593 research outputs found

    Distal unfolding of ferricytochrome C induced by the F82K mutation

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    It is well known that axial coordination of heme iron in mitochondrial cytochrome c has redox-dependent stability. The Met80 heme iron axial ligand in the ferric form of the protein is relatively labile and can be easily replaced by alternative amino acid side chains under non-native conditions induced by alkaline pH, high temperature, or denaturing agents. Here, we showed a redox-dependent destabilization induced in human cytochrome c by substituting Phe82\u2014conserved amino acid and a key actor in cytochrome c intermolecular interactions\u2014with a Lys residue. Introducing a positive charge at position 82 did not significantly affect the structure of ferrous cytochrome c but caused localized unfolding of the distal site in the ferric state. As revealed by1 H NMR fingerprint, the ferric form of the F82K variant had axial coordination resembling the renowned alkaline species, where the detachment of the native Met80 ligand favored the formation of multiple conformations involving distal Lys residues binding to iron, but with more limited overall structural destabilization

    Serum or Plasma (and Which Plasma), That Is the Question

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    Blood derivatives are the biofluids of choice formetabolomic clinical studies since blood can be collected with lowinvasiveness and is rich in biological information. However, the choiceof the blood collection tubes has an undeniable impact on the plasmaand serum metabolic content. Here, we compared the metabolomicand lipoprotein profiles of blood samples collected at the same timeand place from six healthy volunteers but using different collectiontubes (each enrolled volunteer provided multiple blood samples at adistance of a few weeks/months): citrate plasma, EDTA plasma, andserum tubes. All samples were analyzed via nuclear magnetic resonancespectroscopy. Several metabolites showed statistically significantalterations among the three blood matrices, and also metabolites'correlations were shown to be affected. The effects of blood collectiontubes on the lipoproteins'profiles are relevant too, but less marked. Overcoming the issue associated with different blood collectiontubes is pivotal to scale metabolomics and lipoprotein analysis at the level of epidemiological studies based on samples frommulticenter cohorts. We propose a statistical solution, based on regression, that is shown to be efficient in reducing the alterationsinduced by the different collection tubes for both the metabolomic and lipoprotein profile

    Serum or Plasma (and Which Plasma), That Is the Question

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    Oblique stimulated Raman scattering of a short laser pulse in a plasma channel

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    The spatiotemporal evolution of parametric instabilities such as stimulated Raman scattering is studied analytically in time and two spatial dimensions. Initial and boundary conditions are chosen to represent the entrance, propagation, and exit of a laser pulse of finite extent as it progresses through a homogeneous collisional plasma channel. For most scattering angles daughter wave growth is enhanced by lateral reflections within the channel. At late times the two-dimensional interaction approaches a one-dimensional damped mode in which the dissipative loss from lateral transmission of the Stokes wave through the channel boundaries is equivalent to an overall damping of the Stokes amplitudes within the channel

    What do cyclists need to see to avoid single-bicycle crashes?

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    The number of single-bicycle crash victims is substantial in countries with high levels of cycling. To study the role of visual characteristics of the infrastructure, such as pavement markings, in single-bicycle crashes, a study in two steps was conducted. In Study 1, a questionnaire study was conducted among bicycle crash victims (n = 734). Logistic regression was used to study the relationship between the crashes and age, light condition, alcohol use, gaze direction and familiarity with the crash scene. In Study 2, the image degrading and edge detection method (IDED-method) was used to investigate the visual characteristics of 21 of the crash scenes. The results of the studies indicate that crashes, in which the cyclist collided with a bollard or road narrowing or rode off the road, were related to the visual characteristics of bicycle facilities. Edge markings, especially in curves of bicycle tracks, and improved conspicuity of bollards are recommended. Statement of Relevance: Elevated single-bicycle crash numbers are common in countries with high levels of cycling. No research has been conducted on what cyclists need to see to avoid this type of crash. The IDED-method to investigate crash scenes is new and proves to be a powerful tool to quantify 'visual accessibility'. © 2011 Taylor & Francis

    Iron Binding in the Ferroxidase Site of Human Mitochondrial Ferritin

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    Ferritins are nanocage proteins that store iron ions in their central cavity as hydrated ferric oxide biominerals. In mammals, further the L (light) and H (heavy) chains constituting cytoplasmic maxi-ferritins, an additional type of ferritin has been identified, the mitochondrial ferritin (MTF). Human MTF (hMTF) is a functional homopolymeric H-like ferritin performing the ferroxidase activity in its ferroxidase site (FS), in which Fe(II) is oxidized to Fe(III) in the presence of dioxygen. To better investigate its ferroxidase properties, here we performed time-lapse X-ray crystallography analysis of hMTF, providing structural evidence of how iron ions interact with hMTF and of their binding to the FS. Transient iron binding sites, populating the pathway along the cage from the iron entry channel to the catalytic center, were also identified. Furthermore, our kinetic data at variable iron loads indicate that the catalytic iron oxidation reaction occurs via a diferric peroxo intermediate followed by the formation of ferric-oxo species, with significant differences with respect to human H-type ferritin

    Usefulness and acceptability of a standardised orientation and mobility training for partially-sighted older adults using an identification cane

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    <p>Abstract</p> <p>Background</p> <p>Orientation and mobility (O&M) training in using an identification (ID) cane is provided to partially-sighted older adults to facilitate independent functioning and participation in the community. Recently, a protocolised standardised O&M-training in the use of the ID cane was developed in The Netherlands. The purpose of this study is to assess the usefulness and acceptability of both the standardised training and the regular training for participants and O&M-trainers in a randomised controlled trial (NCT00946062).</p> <p>Methods</p> <p>The standardised O&M-training consists of two structured face-to-face sessions and one telephone follow-up, in which, in addition to the regular training, self-management and behavioural change techniques are applied. Questionnaires and interviews were used to collect data on the training’s usefulness, e.g. the population reached, self-reported benefits or achievements, and acceptability, e.g. the performance of the intervention according to protocol and participants’ exposure to and engagement in the training.</p> <p>Results</p> <p>Data was collected from 29 O&M-trainers and 68 participants. Regarding the self-reported benefits, outcomes were comparable for the standardised training and the regular training according the trainers and participants e.g., about 85% of the participants in both groups experienced benefits of the cane and about 70% gained confidence in their capabilities. Participants were actively involved in the standardised training. Nearly 40% of the participants in the standardised training group was not exposed to the training according to protocol regarding the number of sessions scheduled and several intervention elements, such as action planning and contracting.</p> <p>Conclusions</p> <p>The standardised and regular O&M-training showed to be useful and mostly acceptable for the partially-sighted older adults and trainers. Yet, a concern is the deviation from the protocol of the standardised O&M-training by the O&M-trainers regarding distinguishing elements such as action planning. Overall, participants appreciated both trainings and reported benefit.</p
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