The redox-Bohr group associated with iron-sulfur cluster N2 of complex I

Proton pumping respiratory complex I (NADH: ubiquinone oxidoreductase) is a major component of the oxidative phosphorylation system in mitochondria and many bacteria. In mammalian cells it provides 40% of the proton motive force needed to make ATP. Defects in this giant and most complicated membrane-bound enzyme cause numerous human disorders. Yet the mechanism of complex I is still elusive. A group exhibiting redox-linked protonation that is associated with iron-sulfur cluster N2 of complex I has been proposed to act as a central component of the proton pumping machinery. Here we show that a histidine in the 49-kDa subunit that resides near iron-sulfur cluster N2 confers this redox-Bohr effect. Mutating this residue to methionine in complex I from Yarrowia lipolytica resulted in a marked shift of the redox midpoint potential of iron-sulfur cluster N2 to the negative and abolished the redox-Bohr effect. However, the mutation did not significantly affect the catalytic activity of complex I and protons were pumped with an unchanged stoichiometry of 4 H+/2e(-). This finding has significant implications on the discussion about possible proton pumping mechanism for complex I.

On the Problems of Mixing RCTs with Qualitative Research: The Case of the MRC Framework for the Evaluation of Complex Healthcare Interventions

In the perceived hierarchy of research designs, the results from randomized controlled trials are considered to provide the highest level of evidence. Indeed these trials have been upheld as the gold standard in research. The benefits and limitations of the randomized controlled trial as a method of evaluating the effectiveness of healthcare interventions are presented. The article then examines the different levels of complexity within healthcare interventions and the problems this poses in determining effectiveness. In an effort to provide a solution to this problem, the Medical Research Council produced a framework to assist investigators to develop and evaluate complex healthcare interventions. The framework is described with reference to an example of implementing and evaluating protocols for weaning patients in the intensive care unit. The framework is critiqued on the basis that it involves an ambiguous or contradictory ontology, which fails to articulate the relationship between the positivism of randomized controlled trials with the relativism of qualitative approaches. It is concluded that the use of realist strategies in combination with randomized controlled trials provides the most coherent solution to this quandary

Experimental Investigation of Stretch Blow Molding, Part 2: Analysis of Process Variables, Blowing Kinematics, and Bottle Properties

The work presented in this paper takes advantage of newly developed instrumentation suitable for in process monitoring of an industrial stretch blow molding machine. The instrumentation provides blowing pressure and stretch rod force histories along with the kinematics of polymer contact with the mould wall. A Design of Experiments pattern was used to qualitatively relate machine inputs with these process parameters and the thickness distribution of stretch blow molded PET (polyethylene terephtalate) bottles. Material slippage at the mold wall and thickness distribution is also discussed in relation to machine inputs. The key process indicators defined have great potential for use in a closed loop process control system and for validation of process simulations.

Perturbations to trophic interactions and the stability of complex food webs

The pattern of predator-prey interactions is thought to be a key determinant of ecosystem processes and stability. Complex ecological networks are characterized by distributions of interaction strengths that are highly skewed, with many weak and few strong interactors present. Theory suggests that this pattern promotes stability as weak interactors dampen the destabilizing potential of strong interactors. Here, we present an experimental test of this hypothesis and provide empirical evidence that the loss of weak interactors can destabilize communities in nature. We ranked 10 marine consumer species by the strength of their trophic interactions. We removed the strongest and weakest of these interactors from experimental food webs containing >100 species. Extinction of strong interactors produced a dramatic trophic cascade and reduced the temporal stability of key ecosystem process rates, community diversity and resistance to changes in community composition. Loss of weak interactors also proved damaging for our experimental ecosystems, leading to reductions in the temporal and spatial stability of ecosystem process rates, community diversity, and resistance. These results highlight the importance of conserving species to maintain the stabilizing pattern of trophic interactions in nature, even if they are perceived to have weak effects in the system.

