Hydrogen bonding of nitroxide spin labels in membrane proteins

On the basis of experiments at 275 GHz, we reconsider the dependence of the continuous-wave EPR spectra of nitroxide spin-labeled protein sites in sensory- and bacteriorhodopsin on the micro-environment. The high magnetic field provides the resolution necessary to disentangle the effects of hydrogen bonding and polarity. In the gxx region of the 275 GHz EPR spectrum, bands are resolved that derive from spin-label populations carrying no, one or two hydrogen bonds. The gxx value of each population varies hardly from site to site, significantly less than deduced previously from studies at lower microwave frequencies. The fractions of the populations vary strongly, which provides a consistent description of the variation of the average gxx and the average nitrogen-hyperfine interaction Azz from site to site. These variations reflect the difference in the proticity of the micro-environment, and differences in polarity contribute marginally. Concomitant W-band ELDOR- detected NMR experiments on the corresponding nitroxide in perdeuterated water resolve population-specific nitrogen-hyperfine bands, which underlies the interpretation for the proteins

PhyloPat: an updated version of the phylogenetic pattern database contains gene neighborhood

Phylogenetic patterns show the presence or absence of certain genes in a set of full genomes derived from different species. They can also be used to determine sets of genes that occur only in certain evolutionary branches. Previously, we presented a database named PhyloPat which allows the complete Ensembl gene database to be queried using phylogenetic patterns. Here, we describe an updated version of PhyloPat which can be queried by an improved web server. We used a single linkage clustering algorithm to create 241 697 phylogenetic lineages, using all the orthologies provided by Ensembl v49. PhyloPat offers the possibility of querying with binary phylogenetic patterns or regular expressions, or through a phylogenetic tree of the 39 included species. Users can also input a list of Ensembl, EMBL, EntrezGene or HGNC IDs to check which phylogenetic lineage any gene belongs to. A link to the FatiGO web interface has been incorporated in the HTML output. For each gene, the surrounding genes on the chromosome, color coded according to their phylogenetic lineage can be viewed, as well as FASTA files of the peptide sequences of each lineage. Furthermore, lists of omnipresent, polypresent, oligopresent and anticorrelating genes have been included. PhyloPat is freely available at http://www.cmbi.ru.nl/phylopat

Electrochemical synthesis of peroxomonophosphate using boron-doped diamond anodes

A new method for the synthesis of peroxomonophosphate, based on the use of boron-doped diamond electrodes, is described. The amount of oxidant electrogenerated depends on the characteristics of the supporting media (pH and solute concentration) and on the operating conditions (temperature and current density). Results show that the pH, between values of 1 and 5, does not influence either the electrosynthesis of peroxomonophosphate or the chemical stability of the oxidant generated. Conversely, low temperatures are required during the electrosynthesis process to minimize the thermal decomposition of peroxomonophosphate and to guarantee significant oxidant concentration. In addition, a marked influence of both the current density and the initial substrate is observed. This observation can be explained in terms of the contribution of hydroxyl radicals in the oxidation mechanisms that occur on diamond surfaces. In the assays carried out below the water oxidation potential, the generation of hydroxyl radicals did not take place. In these cases, peroxomonophosphate generation occurs through a direct electron transfer and, therefore, at these low current densities lower concentrations are obtained. On the other hand, at higher potentials both direct and hydroxyl radical-mediated mechanisms contribute to the oxidant generation and the process is more efficient. In the same way, the contribution of hydroxyl radicals may also help to explain the significant influence of the substrate concentration. Thus, the coexistence of both phosphate and hydroxyl radicals is required to ensure the generation of significant amounts of peroxomonophosphoric acid

PCR clonality detection in Hodgkin lymphoma

B-cell clonality detection in whole tissue is considered indicative of B-cell non-Hodgkin lymphoma (NHL). We tested frozen tissue of 24 classical Hodgkin lymphomas (cHL) with a varying tumor cell load with the multiplex polymerase chain reaction (PCR) primer sets for IGH and IGK gene rearrangement (BIOMED-2). A clonal population was found in 13 cases with the IGH FR1 and/or FR2/FR3 PCRs. Using the IGK-VJ and IGK-DE PCRs, an additional six cases had a dominant clonal cell population, resulting in a detection rate of 79% in frozen tissue. Of 12 cases, also the formalin-fixed and paraffin-embedded (FFPE) tissue was tested. Surprisingly, in eight of the 12 FFPE cases with acceptable DNA quality (allowing PCR amplification of >200 nt fragments), the IGK multiplex PCRs performed better in detecting clonality (six out of eight clonal IGK rearrangements) than the IGH PCRs (four out of nine clonal rearrangements), despite a rather large amplicon size. There was no evidence of B-cell lymphoma during follow-up of 1 to 6 years and no correlation was found between the presence of a clonal result and Epstein–Barr virus in the tumor cells. Our results indicate that the present routine PCR methods are sensitive enough to detect small numbers of malignant cells in cHL. Therefore, the presence of a clonal B-cell population does not differentiate between cHL and NHL

