EXAFS of the type-1 copper site of rusticyanin

AbstractExtended X-ray absorption fine structure (EXAFS) spectra at the Cu K-edge have been recorded of the oxidized and reduced form at pH 3.5 of rusticyanin, the type-1 or ‘blue’-copper protein from Thiobacillus ferrooxidans. The EXAFS of oxidized rusticyanin is well simulated with models assuming a ligand set of 2 N(His) and 1 S(Cys) at 1.99 and 2.16 Å, respectively. Upon reduction, the average Cu-N ligand distance increases by approx. 0.08Å. For both redox states studied, the fit by the simulation is significantly improved by including a contribution of an additional sulfur ligand at approx. 2.8 Å. From comparison with structural data of other blue-copper proteins, it is concluded that the copper coordination environment is relatively rigid, which may be a clue to its high redox potential

Secondary succession in an experimentally fragmented landscape: community patterns across space and time

Secondary succession reflects, at least in part, community assembly—the sequences of colonizations and extinctions. These processes in turn are expected to be sensitive to the size of the site undergoing assembly and its location relative to source pools. In this paper we describe patterns of succession over 18 years in an experimentally fragmented landscape created in eastern Kansas, USA, in 1984. The design of the experiment permits one to assess the influence of patch size and landscape position on successional dynamics. The general trajectory of succession follows that typical of succession in much of the eastern United States. In the initial years of the study, there was relatively little effect of patch size or distance to sources. Here we show that spatial effects in this system have become increasingly evident with time, as gauged both by repeated-measures ANOVA and ordination techniques. Woody plants have colonized more rapidly (per unit area) on large and nearby patches. Species richness at a local (within-quadrat) scale in general has increased, with slightly greater richness in large than in small patches later in the study. Temporal stability in community composition has generally been greater in large patches. Spatial heterogeneity in community composition has increased during succession, but with different patterns in large and small patches. This long-term experiment suggests that landscape structure influences many aspects of community structure and dynamics during succession, and that such effects become more pronounced with the passage of time

Single wall carbon nanotubes enter cells by endocytosis and not membrane penetration

<p>Abstract</p> <p>Background</p> <p>Carbon nanotubes are increasingly being tested for use in cellular applications. Determining the mode of entry is essential to control and regulate specific interactions with cells, to understand toxicological effects of nanotubes, and to develop nanotube-based cellular technologies. We investigated cellular uptake of Pluronic copolymer-stabilized, purified ~145 nm long single wall carbon nanotubes (SWCNTs) through a series of complementary cellular, cell-mimetic, and in vitro model membrane experiments.</p> <p>Results</p> <p>SWCNTs localized within fluorescently labeled endosomes, and confocal Raman spectroscopy showed a dramatic reduction in SWCNT uptake into cells at 4°C compared with 37°C. These data suggest energy-dependent endocytosis, as shown previously. We also examined the possibility for non-specific physical penetration of SWCNTs through the plasma membrane. Electrochemical impedance spectroscopy and Langmuir monolayer film balance measurements showed that Pluronic-stabilized SWCNTs associated with membranes but did not possess sufficient insertion energy to penetrate through the membrane. SWCNTs associated with vesicles made from plasma membranes but did not rupture the vesicles.</p> <p>Conclusions</p> <p>These measurements, combined, demonstrate that Pluronic-stabilized SWCNTs only enter cells via energy-dependent endocytosis, and association of SWCNTs to membrane likely increases uptake.</p

Single wall carbon nanotubes enter cells by endocytosis and not membrane penetration

<p>Abstract</p> <p>Background</p> <p>Carbon nanotubes are increasingly being tested for use in cellular applications. Determining the mode of entry is essential to control and regulate specific interactions with cells, to understand toxicological effects of nanotubes, and to develop nanotube-based cellular technologies. We investigated cellular uptake of Pluronic copolymer-stabilized, purified ~145 nm long single wall carbon nanotubes (SWCNTs) through a series of complementary cellular, cell-mimetic, and in vitro model membrane experiments.</p> <p>Results</p> <p>SWCNTs localized within fluorescently labeled endosomes, and confocal Raman spectroscopy showed a dramatic reduction in SWCNT uptake into cells at 4°C compared with 37°C. These data suggest energy-dependent endocytosis, as shown previously. We also examined the possibility for non-specific physical penetration of SWCNTs through the plasma membrane. Electrochemical impedance spectroscopy and Langmuir monolayer film balance measurements showed that Pluronic-stabilized SWCNTs associated with membranes but did not possess sufficient insertion energy to penetrate through the membrane. SWCNTs associated with vesicles made from plasma membranes but did not rupture the vesicles.</p> <p>Conclusions</p> <p>These measurements, combined, demonstrate that Pluronic-stabilized SWCNTs only enter cells via energy-dependent endocytosis, and association of SWCNTs to membrane likely increases uptake.</p

