5-HT2A receptor signalling through phospholipase D1 associated with its C-terminal tail

The 5-HT2AR (5-hydroxytryptamine-2A receptor) is a GPCR (G-protein-coupled receptor) that is implicated in the actions of hallucinogens and represents a major target of atypical antipsychotic agents. In addition to its classical signalling though PLC (phospholipase C), the receptor can activate several other pathways, including ARF (ADP-ribosylation factor)-dependent activation of PLD (phospholipase D), which appears to be achieved through a mechanism independent of heterotrimeric G-proteins. In the present study we show that wild-type and inactive constructs of PLD1 (but not PLD2) respectively facilitate and inhibit ARF-dependent PLD signalling by the 5-HT2AR. Furthermore we demonstrate that PLD1 specifically co-immunoprecipitates with the receptor and binds to a distal site in GST (glutathione transferase) fusion protein constructs of its C-terminal tail which is distinct from the ARF-interaction site, thereby suggesting the existence of a functional ARF-PLD signalling complex directly associated with this receptor. This reveals the spatial co-ordination of an important GPCR, transducer and effector into a physical complex that is likely to reinforce the impact of receptor activation on a heterotrimeric G-protein-independent signalling pathway. Signalling of this receptor through such non-canonical pathways may be important to its role in particular disorders

Phospholipase D inhibitors reduce human prostate cancer cell proliferation and colony formation

BACKGROUND: Phospholipases D1 and D2 (PLD1/2) hydrolyse cell membrane glycerophospholipids to generate phosphatidic acid, a signalling lipid, which regulates cell growth and cancer progression through effects on mTOR and PKB/Akt. PLD expression and/or activity is raised in breast, colorectal, gastric, kidney and thyroid carcinomas but its role in prostate cancer (PCa), the major cancer of men in the western world, is unclear. METHODS: PLD1 protein expression in cultured PNT2C2, PNT1A, P4E6, LNCaP, PC3, PC3M, VCaP, 22RV1 cell lines and patient-derived PCa cells was analysed by western blotting. PLD1 protein localisation in normal, benign prostatic hyperplasia (BPH), and castrate-resistant prostate cancer (CRPC) tissue sections and in a PCa tissue microarray (TMA) was examined by immunohistochemistry. PLD activity in PCa tissue was assayed using an Amplex Red method. The effect of PLD inhibitors on PCa cell viability was measured using MTS and colony forming assays. RESULTS: PLD1 protein expression was low in the luminal prostate cell lines (LNCaP, VCaP, 22RV1) compared with basal lines (PC3 and PC3M). PLD1 protein expression was elevated in BPH biopsy tissue relative to normal and PCa samples. In normal and BPH tissue, PLD1 was predominantly detected in basal cells as well in some stromal cells, rather than in luminal cells. In PCa tissue, luminal cells expressed PLD1. In a PCa TMA, the mean peroxidase intensity per DAB-stained Gleason 6 and 7 tissue section was significantly higher than in sections graded Gleason 9. In CRPC tissue, PLD1 was expressed prominently in the stromal compartment, in luminal cells in occasional glands and in an expanding population of cells that co-expressed chromogranin A and neurone-specific enolase. Levels of PLD activity in normal and PCa tissue samples were similar. A specific PLD1 inhibitor markedly reduced the survival of both prostate cell lines and patient-derived PCa cells compared with two dual PLD1/PLD2 inhibitors. Short-term exposure of PCa cells to the same specific PLD1 inhibitor significantly reduced colony formation. CONCLUSIONS: A new specific inhibitor of PLD1, which is well tolerated in mice, reduces PCa cell survival and thus has potential as a novel therapeutic agent to reduce prostate cancer progression. Increased PLD1 expression may contribute to the hyperplasia characteristic of BPH and in the progression of castrate-resistant PCa, where an expanding population of neuroendocrine-like cells express PLD1.British Journal of Cancer advance online publication, 14 November 2017; doi:10.1038/bjc.2017.391 www.bjcancer.com

Interfaces rodoviário-urbanas no processo de produção das cidades: estudo de caso do contorno rodoviário de João Pessoa, PB, Brasil

