Proteomic profiling of glucocorticoid-exposed myogenic cells: Time series assessment of protein translocation and transcription of inactive mRNAs

<p>Abstract</p> <p>Background</p> <p>Prednisone, one of the most highly prescribed drugs, has well characterized effects on gene transcription mediated by the glucocorticoid receptor. These effects are typically occurring on the scale of hours. Prednisone also has a number of non-transcriptional effects (occurring on minutes scale) on protein signaling, yet these are less well studied. We sought to expand the understanding of acute effects of prednisone action on cell signaling using a combination of SILAC strategy and subcellular fractionations from C₂C₁₂myotubes.</p> <p>Results</p> <p><it>De novo </it>translation of proteins was inhibited in both SILAC labeled and unlabeled C₂C₁₂myotubes. Unlabeled cells were exposed to prednisone while SILAC labeled cells remained untreated. After 0, 5, 15, and 30 minutes of prednisone exposure, labeled and unlabeled cells were mixed at 1:1 ratios and fractionated into cytosolic and nuclear fractions. A total of 534 proteins in the cytosol and 626 proteins in the nucleus were identified and quantitated, using 3 or more peptides per protein with peptide based probability ≤ 0.001. We identified significant increases (1.7- to 3.1- fold) in cytoplasmic abundance of 11 ribosomal proteins within 5 minutes of exposure, all of which returned to baseline by 30 min. We hypothesized that these drug-induced acute changes in the subcellular localization of the cell's protein translational machinery could lead to altered translation of quiescent RNAs. To test this, <it>de novo </it>protein synthesis was assayed after 15 minutes of drug exposure. Quantitative fluorography identified 16 2D gel spots showing rapid changes in translation; five of these were identified by MS/MS (pyruvate kinase, annexin A6 isoform A and isoform B, nasopharyngeal epithelium specific protein 1, and isoform 2 of Replication factor C subunit 1), and all showed the 5' terminal oligopyrimidine motifs associated with mRNA sequestration to and from inactive mRNA pools.</p> <p>Conclusion</p> <p>We describe novel approaches of subcellular proteomic profiling and assessment of acute changes on a minute-based time scale. These data expand the current knowledge of acute, non-transcriptional activities of glucocorticoids, including changes in protein subcellular localization, altered translation of quiescent RNA pools, and PKC-mediated cytoskeleton remodeling.</p

Community- and Individual-Level Socioeconomic Status and Breast Cancer Risk: Multilevel Modeling on Cape Cod, Massachusetts

BACKGROUND. Previous research demonstrated increased risk of breast cancer associated with higher socioeconomic status (SES) measured at both the individual and community levels. However, little attention has been paid to simultaneously examining both measures. OBJECTIVES. We evaluated the independent influences of individual and community SES on the risk of breast cancer using case-control data. Because our previous work suggests that associations may be stronger after including a latency period, we also assessed the effect of community-level SES assuming a 10-year latency period. METHODS. We obtained individual education for cases and matched controls diagnosed between 1987 and 1993 on Cape Cod, Massachusetts (USA). We acquired community-level SES from census data for 1980 and 1990. Using SES data at diagnosis and 10 years earlier, we constructed models for breast cancer risk using individual-level SES only, community-level SES only, and a multilevel analysis including both. We adjusted models for other individual-level risk factors. RESULTS. Women with the highest education were at greater risk of developing breast cancer in both 1980 and 1990 [odds ratio (OR) = 1.17 and 1.19, respectively]. Similarly, women living in the highest-SES communities in 1990 had greater risk (OR = 1.30). Results were stronger in the analyses considering a latency period (OR = 1.69). Adjusting for intragroup correlation had little effect on the analyses. CONCLUSIONS. Models including individual- or community-level measures of SES produced associations similar to those observed in previous research. Results for models including both measures are consistent with a contextual effect of SES on risk of breast cancer independent of individual SES.Boston University Center for Interdisciplinary Research in Environmental Exposures and Health; National Institute of Environmental Health (P42ES007381

Attitudes towards the use and acceptance of eHealth technologies : a case study of older adults living with chronic pain and implications for rural healthcare

