Nuclear ARRB1 induces pseudohypoxia and cellular metabolism reprogramming in prostate cancer

Tumour cells sustain their high proliferation rate through metabolic reprogramming, whereby cellular metabolism shifts from oxidative phosphorylation to aerobic glycolysis, even under normal oxygen levels. Hypoxia-inducible factor 1A (HIF1A) is a major regulator of this process, but its activation under normoxic conditions, termed pseudohypoxia, is not well documented. Here, using an integrative approach combining the first genome-wide mapping of chromatin binding for an endocytic adaptor, ARRB1, both in vitro and in vivo with gene expression profiling, we demonstrate that nuclear ARRB1 contributes to this metabolic shift in prostate cancer cells via regulation of HIF1A transcriptional activity under normoxic conditions through regulation of succinate dehydrogenase A (SDHA) and fumarate hydratase (FH) expression. ARRB1-induced pseudohypoxia may facilitate adaptation of cancer cells to growth in the harsh conditions that are frequently encountered within solid tumours. Our study is the first example of an endocytic adaptor protein regulating metabolic pathways. It implicates ARRB1 as a potential tumour promoter in prostate cancer and highlights the importance of metabolic alterations in prostate cancer.

Thiol isomerases negatively regulate the cellular shedding activity of ADAM17

ADAM17 (where ADAM is 'a disintegrin and metalloproteinase') can rapidly modulate cell-surface signalling events by the proteolytic release of soluble forms of proligands for cellular receptors. Many regulatory pathways affect the ADAM17 sheddase activity, but the mechanisms for the activation are still not clear. We have utilized a cell-based ADAM17 assay to show that thiol isomerases, specifically PDI (protein disulfide isomerase), could be responsible for maintaining ADAM17 in an inactive form. Down-regulation of thiol isomerases, by changes in the redox environment (for instance as elicited by phorbol ester modulation of mitochondrial reactive oxygen species) markedly enhanced ADAM17 activation. On the basis of ELISA binding studies with novel fragment antibodies against ADAM17 we propose that isomerization of the disulfide bonds in ADAM17, and the subsequent conformational changes, form the basis for the modulation of ADAM17 activity. The shuffling of disulfide bond patterns in ADAMs has been suggested by a number of recent adamalysin crystal structures, with distinct disulfide bond patterns altering the relative orientations of the domains. Such a mechanism is rapid and reversible, and the role of thiol isomerases should be investigated further as a potential factor in the redox regulation of ADAM17.

A novel fluorescence-based assay for the rapid detection and quantification of cellular deoxyribonucleoside triphosphates

Current methods for measuring deoxyribonucleoside triphosphates (dNTPs) employ reagent and labor-intensive assays utilizing radioisotopes in DNA polymerase-based assays and/or chromatography-based approaches. We have developed a rapid and sensitive 96-well fluorescence-based assay to quantify cellular dNTPs utilizing a standard real-time PCR thermocycler. This assay relies on the principle that incorporation of a limiting dNTP is required for primer-extension and Taq polymerase-mediated 5-3' exonuclease hydrolysis of a dual-quenched fluorophore-labeled probe resulting in fluorescence. The concentration of limiting dNTP is directly proportional to the fluorescence generated. The assay demonstrated excellent linearity (R(2) > 0.99) and can be modified to detect between ∼0.5 and 100 pmol of dNTP. The limits of detection (LOD) and quantification (LOQ) for all dNTPs were defined as <0.77 and <1.3 pmol, respectively. The intra-assay and inter-assay variation coefficients were determined to be <4.6% and <10%, respectively with an accuracy of 100 ± 15% for all dNTPs. The assay quantified intracellular dNTPs with similar results obtained from a validated LC-MS/MS approach and successfully measured quantitative differences in dNTP pools in human cancer cells treated with inhibitors of thymidylate metabolism. This assay has important application in research that investigates the influence of pathological conditions or pharmacological agents on dNTP biosynthesis and regulation.

