Insights into centriole geometry revealed by cryotomography of doublet and triplet centrioles.

Centrioles are cylindrical assemblies comprised of 9 singlet, doublet, or triplet microtubules, essential for the formation of motile and sensory cilia. While the structure of the cilium is being defined at increasing resolution, centriolar structure remains poorly understood. Here, we used electron cryo-tomography to determine the structure of mammalian (triplet) and Drosophila (doublet) centrioles. Mammalian centrioles have two distinct domains: a 200 nm proximal core region connected by A-C linkers, and a distal domain where the C-tubule is incomplete and a pair of novel linkages stabilize the assembly producing a geometry more closely resembling the ciliary axoneme. Drosophila centrioles resemble the mammalian core, but with their doublet microtubules linked through the A tubules. The commonality of core-region length, and the abrupt transition in mammalian centrioles, suggests a conserved length-setting mechanism. The unexpected linker diversity suggests how unique centriolar architectures arise in different tissues and organisms

Limited biomass recovery from gold mining in Amazonian forests

1. Gold mining has rapidly increased across the Amazon Basin in recent years, especially in the Guiana shield, where it is responsible for >90% of total deforestation. However, the ability of forests to recover from gold mining activities remains largely unquantified. 2. Forest inventory plots were installed on recently abandoned mines in two major mining regions in Guyana, and re‐censused 18 months later, to provide the first ground‐based quantification of gold mining impacts on Amazon forest biomass recovery. 3. We found that woody biomass recovery rates on abandoned mining pits and tailing ponds are among the lowest ever recorded for tropical forests, with close to no woody biomass recovery after 3–4 years. 4. On the overburden sites (i.e. areas not mined but where excavated soil is deposited), however, above‐ground biomass recovery rates (0.4–5.4 Mg ha−1 year−1) were within the range of those recorded in other secondary forests across the Neotropics following abandonment of pastures and agricultural lands. 5. Our results suggest that forest recovery is more strongly limited by severe mining‐induced depletion of soil nutrients, especially nitrogen, than by mercury contamination, due to slowing of growth in nutrient‐stripped soils. 6. We estimate that the slow recovery rates in mining pits and ponds currently reduce carbon sequestration across Amazonian secondary forests by ~21,000 t C/year, compared to the carbon that would have accumulated following more traditional land uses such as agriculture or pasture. 7. Synthesis and applications. To achieve large‐scale restoration targets, Guyana and other Amazonian countries will be challenged to remediate previously mined lands. The recovery process is highly dependent on nitrogen availability rather than mercury contamination, affecting woody biomass regrowth. The significant recovery in overburden zones indicates that one potential active remediation strategy to promote biomass recovery may be to backfill mining pits and ponds with excavated soil

Informed consent for clinical trials in acute coronary syndromes and stroke following the European Clinical Trials Directive: investigators' experiences and attitudes

<p>Abstract</p> <p>Background</p> <p>During clinical trials in emergency medicine, providing appropriate oral and written information to a patient is usually a challenge. There is little published information regarding patients' opinions and competence to provide informed consent, nor on physicians' attitudes towards the process. We have investigated the problem of obtaining consent from patients in emergency-setting clinical trials (such as acute coronary syndromes (ACS) and stroke) from a physicians' perspective.</p> <p>Methods</p> <p>A standardised anonymous 14-item questionnaire was distributed to Polish cardiac and stroke centres.</p> <p>Results</p> <p>Two hundred and fourteen informative investigator responses were received. Of these investigators, 73.8% had experience with ACS and 25.2% had experience with acute stroke trials (and 1% with both fields). The complete model of informed consent (embracing all aspects required by Good Clinical Practice (GCP) and law) was used in 53.3% of cases in emergency settings, whereas the legal option of proxy consent was not used at all. While less than 15% of respondents considered written information to have been fully read by patients, 80.4% thought that the amount of information being given to emergency patients is too lengthy. Although there is no legal obligation, more than half of the investigators sought parallel consent (assent) from patients' relatives. Most investigators confirmed that they would adopt the model proposed by the GCP guidelines: abbreviated verbal and written consent in emergency conditions with obligatory "all-embracing" deferred consent to continue the trial once the patient is able to provide it. However, this model would not follow current Polish and European legislation.</p> <p>Conclusion</p> <p>An update of national and European regulations is required to enable implementation of the emergency trial consent model referred to in GCP guidelines.</p

