501 research outputs found

    Dynamic Protein Interactions of the Polycomb Repressive Complex 2 during Differentiation of Pluripotent Cells

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    Polycomb proteins assemble to form complexes with important roles in epigenetic regulation. The Polycomb Repressive Complex 2 (PRC2) modulates the di- and tri-methylation of lysine 27 on histone H3, each of which are associated with gene repression. Although three subunits, EZH1/2, SUZ12, and EED, form the catalytic core of PRC2, a wider group of proteins associate with low stoichiometry. This raises the question of whether dynamic variation of the PRC2 interactome results in alternative forms of the complex during differentiation. Here we compared the physical interactions of PRC2 in undifferentiated and differentiated states of NTERA2 pluripotent embryonic carcinoma cells. Label-free quantitative proteomics was used to assess endogenous immunoprecipitation of the EZH2 and SUZ12 subunits of PRC2. A high stringency data set reflecting the endogenous state of PRC2 was produced that included all previously reported core and associated PRC2 components, and several novel interacting proteins. Comparison of the interactomes obtained in undifferentiated and differentiated cells revealed candidate proteins that were enriched in complexes isolated from one of the two states. For example, SALL4 and ZNF281 associate with PRC2 in pluripotent cells, whereas PCL1 and SMAD3 preferentially associate with PRC2 in differentiating cells. Analysis of the mRNA and protein levels of these factors revealed that their association with PRC2 correlated with their cell state-specific expression. Taken together, we propose that dynamic changes to the PRC2 interactome during differentiation may contribute to directing its activity during cell fate transitions

    Explaining global surface aerosol number concentrations in terms of primary emissions and particle formation

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    We use observations of total particle number concentration at 36 worldwide sites and a global aerosol model to quantify the primary and secondary sources of particle number. We show that emissions of primary particles can reasonably reproduce the spatial pattern of observed condensation nuclei (CN) (R2=0.51) but fail to explain the observed seasonal cycle at many sites (R2=0.1). The modeled CN concentration in the free troposphere is biased low (normalised mean bias, NMB=−88%) unless a secondary source of particles is included, for example from binary homogeneous nucleation of sulfuric acid and water (NMB=−25%). Simulated CN concentrations in the continental boundary layer (BL) are also biased low (NMB=−74%) unless the number emission of anthropogenic primary particles is increased or an empirical BL particle formation mechanism based on sulfuric acid is used. We find that the seasonal CN cycle observed at continental BL sites is better simulated by including a BL particle formation mechanism (R2=0.3) than by increasing the number emission from primary anthropogenic sources (R2=0.18). Using sensitivity tests we derive optimum rate coefficients for this nucleation mechanism, which agree with values derived from detailed case studies at individual sites

    Ranging behaviour of translocated roe deer in a Mediterranean habitat: seasonal and altitudinal influences on home range size and patterns of range use

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    In this study we investigated the causes of seasonal variation in the home ranges of roe deer reintroduced to the Gardunha Mountains (Portugal). From May 2002 to April 2003, 1 year after the animals had been released, we collected data using radio-tracking techniques for five monitored animals (two males and three females). We found differences in the size of home ranges between seasons, with home ranges larger in summer than winter (minimum convex polygon peeled to 95%: summer 409.64 +/- 98.20 ha, winter 116.20 +/- 17.90 ha). This is contrary to evidence from central and northern Europe, where home ranges are typically larger in winter than summer. Moreover, two of the sampled females and one of the males tended to use higher elevations in summer. Comparisons between Mediterranean populations and those in central and northern Europe showed that Mediterranean populations in the winter easily fulfil their needs within a small area, whereas in the hot dry summer a larger area is needed. Furthermore, individuals prefer a higher, cooler mountainous habitat in summer, which is likely to be a means of avoiding warmer temperatures

    ROBERT BURNS AND FRIENDS essays by W. Ormiston Roy Fellows presented to G. Ross Roy