As rodovias desempenham função importante no processo de estruturação das cidades, por facilitar os deslocamentos interurbanos. No entanto, quando absorvidas pela malha urbana de uma cidade, essas rodovias desenvolvem dinâmicas particulares com o espaço intraurbano no qual estão inseridas, com alterações na acessibilidade, na morfologia e no uso e ocupação da terra urbana. Com base nesse contexto, este artigo tem por objetivo analisar as dinâmicas entre a implantação do contorno rodoviário da BR-230 e o processo de produção do espaço intraurbano da cidade de João Pessoa-PB, entre os anos de 1963 e 2013, em termos de morfologia, uso e ocupação do solo urbano. O método empregado foi o da Lógica Evolutiva do Tecido Urbano, que divide a evolução urbana em três estágios: superação de limites, crescimento e combinação/conflitos. O método foi associado a ferramentas analíticas, como a Sintaxe Espacial e softwares de Sistema de Informação Geográfica. Os resultados mostram que a rodovia impulsionou o espraiamento da cidade para o setor sul e produziu uma malha urbana tentacular, onde também se identificou o padrão de crescimento horizontal e fragmentado, alterações na acessibilidade, uso do solo e na segregação socioespacial

Il collo di Lee Miller

Traduzione poetica dal francese di "Il collo di Lee Miller", di Nicole Brossar

Identifying electric vehicle charging styles among consumers: a latent class cluster analysis

The market share of electric vehicles (EVs) is growing rapidly, making it crucial to understand the charging behaviour of current and prospective users. Such understanding is essential for designing policies that positively influence consumers’ charging behaviour and facilitate EV adoption. In this study, we examined the heterogeneity in charging preferences of 994 respondents across Australia using a latent class cluster model that considers indicators of charging behaviour as outcomes of interest. We used sociodemographic characteristics, travel needs, and EV adoption status as covariates to predict class membership. Our findings indicate five segments of consumers with distinct charging preferences: routine-focused frugals, cost-oriented deliberators, range seekers, flexibility seekers, and indifferent late adopters. These segments differ in the importance they attach to charging attributes, their coping strategies with limited battery resources, and their risk attitude. Our results suggest that a uniform approach to EV-related policies is not appropriate, as each consumer segment has unique charging preferences and requirements. Furthermore, the study emphasizes the significance of accounting for charging behaviour heterogeneity in demand modelling, as assumptions in current models may not accurately represent the decision-making of most segments

Public transportation-based crowd-shipping initiatives: Are users willing to participate? Why not?

An emerging stream of Crowd-Shipping (CS) solutions focuses on existing momentum in Public Transportation (PT) to ship viable delivery packages by PT passengers. Few studies have explored the package delivery acceptance behavior of passengers engaged in PT-based CS initiatives while passengers’ behavioral intention to participate (i.e., engage) is not studied. It is requisite that newly introduced CS platforms explore their potential crowdshippers’ behavior on intention to participate and set efficient marketing strategies. Given survey data collected from 2208 PT passengers in Sydney metropolitan area, this study explores the intention of PT passengers as crowd-shippers to participate in PT-based CS initiatives, as well as prohibiting factors in way of participation. Accordingly, a binominal logit model is developed whereby the variables impacting the intention to participate are identified. Then, using an inductive thematic analysis, 917 reasons (text responses) for not participating are scrutinized, and the prohibiting factors are identified and categorized. Considering demographic and socio-economic characteristics of the respondents, the study reveals to what degree passengers with different characteristics are sensitive to prohibiting factors. This research provides several practical insights that can assist in successfully defining, launching, and advertising a new PT-based CS initiative. As a key finding, it is observed that women, full-time employees, elderly, retirees, and low-income PT passengers hardly participate, while the youth, individuals with a positive attitude towards sustainable freight initiatives, and those who experienced working with parcel lockers would participate with a higher probability. Moreover, it is observed that factors relating to time availability/flexibility and physical health condition/importance of passengers are much more important than the compensation level for passengers to accept to participate in PT-based CS initiatives

Modelling interest in co-adoption of electric vehicles and solar photovoltaics in Australia to identify tailored policy needs

Electric vehicles (EVs) and solar photovoltaic systems (PVs) are two technologies that are gaining popularity in households as a means of reducing carbon emissions and improving energy security. However, little is known about the characteristics of households that adopt these technologies jointly. This study investigates the adoption patterns of electric vehicles and solar photovoltaics in Australia. We explain the likelihood of consumers belonging to four distinct groups (those who adopt both PVs and EVs, those who only adopt EVs, those who only adopt PVs, and those who adopt none) based on demographic and attitudinal factors. Using survey data from a representative sample of 2219 Australian heads of households, we found that dwelling ownership, ownership of a home energy management system, gender, and household size were significant predictors of the joint adoption of EVs and PVs. While both pro-environmental and pro-technology attitudes demonstrated a significant role in shaping PV-EV co-adoption patterns, the latter has a much stronger effect than the former. Based on the results, we identified that actions are needed in three key areas to encourage co-adoption: reducing technology adoption constraints associated with living arrangements (such as dwelling type and ownership), providing bundled financial incentives for both technologies, and fostering technology awareness and perceived usefulness among consumers