Subintimal angioplasty of chronic total occlusion in iliac arteries: A safe and durable option

BackgroundTraditionally, aortobifemoral bypass has been the intervention of choice for iliac artery chronic total occlusions (CTOs). However, it is associated with significant morbidity and mortality, limiting its use in high-risk patients. To reduce procedural risk, subintimal angioplasty (SIA) for femoropopliteal CTO has been utilized by many, but few have extended this endovascular technique to treating iliac artery CTOs. We present our experience with 101 successful SIAs for iliac artery CTOs.MethodsA retrospective review of consecutive patients with iliac artery CTOs treated with subintimal angioplasty from June 2000 to January 2009 was completed. Demographic and risk factor data were obtained, along with procedural data. Primary and secondary patency, survival, freedom from claudication, and limb salvage rates were determined by Kaplan-Meier survival analysis. Univariate and multivariate analyses were completed to identify factors adversely affecting primary patency.ResultsOne hundred twenty patients underwent an attempted SIA of an iliac artery CTO, and 101 iliac artery CTOs were successfully treated, giving a technical success rate of 84%. Technical failure was due to the inability to re-enter the lumen in all cases. Indications for intervention were lifestyle-altering claudication in 64 patients (63%) and critical limb ischemia (CLI), in 37 (37%). Eighty-five patients underwent percutaneous SIA, while 11 patients underwent a combined SIA with surgical outflow procedure. Lesions were classified as TransAtlantic InterSociety Consensus (TASC) B, 39 (39%); TASC C, 27 (27%); and TASC D, 35 (35%). In 82 (81%) lesions, stents were deployed with an average of 1.2 (range, 0-3) stents utilized. A re-entry device was used in 14 (14%) lesions. Major complication rate was 3.0%, with a 30-day mortality rate of 1.0%. Primary and secondary patency rates at 1, 2, and 3 years were 86% and 94%, 76% and 92%, and 68% and 80%, respectively. Survival rate was 67% at 5 years, reflecting the poor health of this cohort. Limb salvage for CLI patients at 1 and 5 years was 97% and 95%, respectively. Freedom from claudication at 1 and 3 years was 89% and 73%. Univariate analysis identified hyperlipidemia, coronary artery disease, and prior surgical bypass in treated limb as factors for loss of primary patency; however, on multivariate analysis, no factors remained statistically significant.ConclusionThis study demonstrates that SIA of iliac CTOs is feasible and can be performed safely and effectively, even in high-risk patients. Excellent patency and limb salvage rates can be achieved. In our experience, the safety and durability of SIA makes it an attractive first-line therapy for iliac artery occlusive disease

Measuring the complexity of general practice consultations:development and validation of a complexity measure

Background: The complexity of general practice consultations may be increasing and varies in different settings. A measure of complexity is required to test these hypotheses. Aim: To develop a valid measure of general practice consultation complexity applicable to routine medical records. Design and setting: Delphi study to select potential indicators of complexity followed by a cross-sectional study in English general practices to develop and validate a complexity measure. Method: The online Delphi study over two rounds identified potential indicators of consultation complexity. The cross-sectional study used an age–sex stratified random sample of patients and general practice face-to-face consultations from 2013/2014 in the Clinical Practice Research Datalink. The authors explored independent relationships between each indicator and consultation duration using mixed-effects regression models, and revalidated findings using data from 2017/2018. The proportion of complex consultations in different age–sex groups was assessed. Results: A total of 32 GPs participated in the Delphi study. The Delphi panel endorsed 34 of 45 possible complexity indicators after two rounds. After excluding factors because of low prevalence or confounding, 17 indicators were retained in the cross-sectional study. The study used data from 173 130 patients and 725 616 face-to-face GP consultations. On defining complexity as the presence of any of these 17 factors, 308 370 consultations (42.5%) were found to be complex. Mean duration of complex consultations was 10.49 minutes, compared to 9.64 minutes for non-complex consultations. The proportion of complex consultations was similar in males and females but increased with age. Conclusion: The present consultation complexity measure has face and construct validity. It may be useful for research, management and policy, and for informing decisions about the range of resources needed in different practices

