Comparison of data acquisition methods for the identification and quantification of histone post-translational modifications on a Q Exactive HF hybrid quadrupole Orbitrap mass spectrometer.

RATIONALE: Histone PTMs play key roles in regulating eukaryotic gene expression. Mass spectrometry (MS) has emerged as a powerful method to characterize and quantify histone PTMs as it allows unbiased identification and quantification of multiple histone PTMs including combinations of the modifications present. METHODS: In this study we compared a range of data acquisition methods for the identification and quantification of the histone PTMs using a Q Exactive HF Orbitrap. We compared three different data-dependent analysis (DDA) methods with MS2 resolutions of 120K, 60K, 30K. We also compared a range of data-independent analysis (DIA) methods using MS2 isolation windows of 20 m/z and DIAvw to identify and quantify histone PTMs in Chinese Hamster Ovary (CHO) cells. RESULTS: The increased number of MS2 scans afforded by the lower resolution methods resulted in a higher number of queries, peptide sequence matches (PSMs) and a higher number of peptide proteoforms with a Mascot Ion score greater than 46. No difference in the proportion of peptide proteoforms with Delta scores >17 was observed. Comparing the data acquisition methods increased repeatability in terms of lower CVs afforded by DIA MS1 60K MS2 30K 20m/z isolation windows was observed. CONCLUSION: We observed that DIA which offers advantages in flexibility and identification of isobaric peptide proteoforms performs as well as DDA in the analysis of histone PTMs. We were able to identify 71 modified histone peptides for histone H3 and H4 and quantified 64 across each of the different acquisition methods

Physiological responses to folate overproduction in lactobacillys plantarum WCFS1.

<p>Abstract</p> <p>Background</p> <p>Using a functional genomics approach we addressed the impact of folate overproduction on metabolite formation and gene expression in <it>Lactobacillus plantarum </it>WCFS1. We focused specifically on the mechanism that reduces growth rates in folate-overproducing cells.</p> <p>Results</p> <p>Metabolite formation and gene expression were determined in a folate-overproducing- and wild-type strain. Differential metabolomics analysis of intracellular metabolite pools indicated that the pool sizes of 18 metabolites differed significantly between these strains. The gene expression profile was determined for both strains in pH-regulated chemostat culture and batch culture. Apart from the expected overexpression of the 6 genes of the folate gene cluster, no other genes were found to be differentially expressed both in continuous and batch cultures. The discrepancy between the low transcriptome and metabolome response and the 25% growth rate reduction of the folate overproducing strain was further investigated. Folate production per se could be ruled out as a contributing factor, since in the absence of folate production the growth rate of the overproducer was also reduced by 25%. The higher metabolic costs for DNA and RNA biosynthesis in the folate overproducing strain were also ruled out. However, it was demonstrated that folate-specific mRNAs and proteins constitute 8% and 4% of the total mRNA and protein pool, respectively.</p> <p>Conclusion</p> <p>Folate overproduction leads to very little change in metabolite levels or overall transcript profile, while at the same time the growth rate is reduced drastically. This shows that <it>Lactobacillus plantarum </it>WCFS1 is unable to respond to this growth rate reduction, most likely because the growth-related transcripts and proteins are diluted by the enormous amount of gratuitous folate-related transcripts and proteins.</p

Applying phylogenomics to understand the emergence of Shiga Toxin producing Escherichia coli O157:H7 strains causing severe human disease in the United Kingdom

Shiga Toxin producing Escherichia coli (STEC) O157:H7 is a recently emerged zoonotic pathogen with considerable morbidity. Since the serotype emerged in the 1980s, research has focussed on unravelling the evolutionary events from the E. coli O55:H7 ancestor to the contemporaneous globally dispersed strains. In this study the genomes of over 1000 isolates from human clinical cases and cattle, spanning the history of STEC O157:H7 in the United Kingdom were sequenced. Phylogenetic analysis reveals the ancestry, key acquisition events and global context of the strains. Dated phylogenies estimate the time to the most recent common ancestor of the current circulating global clone to 175 years ago, followed by rapid diversification. We show the acquisition of specific virulence determinates occurred relatively recently and coincides with its recent detection in the human population. Using clinical outcome data from 493 cases of STEC O157:H7 we assess the relative risk of severe disease including HUS from each of the defined clades in the population and show the dramatic effect Shiga toxin complement has on virulence. We describe two strain replacement events that have occurred in the cattle population in the UK over the last 30 years; one resulting in a highly virulent strain that has accounted for the majority of clinical cases in the UK over the last decade. This work highlights the need to understand the selection pressures maintaining Shiga-toxin encoding bacteriophages in the ruminant reservoir and the study affirms the requirement for close surveillance of this pathogen in both ruminant and human populations

Understanding service users’ and therapists’ experiences of pharmacological treatment for sexual preoccupation and/or hypersexuality in incarcerated sex offenders

This research comprises two qualitative studies understanding the experiences of 1) convicted sex offenders voluntarily receiving pharmacological treatment to reduce sexual preoccupation and 2) therapists working with these offenders. The studies form part of a research programme evaluating the use of pharmacological treatment with sexual offenders. In study one, semi-structured interviews were conducted with 13 sexual offenders receiving selective serotonin reuptake inhibitors (SSRIs). In study two, interviews were conducted with eight intervention staff with varying levels of experience of working with offenders taking anti-libidinals. Thematic analysis was used and in study one, two main themes emerged: (i) the impact of the pharmacological treatment on prisoners’ daily functioning; (ii) barriers to compliance/engagement. In study two, three main themes emerged: (i) offenders’ reluctance to engage with pharmacological treatment; (ii) challenges for therapists; (iii) pharmacology: ‘just another piece of the puzzle’. Findings are discussed in relation to practice and future research