Integration of GWAS, CNV and sele ction signature reveals candidate genes for abdominal fat regulation in chickens.

Abstract: Carcass fat content is an economically important trait in commercial chickens. The use of genome-wide high density SNPs may improve the power and resolution to identify QTLs, putative candidate genes and copy number variations (CNVs), for selection programs. The main goal of this study was to identify genomic windows and putative candidate genes for carcass fat content. We checked the overlap of QTL with regions demonstrating signatures of selection and inherited CNVs identified in the same population. A total of 497 42 day-old chickens from the EMBRAPA F2 Chicken Resource Population developed for QTL studies were genotyped with the 600K SNP genotyping array (Affymetrix®), and phenotyped for carcass fat content weight (CFCW) and carcass fat content on a dry matter basis (CFCDM). After quality control, a total of 480 samples and 371,557 SNPs annotated in autosomal chromosomes (GGA1-28) based on Gallus_gallus-5.0 (NCBI) were kept for further analysis. GWAS analyses were performed with GenSel software using BayesB method (π=0.9988) to identify genomic windows associated with CFCW or CFC%. We identified 15 genomic windows associated with CFC% on GGA1, 7, 15, 20 and 28, and from those, we identified two adjacent windows on GGA7 considered as the same QTL explaining 1.31 and 2.18% of the genetic variance for CFCW and CFC%, respectively. This QTL overlapped with one regions previsiouly know to regulate abdominal fat in chickens and the QTL region encompassed two putative candidate genes overlapping with signatures of selection and inherited CNVs. Our findings are helpful to better understand the genetic regulation of fatness in chickens. Resumo: O teor de gordura na carcaça é uma característica economicamente importante em frangos comerciais. O uso de SNPs de alta densidade em todo o genoma pode melhorar o poder e a resolução para identificar QTLs, genes candidatos putativos e variações no número de cópias (CNVs), para programas de seleção. O principal objetivo deste estudo foi identificar janelas genômicas e possíveis genes candidatos para o conteúdo de gordura na carcaça. Verificamos a sobreposição de QTL com regiões demonstrando assinaturas de seleção e CNVs herdadas identificadas na mesma população. Um total de 497 galinhas com 42 dias de idade da EMBRAPA F2 Chicken Resource Population desenvolvidas para estudos QTL foram genotipadas com o arranjo de genótipos SNP 600K (Affymetrix®) e fenotipadas para peso de gordura na carcaça (CFCW) e teor de gordura na carcaça seca. matéria básica (CFCDM). Após o controle de qualidade, um total de 480 amostras e 371.557 SNPs anotados em cromossomos autossômicos (GGA1-28) baseados em Gallus_gallus-5.0 (NCBI) foram mantidos para análise posterior. As análises de GWAS foram realizadas com o software GenSel usando o método de BayesB (π = 0,9988) para identificar janelas genômicas associadas ao CFCW ou CFC%. Foram identificadas 15 janelas genômicas associadas a% CFC em GGA1, 7, 15, 20 e 28 e, a partir delas, identificamos duas janelas adjacentes em GGA7 consideradas como os mesmos QTLs explicando 1,31 e 2,18% da variância genética para CFCW e CFC% , respectivamente. Este QTL se sobrepunha a uma das regiões previsamente conhecidas para regular a gordura abdominal em frangos e a região QTL englobava dois supostos genes candidatos que se sobrepunham com assinaturas de seleção e CNVs herdadas. Nossas descobertas são úteis para entender melhor a regulação genética da gordura em frangos

Simplification and Shift in Cognition of Political Difference: Applying the Geometric Modeling to the Analysis of Semantic Similarity Judgment

