The proton pumping stoichiometry of purified mitochondrial complex I reconstituted into proteoliposomes

NADH:ubiquinone oxidoreductase (complex I) is the largest and most complicated enzyme of aerobic electron transfer. The mechanism how it uses redox energy to pump protons across the bioenergetic membrane is still not understood. Here we determined the pumping stoichiometry of mitochondrial complex I from the strictly aerobic yeast Yarrowia lipolytica. With intact mitochondria, the measured value of 3.8H(->+)/2e(-) indicated that four protons are pumped per NADH oxidized. For purified complex I reconstituted into proteoliposomes we measured a very similar pumping stoichiometry of 3.6H(->+)/2e(-). This is the first demonstration that the proton pump of complex I stayed fully functional after purification of the enzyme. (c) 2006 Elsevier B.V. All rights reserved.

Proton pumping by complex I (NADH : ubiquinone oxidoreductase) from Yarrowia lipolytica reconstituted into proteoliposomes

The mechanism of energy converting NADH:ubiquinone oxidoreductase (complex 1) is Still unknown. A current controversy centers around the question whether electron transport of complex I is always linked to vectorial proton translocation or whether in some organisms the enzyme pumps sodium ions instead. To develop better experimental tools to elucidate its mechanism, we have reconstituted the affinity purified enzyme into proteoliposomes and monitored the generation of Delta pH and Delta psi. We tested several detergents to solubilize the asolectin used for liposome formation. Tightly coupled proteoliposomes containing highly active complex I were obtained by detergent removal with BioBeads after total solubilization or the phospholipids with n-octyl-beta-D-glucopyranoside. We have used dyes to monitor the formation of the two components of the proton motive force, Delta pH and Delta psi, across the liposomal membrane, and analyzed the effects of inhibitors, uncouplers and ionophores on this process. We show that electron transfer of complex I of the lower eukaryote Y. lipolytica is clearly linked to proton translocation. While this study was not specifically designed to demonstrate possible additional sodium translocating properties of complex 1, we did not find indications for primary or secondary Na+ translocation by Y lipolytica complex I. (c) 2005 Elsevier B.V. All rights reserved.

Transient pumping of a Ni-like Ag x-ray laser with a subpicosecond pump pulse in a traveling-wave irradiation geometry

We report what is to our knowledge the first demonstration of a transient x-ray laser pumped by a 350-fs pulse in a traveling-wave irradiation geometry. For a 500-fs pump pulse the traveling-wave irradiation was found to have a strong effect on enhancing the Ni-like silver 4d-4p lasing emission at 13.9 nm. The signal enhancement was significantly less when the pulse duration was lengthened to 1.7 ps. The experimental observations are well reproduced by a simple model when the duration of gain is taken of the order of 15-20 ps. For the 500-fs pulse a gain coefficient of 14.5 cm(-1) was measured for plasma lengths up to 7 mm. Refraction of the amplified photons is believed to be the main cause of the limitation of the effective amplification length. (C) 2000 Optical Society of America.

Travelling wave chirped pulse amplified transient pumping for collisional excitation lasers

A similar to 3 ps travelling wave chirped pulse amplified pulse at 6 x 10(14) W cm(-2) superimposed on similar to 300 ps background pulses is shown to be an efficient method to pump transient collisional excitation X-ray lasers in both Ni-like and Ne-like ions. Measurements of X-ray laser output as a function of plasma length are fitted with results of an amplified spontaneous emission model of the laser output taking account of travelling wave pumping effects. A small signal gain coefficient similar to 42 cm(-1) and a effective gain length product of similar to 18 are measured for the Ni-like Sn laser at 120 Angstrom. Simulations from a hydrodynamic and atomic physics code (EHYBRID) coupled to a ray trace code show that a spatially averaged small signal gain similar to 65 cm(-1) can be obtained in Ne-like Ge provided the optimum pumping pulse arrangement is used. (C) 1999 Elsevier Science B.V. All rights reserved.

Optimization of double pulse pumping for Ni-like Sm x-ray lasers

We report a systematic study of double pulse pumping of the Ni-like Sm x-ray laser at 73 Angstrom, currently the shortest wavelength saturated x-ray laser. It is found that the Sm x-ray laser output can change by orders of magnitude when the intensity ratio of the pumping pulses and their relative delay are varied. Optimum pumping conditions are found and interpreted in terms of a simple model. (C) 1999 American Institute of Physics. [S0021-8979(99)07102-9].

