NAADP regulates human platelet function.

Platelets play a vital role in maintaining haemostasis. Human platelet activation depends on Ca2+ release, leading to cell activation, granule secretion and aggregation. NAADP (nicotinic acid-adenine dinucleotide phosphate) is a Ca2+-releasing second messenger that acts on acidic Ca2+ stores and is used by a number of mammalian systems. In human platelets, NAADP has been shown to release Ca2+ in permeabilized human platelets and contribute to thrombin-mediated platelet activation. In the present study, we have further characterized NAADP-mediated Ca2+ release in human platelets in response to both thrombin and the GPVI (glycoprotein VI)-specific agonist CRP (collagen-related peptide). Using a radioligand-binding assay, we reveal an NAADP-binding site in human platelets, indicative of a platelet NAADP receptor. We also found that NAADP releases loaded 45Ca2+ from intracellular stores and that total platelet Ca2+ release is inhibited by the proton ionophore nigericin. Ned-19, a novel cell-permeant NAADP receptor antagonist, competes for the NAADP-binding site in platelets and can inhibit both thrombin- and CRP-induced Ca2+ release in human platelets. Ned-19 has an inhibitory effect on platelet aggregation, secretion and spreading. In addition, Ned-19 extends the clotting time in whole-blood samples. We conclude that NAADP plays an important role in human platelet function. Furthermore, the development of Ned-19 as an NAADP receptor antagonist provides a potential avenue for platelet-targeted therapy and the regulation of thrombosis.

Tangeretin regulates platelet function through inhibition of phosphoinositide 3-Kinase and cyclic nucleotide signaling

OBJECTIVE: Dietary flavonoids have long been appreciated in reducing cardiovascular disease risk factors, but their mechanisms of action are complex in nature. In this study, the effects of tangeretin, a dietary flavonoid, were explored on platelet function, signaling, and hemostasis. APPROACH AND RESULTS: Tangeretin inhibited agonist-induced human platelet activation in a concentration-dependent manner. It inhibited agonist-induced integrin αIIbβ3 inside-out and outside-in signaling, intracellular calcium mobilization, and granule secretion. Tangeretin also inhibited human platelet adhesion and subsequent thrombus formation on collagen-coated surfaces under arterial flow conditions in vitro and reduced hemostasis in mice. Further characterization to explore the mechanism by which tangeretin inhibits platelet function revealed distinctive effects of platelet signaling. Tangeretin was found to inhibit phosphoinositide 3-kinase-mediated signaling and increase cGMP levels in platelets, although phosphodiesterase activity was unaffected. Consistent with increased cGMP levels, tangeretin increased the phosphorylation of vasodilator-stimulated phosphoprotein at S239. CONCLUSIONS: This study provides support for the ability and mechanisms of action of dietary flavonoids to modulate platelet signaling and function, which may affect the risk of thrombotic disease.

A role for the thiol isomerase. protein ERP5 in platelet function

Formation and rearrangement of disulfide bonds during the correct folding of nascent proteins is modulated by a family of enzymes known as thiol isomerases, which include protein disulfide isomerase (PDI), endoplasmic reticulum protein 5 (ERP5), and ERP57. Recent evidence supports an alternative role for this family of proteins on the surface of cells, where they are involved in receptor 'remodeling and recognition. In platelets, blocking PDI with inhibitory antibodies inhibits a number of platelet activation pathways, including aggregation, secretion, and fibrinogen binding. Analysis of human platelet membrane fractions identified the presence of the thiol isomerase protein ERP5. Further study showed that ERP5 is resident mainly on platelet intracellular membranes, although it is rapidly recruited to the cell, surface in response to a range of platelet agonists. Blocking cell-surface ERP5 using inhibitory antibodies leads to a decrease in platelet aggregation in response to agonists, and a decrease in fibrinogen binding and P-selectin exposure. It is Possible that this is based on the disruption of integrin function, as we observed that ERP5 becomes physically associated with the integrin beta(3) subunit during platelet stimulation. These results provide new insights into the involvement of thiol isomerases and regulation of platelet activation. (C) 2005 by The American Society of Hematology.

Transcription profiling in human platelets reveals LRRFIP1 as a novel protein regulating platelet function

Within the healthy population, there is substantial, heritable, and interindividual variability in the platelet response. We explored whether a proportion of this variability could be accounted for by interindividual variation in gene expression. Through a correlative analysis of genome-wide platelet RNA expression data from 37 subjects representing the normal range of platelet responsiveness within a cohort of 500 subjects, we identified 63 genes in which transcript levels correlated with variation in the platelet response to adenosine diphosphate and/or the collagen-mimetic peptide, cross-linked collagen-related peptide. Many of these encode proteins with no reported function in platelets. An association study of 6 of the 63 genes in 4235 cases and 6379 controls showed a putative association with myocardial infarction for COMMD7 (COMM domain-containing protein 7) and a major deviation from the null hypo thesis for LRRFIP1 [leucine-rich repeat (in FLII) interacting protein 1]. Morpholino-based silencing in Danio rerio identified a modest role for commd7 and a significant effect for lrrfip1 as positive regulators of thrombus formation. Proteomic analysis of human platelet LRRFIP1-interacting proteins indicated that LRRFIP1 functions as a component of the platelet cytoskeleton, where it interacts with the actin-remodeling proteins Flightless-1 and Drebrin. Taken together, these data reveal novel proteins regulating the platelet response.

