Search for axion-like particles using a variable baseline photon regeneration technique

We report the first results of the GammeV experiment, a search for milli-eV mass particles with axion-like couplings to two photons. The search is performed using a "light shining through a wall" technique where incident photons oscillate into new weakly interacting particles that are able to pass through the wall and subsequently regenerate back into detectable photons. The oscillation baseline of the apparatus is variable, thus allowing probes of different values of particle mass. We find no excess of events above background and are able to constrain the two-photon couplings of possible new scalar (pseudoscalar) particles to be less than 3.1x10^{-7} GeV^{-1} (3.5x10^{-7} GeV^{-1}) in the limit of massless particles.Comment: 5 pages, 4 figures. This is the version accepted by PRL and includes updated limit

Physico-chemical foundations underpinning microarray and next-generation sequencing experiments

Hybridization of nucleic acids on solid surfaces is a key process involved in high-throughput technologies such as microarrays and, in some cases, next-generation sequencing (NGS). A physical understanding of the hybridization process helps to determine the accuracy of these technologies. The goal of a widespread research program is to develop reliable transformations between the raw signals reported by the technologies and individual molecular concentrations from an ensemble of nucleic acids. This research has inputs from many areas, from bioinformatics and biostatistics, to theoretical and experimental biochemistry and biophysics, to computer simulations. A group of leading researchers met in Ploen Germany in 2011 to discuss present knowledge and limitations of our physico-chemical understanding of high-throughput nucleic acid technologies. This meeting inspired us to write this summary, which provides an overview of the state-of-the-art approaches based on physico-chemical foundation to modeling of the nucleic acids hybridization process on solid surfaces. In addition, practical application of current knowledge is emphasized

Epigenetic memory in induced pluripotent stem cells.

Somatic cell nuclear transfer and transcription-factor-based reprogramming revert adult cells to an embryonic state, and yield pluripotent stem cells that can generate all tissues. Through different mechanisms and kinetics, these two reprogramming methods reset genomic methylation, an epigenetic modification of DNA that influences gene expression, leading us to hypothesize that the resulting pluripotent stem cells might have different properties. Here we observe that low-passage induced pluripotent stem cells (iPSCs) derived by factor-based reprogramming of adult murine tissues harbour residual DNA methylation signatures characteristic of their somatic tissue of origin, which favours their differentiation along lineages related to the donor cell, while restricting alternative cell fates. Such an 'epigenetic memory' of the donor tissue could be reset by differentiation and serial reprogramming, or by treatment of iPSCs with chromatin-modifying drugs. In contrast, the differentiation and methylation of nuclear-transfer-derived pluripotent stem cells were more similar to classical embryonic stem cells than were iPSCs. Our data indicate that nuclear transfer is more effective at establishing the ground state of pluripotency than factor-based reprogramming, which can leave an epigenetic memory of the tissue of origin that may influence efforts at directed differentiation for applications in disease modelling or treatment

Performance evaluation of commercial miRNA expression array platforms

<p>Abstract</p> <p>Background</p> <p>microRNAs (miRNA) are short, endogenous transcripts that negatively regulate the expression of specific mRNA targets. The relative abundance of miRNAs is linked to function <it>in vivo </it>and miRNA expression patterns are potentially useful signatures for the development of diagnostic, prognostic and therapeutic biomarkers.</p> <p>Finding</p> <p>We compared the performance characteristics of four commercial miRNA array technologies and found that all platforms performed well in separate measures of performance.</p> <p>Conclusions</p> <p>The Ambion and Agilent platforms were more accurate, whereas the Illumina and Exiqon platforms were more specific. Furthermore, the data analysis approach had a large impact on the performance, predominantly by improving precision.</p

R/Bioconductor software for Illumina's Infinium whole-genome genotyping BeadChips

Summary: Illumina produces a number of microarray-based technologies for human genotyping. An Infinium BeadChip is a two-color platform that types between 105 and 106 single nucleotide polymorphisms (SNPs) per sample. Despite being widely used, there is a shortage of open source software to process the raw intensities from this platform into genotype calls. To this end, we have developed the R/Bioconductor package crlmm for analyzing BeadChip data. After careful preprocessing, our software applies the CRLMM algorithm to produce genotype calls, confidence scores and other quality metrics at both the SNP and sample levels. We provide access to the raw summary-level intensity data, allowing users to develop their own methods for genotype calling or copy number analysis if they wish

A search for chameleon particles using a photon regeneration technique

We report the first results from the GammeV search for chameleon particles, which may be created via photon-photon interactions within a strong magnetic field. Chameleons are hypothesized scalar fields that could explain the dark energy problem. We implement a novel technique to create and trap the reflective particles within a jar and to detect them later via their afterglow as they slowly convert back into photons. These measurements provide the first experimental constraints on the couplings of chameleons to photons.Comment: 4 pages, 3 figures, accepted to PRL, minor revisions to introduction and a more quantitative estimate of reflection conditio

Global mRNA expression analysis in myosin II deficient strains of Saccharomyces cerevisiae reveals an impairment of cell integrity functions

