Receptor Activity Modifying Proteins (RAMPs) interact with the VPAC2 Receptor and CRF1 receptors and modulate their function

Background and Purpose Although it is established that the receptor activity modifying proteins (RAMPs) can interact with a number of GPCRs, little is known about the consequences of these interactions. Here the interaction of RAMPs with the glucagon-like peptide 1 receptor (GLP-1 receptor), the human vasoactive intestinal polypeptide/pituitary AC-Activating peptide 2 receptor (VPAC) and the type 1 corticotrophin releasing factor receptor (CRF) has been examined. Experimental Approach GPCRs were co-transfected with RAMPs in HEK 293S and CHO-K1 cells. Cell surface expression of RAMPs and GPCRs was examined by elisa. Where there was evidence for interactions, agonist-stimulated cAMP production, Ca mobilization and GTPγS binding to G, G, G and G were examined. The ability of CRF to stimulate adrenal corticotrophic hormone release in Ramp2 mice was assessed. Key Results The GLP-1 receptor failed to enhance the cell surface expression of any RAMP. VPAC enhanced the cell surface expression of all three RAMPs. CRF enhanced the cell surface expression of RAMP2; the cell surface expression of CRF was also increased. There was no effect on agonist-stimulated cAMP production. However, there was enhanced G-protein coupling in a receptor and agonist-dependent manner. The CRF: RAMP2 complex resulted in enhanced elevation of intracellular calcium to CRF and urocortin 1 but not sauvagine. In Ramp2 mice, there was a loss of responsiveness to CRF. Conclusions and Implications The VPAC and CRF receptors interact with RAMPs. This modulates G-protein coupling in an agonist-specific manner. For CRF, coupling to RAMP2 may be of physiological significance. © 2012 The Authors. British Journal of Pharmacology © 2012 The British Pharmacological Society.

Uncoordinated coexisting IEEE 802.15.4 networks for machine to machine communications

IEEE 802.15.4 standard has been proposed for low power wireless personal area networks. It can be used as an important component in machine to machine (M2M) networks for data collection, monitoring and controlling functions. With an increasing number of machine devices enabled by M2M technology and equipped with 802.15.4 radios, it is likely that multiple 802.15.4 networks may be deployed closely, for example, to collect data for smart metering at residential or enterprise areas. In such scenarios, supporting reliable communications for monitoring and controlling applications is a big challenge. The problem becomes more severe due to the potential hidden terminals when the operations of multiple 802.15.4 networks are uncoordinated. In this paper, we investigate this problem from three typical scenarios and propose an analytic model to reveal how performance of coexisting 802.15.4 networks may be affected by uncoordinated operations under these scenarios. Simulations will be used to validate the analytic model. It is observed that uncoordinated operations may lead to a significant degradation of system performance in M2M applications. With the proposed analytic model, we also investigate the performance limits of the 802.15.4 networks, and the conditions under which coordinated operations may be required to support M2M applications. © 2012 Springer Science + Business Media, LLC.

Receptor activity-modifying protein-dependent effects of mutations in the calcitonin receptor-like receptor:implications for adrenomedullin and calcitonin gene-related peptide pharmacology

Background and Purpose Receptor activity-modifying proteins (RAMPs) define the pharmacology of the calcitonin receptor-like receptor (CLR). The interactions of the different RAMPs with this class B GPCR yield high-affinity calcitonin gene-related peptide (CGRP) or adrenomedullin (AM) receptors. However, the mechanism for this is unclear. Experimental Approach Guided by receptor models, we mutated residues in the N-terminal helix of CLR, RAMP2 and RAMP3 hypothesized to be involved in peptide interactions. These were assayed for cAMP production with AM, AM2 and CGRP together with their cell surface expression. Binding studies were also conducted for selected mutants. Key Results An important domain for peptide interactions on CLR from I32 to I52 was defined. Although I41 was universally important for binding and receptor function, the role of other residues depended on both ligand and RAMP. Peptide binding to CLR/RAMP3 involved a more restricted range of residues than that to CLR/RAMP1 or CLR/RAMP2. E101 of RAMP2 had a major role in AM interactions, and F111/W84 of RAMP2/3 was important with each peptide. Conclusions and Implications RAMP-dependent effects of CLR mutations suggest that the different RAMPs control accessibility of peptides to binding residues situated on the CLR N-terminus. RAMP3 appears to alter the role of specific residues at the CLR-RAMP interface compared with RAMP1 and RAMP2. © 2013 The Authors. British Journal of Pharmacology published by John Wiley &. Sons Ltd on behalf of The British Pharmacological Society.

