784 research outputs found

    Flow Cytometry for Identification of PRDM1-eYFP Transgenic Mice

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    The goal of this study was to determine if flow cytometry could be used to identify transgene carrier mice [B6.Cg-Tg(Prdm1-EYFP)1Mnz/J] by their expression of the enhanced yellow fluorescent protein (eYFP) in peripheral blood lymphocytes. In these mice, eYFP expression is under the control of the Prdm1 gene promoter. Littermates were identified as being either wild type or transgene carriers using a polymerase chain reaction (PCR) analysis of tail tissue. Peripheral blood leukocytes were isolated from each of the mice, and both the percent and mean fluorescence intensity (MFI) were determined for eYFP expression by lymphocytes. We found that blood lymphocytes from eYFP transgene carrier mice contained an average of 1.54% eYFP+ cells. In contrast, wild type animals contained significantly fewer (P \u3c 0.05) with an average of 0.18%. The percent eYFP- and eYFP+ data ranges did not overlap. The average MFI values for these groups were also significantly different, but their data ranges overlapped. The PCR process performed in our laboratory required approximately 20 hours over three days to complete with an estimated per animal cost of 16.00.Incontrast,theflowcytometryprocedurerequiredapproximatelysixhourswithinonedaytocompletewithanestimatedperanimalcostof16.00. In contrast, the flow cytometry procedure required approximately six hours within one day to complete with an estimated per animal cost of 7.85. We conclude that flow cytometry is an adequate and less costly method for identifying eYFP transgene carrier mice

    Scottish theme towns: have new identities enhanced development?

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    Three small towns in southwest Scotland have recently been branded as distinct theme towns, based on books, artists and food. This is an attempt to make them more attractive to visitors and thereby improve their economy. The objective of this research is to establish whether the new identities possessed by the towns have enhanced their development. It is argued, using data reviewing the past decade, that they have all developed, albeit at different rates, in terms of the economy and culture. Moreover, it is maintained that social capital has been enhanced and is a factor whose importance has been under-appreciated by planners and observers of this type of process. The relevance of the new identity to the pre-branding identity is also seen as a factor in successful development and ideas of authenticity and heritage are brought to bear on the relationship

    STAT3 Impairs STAT5 Activation in the Development of IL-9-Secreting T Cells

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    Th cell subsets develop in response to multiple activating signals, including the cytokine environment. IL-9-secreting T cells develop in response to the combination of IL-4 and TGF-ÎČ, although they clearly require other cytokine signals, leading to the activation of transcription factors including STAT5. In Th17 cells, there is a molecular antagonism of STAT5 with STAT3 signaling, although whether this paradigm exists in other Th subsets is not clear. In this paper, we demonstrate that STAT3 attenuates the ability of STAT5 to promote the development of IL-9-secreting T cells. We demonstrate that production of IL-9 is increased in the absence of STAT3 and cytokines that result in a sustained activation of STAT3, including IL-6, have the greatest potency in repressing IL-9 production in a STAT3-dependent manner. Increased IL-9 production in the absence of STAT3 correlates with increased endogenous IL-2 production and STAT5 activation, and blocking IL-2 responses eliminates the difference in IL-9 production between wild-type and STAT3-deficient T cells. Moreover, transduction of developing Th9 cells with a constitutively active STAT5 eliminates the ability of IL-6 to reduce IL-9 production. Thus, STAT3 functions as a negative regulator of IL-9 production through attenuation of STAT5 activation and function

    The ETS family transcription factors Etv5 and PU.1 function in parallel to promote Th9 cell development

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    The IL-9-secreting Th9 subset of CD4 T helper cells develop in response to an environment containing IL-4 and TGFÎČ, promoting allergic disease, autoimmunity, and resistance to pathogens. We previously identified a requirement for the ETS family transcription factor PU.1 in Th9 development. In this report we demonstrate that the ETS transcription factor ETV5 promotes IL-9 production in Th9 cells by binding and recruiting histone acetyltransferases to the Il9 locus at sites distinct from PU.1. In cells that are deficient in both PU.1 and ETV5 there is lower IL-9 production than in cells lacking either factor alone. In vivo loss of PU.1 and ETV5 in T cells results in distinct affects on allergic inflammation in the lung, suggesting that these factors function in parallel. Together, these data define a role for ETV5 in Th9 development and extend the paradigm of related transcription factors having complementary functions during differentiation

    PU.1 expression in T follicular helper cells limits CD40L-dependent germinal center B cell development.

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    PU.1 is an ETS family transcription factor important for the development of multiple hematopoietic cell lineages. Previous work demonstrated a critical role for PU.1 in promoting Th9 development, and in limiting Th2 cytokine production. Whether PU.1 has functions in other T helper lineages is not clear. In this report we examined the effects of ectopic expression of PU.1 in CD4+T cells and observed decreased expression of genes involved with the function of T follicular helper (Tfh) cells, including Il21 and Tnfsf5 (encoding CD40L). T cells from conditional mutant mice that lack expression of PU.1 in T cells (Sfpi1lck−/−) demonstrated increased production of CD40L and IL-21 in vitro. Following adjuvant-dependent or adjuvant-independent immunization, we observed that Sfpi1lck−/− mice had increased numbers of Tfh cells, increased germinal center B cells, and increased antibody production in vivo. This correlated with increased expression of IL-21 and CD40L in Tfh cells from Sfpi1lck−/− mice, compared to control mice. Finally, although blockade of IL-21 did not affect germinal center B cells in Sfpi1lck−/− mice, anti-CD40L treatment of immunized Sfpi1lck−/− mice decreased germinal center B cell numbers and antigen-specific immunoglobulin concentrations. Together, these data indicate an inhibitory role of PU.1 in the function of T follicular helper cells, germinal centers, and Tfh-dependent humoral immunity

