ALBERT

Beschreibung: Measurements of ground deformation can be used to identify and interpret geophysical processes occurring at volcanoes. Most studies rely on a single geodetic technique, or fit a geophysical model to the results of multiple geodetic techniques. Here we present a methodology that combines GPS, Total Station measurements and InSAR into a single reference frame to produce an integrated 3-D geodetic velocity surface without any prior geophysical assumptions. The methodology consists of five steps: design of the network, acquisition and processing of the data, spatial integration of the measurements, time series computation and finally the integration of spatial and temporal measurements. The most significant improvements of this method are (1) the reduction of the required field time, (2) the unambiguous detection of outliers, (3) an increased measurement accuracy and (4) the construction of a 3-D geodetic velocity field. We apply this methodology to ongoing motion on Arenal's western flank. Integration of multiple measurement techniques at Arenal volcano revealed a deformation field that is more complex than that described by individual geodetic techniques, yet remains consistent with previous studies. This approach can be applied to volcano monitoring worldwide and has the potential to be extended to incorporate other geodetic techniques and to study transient deformation.

Beschreibung: Combinatorial transcription factor (TF) binding is essential for cell-type-specific gene regulation. However, much remains to be learned about the mechanisms of TF interactions, including to what extent constrained spacing and orientation of interacting TFs are critical for regulatory element activity. To examine the relative prevalence of the ‘enhanceosome’ versus the ‘TF collective’ model of combinatorial TF binding, a comprehensive analysis of TF binding site sequences in large scale datasets is necessary. We developed a motif-pair discovery pipeline to identify motif co-occurrences with preferential distance(s) between motifs in TF-bound regions. Utilizing a compendium of 289 mouse haematopoietic TF ChIP-seq datasets, we demonstrate that haematopoietic-related motif-pairs commonly occur with highly conserved constrained spacing and orientation between motifs. Furthermore, motif clustering revealed specific associations for both heterotypic and homotypic motif-pairs with particular haematopoietic cell types. We also showed that disrupting the spacing between motif-pairs significantly affects transcriptional activity in a well-known motif-pair—E-box and GATA, and in two previously unknown motif-pairs with constrained spacing—Ets and Homeobox as well as Ets and E-box. In this study, we provide evidence for widespread sequence-specific TF pair interaction with DNA that conforms to the ‘enhanceosome’ model, and furthermore identify associations between specific haematopoietic cell-types and motif-pairs.

Beschreibung: In autumn 2012, the new release 05 (RL05) of monthly geopotencial spherical harmonics Stokes coefficients (SC) from Gravity Recovery and Climate Experiment (GRACE) mission was published. This release reduces the noise in high degree and order SC, but they still need to be filtered. One of the most common filtering processing is the combination of decorrelation and Gaussian filters. Both of them are parameters dependent and must be tuned by the users. Previous studies have analyzed the parameters choice for the RL05 GRACE data for oceanic applications, and for RL04 data for global application. This study updates the latter for RL05 data extending the statistics analysis. The choice of the parameters of the decorrelation filter has been optimized to: (1) balance the noise reduction and the geophysical signal attenuation produced by the filtering process; (2) minimize the differences between GRACE and model-based data and (3) maximize the ratio of variability between continents and oceans. The Gaussian filter has been optimized following the latter criteria. Besides, an anisotropic filter, the fan filter, has been analyzed as an alternative to the Gauss filter, producing better statistics.

Beschreibung: Homologous non-coding RNAs frequently exhibit domain insertions, where a branch of secondary structure is inserted in a sequence with respect to its homologs. Dynamic programming algorithms for common secondary structure prediction of multiple RNA homologs, however, do not account for these domain insertions. This paper introduces a novel dynamic programming algorithm methodology that explicitly accounts for the possibility of inserted domains when predicting common RNA secondary structures. The algorithm is implemented as Dynalign II, an update to the Dynalign software package for predicting the common secondary structure of two RNA homologs. This update is accomplished with negligible increase in computational cost. Benchmarks on ncRNA families with domain insertions validate the method. Over base pairs occurring in inserted domains, Dynalign II improves accuracy over Dynalign, attaining 80.8% sensitivity (compared with 14.4% for Dynalign) and 91.4% positive predictive value (PPV) for tRNA; 66.5% sensitivity (compared with 38.9% for Dynalign) and 57.0% PPV for RNase P RNA; and 50.1% sensitivity (compared with 24.3% for Dynalign) and 58.5% PPV for SRP RNA. Compared with Dynalign, Dynalign II also exhibits statistically significant improvements in overall sensitivity and PPV. Dynalign II is available as a component of RNAstructure, which can be downloaded from http://rna.urmc.rochester.edu/RNAstructure.html .