Acknowledgements The research described here is supported by the award made by the RCUK Digital Economy programme to the dot.rural Digital Economy Hub; award reference: EP/G066051/1. MC’s time writing the paper is funded by the Scottish Government’s Rural and Environmental Science and Analytical Services Division (RESAS) under Theme 8 ‘Vibrant Rural Communities’ of the Food, Land and People Programme (2011–2016). MC is also an Honorary Research Fellow at the Division of Applied Health Sciences, University of Aberdeen. The input of other members of the TOPS research team, Alastair Mort, Fiona Williams, Sophie Corbett, Phil Wilson and Paul MacNamee who contributed to be wider study and discussed preliminary findings reported here with the authors of the paper is acknowledged. We acknowledge the feedback on earlier versions of this paper provided by members of the Trans-Atlantic Rural Research Network, especially Stefanie Doebler and Carmen Hubbard. We also thank Deb Roberts for her comments.Peer reviewedPublisher PD

Quasi-particle interference and superconducting gap in a high-temperature superconductor Ca2-xNaxCuO2Cl2

High-transition-temperature (high-Tc) superconductivity is ubiquitous in the cuprates containing CuO2 planes but each cuprate has its own character. The study of the material dependence of the d-wave superconducting gap (SG) should provide important insights into the mechanism of high-Tc. However, because of the 'pseudogap' phenomenon, it is often unclear whether the energy gaps observed by spectroscopic techniques really represent the SG. Here, we report spectroscopic imaging scanning tunneling microscopy (SI-STM) studies of nearly-optimally-doped Ca2-xNaxCuO2Cl2 (Na-CCOC) with Tc = 25 ~ 28 K. They enable us to observe the quasi-particle interference (QPI) effect in this material, through which unambiguous new information on the SG is obtained. The analysis of QPI in Na-CCOC reveals that the SG dispersion near the gap node is almost identical to that of Bi2Sr2CaCu2Oy (Bi2212) at the same doping level, while Tc of Bi2212 is 3 times higher than that of Na-CCOC. We also find that SG in Na-CCOC is confined in narrower energy and momentum ranges than Bi2212. This explains at least in part the remarkable material dependence of TcComment: 13pages, 4fig

Inflammasome and IL-1β-Mediated Disorders

The NLRP3 inflammasome is an intracellular complex that regulates the release of proinflammatory cytokines such as interleukin-1β in response to exogenous pathogens and endogenous danger signals. Evidence from studies involving human genetics, human ex vivo mononuclear cell responses, and in vivo and in vitro murine models confirms the importance of the inflammasome and interleukin-1β in the pathogenesis of several inherited and complex diseases. The availability of several effective interleukin-1β targeted therapies has allowed for successful proof-of-concept studies in several of these disorders. However, many other diseases are likely to be mediated by the inflammasome and interleukin-1β, providing additional targets in the future

Structure formation in active networks

Structure formation and constant reorganization of the actin cytoskeleton are key requirements for the function of living cells. Here we show that a minimal reconstituted system consisting of actin filaments, crosslinking molecules and molecular-motor filaments exhibits a generic mechanism of structure formation, characterized by a broad distribution of cluster sizes. We demonstrate that the growth of the structures depends on the intricate balance between crosslinker-induced stabilization and simultaneous destabilization by molecular motors, a mechanism analogous to nucleation and growth in passive systems. We also show that the intricate interplay between force generation, coarsening and connectivity is responsible for the highly dynamic process of structure formation in this heterogeneous active gel, and that these competing mechanisms result in anomalous transport, reminiscent of intracellular dynamics

Differential pain response at local and remote muscle sites following aerobic cycling exercise at mild and moderate intensity

Physical exercise has been shown to inhibit experimental pain response in the post-exercise period. Modulation of the pain system may be differentiated between muscle sites engaging in contractile activity. The purpose of this study was to assess the pain response at remote and local muscle sites following aerobic exercise at different work intensities. Participants included 10 healthy and physically active males (mean age ± SD, 21.2 ± 3.4). Somatic pressure pain threshold (PPT) at the rectus femoris (local) and brachioradialis (remote) muscle site was measured at before (Pre), 5 min after (Post1), and 15 min after (Post2) aerobic cycling exercise at 70 and 30 % of peak oxygen uptake (VO(2peak)) performed on different occasions in a counterbalanced order, separated by minimum of 3 days interval. Repeated measures ANOVA for PPT reveals significant main effect for time (f = 3.581, p = 0.049, observed power = 0.588) and muscle site (f = 17.931, p = 0.002, observed power = 0.963). There was a significant interaction shown for exercise intensity by time (f = 11.390, p = 0.012, observed power = 0.790). PPT at rectus femoris following cycling exercise at 70 % of VO(2peak) reveals a significant increase between Pre-Post1 (p = 0.040). PPT for rectus femoris following cycling exercise at 30 % of VO(2peak) revealed a significant decrease between Pre-Post1 (p = 0.026) and Pre-Post2 (p = 0.008). The PPT for brachioradialis following cycling exercise at 30 % of VO(2peak) revealed a significant decrease between Pre-Post1 (p = 0.011) and Pre-Post2 (p = 0.005). These results show that aerobic exercise increases PPT locally at the exercise muscle site following exercise at 70 % of VO(2peak) but reduces PPT following exercise at 30 % of VO(2peak)