    The Genomic Signature of Crop-Wild Introgression in Maize

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    The evolutionary significance of hybridization and subsequent introgression has long been appreciated, but evaluation of the genome-wide effects of these phenomena has only recently become possible. Crop-wild study systems represent ideal opportunities to examine evolution through hybridization. For example, maize and the conspecific wild teosinte Zea mays ssp. mexicana, (hereafter, mexicana) are known to hybridize in the fields of highland Mexico. Despite widespread evidence of gene flow, maize and mexicana maintain distinct morphologies and have done so in sympatry for thousands of years. Neither the genomic extent nor the evolutionary importance of introgression between these taxa is understood. In this study we assessed patterns of genome-wide introgression based on 39,029 single nucleotide polymorphisms genotyped in 189 individuals from nine sympatric maize-mexicana populations and reference allopatric populations. While portions of the maize and mexicana genomes were particularly resistant to introgression (notably near known cross-incompatibility and domestication loci), we detected widespread evidence for introgression in both directions of gene flow. Through further characterization of these regions and preliminary growth chamber experiments, we found evidence suggestive of the incorporation of adaptive mexicana alleles into maize during its expansion to the highlands of central Mexico. In contrast, very little evidence was found for adaptive introgression from maize to mexicana. The methods we have applied here can be replicated widely, and such analyses have the potential to greatly informing our understanding of evolution through introgressive hybridization. Crop species, due to their exceptional genomic resources and frequent histories of spread into sympatry with relatives, should be particularly influential in these studies

    The Paranormal is (Still) Normal: The Sociological Implications of a Survey of Paranormal Experiences in Great Britain

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    Historically, there has been limited sociological interest in the paranormal and no systematic study of reported paranormal experiences. There are also few medium-to-large-scale survey results with nationally representative populations focusing on paranormal experiences. This paper provides details of an exploratory survey conducted in 2009 with a nationally representative sample of 4,096 adults aged 16 years and over across Great Britain. Our findings show that 37% of British adults report at least one paranormal experience and that women, those who are middle-aged or individuals resident in the South West are more likely to report such experiences. These results establish incidence levels of reported paranormal experiences in contemporary Britain. We argue also that they merit a more sustained sociological consideration of the paranormal. In this respect we renew and update the robust justification and call for serious research positioning the paranormal as a social phenomenon, originally proposed well over thirty years ago by Greeley (1975)

    Supernova Simulations and Strategies For the Dark Energy Survey

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    We present an analysis of supernova light curves simulated for the upcoming Dark Energy Survey (DES) supernova search. The simulations employ a code suite that generates and fits realistic light curves in order to obtain distance modulus/redshift pairs that are passed to a cosmology fitter. We investigated several different survey strategies including field selection, supernova selection biases, and photometric redshift measurements. Using the results of this study, we chose a 30 square degree search area in the griz filter set. We forecast 1) that this survey will provide a homogeneous sample of up to 4000 Type Ia supernovae in the redshift range 0.05<z<1.2, and 2) that the increased red efficiency of the DES camera will significantly improve high-redshift color measurements. The redshift of each supernova with an identified host galaxy will be obtained from spectroscopic observations of the host. A supernova spectrum will be obtained for a subset of the sample, which will be utilized for control studies. In addition, we have investigated the use of combined photometric redshifts taking into account data from both the host and supernova. We have investigated and estimated the likely contamination from core-collapse supernovae based on photometric identification, and have found that a Type Ia supernova sample purity of up to 98% is obtainable given specific assumptions. Furthermore, we present systematic uncertainties due to sample purity, photometric calibration, dust extinction priors, filter-centroid shifts, and inter-calibration. We conclude by estimating the uncertainty on the cosmological parameters that will be measured from the DES supernova data.Comment: 46 pages, 30 figures, resubmitted to ApJ as Revision 2 (final author revision), which has subtle editorial differences compared to the published paper (ApJ, 753, 152). Note that this posting includes PDF only due to a bug in either the latex macros or the arXiv submission system. The source files are available in the DES document database: http://des-docdb.fnal.gov/cgi-bin/ShowDocument?docid=624

    Gapless Assembly of Maize Chromosomes Using Long-Read Technologies

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    Creating gapless telomere-to-telomere assemblies of complex genomes is one of the ultimate challenges in genomics. We use two independent assemblies and an optical map-based merging pipeline to produce a maize genome (B73-Ab10) composed of 63 contigs and a contig N50 of 162 Mb. This genome includes gapless assemblies of chromosome 3 (236 Mb) and chromosome 9 (162 Mb), and 53 Mb of the Ab10 meiotic drive haplotype. The data also reveal the internal structure of seven centromeres and five heterochromatic knobs, showing that the major tandem repeat arrays (CentC, knob180, and TR-1) are discontinuous and frequently interspersed with retroelements
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