Mapping interaction sites within the N-terminus of the calcitonin gene-related peptide receptor; the role of residues 23-60 of the calcitonin receptor-like receptor

The calcitonin receptor-like receptor (CLR) acts as a receptor for the calcitonin gene-related peptide (CGRP) but in order to recognize CGRP, it must form a complex with an accessory protein, receptor activity modifying protein 1 (RAMP1). Identifying the protein/protein and protein/ligand interfaces in this unusual complex would aid drug design. The role of the extreme N-terminus of CLR (Glu23-Ala60) was examined by an alanine scan and the results were interpreted with the help of a molecular model. The potency of CGRP at stimulating cAMP production was reduced at Leu41Ala, Gln45Ala, Cys48Ala and Tyr49Ala; furthermore, CGRP-induced receptor internalization at all of these receptors was also impaired. Ile32Ala, Gly35Ala and Thr37Ala all increased CGRP potency. CGRP specific binding was abolished at Leu41Ala, Ala44Leu, Cys48Ala and Tyr49Ala. There was significant impairment of cell surface expression of Gln45Ala, Cys48Ala and Tyr49Ala. Cys48 takes part in a highly conserved disulfide bond and is probably needed for correct folding of CLR. The model suggests that Gln45 and Tyr49 mediate their effects by interacting with RAMP1 whereas Leu41 and Ala44 are likely to be involved in binding CGRP. Ile32, Gly35 and Thr37 form a separate cluster of residues which modulate CGRP binding. The results from this study may be applicable to other family B GPCRs which can associate with RAMPs.

Lithological mapping of Northwest Argentina with remote sensing data using tonal, textural and contextual features

Tonal, textural and contextual properties are used in manual photointerpretation of remotely sensed data. This study has used these three attributes to produce a lithological map of semi arid northwest Argentina by semi automatic computer classification procedures of remotely sensed data. Three different types of satellite data were investigated, these were LANDSAT MSS, TM and SIR-A imagery. Supervised classification procedures using tonal features only produced poor classification results. LANDSAT MSS produced classification accuracies in the range of 40 to 60%, while accuracies of 50 to 70% were achieved using LANDSAT TM data. The addition of SIR-A data produced increases in the classification accuracy. The increased classification accuracy of TM over the MSS is because of the better discrimination of geological materials afforded by the middle infra red bands of the TM sensor. The maximum likelihood classifier consistently produced classification accuracies 10 to 15% higher than either the minimum distance to means or decision tree classifier, this improved accuracy was obtained at the cost of greatly increased processing time. A new type of classifier the spectral shape classifier, which is computationally as fast as a minimum distance to means classifier is described. However, the results for this classifier were disappointing, being lower in most cases than the minimum distance or decision tree procedures. The classification results using only tonal features were felt to be unacceptably poor, therefore textural attributes were investigated. Texture is an important attribute used by photogeologists to discriminate lithology. In the case of TM data, texture measures were found to increase the classification accuracy by up to 15%. However, in the case of the LANDSAT MSS data the use of texture measures did not provide any significant increase in the accuracy of classification. For TM data, it was found that second order texture, especially the SGLDM based measures, produced highest classification accuracy. Contextual post processing was found to increase classification accuracy and improve the visual appearance of classified output by removing isolated misclassified pixels which tend to clutter classified images. Simple contextual features, such as mode filters were found to out perform more complex features such as gravitational filter or minimal area replacement methods. Generally the larger the size of the filter, the greater the increase in the accuracy. Production rules were used to build a knowledge based system which used tonal and textural features to identify sedimentary lithologies in each of the two test sites. The knowledge based system was able to identify six out of ten lithologies correctly.

Open-source mapping and services for Web-based land-cover validation

Monitoring land-cover changes on sites of conservation importance allows environmental problems to be detected, solutions to be developed and the effectiveness of actions to be assessed. However, the remoteness of many sites or a lack of resources means these data are frequently not available. Remote sensing may provide a solution, but large-scale mapping and change detection may not be appropriate, necessitating site-level assessments. These need to be easy to undertake, rapid and cheap. We present an example of a Web-based solution based on free and open-source software and standards (including PostGIS, OpenLayers, Web Map Services, Web Feature Services and GeoServer) to support assessments of land-cover change (and validation of global land-cover maps). Authorised users are provided with means to assess land-cover visually and may optionally provide uncertainty information at various levels: from a general rating of their confidence in an assessment to a quantification of the proportions of land-cover types within a reference area. Versions of this tool have been developed for the TREES-3 initiative (Simonetti, Beuchle and Eva, 2011). This monitors tropical land-cover change through ground-truthing at latitude / longitude degree confluence points, and for monitoring of change within and around Important Bird Areas (IBAs) by Birdlife International and the Royal Society for the Protection of Birds (RSPB). In this paper we present results from the second of these applications. We also present further details on the potential use of the land-cover change assessment tool on sites of recognised conservation importance, in combination with NDVI and other time series data from the eStation (a system for receiving, processing and disseminating environmental data). We show how the tool can be used to increase the usability of earth observation data by local stakeholders and experts, and assist in evaluating the impact of protection regimes on land-cover change.

Integrated Omics for the global mapping of biomolecules in LDL

Circulating low density lipoproteins (LDL) are thought to play a crucial role in the onset and development of atherosclerosis, though the detailed molecular mechanisms responsible for their biological effects remain controversial. The complexity of biomolecules (lipids, glycans and protein) and structural features (isoforms and chemical modifications) found in LDL particles hampers the complete understanding of the mechanism underlying its atherogenicity. For this reason the screening of LDL for features discriminative of a particular pathology in search of biomarkers is of high importance. Three major biomolecule classes (lipids, protein and glycans) in LDL particles were screened using mass spectrometry coupled to liquid chromatography. Dual-polarity screening resulted in good lipidome coverage, identifying over 300 lipid species from 12 lipid sub-classes. Multivariate analysis was used to investigate potential discriminators in the individual lipid sub-classes for different study groups (age, gender, pathology). Additionally, the high protein sequence coverage of ApoB-100 routinely achieved (≥70%) assisted in the search for protein modifications correlating to aging and pathology. The large size and complexity of the datasets required the use of chemometric methods (Partial Least Square-Discriminant Analysis, PLS-DA) for their analysis and for the identification of ions that discriminate between study groups. The peptide profile from enzymatically digested ApoB-100 can be correlated with the high structural complexity of lipids associated with ApoB-100 using exploratory data analysis. In addition, using targeted scanning modes, glycosylation sites within neutral and acidic sugar residues in ApoB-100 are also being explored. Together or individually, knowledge of the profiles and modifications of the major biomolecules in LDL particles will contribute towards an in-depth understanding, will help to map the structural features that contribute to the atherogenicity of LDL, and may allow identification of reliable, pathology-specific biomarkers. This research was supported by a Marie Curie Intra-European Fellowship within the 7th European Community Framework Program (IEF 255076). Work of A. Rudnitskaya was supported by Portuguese Science and Technology Foundation, through the European Social Fund (ESF) and "Programa Operacional Potencial Humano - POPH".