Proteinase-activated receptors, targets for kallikrein signaling

Serine proteinases like thrombin can signal to cells by the cleavage/activation of proteinase-activated receptors (PARs). Although thrombin is a recognized physiological activator of PAR(1) and PAR(4), the endogenous enzymes responsible for activating PAR(2) in settings other than the gastrointestinal system, where trypsin can activate PAR(2), are unknown. We tested the hypothesis that the human tissue kallikrein (hK) family of proteinases regulates PAR signaling by using the following: 1) a high pressure liquid chromatography (HPLC)-mass spectral analysis of the cleavage products yielded upon incubation of hK5, -6, and -14 with synthetic PAR N-terminal peptide sequences representing the cleavage/activation motifs of PAR(1), PAR(2), and PAR(4); 2) PAR-dependent calcium signaling responses in cells expressing PAR(1), PAR(2), and PAR(4) and in human platelets; 3) a vascular ring vasorelaxation assay; and 4) a PAR(4)-dependent rat and human platelet aggregation assay. We found that hK5, -6, and -14 all yielded PAR peptide cleavage sequences consistent with either receptor activation or inactivation/disarming. Furthermore, hK14 was able to activate PAR(1), PAR(2), and PAR(4) and to disarm/inhibit PAR(1). Although hK5 and -6 were also able to activate PAR(2), they failed to cause PAR(4)-dependent aggregation of rat and human platelets, although hK14 did. Furthermore, the relative potencies and maximum effects of hK14 and -6 to activate PAR(2)-mediated calcium signaling differed. Our data indicate that in physiological settings, hKs may represent important endogenous regulators of the PARs and that different hKs can have differential actions on PAR(1), PAR(2), and PAR(4).

Human geographies of climate change: landscape, temporality and lay knowledges.

In this paper we bring together work on landscape, temporality and lay knowledges to propose new ways of understanding climate change. A focus on the familiar landscapes of everyday life offers an opportunity to examine how climate change could be researched as a relational phenomenon, understood on a local level, with distinctive spatialities and temporalities. Climate change can be observed in relation to landscape but also felt, sensed, apprehended emotionally as part of the fabric of everyday life in which acceptance, denial, resignation and action co-exist as personal and social responses to the local manifestations of a global problem.

Future directions for doll play narrative research: a commentary

Past research into doll play narratives has been productive in elucidating children's inner experiences, their determinants, and their role in child behaviour problems. The current volume takes this work forward in several directions: first, it indicates the value of designing story stems and coding schemes to address more specific questions about the developmental process of specific syndromes. Second, contributions demonstrate the "added value" provided by children's narratives, over and above information derived from other sources. Third, this recent research enhances our understanding of the role of parental representations and states of mind in influencing children's narratives; how these may come to influence child functioning via co-constructed parent-child dialogues is an important area for future research. Finally, possibilities of extending the clinical utility of doll play narratives are explored.

The "Fair trade nation": market-oriented development in devolved European regions

Scholars have largely ignored the roles played by government and public sector institutions in the fair trade movement. This article addresses the knowledge gap through examining government involvement in fair trade networks in the context of European devolution and the localization of international development action. Proposing a relational view of fair trade networks, and considering the Fair Trade Nation as a social category for development, it highlights how power sources outside the centralized nation-state permit a political community to associate itself with fair trade. Research from Wales demonstrates that government acts in a leadership role rather than as regulator, conferring political voice and finance while enhancing its international credentials and contributing to the politics of nation-building. Our conclusion is cautious; campaigners celebrate political commitment to fair trade embodied within the category of the Fair Trade Nation, but evidence suggests that government reliance on the market as a vehicle for decentralized development action is limited by how the Fair Trade Nation is currently executed.

Motivations behind farmers’ pesticide use in Bangladesh rice farming

This paper addresses the motivations behind farmers’ pesticide use in two regions of Bangladesh. The paper considers farmers’ knowledge of arthropods and their perceptions about pests and pest damage, and identifies why many farmers do not use recommended pest management practices. We propose that using the novel approach of classifying farmers according to their motivations and constraints rather than observed pesticide use can improve training approaches and increase farmers’ uptake and retention of more appropriate integrated pest management technologies.

The neolithisation of Liguria (NW Italy): an environmental archaeological and palaeoenvironmental perspective

The archaeological evidence compiled for Liguria has enabled the formulation of a comprehensive model of Neolithic social, technological and economic development (∼7800–5700 cal yrs BP). The model indicates that during the Early and Middle Neolithic (∼7800–6300 cal yrs BP; ‘Impressed Ware’ and ‘Square Mouthed’ pottery cultures) human activity mainly focussed on low (coastal) and mid-altitude areas. By the Late Neolithic (∼6300–5700 cal yrs BP; ‘Chassey’ culture) farming practices were taking place over a wider range of altitudes and involved transhumant pastoralism. Complementary environmental archaeological and palaeoecological records from caves, open-air sites, lakes and mires indicate that human activities had a more significant impact on the environment than previously thought. This included clearance, especially Abies, Ulmus, Fraxinus and Tilia, and woodland utilisation and management (e.g. leaf foddering), as well as cereal cultivation and animal husbandry. The influence of Middle Holocene climatic changes, especially from ∼7800 cal yrs BP, on the direction of vegetation changes and socio-economic developments during the Neolithic remain uncertain.