Structural basis for receptor activity-modifying protein-dependent selective peptide recognition by a G protein-coupled receptor

Association of receptor activity-modifying proteins (RAMP1-3) with the G protein-coupled receptor (GPCR) calcitonin receptor-like receptor (CLR) enables selective recognition of the peptides calcitonin gene-related peptide (CGRP) and adrenomedullin (AM) that have diverse functions in the cardiovascular and lymphatic systems. How peptides selectively bind GPCR:RAMP complexes is unknown. We report crystal structures of CGRP analog-bound CLR:RAMP1 and AM-bound CLR:RAMP2 extracellular domain heterodimers at 2.5 and 1.8 Å resolutions, respectively. The peptides similarly occupy a shared binding site on CLR with conformations characterized by a β-turn structure near their C termini rather than the α-helical structure common to peptides that bind related GPCRs. The RAMPs augment the binding site with distinct contacts to the variable C-terminal peptide residues and elicit subtly different CLR conformations. The structures and accompanying pharmacology data reveal how a class of accessory membrane proteins modulate ligand binding of a GPCR and may inform drug development targeting CLR:RAMP complexes.

Connection design without accurate LV feeder load data:an argument for LV monitoring

Since privatisation, maintenance of DNO LV feeder maximum demand information has gradually demised in some Utility Areas, and it is postulated that lack of knowledge about 11kV and LV electrical networks is resulting in a less economical and energy efficient Network as a whole. In an attempt to quantify the negative impact, this paper examines ten postulated new connection scenarios for a set of real LV load readings, in order to find the difference in design solutions when LV load readings were and were not known. The load profiles of the substations were examined in order to explore the utilisation profile. It was found that in 70% of the scenarios explored, significant cost differences were found. These cost differences varied by an average of 1000%, between schemes designed with and without load readings. Obviously, over designing a system and therefore operating more, underutilised transformers becomes less financially beneficial and less energy efficient. The paper concludes that new connection design is improved in terms of cost when carried out based on known LV load information and enhances the case for regular maximum feeder demand information and/or metering of LV feeders. © 2013 IEEE.

Receptor activity-modifying proteins; multifunctional G protein-coupled receptor accessory proteins

Receptor activity-modifying proteins (RAMPs) are single pass membrane proteins initially identified by their ability to determine the pharmacology of the calcitonin receptor-like receptor (CLR), a family B G protein-coupled receptor (GPCR). It is now known that RAMPs can interact with a much wider range of GPCRs. This review considers recent developments on the structure of the complexes formed between the extracellular domains (ECDs) of CLR and RAMP1 or RAMP2 as these provide insights as to how the RAMPs direct ligand binding. The range of RAMP interactions is also considered; RAMPs can interact with numerous family B GPCRs as well as examples of family A and family C GPCRs. They influence receptor expression at the cell surface, trafficking, ligand binding and G protein coupling. The GPCR-RAMP interface offers opportunities for drug targeting, illustrated by examples of drugs developed for migraine.

Receptor activity-modifying proteins 2 and 3 generate adrenomedullin receptor subtypes with distinct molecular properties

Adrenomedullin (AM) is a peptide hormone with numerous effects in the vascular systems. AM signals through the AM1 and AM2 receptors formed by the obligate heterodimerization of a G protein-coupled receptor, the calcitonin receptor-like receptor (CLR), and receptor activity-modifying proteins (RAMP) 2 and 3, respectively. These different CLR-RAMP interactions yield discrete receptor pharmacology and physiological effects. The effective design of therapeutics that target the individual AM receptors is dependent on understanding the molecular details of the effects of RAMPs on CLR. To understand the role of RAMPs 2 and 3 on the activation and conformation of the CLR subunit of AM receptors we mutated 68 individual amino acids in the juxtamembrane region of CLR, a key region for activation of AM receptors and determined the effects on cAMP signalling. Sixteen CLR mutations had differential effects between the AM1 and AM2 receptors. Accompanying this, independent molecular modelling of the full-length AM-bound AM1 and AM2 receptors predicted differences in the binding pocket, and differences in the electrostatic potential of the two AM receptors. Druggability analysis indicated unique features that could be used to develop selective small molecule ligands for each receptor. The interaction of RAMP2 or RAMP3 with CLR induces conformational variation in the juxtamembrane region, yielding distinct binding pockets, probably via an allosteric mechanism. These subtype-specific differences have implications for the design of therapeutics aimed at specific AM receptors and for understanding the mechanisms by which accessory proteins affect G protein-coupled receptor function.