A chromatin-independent role of Polycomb-like 1 to stabilize p53 and promote cellular quiescence

Polycomb-like proteins 1-3 (PCL1-3) are substoichiometric components of the Polycomb-repressive complex 2 (PRC2) that are essential for association of the complex with chromatin. However, it remains unclear why three proteins with such apparent functional redundancy exist in mammals. Here we characterize their divergent roles in both positively and negatively regulating cellular proliferation. We show that while PCL2 and PCL3 are E2F-regulated genes expressed in proliferating cells, PCL1 is a p53 target gene predominantly expressed in quiescent cells. Ectopic expression of any PCL protein recruits PRC2 to repress the INK4A gene; however, only PCL2 and PCL3 confer an INK4A-dependent proliferative advantage. Remarkably, PCL1 has evolved a PRC2- and chromatin-independent function to negatively regulate proliferation. We show that PCL1 binds to and stabilizes p53 to induce cellular quiescence. Moreover, depletion of PCL1 phenocopies the defects in maintaining cellular quiescence associated with p53 loss. This newly evolved function is achieved by the binding of the PCL1 N-terminal PHD domain to the C-terminal domain of p53 through two unique serine residues, which were acquired during recent vertebrate evolution. This study illustrates the functional bifurcation of PCL proteins, which act in both a chromatin-dependent and a chromatin-independent manner to regulate the INK4A and p53 pathways.

A Tractable Approach to Uplink Spectral Efficiency of Two-Tier Massive MIMO Cellular HetNets

This letter investigates the uplink spectral efficiency (SE) of a two-tier cellular network, where massive multiple-input multiple-output macro base stations are overlaid with dense small cells. Macro user equipments (MUEs) and small cells with single user equipment uniformly scattered are modeled as two independent homogeneous Poisson point processes. By applying stochastic geometry, we analyze the SE of the multiuser uplink at a macro base station that employs a zero-forcing detector and we obtain a novel lower bound as well as its approximation. According to the simple and near-exact analytical expression, we observe that the ideal way to improve the SE is by increasing the MUE density and the base station antennas synchronously rather than increasing them individually. Furthermore, a large value of path loss exponent has a positive effect on the SE due to the reduced aggregated interference.

Fading Characteristics of Body–to–Body Channels Subject to Vehicular Traffic Conditions at 2.45 GHz

In this paper we investigate the small-scale fading characteristics of body-to-body communications channels in an urban environment at 2.45 GHz. The experiments considered body-to-body channels between devices positioned on two persons on either side of a busy road. The Ricean-K factors estimated from the measurements suggest that a significant dominant component existed in the majority of the channels.

Vehicular Traffic Intersecting Body-to-Body Communications Channels at 2.45 GHz

In this paper we investigate the effects of vehicular traffic on body-to-body (B2B) communications channels in an urban environment at 2.45 GHz. In particular, the impact of differing vehicle types passing in the vicinity of a B2B link are investigated for different body orientations relative to one another at the side of a busy urban street. Initial findings suggest that the average disturbance in a B2B channel can last for 2 seconds and depending on the vehicle size, fades in excess of 40 dB can occur. The body orientations are shown to be a significant factor on the effects of vehicular traffic on the B2B channel.

Effects of defined mixtures of persistent organic pollutants (POPs) on multiple cellular responses in the human hepatocarcinoma cell line, HepG2, using high content analysis screening

