Ligand- and cell-dependent determinants of internalization and cAMP modulation by delta opioid receptor (DOR) agonists

Signaling bias refers to G protein-coupled receptor ligand ability to preferentially activate one type of signal over another. Bias to evoke signaling as opposed to sequestration has been proposed as a predictor of opioid ligand potential for generating tolerance. Here we measured whether delta opioid receptor agonists preferentially inhibited cyclase activity over internalization in HEK cells. Efficacy (τ) and affinity (KA) values were estimated from functional data and bias was calculated from efficiency coefficients (log τ/KA). This approach better represented the data as compared to alternative methods that estimate bias exclusively from τ values. Log (τ/KA) coefficients indicated that SNC-80 and UFP-512 promoted cyclase inhibition more efficiently than DOR internalization as compared to DPDPE (bias factor for SNC-80: 50 and for UFP-512: 132). Molecular determinants of internalization were different in HEK293 cells and neurons with βarrs contributing to internalization in both cell types, while PKC and GRK2 activities were only involved in neurons. Rank orders of ligand ability to engage different internalization mechanisms in neurons were compared to rank order of E _max values for cyclase assays in HEK cells. Comparison revealed a significant reversal in rank order for cyclase E _max values and βarr-dependent internalization in neurons, indicating that these responses were ligand-specific. Despite this evidence, and because kinases involved in internalization were not the same across cellular backgrounds, it is not possible to assert if the magnitude and nature of bias revealed by rank orders of maximal responses is the same as the one measured in HEK cells.     

A multiperspective systems-based framework for decision support systems design

This paper reviews the dominant approaches to the design of decision support systems with reference to recent developments in information technology. The complexities and difficulties of the various developments are also discussed and their implications explored. A multiperspective, systems-based framework is presented which aims to lessen these difficulties. The framework is based on the theory and concepts of critical systems thinking.     

Warm tropical ocean surface and global anoxia during the mid-Cretaceous period

The middle of the Cretaceous period (about 120 to 80 Myr ago) was a time of unusually warm polar temperatures, repeated reef-drowning in the tropics and a series of oceanic anoxic events (OAEs) that promoted both the widespread deposition of organic-carbon-rich marine sediments and high biological turnover. The cause of the warm temperatures is unproven but widely attributed to high levels of atmospheric greenhouse gases such as carbon dioxide. In contrast, there is no consensus on the climatic causes and effects of the OAEs, with both high biological productivity and ocean 'stagnation' being invoked as the cause of ocean anoxia. Here we show, using stable isotope records from multiple species of well-preserved foraminifera, that the thermal structure of surface waters in the western tropical Atlantic Ocean underwent pronounced variability about 100 Myr ago, with maximum sea surface temperatures 3-5 degrees C warmer than today. This variability culminated in a collapse of upper-ocean stratification during OAE-1d (the 'Breistroffer' event), a globally significant period of organic-carbon burial that we show to have fundamental, stratigraphically valuable, geochemical similarities to the main OAEs of the Mesozoic era. Our records are consistent with greenhouse forcing being responsible for the warm temperatures, but are inconsistent both with explanations for OAEs based on ocean stagnation, and with the traditional view (reviewed in ref. 12) that past warm periods were more stable than today's climate.     

Doping semiconductor nanocrystals

Doping--the intentional introduction of impurities into a material--is fundamental to controlling the properties of bulk semiconductors. This has stimulated similar efforts to dope semiconductor nanocrystals. Despite some successes, many of these efforts have failed, for reasons that remain unclear. For example, Mn can be incorporated into nanocrystals of CdS and ZnSe (refs 7-9), but not into CdSe (ref. 12)--despite comparable bulk solubilities of near 50 per cent. These difficulties, which have hindered development of new nanocrystalline materials, are often attributed to 'self-purification', an allegedly intrinsic mechanism whereby impurities are expelled. Here we show instead that the underlying mechanism that controls doping is the initial adsorption of impurities on the nanocrystal surface during growth. We find that adsorption--and therefore doping efficiency--is determined by three main factors: surface morphology, nanocrystal shape, and surfactants in the growth solution. Calculated Mn adsorption energies and equilibrium shapes for several nanocrystals lead to specific doping predictions. These are confirmed by measuring how the Mn concentration in ZnSe varies with nanocrystal size and shape. Finally, we use our predictions to incorporate Mn into previously undopable CdSe nanocrystals. This success establishes that earlier difficulties with doping are not intrinsic, and suggests that a variety of doped nanocrystals--for applications from solar cells to spintronics--can be anticipated.     

Oxidation state of the active-site cysteine in protein tyrosine phosphatase 1B

Protein tyrosine phosphatases regulate signal transduction pathways involving tyrosine phosphorylation and have been implicated in the development of cancer, diabetes, rheumatoid arthritis and hypertension. Increasing evidence suggests that the cellular redox state is involved in regulating tyrosine phosphatase activity through the reversible oxidization of the catalytic cysteine to sulphenic acid (Cys-SOH). But how further oxidation to the irreversible sulphinic (Cys-SO2H) and sulphonic (Cys-SO3H) forms is prevented remains unclear. Here we report the crystal structures of the regulatory sulphenic and irreversible sulphinic and sulphonic acids of protein tyrosine phosphatase 1B (PTP1B), an important enzyme in the negative regulation of the insulin receptor and a therapeutic target in type II diabetes and obesity. We also identify a sulphenyl-amide species that is formed through oxidation of its catalytic cysteine. Formation of the sulphenyl-amide causes large changes in the PTP1B active site, which are reversible by reduction with the cellular reducing agent glutathione. The sulphenyl-amide is a protective intermediate in the oxidative inhibition of PTP1B. In addition, it may facilitate reactivation of PTP1B by biological thiols and signal a unique state of the protein.