Climate change 2014 : impacts, adaptation, and vulnerability

Current and future climate-related drivers of risk for small islands during the 21st century include sea level rise (SLR), tropical and extratropical cyclones, increasing air and sea surface temperatures, and changing rainfall patterns (high confidence; robust evidence, high agreement). Current impacts associated with these changes confirm findings reported on small islands from the Fourth Assessment Report (AR4) and previous IPCC assessments. The future risks associated with these drivers include loss of adaptive capacity and ecosystem services critical to lives and livelihoods in small islands.peer-reviewe

Red and green loops help uncover missing feedbacks in a coral reef social–ecological system

Social–ecological systems (SES) exhibit complex cause‐and‐effect relationships. Capturing, interpreting, and responding to signals that indicate changes in ecosystems is key for sustainable management in SES. Breaks in this signal–response chain, when feedbacks are missing, will allow change to continue until a point when abrupt ecological surprises may occur. In these situations, societies and local ecosystems can often become uncoupled. In this paper, we demonstrate how the red loop–green loop (RL–GL) concept can be used to uncover missing feedbacks and to better understand past social–ecological dynamics. Reinstating these feedbacks in order to recouple the SES may ultimately create more sustainable systems on local scales. The RL–GL concept can uncover missing feedbacks through the characterization of SES dynamics along a spectrum of human resource dependence. Drawing on diverse qualitative and quantitative data sources, we classify SES dynamics throughout the history of Jamaican coral reefs along the RL–GL spectrum. We uncover missing feedbacks in red‐loop and red‐trap scenarios from around the year 600 until now. The Jamaican coral reef SES dynamics have moved between all four dynamic states described in the RL–GL concept: green loop, green trap, red loop and red trap. We then propose mechanisms to guide the current unsustainable red traps back to more sustainable green loops, involving mechanisms of seafood trade and ecological monitoring. By gradually moving away from seafood exports, Jamaica may be able to return to green‐loop dynamics between the local society and their locally sourced seafood. We discuss the potential benefits and drawbacks of this proposed intervention and give indications of why an export ban may insure against future missing feedbacks and could prolong the sustainability of the Jamaican coral reef ecosystem. Our approach demonstrates how the RL–GL approach can uncover missing feedbacks in a coral reef SES, a way the concept has not been used before. We advocate for how the RL–GL concept in a feedback setting can be used to synthesize various types of data and to gain an understanding of past, present and future sustainability that can be applied in diverse social–ecological settings

Three-dimensional structural characterization of centrosomes from early Drosophila embryos.

An understanding of the mechanism and structure of microtubule (MT)-nucleating sites within the pericentriolar material (PCM) of the centrosome has been elusive. This is partly due to the difficulty in obtaining large quantities of centrosomes for analysis, as well as to the problem of attaining interpretable structural data with conventional EM techniques. We describe a protocol for isolating a large quantity of functional centrosomes from early Drosophila embryos. Using automated electron tomography, we have begun a three-dimensional structural characterization of these intact centrosomes with and without regrown MTs. Reconstructions of the centrosomes to approximately 6-8 nm resolution revealed no large structures at the minus ends of MTs, suggesting that if MT-nucleating material physically contacts the MTs, it must conform closely to the shape of the minus end. While many MTs originate near the centrioles, MT minus ends were found throughout the PCM, and even close to its outer boundary. The MTs criss-crossed the PCM, suggesting that nucleating sites are oriented in many different directions. Reconstructions of centrosomes without MTs suggest that there is a reorganization of the PCM upon MT regrowth; moreover, ring-like structures that have a similar diameter as MTs are apparent in the PCM of centrosomes without MTs, and may be MT-nucleating sites

Synthesis of Functionalized 2-(4-Hydroxyphenyl)-3-methylbenzofuran Allosteric Modulators of Hsp90 Activity

Hsp90 is a molecular chaperone that plays a pivotal role in the cell life cycle. ATP-regulated internal dynamics are critical to Hsp90 function and we recently demonstrated that these dynamics can be modulated in an allosteric fashion; the protein C-terminal domain (CTD) can be effectively targeted with a family of 2-phenyl-benzofuran derivatives. Here we describe the expansion of the initial library, reporting 28 new derivatives that explore the chemical space at opposite ends of the benzofuran scaffold. Interactions of the compounds with a full-length protein homolog were explored by Saturation Transfer Difference (STD) NMR spectroscopy. In this context we also report the interaction epitope of Novobiocin, a known CTD inhibitor