Beschreibung: Genome-wide assessment of protein–DNA interaction by chromatin immunoprecipitation followed by massive parallel sequencing (ChIP-seq) is a key technology for studying transcription factor (TF) localization and regulation of gene expression. Signal-to-noise-ratio and signal specificity in ChIP-seq studies depend on many variables, including antibody affinity and specificity. Thus far, efforts to improve antibody reagents for ChIP-seq experiments have focused mainly on generating higher quality antibodies. Here we introduce KOIN (knockout implemented normalization) as a novel strategy to increase signal specificity and reduce noise by using TF knockout mice as a critical control for ChIP-seq data experiments. Additionally, KOIN can identify ‘hyper ChIPable regions’ as another source of false-positive signals. As the use of the KOIN algorithm reduces false-positive results and thereby prevents misinterpretation of ChIP-seq data, it should be considered as the gold standard for future ChIP-seq analyses, particularly when developing ChIP-assays with novel antibody reagents.

Beschreibung: DNA methylation is an important epigenetic modification that has essential roles in cellular processes including gene regulation, development and disease and is widely dysregulated in most types of cancer. Recent advances in sequencing technology have enabled the measurement of DNA methylation at single nucleotide resolution through methods such as whole-genome bisulfite sequencing and reduced representation bisulfite sequencing. In DNA methylation studies, a key task is to identify differences under distinct biological contexts, for example, between tumor and normal tissue. A challenge in sequencing studies is that the number of biological replicates is often limited by the costs of sequencing. The small number of replicates leads to unstable variance estimation, which can reduce accuracy to detect differentially methylated loci (DML). Here we propose a novel statistical method to detect DML when comparing two treatment groups. The sequencing counts are described by a lognormal-beta-binomial hierarchical model, which provides a basis for information sharing across different CpG sites. A Wald test is developed for hypothesis testing at each CpG site. Simulation results show that the proposed method yields improved DML detection compared to existing methods, particularly when the number of replicates is low. The proposed method is implemented in the Bioconductor package DSS.

Beschreibung: Determining the taxonomic affiliation of sequences assembled from metagenomes remains a major bottleneck that affects research across the fields of environmental, clinical and evolutionary microbiology. Here, we introduce MyTaxa, a homology-based bioinformatics framework to classify metagenomic and genomic sequences with unprecedented accuracy. The distinguishing aspect of MyTaxa is that it employs all genes present in an unknown sequence as classifiers, weighting each gene based on its (predetermined) classifying power at a given taxonomic level and frequency of horizontal gene transfer. MyTaxa also implements a novel classification scheme based on the genome-aggregate average amino acid identity concept to determine the degree of novelty of sequences representing uncharacterized taxa, i.e. whether they represent novel species, genera or phyla. Application of MyTaxa on in silico generated (mock) and real metagenomes of varied read length (100–2000 bp) revealed that it correctly classified at least 5% more sequences than any other tool. The analysis also showed that ~10% of the assembled sequences from human gut metagenomes represent novel species with no sequenced representatives, several of which were highly abundant in situ such as members of the Prevotella genus. Thus, MyTaxa can find several important applications in microbial identification and diversity studies.

