Development of Functional Microfold (M) Cells from Intestinal Stem Cells in Primary Human Enteroids.

Background & aimsIntestinal microfold (M) cells are specialized epithelial cells that act as gatekeepers of luminal antigens in the intestinal tract. They play a critical role in the intestinal mucosal immune response through transport of viruses, bacteria and other particles and antigens across the epithelium to immune cells within Peyer's patch regions and other mucosal sites. Recent studies in mice have demonstrated that M cells are generated from Lgr5+ intestinal stem cells (ISCs), and that infection with Salmonella enterica serovar Typhimurium increases M cell formation. However, it is not known whether and how these findings apply to primary human small intestinal epithelium propagated in an in vitro setting.MethodsHuman intestinal crypts were grown as monolayers with growth factors and treated with recombinant RANKL, and assessed for mRNA transcripts, immunofluorescence and uptake of microparticles and S. Typhimurium.ResultsFunctional M cells were generated by short-term culture of freshly isolated human intestinal crypts in a dose- and time-dependent fashion. RANKL stimulation of the monolayer cultures caused dramatic induction of the M cell-specific markers, SPIB, and Glycoprotein-2 (GP2) in a process primed by canonical WNT signaling. Confocal microscopy demonstrated a pseudopod phenotype of GP2-positive M cells that preferentially take up microparticles. Furthermore, infection of the M cell-enriched cultures with the M cell-tropic enteric pathogen, S. Typhimurium, led to preferential association of the bacteria with M cells, particularly at lower inoculum sizes. Larger inocula caused rapid induction of M cells.ConclusionsHuman intestinal crypts containing ISCs can be cultured and differentiate into an epithelial layer with functional M cells with characteristic morphological and functional properties. This study is the first to demonstrate that M cells can be induced to form from primary human intestinal epithelium, and that S. Typhimurium preferentially infect these cells in an in vitro setting. We anticipate that this model can be used to generate large numbers of M cells for further functional studies of these key cells of intestinal immune induction and their impact on controlling enteric pathogens and the intestinal microbiome

First results of the ROSEBUD Dark Matter experiment

Rare Objects SEarch with Bolometers UndergrounD) is an experiment which attempts to detect low mass Weak Interacting Massive Particles (WIMPs) through their elastic scattering off Al and O nuclei. It consists of three small sapphire bolometers (of a total mass of 100 g) with NTD-Ge sensors in a dilution refrigerator operating at 20 mK in the Canfranc Underground Laboratory. We report in this paper the results of several runs (of about 10 days each) with successively improved energy thresholds, and the progressive background reduction obtained by improvement of the radiopurity of the components and subsequent modifications in the experimental assembly, including the addition of old lead shields. Mid-term plans and perspectives of the experiment are also presented.Comment: 14 pages, 8 figures, submitted to Astroparticle Physic

How an ancient, salt-tolerant fruit crop, Ficus carica L., copes with salinity: a transcriptome analysis

Although Ficus carica L. (fig) is one of the most resistant fruit tree species to salinity, no comprehensive studies are currently available on its molecular responses to salinity. Here we report a transcriptome analysis of F. carica cv. Dottato exposed to 100 mM sodium chloride for 7 weeks, where RNA-seq analysis was performed on leaf samples at 24 and 48 days after the beginning of salinization; a genomederived fig transcriptome was used as a reference. At day 24, 224 transcripts were significantly upregulated and 585 were down-regulated, while at day 48, 409 genes were activated and 285 genes were repressed. Relatively small transcriptome changes were observed after 24 days of salt treatment, showing that fig plants initially tolerate salt stress. However, after an early down-regulation of some cell functions, major transcriptome changes were observed after 48 days of salinity. Seven weeks of 100 mM NaCl dramatically changed the repertoire of expressed genes, leading to activation or reactivation of many cell functions. We also identified salt-regulated genes, some of which had not been previously reported to be involved in plant salinity responses. These genes could be potential targets for the selection of favourable genotypes, through breeding or biotechnology, to improve salt tolerance in fig or other crops

Recent results from the canfranc dark matter search with germanium detectors

Two germanium detectors are currently operating in the Canfranc Underground Laboratory at 2450 m.w.e looking for WIMP dark matter. One is a 2 kg 76Ge IGEX detector (RG-2) which has an energy threshold of 4 keV and a low-energy background rate of about 0.3 c/keV/kg/day. The other is a small (234 g) natural abundance Ge detector (COSME), of low energy threshold (2.5 keV) and an energy resolution of 0.4 keV at 10 keV which is looking for WIMPs and for solar axions. The analysis of 73 kg-days of data taken by COSME in a search for solar axions via their photon Primakoff conversion and Bragg scattering in the Ge crystal yields a 95% C.L. limit for the axion-photon coupling g < 2.8 10^-9 GeV^-1. These data, analyzed for WIMP searches provide an exclusion plot for WIMP-nucleon spin-independent interaction which improves previous plots in the low mass region. On the other hand, the exclusion plot derived from the 60 kg-days of data from the RG-2 IGEX detector improves the exclusion limits derived from other ionization (non thermal) germanium detector experiments in the region of WIMP masses from 30 to 100 GeV recently singled out by the reported DAMA annual modulation effect.Comment: 6 pages, talk given at IDM2000, York, September 200

Talk the talk, walk the walk: Defining Critical Race Theory in research

Over the last decade there has been a noticeable growth in published works citing Critical Race Theory (CRT). This has led to a growth in interest in the UK of practical research projects utilising CRT as their framework. It is clear that research on 'race' is an emerging topic of study. What is less visible is a debate on how CRT is positioned in relation to methodic practice, substantive theory and epistemological underpinnings. The efficacy of categories of data gathering tools, both traditional and non-traditional is a discussion point here to explore the complexities underpinning decisions to advocate a CRT framework. Notwithstanding intersectional issues, a CRT methodology is recognisable by how philosophical, political and ethical questions are established and maintained in relation to racialised problematics. This paper examines these tensions in establishing CRT methodologies and explores some of the essential criteria for researchers to consider in utilising a CRT framework. © 2012 Copyright Taylor and Francis Group, LLC

The IGEX experiment revisited: a response to the critique of Klapdor-Kleingrothaus,Dietz, and Krivosheina

This paper is a response to the article "Critical View to" the IGEX neutrinoless double-beta decay experiment..."published in Phys. Rev.D, Volume 65 (2002) 092007," by H.V.Klapdor-Kleingrothaus, A. Dietz, and I.V.Krivosheina, published as preprint hep-ph/0403056. The criticisms are confronted, and the questions raised are answered. We demonstrate that the lower limit quoted by IGEX, for the half life of Ge-76 neutrinoless double beta decay, 1.57x10**25 y, is correct and that there was no "arithmetical error"-as claimed in the " Critical Review" article