Perceiving differences by means of spatial analogies is intrinsic to human cognition. Multi-dimensional scaling (MDS) analysis based on Minkowski geometry has been used primarily on data on sensory similarity judgments, leaving judgments on abstractive differences unanalyzed. Indeed, analysts have failed to find appropriate experimental or real-life data in this regard. Our MDS analysis used survey data on political scientists' judgments of the similarities and differences between political positions expressed in terms of distance. Both distance smoothing and majorization techniques were applied to a three-way dataset of similarity judgments provided by at least seven experts on at least five parties' positions on at least seven policies (i.e., originally yielding 245 dimensions) to substantially reduce the risk of local minima. The analysis found two dimensions, which were sufficient for mapping differences, and fit the city-block dimensions better than the Euclidean metric in all datasets obtained from 13 countries. Most city-block dimensions were highly correlated with the simplified criterion (i.e., the left–right ideology) for differences that are actually used in real politics. The isometry of the city-block and dominance metrics in two-dimensional space carries further implications. More specifically, individuals may pay attention to two dimensions (if represented in the city-block metric) or focus on a single dimension (if represented in the dominance metric) when judging differences between the same objects. Switching between metrics may be expected to occur during cognitive processing as frequently as the apparent discontinuities and shifts in human attention that may underlie changing judgments in real situations occur. Consequently, the result has extended strong support for the validity of the geometric models to represent an important social cognition, i.e., the one of political differences, which is deeply rooted in human nature

Third Report on Chicken Genes and Chromosomes 2015

Following on from the First Report on Chicken Genes and Chromosomes [Schmid et al., 2000] and the Second Report in 2005 [Schmid et al., 2005], we are pleased to publish this long-awaited Third Report on the latest developments in chicken genomics. The First Report highlighted the availability of genetic and physical maps, while the Second Report was published as the chicken genome sequence was released. This report comes at a time of huge technological advances (particularly in sequencing methodologies) which have allowed us to examine the chicken genome in detail not possible until now. This has also heralded an explosion in avian genomics, with the current availability of more than 48 bird genomes [Zhang G et al., 2014b; Eöry et al., 2015], with many more planned

Phonons in Slow Motion: Dispersion Relations in Ultra-Thin Si Membranes

We report the changes in dispersion relations of hypersonic acoustic phonons in free-standing silicon membranes as thin as \sim 8 nm. We observe a reduction of the phase and group velocities of the fundamental flexural mode by more than one order of magnitude compared to bulk values. The modification of the dispersion relation in nanostructures has important consequences for noise control in nano and micro-electromechanical systems (MEMS/NEMS) as well as opto-mechanical devices.Comment: 5 page

Two new rapid SNP-typing methods for classifying Mycobacterium tuberculosis complex into the main phylogenetic lineages

There is increasing evidence that strain variation in Mycobacterium tuberculosis complex (MTBC) might influence the outcome of tuberculosis infection and disease. To assess genotype-phenotype associations, phylogenetically robust molecular markers and appropriate genotyping tools are required. Most current genotyping methods for MTBC are based on mobile or repetitive DNA elements. Because these elements are prone to convergent evolution, the corresponding genotyping techniques are suboptimal for phylogenetic studies and strain classification. By contrast, single nucleotide polymorphisms (SNP) are ideal markers for classifying MTBC into phylogenetic lineages, as they exhibit very low degrees of homoplasy. In this study, we developed two complementary SNP-based genotyping methods to classify strains into the six main human-associated lineages of MTBC, the 'Beijing' sublineage, and the clade comprising Mycobacterium bovis and Mycobacterium caprae. Phylogenetically informative SNPs were obtained from 22 MTBC whole-genome sequences. The first assay, referred to as MOL-PCR, is a ligation-dependent PCR with signal detection by fluorescent microspheres and a Luminex flow cytometer, which simultaneously interrogates eight SNPs. The second assay is based on six individual TaqMan real-time PCR assays for singleplex SNP-typing. We compared MOL-PCR and TaqMan results in two panels of clinical MTBC isolates. Both methods agreed fully when assigning 36 well-characterized strains into the main phylogenetic lineages. The sensitivity in allele-calling was 98.6% and 98.8% for MOL-PCR and TaqMan, respectively. Typing of an additional panel of 78 unknown clinical isolates revealed 99.2% and 100% sensitivity in allele-calling, respectively, and 100% agreement in lineage assignment between both methods. While MOL-PCR and TaqMan are both highly sensitive and specific, MOL-PCR is ideal for classification of isolates with no previous information, whereas TaqMan is faster for confirmation. Furthermore, both methods are rapid, flexible and comparably inexpensive