Beschreibung: Increasing numbers of protein structures are solved each year, but many of these structures belong to proteins whose sequences are homologous to sequences in the Protein Data Bank. Nevertheless, the structures of homologous proteins belonging to the same family contain useful information because functionally important residues are expected to preserve physico-chemical, structural and energetic features. This information forms the basis of our method, which detects RNA-binding residues of a given RNA-binding protein as those residues that preserve physico-chemical, structural and energetic features in its homologs. Tests on 81 RNA-bound and 35 RNA-free protein structures showed that our method yields a higher fraction of true RNA-binding residues (higher precision) than two structure-based and two sequence-based machine-learning methods. Because the method requires no training data set and has no parameters, its precision does not degrade when applied to ‘novel’ protein sequences unlike methods that are parameterized for a given training data set. It was used to predict the ‘unknown’ RNA-binding residues in the C-terminal RNA-binding domain of human CPEB3. The two predicted residues, F430 and F474, were experimentally verified to bind RNA, in particular F430, whose mutation to alanine or asparagine nearly abolished RNA binding. The method has been implemented in a webserver called DR_bind1, which is freely available with no login requirement at http://drbind.limlab.ibms.sinica.edu.tw .

Beschreibung: Epigenetic regulation of gene expression involves, besides DNA and histone modifications, the relative positioning of DNA sequences within the nucleus. To trace specific DNA sequences in living cells, we used programmable sequence-specific DNA binding of designer transcription activator-like effectors (dTALEs). We designed a recombinant dTALE (msTALE) with variable repeat domains to specifically bind a 19-bp target sequence of major satellite DNA. The msTALE was fused with green fluorescent protein (GFP) and stably expressed in mouse embryonic stem cells. Hybridization with a major satellite probe (3D-fluorescent in situ hybridization) and co-staining for known cellular structures confirmed in vivo binding of the GFP-msTALE to major satellite DNA present at nuclear chromocenters. Dual tracing of major satellite DNA and the replication machinery throughout S-phase showed co-localization during mid to late S-phase, directly demonstrating the late replication timing of major satellite DNA. Fluorescence bleaching experiments indicated a relatively stable but still dynamic binding, with mean residence times in the range of minutes. Fluorescently labeled dTALEs open new perspectives to target and trace DNA sequences and to monitor dynamic changes in subnuclear positioning as well as interactions with functional nuclear structures during cell cycle progression and cellular differentiation.

Beschreibung: Coupling bisulfite conversion with next-generation sequencing (Bisulfite-seq) enables genome-wide measurement of DNA methylation, but poses unique challenges for mapping. However, despite a proliferation of Bisulfite-seq mapping tools, no systematic comparison of their genomic coverage and quantitative accuracy has been reported. We sequenced bisulfite-converted DNA from two tissues from each of two healthy human adults and systematically compared five widely used Bisulfite-seq mapping algorithms: Bismark, BSMAP, Pash, BatMeth and BS Seeker. We evaluated their computational speed and genomic coverage and verified their percentage methylation estimates. With the exception of BatMeth, all mappers covered 〉70% of CpG sites genome-wide and yielded highly concordant estimates of percentage methylation ( r 2 ≥ 0.95). Fourfold variation in mapping time was found between BSMAP (fastest) and Pash (slowest). In each library, 8–12% of genomic regions covered by Bismark and Pash were not covered by BSMAP. An experiment using simulated reads confirmed that Pash has an exceptional ability to uniquely map reads in genomic regions of structural variation. Independent verification by bisulfite pyrosequencing generally confirmed the percentage methylation estimates by the mappers. Of these algorithms, Bismark provides an attractive combination of processing speed, genomic coverage and quantitative accuracy, whereas Pash offers considerably higher genomic coverage.

Beschreibung: Three-dimensional organization of chromatin is fundamental for transcriptional regulation. Tissue-specific transcriptional programs are orchestrated by transcription factors and epigenetic regulators. The RUNX2 transcription factor is required for differentiation of precursor cells into mature osteoblasts. Although organization and control of the bone-specific Runx2-P1 promoter have been studied extensively, long-range regulation has not been explored. In this study, we investigated higher-order organization of the Runx2-P1 promoter during osteoblast differentiation. Mining the ENCODE database revealed interactions between Runx2-P1 and  Supt3h promoters in several non-mesenchymal human cell lines. Supt3h is a ubiquitously expressed gene located within the first intron of Runx2 . These two genes show shared synteny across species from humans to sponges. Chromosome conformation capture analysis in the murine pre-osteoblastic MC3T3-E1 cell line revealed increased contact frequency between Runx2-P1 and Supt3h promoters during differentiation. This increase was accompanied by enhanced DNaseI hypersensitivity along with RUNX2 and CTCF binding at the Supt3h promoter. Furthermore, interplasmid-3C and luciferase reporter assays showed that the Supt3h promoter can modulate Runx2-P1 activity via direct association. Taken together, our data demonstrate physical proximity between Runx2-P1 and Supt3h promoters, consistent with their syntenic nature. Importantly, we identify the Supt3h promoter as a potential regulator of the bone-specific Runx2-P1 promoter .

