New Structural and Functional Contexts of the Dx[DN]xDG Linear Motif: Insights into Evolution of Calcium-Binding Proteins

Binding of calcium ions (Ca2+) to proteins can have profound effects on their structure and function. Common roles of calcium binding include structure stabilization and regulation of activity. It is known that diverse families – EF-hands being one of at least twelve – use a Dx[DN]xDG linear motif to bind calcium in near-identical fashion. Here, four novel structural contexts for the motif are described. Existing experimental data for one of them, a thermophilic archaeal subtilisin, demonstrate for the first time a role for Dx[DN]xDG-bound calcium in protein folding. An integrin-like embedding of the motif in the blade of a β-propeller fold – here named the calcium blade – is discovered in structures of bacterial and fungal proteins. Furthermore, sensitive database searches suggest a common origin for the calcium blade in β-propeller structures of different sizes and a pan-kingdom distribution of these proteins. Factors favouring the multiple convergent evolution of the motif appear to include its general Asp-richness, the regular spacing of the Asp residues and the fact that change of Asp into Gly and vice versa can occur though a single nucleotide change. Among the known structural contexts for the Dx[DN]xDG motif, only the calcium blade and the EF-hand are currently found intracellularly in large numbers, perhaps because the higher extracellular concentration of Ca2+ allows for easier fixing of newly evolved motifs that have acquired useful functions. The analysis presented here will inform ongoing efforts toward prediction of similar calcium-binding motifs from sequence information alone

Development and evaluation of an instrument for the critical appraisal of randomized controlled trials of natural products

<p>Abstract</p> <p>Background</p> <p>The efficacy of natural products (NPs) is being evaluated using randomized controlled trials (RCTs) with increasing frequency, yet a search of the literature did not identify a widely accepted critical appraisal instrument developed specifically for use with NPs. The purpose of this project was to develop and evaluate a critical appraisal instrument that is sufficiently rigorous to be used in evaluating RCTs of conventional medicines, and also has a section specific for use with single entity NPs, including herbs and natural sourced chemicals.</p> <p>Methods</p> <p>Three phases of the project included: 1) using experts and a Delphi process to reach consensus on a list of items essential in describing the identity of an NP; 2) compiling a list of non-NP items important for evaluating the quality of an RCT using systematic review methodology to identify published instruments and then compiling item categories that were part of a validated instrument and/or had empirical evidence to support their inclusion and 3) conducting a field test to compare the new instrument to a published instrument for usefulness in evaluating the quality of 3 RCTs of a NP and in applying results to practice.</p> <p>Results</p> <p>Two Delphi rounds resulted in a list of 15 items essential in describing NPs. Seventeen item categories fitting inclusion criteria were identified from published instruments for conventional medicines. The new assessment instrument was assembled based on content of the two lists and the addition of a Reviewer's Conclusion section. The field test of the new instrument showed good criterion validity. Participants found it useful in translating evidence from RCTs to practice.</p> <p>Conclusion</p> <p>A new instrument for the critical appraisal of RCTs of NPs was developed and tested. The instrument is distinct from other available assessment instruments for RCTs of NPs in its systematic development and validation. The instrument is ready to be used by pharmacy students, health care practitioners and academics and will continue to be refined as required.</p

The molecular logic of endocannabinoid signalling

The endocannabinoids are a family of lipid messengers that engage the cell surface receptors that are targeted by Δ9-tetrahydrocannabinol, the active principle in marijuana (Cannabis). They are made on demand through cleavage of membrane precursors and are involved in various short-range signalling processes. In the brain, they combine with CB1 cannabinoid receptors on axon terminals to regulate ion channel activity and neurotransmitter release. Their ability to modulate synaptic efficacy has a wide range of functional consequences and provides unique therapeutic possibilities. © 2003, Nature Publishing Group. All rights reserved

Comparative Assessment of Goods and Services Provided by Grazing Regulation and Reforestation in Degraded Mediterranean Rangelands

Several management actions are applied to restore ecosystem services in degraded Mediterranean rangelands, which range from adjusting the grazing pressure to the removal of grazers and pine plantations. Four such actions were assessed in Quercus coccifera L. shrublands in northern Greece: (i) moderate grazing by goats and sheep; (ii) no grazing; (iii) no grazing plus pine (Pinus pinaster Aiton) plantation in forest gaps (gap reforestation); and (iv) no grazing plus full reforestation of shrubland areas, also with P. pinaster. In addition, heavy grazing was also assessed to serve as a control action. We comparatively assessed the impact of these actions on key provisioning, regulating and supporting ecosystem services by using ground-based indicators. Depending on the ecosystem service considered, the management actions were ranked differently. However, the overall provision of services was particularly favoured under moderate and no grazing management options, with moderate grazing outranking any other action in provisioning services and the no grazing action presenting the most balanced provision of services. Pine reforestations largely contributed to water and soil conservation and C sequestration but had a negative impact on plant diversity when implemented at the expense of removing natural vegetation in the area. Heavy grazing had the lowest provision of ecosystem services. It is concluded that degraded rangelands can be restored by moderating the grazing pressure rather than completely banning livestock grazing or converting them into pine plantations.This research is part of the European Union’s Seventh Framework Programme (FP7/2007-2013) and Support Action PRACTICE (Prevention and Restoration Actions to Combat Desertification: An Integrative assessment—grant agreement no. 226828). The research of S. B., A. G. M. and V. R. V. has received funding from the projects CASCADE (funded by EU-FP7; grant agreement no. 283068), GRACCIE (CSD2007-00067, funded by the Spanish Ministry of Innovation and Science, Consolider-Ingenio 2010 Program) and SFUN (CGL2011-30515-C02-00, funded by the Spanish Ministry of Economy and Competitiveness)