Patient consultation rate and clinical and NHS outcomes:A cross-sectional analysis of English primary care data from 2.7 million patients in 238 practices

Supplementary results. This file contains additional tables presenting results from the analyses which could not be included in the main manuscript. (DOCX 29 kb

Particularized protection: UNSC mandates and the protection of civilians in armed conflict

The protection of civilians at risk in armed conflict has, since the late 1990s, become institutionalized at the United Nations (UN), gaining acceptance as a normative rationale for UN peacekeeping. However, the bulk of civilians in need of protection in armed conflict are unlikely to attain it. The article develops an argument on ‘particularized protection’ - particularized in that UN Security Council (SC) mandates are formulated and adjusted over time to direct mission protection to specific subsets of civilian populations, that is, those relevant to the UN itself, the host state, other states, NGOs and the media, leaving most local civilians receiving little effective protection. Particularized protection, we argue, is a result of the institutional dynamics involving actors producing mandates - the UNSC - and those providing protection - peacekeeping missions - whereby mandates are specified to direct mission protection to selected, particularized groups. We demonstrate these dynamics in two cases, Côte d’Ivoire and Somalia

Where's all the 'good' sports journalism? Sports media research, the sociology of sport, and the question of quality sports reporting

Across newsrooms and journalism schools, questions as to what constitutes or ‘counts’ as excellent reporting are currently inciting much debate. Among the various frameworks being put forward to describe and encourage ‘excellent’ journalism in its various forms, sport is seldom mentioned – a legacy perhaps of its perennial dismissal as trivial subject matter. This essay grew from our curiosity as to whether the reverse was also true: that is, whether and what those who study sports journalism and sports media – in particular sociologists of sport – have contributed to understandings of ‘best’ and even excellent journalistic practice. We identified and analysed 376 articles from eight leading scholarly journals that feature sports media research with the aim of examining instances where ‘excellent’ sports reporting was either highlighted, described or advocated. After outlining the major themes that emerged from this analysis, we reflect on why so few of the sampled articles explicitly advise on what best practice sports journalism might look like – especially when it comes to coverage of the sport-related social issues that sociologists of sport tend to focus on – and why so little theoretical attention has been afforded to the question of excellent sports journalism more generally. While there are good sociological reasons for focusing on problematic sports reporting, on structural and systemic issues in which media are implicated, and on producing alternatives to hegemonic sports media, we conclude that it is high time for instances of excellent sports journalism to be afforded the theoretical and empirical attention long granted to their ‘bad’ journalistic counterparts

Consistent phenological shifts in the making of a biodiversity hotspot: the Cape flora

Background The best documented survival responses of organisms to past climate change on short (glacial-interglacial) timescales are distributional shifts. Despite ample evidence on such timescales for local adaptations of populations at specific sites, the long-term impacts of such changes on evolutionary significant units in response to past climatic change have been little documented. Here we use phylogenies to reconstruct changes in distribution and flowering ecology of the Cape flora - South Africa's biodiversity hotspot - through a period of past (Neogene and Quaternary) changes in the seasonality of rainfall over a timescale of several million years. Results Forty-three distributional and phenological shifts consistent with past climatic change occur across the flora, and a comparable number of clades underwent adaptive changes in their flowering phenology (9 clades; half of the clades investigated) as underwent distributional shifts (12 clades; two thirds of the clades investigated). Of extant Cape angiosperm species, 14-41% have been contributed by lineages that show distributional shifts consistent with past climate change, yet a similar proportion (14-55%) arose from lineages that shifted flowering phenology. Conclusions Adaptive changes in ecology at the scale we uncover in the Cape and consistent with past climatic change have not been documented for other floras. Shifts in climate tolerance appear to have been more important in this flora than is currently appreciated, and lineages that underwent such shifts went on to contribute a high proportion of the flora's extant species diversity. That shifts in phenology, on an evolutionary timescale and on such a scale, have not yet been detected for other floras is likely a result of the method used; shifts in flowering phenology cannot be detected in the fossil record