Beschreibung: Nanotechnology and synthetic biology currently constitute one of the most innovative, interdisciplinary fields of research, poised to radically transform society in the 21st century. This paper concerns the synthetic design of ribonucleic acid molecules, using our recent algorithm, RNAiFold , which can determine all RNA sequences whose minimum free energy secondary structure is a user-specified target structure. Using RNAiFold , we design ten cis -cleaving hammerhead ribozymes, all of which are shown to be functional by a cleavage assay. We additionally use RNAiFold to design a functional cis -cleaving hammerhead as a modular unit of a synthetic larger RNA. Analysis of kinetics on this small set of hammerheads suggests that cleavage rate of computationally designed ribozymes may be correlated with positional entropy, ensemble defect, structural flexibility/rigidity and related measures. Artificial ribozymes have been designed in the past either manually or by SELEX (Systematic Evolution of Ligands by Exponential Enrichment); however, this appears to be the first purely computational design and experimental validation of novel functional ribozymes. RNAiFold is available at http://bioinformatics.bc.edu/clotelab/RNAiFold/ .

Beschreibung: The paper in question by Van Camp and co-authors [MVC] challenges previous work showing that ground gravity data arising from hydrology can provide a consistent signal for the comparison with satellite gravity data. The data sets used are similar to those used previously, that is, the gravity field as measured by the GRACE satellites versus ground-based data from superconducting gravimeters (SGs) over the same continental area, in this case Central Europe. One of the main impediments in this paper is the presentation that is frequently confusing and misleading as to what the data analysis really shows, for example, the irregular treatment of annual components that are first subtracted then reappear in the analysis. More importantly, we disagree on specific points. Two calculations are included in our comment to illustrate where we believe that the processing in [MVC] paper is deficient. The first deals with their erroneous treatment of the global hydrology using a truncated spherical harmonic approach which explains almost a factor 2 error in their computation of the loading. The second shows the effect of making the wrong assumption in the GRACE/hydrology/surface gravity comparison by inverting the whole of the hydrology loading for underground stations. We also challenge their claims that empirical orthogonal function techniques cannot be done in the presence of periodic components, and that SG data cannot be corrected for comparisons with GRACE data. The main conclusion of their paper, that there is little coherence between ground gravity stations and this invalidates GRACE comparisons, is therefore questionable. There is nothing in [MVC] that contradicts any of the previous papers that have shown clearly a strong relation between seasonal signals obtained from both ground gravity and GRACE satellite data.

