Cl)2]+, [Fe2+L9(Cl)]+, and [Fe2+L10(Cl)]+) were compared to identify the effect of structural and electronic properties on catalytic efficiency. Specifically, catalyst complexes were compared to evaluate the effect of five properties on catalyst reaction yields: (1) the coordination requirements of the catalyst, (2) redox half-potential of each complex, (3) topological constraint/rigidity, (4) N atom modification(s) increasing oxidative stability of the complex, and (5) geometric parameters. The need for two labile cis-coordination sites was confirmed based on a 42% decrease in catalytic reaction yield observed when complexes containing pentadentate ligands were used in place of complexes with tetradentate ligands. A strong correlation between iron(III/II) redox potential and catalytic reaction yields was also observed, with [Fe2+L6(Cl)2] providing the highest yield (81%, −405 mV). A Lorentzian fitting of redox potential versus yields predicts that these catalysts can undergo more fine-tuning to further increase yields. Interestingly, the remaining properties explored did not show a direct, strong relationship to catalytic reaction yields. Altogether, these results show that modifications to the ligand scaffold using fundamental concepts of inorganic coordination chemistry can be used to control the catalytic activity of macrocyclic iron complexes by controlling redox chemistry of the iron center. Furthermore, the data provide direction for the design of improved catalysts for this reaction and strategies to understand the impact of a ligand scaffold on catalytic activity of other reactions