THE CHEMISTRY OF COMPLEX-MOLECULES IN INTERSTELLAR CLOUDS

This paper is concerned with the chemical evolution of large molecules in interstellar clouds. We consider the chemistry and ionisation balance of large polycyclic aromatic hydrocarbon (PAH) type molecules in diffuse clouds and show that certain PAH molecules can be doubly ionised by the interstellar ultraviolet radiation field. If recombination of the dications so produced with electrons is dissociative rather than radiative, then PAHs are rapidly destroyed. PAHs which can only be singly ionised have much smaller recombination energies and can be long lasting in these regions. This type of property may be very important in selecting the PAH species which can populate the general interstellar medium and account for certain of the diffuse bands observed in optical spectra. Destruction of PAH molecules via formation of dications may be responsible for the weakening of the diffuse bands observed in regions of high UV flux.

Roche tomography of cataclysmic variables - V. A high-latitude star-spot on RU Pegasi

We present Roche tomograms of the secondary star in the dwarf nova system RU Pegasi derived from blue and red arm ISIS data taken on the 4.2-m William Herschel Telescope. We have applied the entropy landscape technique to determine the system parameters and obtained component masses of M1 = 1.06 Msun, M2 = 0.96 Msun, an orbital inclination angle of i = 43 degrees, and an optimal systemic velocity of gamma = 7 km/s. These are in good agreement with previously published values. Our Roche tomograms of the secondary star show prominent irradiation of the inner Lagrangian point due to illumination by the disc and/or bright spot, which may have been enhanced as RU Peg was in outburst at the time of our observations.We find that this irradiation pattern is axi-symmetric and confined to regions of the star which have a direct view of the accretion regions. This is in contrast to previous attempts to map RU Peg which suggested that the irradiation pattern was non-symmetric and extended beyond the terminator. We also detect additional inhomogeneities in the surface distribution of stellar atomic absorption that we ascribe to the presence of a large star-spot. This spot is centred at a latitude of about 82 degrees and covers approximately 4 per cent of the total surface area of the secondary. In keeping with the high latitude spots mapped on the cataclysmic variables AE Aqr and BV Cen, the spot on RU Peg also appears slightly shifted towards the trailing hemisphere of the star. Finally, we speculate that early mapping attempts which indicated non-symmetric irradiation patterns which extended beyond the terminator of CV donors could possibly be explained by a superposition of symmetric heating and a large spot.

Contribution of process variables to the entrapment efficiency of propranolol hydrochloride within ethylcellulose microspheres prepared by the solvent evaporation method as evaluated using a factorial design

The effects of the process variables, pH of aqueous phase, rate of addition of organic, polymeric, drug-containing phase to aqueous phase, organic:aqueous phase volume ratio and aqueous phase temperature on the entrapment of propranolol hydrochloride in ethylcellulose (N4) microspheres prepared by the solvent evaporation method were examined using a factorial design. The observed range of drug entrapment was 1.43 +/- 0.02%w/w (pH 6, 25 degrees C, phase volume ratio 1:10, fast rate of addition) to 16.63 +/- 0.92%w/w (pH 9, 33 degrees C, phase volume ratio 1:10, slow rate of addition) which corresponded to mean entrapment efficiencies of 2.86 and 33.26, respectively. Increased pH, increased temperature and decreased rate of addition significantly enhanced entrapment efficiency. However, organic:aqueous phase volume ratio did not significantly affect drug entrapment. Statistical interactions were observed between pH and rate of addition, pH and temperature, and temperature and rate of addition. The observed interactions involving pH are suggested to be due to the abilities of increased temperature and slow rate of addition to sufficiently enhance the solubility of dichloromethane in the aqueous phase, which at pH 9, but not pH 6, allows partial polymer precipitation prior to drug partitioning into the aqueous phase. The interaction between temperature and rate of addition is due to the relative lack of effect of increased temperature on drug entrapment following slow rate of addition of the organic phase. In comparison to the effects of pH on drug entrapment, the contributions of the other physical factors examined were limited.