Beschreibung: The influence of changes in surface ice-mass redistribution and associated viscoelastic response of the Earth, known as glacial isostatic adjustment (GIA), on the Earth's rotational dynamics has long been known. Equally important is the effect of the changes in the rotational dynamics on the viscoelastic deformation of the Earth. This signal, known as the rotational feedback, or more precisely, the rotational feedback on the sea level equation, has been mathematically described by the sea level equation extended for the term that is proportional to perturbation in the centrifugal potential and the second-degree tidal Love number. The perturbation in the centrifugal force due to changes in the Earth's rotational dynamics enters not only into the sea level equation, but also into the conservation law of linear momentum such that the internal viscoelastic force, the perturbation in the gravitational force and the perturbation in the centrifugal force are in balance. Adding the centrifugal-force perturbation to the linear-momentum balance creates an additional rotational feedback on the viscoelastic deformations of the Earth. We term this feedback mechanism, which is studied in this paper, as the rotational feedback on the linear-momentum balance. We extend both the time-domain method for modelling the GIA response of laterally heterogeneous earth models developed by Martinec and the traditional Laplace-domain method for modelling the GIA-induced rotational response to surface loading by considering the rotational feedback on linear-momentum balance. The correctness of the mathematical extensions of the methods is validated numerically by comparing the polar-motion response to the GIA process and the rotationally induced degree 2 and order 1 spherical harmonic component of the surface vertical displacement and gravity field. We present the difference between the case where the rotational feedback on linear-momentum balance is considered against that where it is not. Numerical simulations show that the resulting difference in radial displacement and sea level change between these situations since the Last Glacial Maximum reaches values of ±25 and ±1.8 m, respectively. Furthermore, the surface deformation pattern is modified by up to 10 per cent in areas of former or ongoing glaciation, but by up to 50 per cent at the bottom of the southern Indian ocean. This also results in the movement of coastlines during the last deglaciation to differ between the two cases due to the difference in the ocean loading, which is seen for instance in the area around Hudson Bay, Canada and along the Chinese, Australian or Argentinian coastlines.

Beschreibung: Passive interferometry technology is based on the relation between the reflection and the transmission responses of the subsurface. The transmission response can be received at surface in the presence of the ambient noise source in the subsurface with the cross-correlation (CC) or multidimensional deconvolution methods. We investigate the feasibility of electromagnetic (EM) wave passive interferometry with CC method. We design a 2-D finite-difference time domain (FDTD) algorithm to simulate the long-duration ground penetrating radar (GPR) measurements with random distribution of passive EM sources. The noise sources have random duration time, waveform and spatial distribution. We test the FDTD GPR passive interferometry code with above source characteristics and apply the method to light non-aqueous phase liquid (LNAPL) monitoring. Based on the model simulation data, by using common midpoint velocity analysis and normal move out correction to process the interferometry retrieve record, we can accurately obtain the dynamic changing characteristics of the target's permittivity. The LNAPL dynamic leakage model can be imaged as well. The synthetic results demonstrate that the GPR passive interferometry is feasible in subsurface LNAPL monitoring. Our work provides a foundation for a passive interferometry field application using GPR.

Beschreibung: 3-D electrical resistivity surveys and inversion models are required to accurately resolve structures in areas with very complex geology where 2-D models might suffer from artefacts. Many 3-D surveys use a grid where the number of electrodes along one direction ( x ) is much greater than in the perpendicular direction ( y ). Frequently, due to limitations in the number of independent electrodes in the multi-electrode system, the surveys use a roll-along system with a small number of parallel survey lines aligned along the x -direction. The ‘Compare R’ array optimization method previously used for 2-D surveys is adapted for such 3-D surveys. Offset versions of the inline arrays used in 2-D surveys are included in the number of possible arrays (the comprehensive data set) to improve the sensitivity to structures in between the lines. The array geometric factor and its relative error are used to filter out potentially unstable arrays in the construction of the comprehensive data set. Comparisons of the conventional (consisting of dipole-dipole and Wenner–Schlumberger arrays) and optimized arrays are made using a synthetic model and experimental measurements in a tank. The tests show that structures located between the lines are better resolved with the optimized arrays. The optimized arrays also have significantly better depth resolution compared to the conventional arrays.

Beschreibung: We apply a reversible-jump Markov chain Monte Carlo method to sample the Bayesian posterior model probability density function of 2-D seafloor resistivity as constrained by marine controlled source electromagnetic data. This density function of earth models conveys information on which parts of the model space are illuminated by the data. Whereas conventional gradient-based inversion approaches require subjective regularization choices to stabilize this highly non-linear and non-unique inverse problem and provide only a single solution with no model uncertainty information, the method we use entirely avoids model regularization. The result of our approach is an ensemble of models that can be visualized and queried to provide meaningful information about the sensitivity of the data to the subsurface, and the level of resolution of model parameters. We represent models in 2-D using a Voronoi cell parametrization. To make the 2-D problem practical, we use a source–receiver common midpoint approximation with 1-D forward modelling. Our algorithm is transdimensional and self-parametrizing where the number of resistivity cells within a 2-D depth section is variable, as are their positions and geometries. Two synthetic studies demonstrate the algorithm's use in the appraisal of a thin, segmented, resistive reservoir which makes for a challenging exploration target. As a demonstration example, we apply our method to survey data collected over the Scarborough gas field on the Northwest Australian shelf.