Persistent organic pollutants (POPs) are toxic substances, highly resistant to environmental degradation, which can bio-accumulate and have long-range atmospheric transport potential. Most studies focus on single compound effects, however as humans are exposed to several POPs simultaneously, investigating exposure effects of real life POP mixtures on human health is necessary. A defined mixture of POPs was used, where the compound concentration reflected its contribution to the levels seen in Scandinavian human serum (total mix). Several sub mixtures representing different classes of POP were also constructed. The perfluorinated (PFC) mixture contained six perfluorinated compounds, brominated (Br) mixture contained seven brominated compounds, chlorinated (Cl) mixture contained polychlorinated biphenyls and also p,p'-dichlorodiphenyldichloroethylene, hexachlorobenzene, three chlordanes, three hexachlorocyclohexanes and dieldrin. Human hepatocarcinoma (HepG2) cells were used for 2h and 48h exposures to the seven mixtures and analysis on a CellInsight™ NXT High Content Screening platform. Multiple cytotoxic endpoints were investigated: cell number, nuclear intensity and area, mitochondrial mass and membrane potential (MMP) and reactive oxygen species (ROS). Both the Br and Cl mixtures induced ROS production but did not lead to apoptosis. The PFC mixture induced the ROS production and likely induced cell apoptosis accompanied by the dissipation of MMP. Synergistic effects were evident for ROS induction when cells were exposed to the PFC+Br mixture. No significant effects were detected in the Br+Cl, PFC+Cl or total mixtures, which contain the same concentrations of chlorinated compounds as the Cl mixture plus additional compounds; highlighting the need for further exploration of POP mixtures in risk assessment.

The effect of niobium and tantalum on physicochemical and catalytic properties of silver and platinum catalysts based on MCF mesoporous cellular foams

MCF, NbMCF and TaMCF Mesostructured Cellular Foams were used as supports for platinum and silver (1 wt%). Metallic and bimetallic catalysts were prepared by grafting of metal species on APTMS (3-aminopropyltrimethoxysilane) and MPTMS (2-mercaptopropyltrimethoxysilane) functionalized supports. Characterizations by X-ray diffraction (XRD), ultraviolet–visible (UV–Vis) spectroscopy, X-ray photoelectron spectroscopy (XPS), X-ray fluorescence (XRF) spectroscopy, and in situ Fourier Transform Infrared (FTIR) spectroscopy allowed to monitor the oxidation state of metals and surface properties of the catalysts, in particular the formation of bimetallic phases and the strong metal–support interactions. It was evidenced that the functionalization agent (APTMS or MPTMS) influenced the metals dispersion, the type of bimetallic species and Nb/Ta interaction with Pt/Ag. Strong Nb–Ag interaction led to the reduction of niobium in the support and oxidation of silver. MPTMS interacted at first with Pt to form Pt–Ag ensembles highly active in CH3OH oxidation. The effect of Pt particle size and platinum–silver interaction on methanol oxidation was also considered. The nature of the functionalization agent strongly influenced the species formed on the surface during reaction with methanol and determined the catalytic activity and selectivity.

Protocol for a pilot randomised controlled trial of an intervention to increase the use of traffic light food labelling in UK shoppers (the FLICC trial)

Background: Traffic light labelling of foods—a system that incorporates a colour-coded assessment of the level of total fat, saturated fat, sugar and salt on the front of packaged foods—has been recommended by the UK Government and is currently in use or being phased in by many UK manufacturers and retailers. This paper describes a protocol for a pilot randomised controlled trial of an intervention designed to increase the use of traffic light labelling during real-life food purchase decisions.

Methods/design: The objectives of this two-arm randomised controlled pilot trial are to assess recruitment, retention and data completion rates, to generate potential effect size estimates to inform sample size calculations for the main trial and to assess the feasibility of conducting such a trial. Participants will be recruited by email from a loyalty card database of a UK supermarket chain. Eligible participants will be over 18 and regular shoppers who frequently purchase ready meals or pizzas. The intervention is informed by a review of previous interventions encouraging the use of nutrition labelling and the broader behaviour change literature. It is designed to impact on mechanisms affecting belief and behavioural intention formation as well as those associated with planning and goal setting and the adoption and maintenance of the behaviour of interest, namely traffic light label use during purchases of ready meals and pizzas. Data will be collected using electronic sales data via supermarket loyalty cards and web-based questionnaires and will be used to estimate the effect of the intervention on the nutrition profile of purchased ready meals and pizzas and the behavioural mechanisms associated with label use. Data collection will take place over 48 weeks. A process evaluation including semi-structured interviews and web analytics will be conducted to assess feasibility of a full trial.