Detection of Mixing Dynamics During Pumping of a Flooded Coal Mine

In complex hydrogeological environments the effective management of groundwater quality problems by pump-and-treat operations can be most confidently achieved if the mixing dynamics induced within the aquifer by pumping are well understood. The utility of isotopic environmental tracers (C-, H-, O-, S-stable isotopic analyses and age indicators—14C, 3H) for this purpose is illustrated by the analysis of a pumping test in an abstraction borehole drilled into flooded, abandoned coal mineworkings at Deerplay (Lancashire, UK). Interpretation of the isotope data was undertaken conjunctively with that of major ion hydrochemistry, and interpreted in the context of the particular hydraulic setting of flooded mineworkings to identify the sources and mixing of water qualities in the groundwater system. Initial pumping showed breakdown of initial water quality stratification in the borehole, and gave evidence for distinctive isotopic signatures (d34S(SO4) ~= -1.6‰, d18O(SO4) ~= +15‰) associated with primary oxidation of pyrite in the zone of water table fluctuation—the first time this phenomenon has been successfully characterized by these isotopes in a flooded mine system. The overall aim of the test pumping—to replace an uncontrolled outflow from a mine entrance in an inconvenient location with a pumped discharge on a site where treatment could be provided—was swiftly achieved. Environmental tracing data illustrated the benefits of pumping as little as possible to attain this aim, as higher rates of pumping induced in-mixing of poorer quality waters from more distant old workings, and/or renewed pyrite oxidation in the shallow subsurface.

Symmetrical Solutions, Asymmetrical Realities: Beyond the Politics of Paralysis

The historic significance of the Good Friday Agreement and its role in ending organized political violence is acknowledged at the outset. The article then goes on to probe the roots of the political paralysis built into the architecture of the Agreement that are predicated on a misplaced political and cultural symmetry between the “two communities.” It is suggested that the institutionalized relationship between Northern Ireland and the rest of the U.K. facilitates a cross-party, populist, socio-economic consensus among the nationalist and unionist political parties on the welfare state, taxation and maintaining the massive British subvention to the region. This in turn allows them to concentrate on a divisive culturalist politics, i.e., on antagonistic forms of cultural and identity politics over such issues as flags, parades, and the legacy of the “Troubles” which spills over into gridlock into many areas of regional administration. The article argues for a much broader understanding of culture and identity rooted in the different, if overlapping and interdependent, material realities of both communities while challenging the idea of two cultures/identities as fixed, mutually exclusive, non-negotiable and mutually antagonistic. It then focuses on the importance of Belfast as a key arena which will determine the long-term prospects of an alternative and more constructive form of politics, and enable a fuller recognition of the fundamental asymmetries and inter-dependence between the “two communities.” In the long run, this involves re-defining and reconstructing what is meant by the “Union” and a “United Ireland.”

Asymmetrical balance of power in preadolescents' mutual friendships

Dyadic friendShip' asymmetry was examined in relation to gender, friendship quality, and friendship status. Sixty-nine grade five children and their mutual, same-sex, friends participated in a laboratory session comprised ofvarious activities and completed questionnaires to identify mutual friends and evaluate friendship quality. Asymmetry of power was assessed observationally during an origami task. Variations in balance of power were evident in children's friendships. Boys' dyads had significantly greater levels of asymmetry than girls' dyads. Regardless of gender, asymmetry was associated with lower friendship quality, particularly as indexed by validation and caring and conflict resolution. Asymmetry was unrelated to differences in friendship status. Furthermore, relative individual power within the friendship was not related to individual perceptions offriendship quality. The implications ofthese findings in the theoretical and empirical literature were considered and suggestions for future research were made.

Simple linear asymmetrical complexes of silver(I): NC-Ag-NH3 and Br-Ag-NH3

The compounds Ag(CN)(NH3) and Ag(Br)(NH3) are remarkable in that they form solids containing the simple molecular units NC-Ag-NH3 and Br-Ag-NH3, rather than extended solids, and are the first examples of simple linear asymmetric complexes of silver(I).