<p>Abstract</p> <p>Background</p> <p>The <it>Saccharomyces cerevisiae MYO1 </it>gene encodes the myosin II heavy chain (Myo1p), a protein required for normal cytokinesis in budding yeast. Myo1p deficiency in yeast (<it>myo1Δ</it>) causes a cell separation defect characterized by the formation of attached cells, yet it also causes abnormal budding patterns, formation of enlarged and elongated cells, increased osmotic sensitivity, delocalized chitin deposition, increased chitin synthesis, and hypersensitivity to the chitin synthase III inhibitor Nikkomycin Z. To determine how differential expression of genes is related to these diverse cell wall phenotypes, we analyzed the global mRNA expression profile of <it>myo1Δ </it>strains.</p> <p>Results</p> <p>Global mRNA expression profiles of <it>myo1Δ </it>strains and their corresponding wild type controls were obtained by hybridization to yeast oligonucleotide microarrays. Results for selected genes were confirmed by real time RT-PCR. A total of 547 differentially expressed genes (p ≤ 0.01) were identified with 263 up regulated and 284 down regulated genes in the <it>myo1Δ </it>strains. Gene set enrichment analysis revealed the significant over-representation of genes in the protein biosynthesis and stress response categories. The <it>SLT2/MPK1 </it>gene was up regulated in the microarray, and a <it>myo1Δslt2Δ </it>double mutant was non-viable. Overexpression of ribosomal protein genes <it>RPL30 </it>and <it>RPS31 </it>suppressed the hypersensitivity to Nikkomycin Z and increased the levels of phosphorylated Slt2p in <it>myo1Δ </it>strains. Increased levels of phosphorylated Slt2p were also observed in wild type strains under these conditions.</p> <p>Conclusion</p> <p>Following this analysis of global mRNA expression in yeast <it>myo1Δ </it>strains, we conclude that 547 genes were differentially regulated in <it>myo1Δ </it>strains and that the stress response and protein biosynthesis gene categories were coordinately regulated in this mutant. The <it>SLT2/MPK1 </it>gene was confirmed to be essential for <it>myo1Δ </it>strain viability, supporting that the up regulated stress response genes are regulated by the <it>PKC1 </it>cell integrity pathway. Suppression of Nikkomycin Z hypersensitivity together with Slt2p phosphorylation was caused by the overexpression of ribosomal protein genes <it>RPL30 </it>and <it>RPS31</it>. These ribosomal protein mRNAs were down regulated in the <it>myo1Δ </it>arrays, suggesting that down regulation of ribosomal biogenesis may affect cell integrity in <it>myo1Δ </it>strains.</p

The influence of feature selection methods on accuracy, stability and interpretability of molecular signatures

Motivation: Biomarker discovery from high-dimensional data is a crucial problem with enormous applications in biology and medicine. It is also extremely challenging from a statistical viewpoint, but surprisingly few studies have investigated the relative strengths and weaknesses of the plethora of existing feature selection methods. Methods: We compare 32 feature selection methods on 4 public gene expression datasets for breast cancer prognosis, in terms of predictive performance, stability and functional interpretability of the signatures they produce. Results: We observe that the feature selection method has a significant influence on the accuracy, stability and interpretability of signatures. Simple filter methods generally outperform more complex embedded or wrapper methods, and ensemble feature selection has generally no positive effect. Overall a simple Student's t-test seems to provide the best results. Availability: Code and data are publicly available at http://cbio.ensmp.fr/~ahaury/

Genome-scale DNA methylation mapping of clinical samples at single-nucleotide resolution

August 1, 2010Bisulfite sequencing measures absolute levels of DNA methylation at single-nucleotide resolution, providing a robust platform for molecular diagnostics. Here, we optimize bisulfite sequencing for genome-scale analysis of clinical samples. Specifically, we outline how restriction digestion targets bisulfite sequencing to hotspots of epigenetic regulation; we show that 30ng of DNA are sufficient for genome-scale analysis; we demonstrate that our protocol works well on formalinfixed, paraffin-embedded (FFPE) samples; and we describe a statistical method for assessing significance of altered DNA methylation patterns.National Institutes of Health (U.S.) (Grant R01HG004401)National Institutes of Health (U.S.) (Grant U54HG03067)National Institutes of Health (U.S.) (Grant U01ES017155

Randomization in Laboratory Procedure Is Key to Obtaining Reproducible Microarray Results

The quality of gene expression microarray data has improved dramatically since the first arrays were introduced in the late 1990s. However, the reproducibility of data generated at multiple laboratory sites remains a matter of concern, especially for scientists who are attempting to combine and analyze data from public repositories. We have carried out a study in which a common set of RNA samples was assayed five times in four different laboratories using Affymetrix GeneChip arrays. We observed dramatic differences in the results across laboratories and identified batch effects in array processing as one of the primary causes for these differences. When batch processing of samples is confounded with experimental factors of interest it is not possible to separate their effects, and lists of differentially expressed genes may include many artifacts. This study demonstrates the substantial impact of sample processing on microarray analysis results and underscores the need for randomization in the laboratory as a means to avoid confounding of biological factors with procedural effects