Discussion: The design of the pilot trial allows for efficient recruitment and data collection. The intervention could be generalised to a wider population if shown to be feasible in the main trial.

Secure D2D Communication in Large-Scale Cognitive Cellular Networks: A Wireless Power Transfer Model

In this paper, we investigate secure device-to-device (D2D) communication in energy harvesting large-scale cognitive cellular networks. The energy constrained D2D transmitter harvests energy from multiantenna equipped power beacons (PBs), and communicates with the corresponding receiver using the spectrum of the primary base stations (BSs). We introduce a power transfer model and an information signal model to enable wireless energy harvesting and secure information transmission. In the power transfer model, three wireless power transfer (WPT) policies are proposed: 1) co-operative power beacons (CPB) power transfer, 2) best power beacon (BPB) power transfer, and 3) nearest power beacon (NPB) power transfer. To characterize the power transfer reliability of the proposed three policies, we derive new expressions for the exact power outage probability. Moreover, the analysis of the power outage probability is extended to the case when PBs are equipped with large antenna arrays. In the information signal model, we present a new comparative framework with two receiver selection schemes: 1) best receiver selection (BRS), where the receiver with the strongest channel is selected; and 2) nearest receiver selection (NRS), where the nearest receiver is selected. To assess the secrecy performance, we derive new analytical expressions for the secrecy outage probability and the secrecy throughput considering the two receiver selection schemes using the proposed WPT policies. We presented Monte carlo simulation results to corroborate our analysis and show: 1) secrecy performance improves with increasing densities of PBs and D2D receivers due to larger multiuser diversity gain; 2) CPB achieves better secrecy performance than BPB and NPB but consumes more power; and 3) BRS achieves better secrecy performance than NRS but demands more instantaneous feedback and overhead. A pivotal conclusion- is reached that with increasing number of antennas at PBs, NPB offers a comparable secrecy performance to that of BPB but with a lower complexity.

Traffic gap judgment in people with significant peripheral field loss.

PURPOSE: Subjects with significant peripheral field loss (PFL) self report difficulty in street crossing. In this study, we compared the traffic gap judgment ability of fully sighted and PFL subjects to determine whether accuracy in identifying crossable gaps was adversely affected because of field loss. Moreover, we explored the contribution of visual and nonvisual factors to traffic gap judgment ability. METHODS: Eight subjects with significant PFL as a result of advanced retinitis pigmentosa or glaucoma with binocular visual field <20 degrees and five age-matched normals (NV) were recruited. All subjects were required to judge when they perceived it was safe to cross at a 2-way 4-lane street while they stood on the curb. Eye movements were recorded by an eye tracker as the subjects performed the decision task. Movies of the eye-on-scene were made offline and fixation patterns were classified into either relevant or irrelevant. Subjects' street-crossing behavior, habitual approach to street crossing, and perceived difficulties were assessed. RESULTS: Compared with normal vision (NV) subjects, the PFL subjects identified 12% fewer crossable gaps while making 23% more errors by identifying a gap as crossable when it was too short (p < 0.05). The differences in traffic gap judgment ability of the PFL subjects might be explained by the significantly smaller fixation area (p = 0.006) and fewer fixations distributed to the relevant tasks (p = 0.001). The subjects' habitual approach to street crossing and perceived difficulties in street crossing (r > 0.60) were significantly correlated with traffic gap judgment performance. CONCLUSIONS: As a consequence of significant field loss, limited visual information about the traffic environment can be acquired, resulting in significantly reduced performance in judging safe crossable gaps. This poor traffic gap judgment ability in the PFL subjects raises important concerns for their safety when attempting to cross the street.