Exaggerated CpH methylation in the autism-affected brain.

BackgroundThe etiology of autism, a complex, heritable, neurodevelopmental disorder, remains largely unexplained. Given the unexplained risk and recent evidence supporting a role for epigenetic mechanisms in the development of autism, we explored the role of CpG and CpH (H = A, C, or T) methylation within the autism-affected cortical brain tissue.MethodsReduced representation bisulfite sequencing (RRBS) was completed, and analysis was carried out in 63 post-mortem cortical brain samples (Brodmann area 19) from 29 autism-affected and 34 control individuals. Analyses to identify single sites that were differentially methylated and to identify any global methylation alterations at either CpG or CpH sites throughout the genome were carried out.ResultsWe report that while no individual site or region of methylation was significantly associated with autism after multi-test correction, methylated CpH dinucleotides were markedly enriched in autism-affected brains (~2-fold enrichment at p < 0.05 cutoff, p = 0.002).ConclusionsThese results further implicate epigenetic alterations in pathobiological mechanisms that underlie autism

RNA-Seq optimization with eQTL gold standards.

BackgroundRNA-Sequencing (RNA-Seq) experiments have been optimized for library preparation, mapping, and gene expression estimation. These methods, however, have revealed weaknesses in the next stages of analysis of differential expression, with results sensitive to systematic sample stratification or, in more extreme cases, to outliers. Further, a method to assess normalization and adjustment measures imposed on the data is lacking.ResultsTo address these issues, we utilize previously published eQTLs as a novel gold standard at the center of a framework that integrates DNA genotypes and RNA-Seq data to optimize analysis and aid in the understanding of genetic variation and gene expression. After detecting sample contamination and sequencing outliers in RNA-Seq data, a set of previously published brain eQTLs was used to determine if sample outlier removal was appropriate. Improved replication of known eQTLs supported removal of these samples in downstream analyses. eQTL replication was further employed to assess normalization methods, covariate inclusion, and gene annotation. This method was validated in an independent RNA-Seq blood data set from the GTEx project and a tissue-appropriate set of eQTLs. eQTL replication in both data sets highlights the necessity of accounting for unknown covariates in RNA-Seq data analysis.ConclusionAs each RNA-Seq experiment is unique with its own experiment-specific limitations, we offer an easily-implementable method that uses the replication of known eQTLs to guide each step in one's data analysis pipeline. In the two data sets presented herein, we highlight not only the necessity of careful outlier detection but also the need to account for unknown covariates in RNA-Seq experiments

The Chicken Yolk Sac IgY Receptor, a Functional Equivalent of the Mammalian MHC-Related Fc Receptor, Is a Phospholipase A2 Receptor Homolog

AbstractIn mammals, IgG is transferred from mother to young by the MHC-related receptor FcRn, which binds IgG in acidic endosomes and releases it at basic pH into blood. Maternal IgY, the avian counterpart of IgG, is transferred to embryos across yolk sac membranes. We affinity-purified the chicken yolk sac IgY receptor (FcRY) and sequenced its gene. FcRY is unrelated to MHC molecules but is a homolog of the mammalian phospholipase A2 receptor. Analytical ultracentrifugation and truncation experiments suggest that FcRY forms a compact structure containing an IgY binding site at acidic pH but undergoes a conformational change at basic pH that disrupts the site. FcRY is thus unrelated to mammalian FcRn in both its structure and mechanism for pH-dependent binding, illustrating distinct routes utilized by evolution to transfer antibodies

Comparative Gene Expression Profiling of Benign and Malignant Lesions Reveals Candidate Therapeutic Compounds for Leiomyosarcoma

Leiomyosarcoma (LMS) is a malignant, soft-tissue tumor for which few effective therapies exist. Previously, we showed that there are three molecular subtypes of LMS. Here, we analyzed genes differentially expressed in each of the three LMS subtypes as compared to benign leiomyomas and then used the Connectivity Map (cmap) to calculate enrichment scores for the 1309 cmap drugs in order to identify candidate molecules with the potential to induce a benign, leiomyoma-like phenotype in LMS cells. 11 drugs were selected and tested for their ability to inhibit the growth of three human LMS cell lines. We identified two drugs with in vitro efficacy against LMS, one of which had a strongly negative enrichment score (Cantharidin) and the other of which had a strongly positive enrichment score (MG-132). Given MG-132's strong inhibitory effect on LMS cell viability, we hypothesized that LMS cells may be sensitive to treatment with other proteasome inhibitors and demonstrated that bortezomib, a clinically-approved proteasome inhibitor not included in the original cmap screen, potently inhibited the viability of the LMS cell lines. These findings suggest that systematically linking LMS subtype-specific expression signatures with drug-associated expression profiles represents a promising approach for the identification of new drugs for LMS

Collaborative development of the Arrowsmith two node search interface designed for laboratory investigators.