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    Robert Burns & Friends essays by W. Ormiston Roy Fellows presented to G. Ross Roy edited by Patrick Scott and Kenneth Simpson This volume of essays about the Scottish poet Robert Burns (1759-1796) pays tribute to the distinguished Burns scholar G. Ross Roy. Subjects covered include writers who influenced Burns; aspects of the writing of Burns and that of his friends and contemporaries; and Burns\u27s influence on later writers. The volume also includes essays on Ross Roy\u27s own accomplishments and on the Burns collection he built (now at the University of South Carolina), together with a checklist of his published writings. G. Ross Roy, Distinguished Professor Emeritus of English and Comparative Literature, founded the journal Studies in Scottish Literature in 1963, and as its editor for nearly fifty years he has had a central role in establishing international academic recognition for the field. His own scholarly work includes the standard Letters of Robert Burns (2 vols., Clarendon Press, 1985). His contributions to Scottish literature have earned him honorary doctorates from the Universities of Edinburgh (2002) and Glasgow (2009). The contributors are all former W. Ormiston Roy Visiting Fellows at the University of South Carolina. This book is also available in a print edition (ISBN: 978-1439270974) through the usual on-line vendors. It is not available for direct purchase from the editors or the University of South Carolina

    TLR7-mediated skin inflammation remotely triggers chemokine expression and leukocyte accumulation in the brain

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    Background: The relationship between the brain and the immune system has become increasingly topical as, although it is immune-specialised, the CNS is not free from the influences of the immune system. Recent data indicate that peripheral immune stimulation can significantly affect the CNS. But the mechanisms underpinning this relationship remain unclear. The standard approach to understanding this relationship has relied on systemic immune activation using bacterial components, finding that immune mediators, such as cytokines, can have a significant effect on brain function and behaviour. More rarely have studies used disease models that are representative of human disorders. Methods: Here we use a well-characterised animal model of psoriasis-like skin inflammation—imiquimod—to investigate the effects of tissue-specific peripheral inflammation on the brain. We used full genome array, flow cytometry analysis of immune cell infiltration, doublecortin staining for neural precursor cells and a behavioural read-out exploiting natural burrowing behaviour. Results: We found that a number of genes are upregulated in the brain following treatment, amongst which is a subset of inflammatory chemokines (CCL3, CCL5, CCL9, CXCL10, CXCL13, CXCL16 and CCR5). Strikingly, this model induced the infiltration of a number of immune cell subsets into the brain parenchyma, including T cells, NK cells and myeloid cells, along with a reduction in neurogenesis and a suppression of burrowing activity. Conclusions: These findings demonstrate that cutaneous, peripheral immune stimulation is associated with significant leukocyte infiltration into the brain and suggest that chemokines may be amongst the key mediators driving this response

    Detection and identification of human Plasmodium species with real-time quantitative nucleic acid sequence-based amplification

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    BACKGROUND: Decisions concerning malaria treatment depend on species identification causing disease. Microscopy is most frequently used, but at low parasitaemia (<20 parasites/μl) the technique becomes less sensitive and time consuming. Rapid diagnostic tests based on Plasmodium antigen detection do often not allow for species discrimination as microscopy does, but also become insensitive at <100 parasites/μl. METHODS: This paper reports the development of a sensitive and specific real-time Quantitative Nucleic Acid Sequence Based Amplification (real-time QT-NASBA) assays, based on the small-subunit 18S rRNA gene, to identify the four human Plasmodium species. RESULTS: The lower detection limit of the assay is 100 – 1000 molecules in vitro RNA for all species, which corresponds to 0.01 – 0.1 parasite per diagnostic sample (i.e. 50 μl of processed blood). The real-time QT-NASBA was further evaluated using 79 clinical samples from malaria patients: i.e. 11 Plasmodium. falciparum, 37 Plasmodium vivax, seven Plasmodium malariae, four Plasmodium ovale and 20 mixed infections. The initial diagnosis of 69 out of the 79 samples was confirmed with the developed real-time QT-NASBA. Re-analysis of seven available original slides resolved five mismatches. Three of those were initially identified as P. malariae mono-infection, but after re-reading the slides P. falciparum was found, confirming the real-time QT-NASBA result. The other two slides were of poor quality not allowing true species identification. The remaining five discordant results could not be explained by microscopy, but may be due to extreme low numbers of parasites present in the samples. In addition, 12 Plasmodium berghei isolates from mice and 20 blood samples from healthy donors did not show any reaction in the assay. CONCLUSION: Real-time QT-NASBA is a very sensitive and specific technique with a detection limit of 0.1 Plasmodium parasite per diagnostic sample (50 μl of blood) and can be used for the detection, identification and quantitative measurement of low parasitaemia of Plasmodium species, thus making it an effective tool for diagnostic purposes and useful for epidemiological and drug studies
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