Development of operational performance models for truck-lane restrictions on freeway corridors

Highways are generally designed to serve a mixed traffic flow that consists of passenger cars, trucks, buses, recreational vehicles, etc. The fact that the impacts of these different vehicle types are not uniform creates problems in highway operations and safety. A common approach to reducing the impacts of truck traffic on freeways has been to restrict trucks to certain lane(s) to minimize the interaction between trucks and other vehicles and to compensate for their differences in operational characteristics. ^ The performance of different truck lane restriction alternatives differs under different traffic and geometric conditions. Thus, a good estimate of the operational performance of different truck lane restriction alternatives under prevailing conditions is needed to help make informed decisions on truck lane restriction alternatives. This study develops operational performance models that can be applied to help identify the most operationally efficient truck lane restriction alternative on a freeway under prevailing conditions. The operational performance measures examined in this study include average speed, throughput, speed difference, and lane changes. Prevailing conditions include number of lanes, interchange density, free-flow speeds, volumes, truck percentages, and ramp volumes. ^ Recognizing the difficulty of collecting sufficient data for an empirical modeling procedure that involves a high number of variables, the simulation approach was used to estimate the performance values for various truck lane restriction alternatives under various scenarios. Both the CORSIM and VISSIM simulation models were examined for their ability to model truck lane restrictions. Due to a major problem found in the CORSIM model for truck lane modeling, the VISSIM model was adopted as the simulator for this study. ^ The VISSIM model was calibrated mainly to replicate the capacity given in the 2000 Highway Capacity Manual (HCM) for various free-flow speeds under the ideal basic freeway section conditions. Non-linear regression models for average speed, throughput, average number of lane changes, and speed difference between the lane groups were developed. Based on the performance models developed, a simple decision procedure was recommended to select the desired truck lane restriction alternative for prevailing conditions. ^

A theory of constraints service systems improvement method: Case of the airline turnaround problem

This dissertation develops a process improvement method for service operations based on the Theory of Constraints (TOC), a management philosophy that has been shown to be effective in manufacturing for decreasing WIP and improving throughput. While TOC has enjoyed much attention and success in the manufacturing arena, its application to services in general has been limited. The contribution to industry and knowledge is a method for improving global performance measures based on TOC principles. The method proposed in this dissertation will be tested using discrete event simulation based on the scenario of the service factory of airline turnaround operations. To evaluate the method, a simulation model of aircraft turn operations of a U.S. based carrier was made and validated using actual data from airline operations. The model was then adjusted to reflect an application of the Theory of Constraints for determining how to deploy the scarce resource of ramp workers. The results indicate that, given slight modifications to TOC terminology and the development of a method for constraint identification, the Theory of Constraints can be applied with success to services. Bottlenecks in services must be defined as those processes for which the process rates and amount of work remaining are such that completing the process will not be possible without an increase in the process rate. The bottleneck ratio is used to determine to what degree a process is a constraint. Simulation results also suggest that redefining performance measures to reflect a global business perspective of reducing costs related to specific flights versus the operational local optimum approach of turning all aircraft quickly results in significant savings to the company. Savings to the annual operating costs of the airline were simulated to equal 30% of possible current expenses for misconnecting passengers with a modest increase in utilization of the workers through a more efficient heuristic of deploying them to the highest priority tasks. This dissertation contributes to the literature on service operations by describing a dynamic, adaptive dispatch approach to manage service factory operations similar to airline turnaround operations using the management philosophy of the Theory of Constraints.

Long-Term Effects of Adding Nutrients to an Oligotrophic Coastal Environment

Management of ecological disturbances requires an understanding of the time scale and dynamics of community responses to disturbance events. To characterize long-term seagrass bed responses to nutrient enrichment, we established six study sites in Florida Bay, USA. In 24 plots (0.25 m2) at each site, we regularly added nitrogen (N) and phosphorus (P) in a factorial design for 7 years. Five of the six sites exhibited strong P limitation. Over the first 2 years, P enrichment increased Thalassia testudinum cover in the three most P-limited sites. After 3 years, Halodule wrightii began to colonize many of the P-addition plots, but the degree of colonization was variable among sites, possibly due to differences in the supply of viable propagules. Thalassia increased its allocation to aboveground tissue in response to P enrichment; Halodule increased in total biomass but did not appear to change its aboveground: belowground tissue allocation. Nutrient enrichment did not cause macroalgal or epiphytic overgrowth of the seagrass. Nitrogen retention in the study plots was variable but relatively low, whereas phosphorus retention was very high, often exceeding 100% of the P added as fertilizer over the course of our experiments. Phosphorus retentions exceeding 100% may have been facilitated by increases in Thalassia aboveground biomass, which promoted the settlement of suspended particulate matter containing phosphorus. Our study demonstrated that lowintensity press disturbance events such as phosphorus enrichment can initiate a slow, ramped successional process that may alter community structure over many years.