Arrowsmith is a unique computer-assisted strategy designed to assist investigators in detecting biologically-relevant connections between two disparate sets of articles in Medline. This paper describes how an inter-institutional consortium of neuroscientists used the UIC Arrowsmith web interface http://arrowsmith.psych.uic.edu in their daily work and guided the development, refinement and expansion of the system into a suite of tools intended for use by the wider scientific community

Using M Dwarf Spectra to Map Extinction in the Local Galaxy

We use spectra of more than 56,000 M dwarfs from the Sloan Digital Sky Survey (SDSS) to create a high-latitude extinction map of the local Galaxy. Our technique compares spectra from the stars in the SDSS Data Release 7 M dwarf sample in low-extinction lines of sight, as determined by Schlegel, Finkbeiner, & Davis, to other SDSS M dwarf spectra in order to derive improved distance estimates and accurate line-of-sight extinctions. Unlike most previous studies, which have used a two-color method to determine extinction, we fit extinction curves to fluxes across the entire spectral range from 5700 to 9200 {\AA} for every star in our sample. Our result is an extinction map that extends from a few tens of pc to approximately 2 kpc away from the sun. We also use a similar technique to create a map of R_V values within approximately 1 kpc of the sun, and find they are consistent with the widely accepted diffuse interstellar medium value of 3.1. Using our extinction data, we derive a dust scale height for the local galaxy of 119\pm15 parsecs and find evidence for a local dust cavity.Comment: 11 pages, 14 figures, accepted for publication in A

Dynamic and redundant regulation of LRRK2 and LRRK1 expression

<p>Abstract</p> <p>Background</p> <p>Mutations within the <it>leucine-rich repeat kinase 2 </it>(<it>LRRK2</it>) gene account for a significant proportion of autosomal-dominant and some late-onset sporadic Parkinson's disease. Elucidation of LRRK2 protein function in health and disease provides an opportunity for deciphering molecular pathways important in neurodegeneration. In mammals, LRRK1 and LRRK2 protein comprise a unique family encoding a GTPase domain that controls intrinsic kinase activity. The expression profiles of the murine LRRK proteins have not been fully described and insufficiently characterized antibodies have produced conflicting results in the literature.</p> <p>Results</p> <p>Herein, we comprehensively evaluate twenty-one commercially available antibodies to the LRRK2 protein using mouse <it>LRRK2 </it>and human <it>LRRK2 </it>expression vectors, wild-type and <it>LRRK2</it>-null mouse brain lysates and human brain lysates. Eleven antibodies detect over-expressed human LRRK2 while four antibodies detect endogenous human LRRK2. In contrast, two antibodies recognize over-expressed mouse LRRK2 and one antibody detected endogenous mouse LRRK2. LRRK2 protein resides in both soluble and detergent soluble protein fractions. <it>LRRK2 </it>and the related <it>LRRK1 </it>genes encode low levels of expressed mRNA species corresponding to low levels of protein both during development and in adulthood with largely redundant expression profiles.</p> <p>Conclusion</p> <p>Despite previously published results, commercially available antibodies generally fail to recognize endogenous mouse LRRK2 protein; however, several antibodies retain the ability to detect over-expressed mouse LRRK2 protein. Over half of the commercially available antibodies tested detect over-expressed human LRRK2 protein and some have sufficient specificity to detect endogenous LRRK2 in human brain. The mammalian LRRK proteins are developmentally regulated in several tissues and coordinated expression suggest possible redundancy in the function between <it>LRRK1 </it>and <it>LRRK2</it>.</p

Sex ratios in the rodent malaria parasite, Plasmodium chabaudi

The sex ratios of malaria and related Apicomplexan parasites play a major role in transmission success. Here, we address 2 fundamental issues in the sex ratios of the rodent malaria parasite, Plasmodium chabaudi. First we test the accuracy of empirical methods for estimating sex ratios in malaria parasites, and show that sex ratios made with standard thin smears may overestimate the proportion of female gametocytes. Secondly, we test whether the mortality rate differs between male and female gametocytes, as assumed by sex ratio theory. Conventional application of sex ratio theory to malaria parasites assumes that the primary sex ratio can be accurately determined from mature gametocytes circulating in the peripheral circulation. We stopped gametocyte production with chloroquine in order to study a cohort of gametocytes in vitro. The mortality rate was significantly higher for female gametocytes, with an average half-life of 8 h for female gametocytes and 16 h for male gametocytes

Dark Matter and Stellar Mass in the Luminous Regions of Disk Galaxies

We investigate the correlations among stellar mass (M_*), disk scale length (R_d), and rotation velocity at 2.2 disk scale lengths (V_2.2) for a sample of 81 disk-dominated galaxies (disk/total >= 0.9) selected from the SDSS. We measure V_2.2 from long-slit H-alpha rotation curves and infer M_* from galaxy i-band luminosities (L_i) and g-r colors. We find logarithmic slopes of 2.60+/-0.13 and 3.05+/-0.12 for the L_i-V_2.2 and M_*-V_2.2 relations, somewhat shallower than most previous studies, with intrinsic scatter of 0.13 dex and 0.16 dex. Our direct estimates of the total-to-stellar mass ratio within 2.2R_d, assuming a Kroupa IMF, yield a median ratio of 2.4 for M_*>10^10 Msun and 4.4 for M_*=10^9-10^10 Msun, with large scatter at a given M_* and R_d. The typical ratio of the rotation speed predicted for the stellar disk alone to the observed rotation speed at 2.2R_d is ~0.65. The distribution of R_d at fixed M_* is broad, but we find no correlation between disk size and the residual from the M_*-V_2.2 relation, implying that this relation is an approximately edge-on view of the disk galaxy fundamental plane. Independent of the assumed IMF, this result implies that stellar disks do not, on average, dominate the mass within 2.2R_d. We discuss our results in the context of infall models of disk formation in cold dark matter halos. A model with a disk-to-halo mass ratio m_d=0.05 provides a reasonable match to the R_d-M_* distribution for spin parameters \lambda ranging from ~0.04-0.08, and it yields a reasonable match to the mean M_*-V_2.2 relation. A model with m_d=0.1 predicts overly strong correlations between disk size and M_*-V_2.2 residual. Explaining the wide range of halo-to-disk mass ratios within 2.2R_d requires significant scatter in m_d values, with systematically lower m_d for galaxies with lower $M_*$.Comment: 18 pages, 2 tables, 7 figures, Accepted to ApJ, Table 1 updated, otherwise minor change