Globally Continuous and Non-Markovian Crowd Activity Analysis from Videos

Automatically recognizing activities in video is a classic problem in vision and helps to understand behaviors, describe scenes and detect anomalies. We propose an unsupervised method for such purposes. Given video data, we discover recurring activity patterns that appear, peak, wane and disappear over time. By using non-parametric Bayesian methods, we learn coupled spatial and temporal patterns with minimum prior knowledge. To model the temporal changes of patterns, previous works compute Markovian progressions or locally continuous motifs whereas we model time in a globally continuous and non-Markovian way. Visually, the patterns depict flows of major activities. Temporally, each pattern has its own unique appearance-disappearance cycles. To compute compact pattern representations, we also propose a hybrid sampling method. By combining these patterns with detailed environment information, we interpret the semantics of activities and report anomalies. Also, our method fits data better and detects anomalies that were difficult to detect previously

Imaging the Two Gaps of the High-TC Superconductor Pb-Bi2Sr2CuO6+x

The nature of the pseudogap state, observed above the superconducting transition temperature TC in many high temperature superconductors, is the center of much debate. Recently, this discussion has focused on the number of energy gaps in these materials. Some experiments indicate a single energy gap, implying that the pseudogap is a precursor state. Others indicate two, suggesting that it is a competing or coexisting phase. Here we report on temperature dependent scanning tunneling spectroscopy of Pb-Bi2Sr2CuO6+x. We have found a new, narrow, homogeneous gap that vanishes near TC, superimposed on the typically observed, inhomogeneous, broad gap, which is only weakly temperature dependent. These results not only support the two gap picture, but also explain previously troubling differences between scanning tunneling microscopy and other experimental measurements.Comment: 6 page

Influence of family and friend smoking on intentions to smoke and smoking-related attitudes and refusal self-efficacy among 9-10 year old children from deprived neighbourhoods: a cross-sectional study.

BACKGROUND: Smoking often starts in early adolescence and addiction can occur rapidly. For effective smoking prevention there is a need to identify at risk groups of preadolescent children and whether gender-specific intervention components are necessary. This study aimed to examine associations between mother, father, sibling and friend smoking and cognitive vulnerability to smoking among preadolescent children living in deprived neighbourhoods. METHODS: Cross-sectional data was collected from 9-10 year old children (n =1143; 50.7% girls; 85.6% White British) from 43 primary schools in Merseyside, England. Children completed a questionnaire that assessed their smoking-related behaviour, intentions, attitudes, and refusal self-efficacy, as well as parent, sibling and friend smoking. Data for boys and girls were analysed separately using multilevel linear and logistic regression models, adjusting for individual cognitions and school and deprivation level. RESULTS: Compared to girls, boys had lower non-smoking intentions (P = 0.02), refusal self-efficacy (P = 0.04) and were less likely to agree that smoking is 'definitely' bad for health (P < 0.01). Friend smoking was negatively associated with non-smoking intentions in girls (P < 0.01) and boys (P < 0.01), and with refusal self-efficacy in girls (P < 0.01). Sibling smoking was negatively associated with non-smoking intentions in girls (P < 0.01) but a positive association was found in boys (P = 0.02). Boys who had a smoking friend were less likely to 'definitely' believe that the smoke from other people's cigarettes is harmful (OR 0.57, 95% CI: 0.35 to 0.91, P = 0.02). Further, boys with a smoking friend (OR 0.38, 95% CI: 0.21 to 0.69, P < 0.01) or a smoking sibling (OR 0.45, 95% CI: 0.21 to 0.98) were less likely to 'definitely' believe that smoking is bad for health. CONCLUSION: This study indicates that sibling and friend smoking may represent important influences on 9-10 year old children's cognitive vulnerability toward smoking. Whilst some differential findings by gender were observed, these may not be sufficient to warrant separate prevention interventions. However, further research is needed