Developing a high density platinum/alumina hermetic feedthrough

Typically, hermetic feedthroughs for implantable devices, such as pacemakers, use a alumina ceramic insulator brazed to a platinum wire pin. This combination of material has a long history in implantable devices and has been approved by the FDA for implantable hermetic feedthroughs. The growing demand for increased input/output (I/O) hermetic feedthroughs for implantable neural stimulator applications could be addressed by developing a new, cofired platinum/alumina multilayer ceramic technology in a configuration that supports 300 plus I/Os, which is not commercially available. Seven platinum powders with different particle sizes were used to develop different conductive cofire inks to control the densification mismatch between platinum and alumina. Firing profile (ramp rate, burn- out and holding times) and firing atmosphere and concentrations (hydrogen (wet/dry), air, neutral, vacuum) were also optimized. Platinum and alumina exhibit the alloy formation reaction in a reduced atmosphere. Formation of any compound can increase the bonding of the metal/ceramic interface, resulting in enhanced hermeticity. The feedthrough fabricated in a reduced atmosphere demonstrated significantly superior performance than that of other atmospheres. A composite structure of tungsten/platinum ratios graded thru the via structure (pure W, 50/50 W/Pt, 80/20 Pt/W and pure Pt) exhibited the best performance in comparison to the performance of other materials used for ink metallization. Studies on the high temperature reaction of platinum and alumina, previously unreported, showed that, at low temperatures in reduced atmosphere, Pt 3Al or Pt8Al21 with a tetragonal structure would be formed. Cubic Pt3Al is formed upon heating the sample to temperatures above 1350 °C. This cubic structure is the equilibrium state of Pt-Al alloy at high temperatures. The alumina dissolves into the platinum ink and is redeposited as a surface coating. This was observed on both cofired samples and pure platinum thin films coated on a 99.6 Wt% alumina and fired at 1550 °C. Different mechanisms are proposed to describe this behavior based on the size of the platinum particle

Hospitality IT: What Does the Future Hold?

The impact of information technology (IT) is far-reaching and driving dramatic shifts in business paradigms. Trends suggest greater adoption of IT will continue and develop at accelerating rates. Hence, hotel operators and executives must learn how to embrace IT and capitalize on the many capabilities it has to offer while minimizing the threats. The authors attempt to provide a sense of focus and a roadmap to help hoteliers understand the issues, see the future, and find an appropriate on ramp to the information superhighway.

Uso da cinza do bagaço da cana-de-açúcar como matéria-prima na substituição dos fundentes para aplicações em porcelanato

The present study aims to evaluate the potential use of bagasse ash from sugar cane (CBC) as a flux, replacing phyllite and/or feldspar in standard industrial mass production of enameled porcelain, verifying the possibility of the CBC contribute to the overall reduction of the coefficient of thermal expansion of the ceramic mass. To this end, as a result of the research, we characterized the raw material components of the standard mass (clay, phyllite, kaolin, feldspar, quartz and talc) and the residue of BCC, by testing by XRF, XRD, AG, DTA and ATG. Specimens (CDP) were manufactured in the dimensions of 100 mm x 50 mm x 8 mm in uniaxial matrix under compaction pressure of 33 MPa, assembled in batches of 3 units subsequently sintered at temperatures of 1150°C to 1210°C by varying the Rating Scale at 10°C, heating and cooling ramp of 50°C/min and 25°C/min, with levels of 1 min, 3 min, 5 min, 8 min, 10 min, 15 min, 30 min and 60 min. analyzing the results of the physical properties of water absorption (WA), linear firing shrinkage (LFS), dilatometric analysis (DTA), flexural strain (SFT) and SEM of the sintered bodies in order to verify the adequacy of CDP to ISO 13006, ISO 10545, NBR 13816 standards; NBR 13817 and NBR 13818. The study showed that the formulations that best suit the requirements of the standards are:. G4 - which was applied in 10% of replacing the CBC phyllite, sintering temperature 1210 ° C for 10 min and porch, and F3 - with application of 7.5% of CBC to replace the feldspar in the sintering temperatures of 1190°C, 1200°C and 1210°C for 10 min and porch. These formulations showed better performance regarding the formation of primary and secondary mullite, with considerable reduction of cracks and pores, meeting the prerequisites of standards for glazed porcelain. The results shows that the use of the CBC as a flux in the preparation of porcelain mass meets standard parameters for the manufacture of the product, and thereby can reduce environmental impact and the cost of production. Therefore, it is recommended to use this residue in the ceramics industry, due to its industrial, commercial and collaborative viability for sustainability.

Approche biophysique à l'étude des interactions allostériques entre les récepteurs et les protéines G

Mémoire numérisé par la Direction des bibliothèques de l'Université de Montréal.

An investigation into the tension arising between natural language and mathematical language experienced by mechanical engineering students

This investigation is grounded within the concept of embodied cognition where the mind is considered to be part of a biological system. A first year undergraduate Mechanical Engineering cohort of students was tasked with explaining the behaviour of three balls of different masses being rolled down a ramp. The explanations given by the students highlighted the cognitive conflict between the everyday interpretation of the word energy and its mathematical use. The results showed that even after many years of schooling, students found it challenging to interpret the mathematics they had learned and relied upon pseudo-scientific notions to account for the behaviour of the balls.