Characterization of a Mixed Methanotrophic Culture Capable of Chloroethylene Degradation

A consortium of methanotrophs cultured from the St. Joseph's aquifer in Schoolcraft, MI, was found to exhibit similar methane consumption rates as pure cultures of methanotrophs. The methanotrophic consortium resides within a portion of the aquifer contaminated with a mixed waste plume of perchloroethylene (PCE) and its reductive dechlorination products from natural attenuation, trichloroethylene (TCE), cis-dichloroethylene (c-DCE), and vinyl chloride (VC). Oxidation kinetics for TCE, c-DCE, and VC were measured for the mixed methanotroph consortium and compared to reported rate parameters for degradation of these chloroethylene compounds by pure methanotrophic cultures. The results demonstrate that the kinetics of chloroethylene oxidation by the Schoolcraft methanotroph population mimic the degradation rates of pure methanotrophic cultures that primarily express particulate methane monooxygenase (pMMO). Molecular and biochemical analyses confirmed that sMMO was not being expressed by these cells. Rather, using competitive reverse transcriptionpolymerase chain reaction, pmoA, a gene encoding one of the polypeptides of the pMMO was found at a level of (1.57 ± 0.10) × 10–17 mol pmoA mRNA/g wet soil in soil slurries and (2.65 ± 0.43) × 10–17 mol pmoA mRNA/μl in groundwater. No expression of mmoX, a gene encoding one of the polypeptides of the sMMO, was detected.Peer Reviewedhttp://deepblue.lib.umich.edu/bitstream/2027.42/63398/1/ees.2005.22.177.pd

Quantization and Compressive Sensing

Quantization is an essential step in digitizing signals, and, therefore, an indispensable component of any modern acquisition system. This book chapter explores the interaction of quantization and compressive sensing and examines practical quantization strategies for compressive acquisition systems. Specifically, we first provide a brief overview of quantization and examine fundamental performance bounds applicable to any quantization approach. Next, we consider several forms of scalar quantizers, namely uniform, non-uniform, and 1-bit. We provide performance bounds and fundamental analysis, as well as practical quantizer designs and reconstruction algorithms that account for quantization. Furthermore, we provide an overview of Sigma-Delta ($\Sigma\Delta$) quantization in the compressed sensing context, and also discuss implementation issues, recovery algorithms and performance bounds. As we demonstrate, proper accounting for quantization and careful quantizer design has significant impact in the performance of a compressive acquisition system.Comment: 35 pages, 20 figures, to appear in Springer book "Compressed Sensing and Its Applications", 201

Detector Description and Performance for the First Coincidence Observations between LIGO and GEO

For 17 days in August and September 2002, the LIGO and GEO interferometer gravitational wave detectors were operated in coincidence to produce their first data for scientific analysis. Although the detectors were still far from their design sensitivity levels, the data can be used to place better upper limits on the flux of gravitational waves incident on the earth than previous direct measurements. This paper describes the instruments and the data in some detail, as a companion to analysis papers based on the first data.Comment: 41 pages, 9 figures 17 Sept 03: author list amended, minor editorial change

Search for Gravitational Waves from Primordial Black Hole Binary Coalescences in the Galactic Halo

We use data from the second science run of the LIGO gravitational-wave detectors to search for the gravitational waves from primordial black hole (PBH) binary coalescence with component masses in the range 0.2--$1.0 M_\odot$. The analysis requires a signal to be found in the data from both LIGO observatories, according to a set of coincidence criteria. No inspiral signals were found. Assuming a spherical halo with core radius 5 kpc extending to 50 kpc containing non-spinning black holes with masses in the range 0.2--$1.0 M_\odot$, we place an observational upper limit on the rate of PBH coalescence of 63 per year per Milky Way halo (MWH) with 90% confidence.Comment: 7 pages, 4 figures, to be submitted to Phys. Rev.

The backward-bending commute times of married women with household responsibility

The purpose of this paper is to examine theoretically and empirically whether the commute times of married women follow a backward-bending pattern with respect to wage rates. The existing literature has shown that married women tend to choose short commutes because of their relatively low wages combined with comparatively heavy household responsibilities. However, a workleisure model, which includes the simultaneous decision wives take regarding commute times and wage rates, suggests that married women employed in highly paid positions also undertake short commutes, while married women with wage rates in the middle range choose long commutes. These results suggest that the commute times of married women display a backward-bending pattern. Applying an instrumental variable strategy that accounts for the endogeneity of wage rates, the empirical results for employed married women in Japan appear to support this nding. Moreover, one of our results suggests that highly paid married women can still secure greater leisure time with short commutes, despite retaining a heavy load of domestic responsibilities.Working Paper, No.234, 2008.9.1版http://hdl.handle.net/10110/254

История развития физической культуры и спорта на Урале в дореволюционный период

На сегодняшний день становится чрезвычайно актуальным рассмотрение феномена физической культуры и спорта сквозь призму принципа историзма. Существует еще много неизвестного в истории физической культуры, что требует переоценки событий, фактов с позиции современност