Beschreibung: Compressional and shear wave seismic measurements were performed in an old railway tunnel and in galleries excavated in a 250-m-thick Toarcian claystone formation in the Tournemire experimental station (France). Three component (3C) geophones and three orthogonal orientations of the vibroseismic force source were used. Additionally, vertical seismic profiling (VSP) measurements were recorded with a 3C borehole geophone, a hydrophone and a microphone in a 159 m deep borehole (ID180) in the tunnel. The seismic data show that Toarcian claystone has strong transverse isotropy (TI) with a vertical symmetry axis. The qP , SH and qSV wave propagation velocities in horizontal directions—the plane of isotropy of the TI medium—are measured as 3550, 1850 and 1290 m s –1 , respectively. The zero-offset VSP reveals that only one shear wave propagates in the vertical (depth) direction and the P - and S -wave velocities are 3100 and 1375 m s –1 , respectively. Four elastic moduli of the TI medium are determined from the seismic velocities and from the bulk density of 2.53 g cm –3 : c 11 = 31.9 GPa, c 33 = 24.3 GPa, c 44 = 4.5 GPa and c 66 = 8.7 GPa. A walkaway VSP with the borehole geophone at 50 m depth in borehole ID180 and shot points in the galleries leads to oblique seismic ray paths which allow us to determine the fifth elastic modulus of the TI medium to c 13 = 16 GPa. The tube wave recorded by a hydrophone in the water filled lower part of the borehole propagates with 1350 m s –1 , which confirms the estimate of the elastic constant c 66 . The analysis of body wave and surface wave data from a seismic experiment in Galerie Est shows reflections from several fracture zones in the gallery floor. The thickness of the excavation damaged zone (EDZ) in the floor of Galerie Est is estimated to 0.7 m.

Beschreibung: During megathrust earthquakes, great ruptures are accompanied by large scale mass redistribution inside the solid Earth and by ocean mass redistribution due to bathymetry changes. These large scale mass displacements can be detected using the monthly gravity maps of the GRACE satellite mission. In recent years it has become increasingly common to use the long wavelength changes in the Earth's gravity field observed by GRACE to infer seismic source properties for large megathrust earthquakes. An important advantage of space gravimetry is that it is independent from the availability of land for its measurements. This is relevant for observation of megathrust earthquakes, which occur mostly offshore, such as the $M_{\text{w}} \sim 9$ 2004 Sumatra–Andaman, 2010 Maule (Chile) and 2011 Tohoku-Oki (Japan) events. In Broerse et al. , we examined the effect of the presence of an ocean above the rupture on long wavelength gravity changes and showed it to be of the first order. Here we revisit the implementation of an ocean layer through the sea level equation and compare the results with approximated methods that have been used in the literature. One of the simplifications usually lies in the assumption of a globally uniform ocean layer. We show that especially in the case of the 2010 Maule earthquake, due to the closeness of the South American continent, the uniform ocean assumption is not valid and causes errors up to 57 per cent for modelled peak geoid height changes (expressed at a spherical harmonic truncation degree of 40). In addition, we show that when a large amount of slip occurs close to the trench, horizontal motions of the ocean floor play a mayor role in the ocean contribution to gravity changes. Using a slip model of the 2011 Tohoku-Oki earthquake that places the majority of slip close to the surface, the peak value in geoid height change increases by 50 per cent due to horizontal ocean floor motion. Furthermore, we test the influence of the maximum spherical harmonic degree at which the sea level equation is performed for sea level changes occurring along coastlines, which shows to be important for relative sea level changes occurring along the shore. Finally, we demonstrate that ocean floor loading, self-gravitation of water and conservation of water mass are of second order importance for coseismic gravity changes. When GRACE observations are used to determine earthquake parameters such as seismic moment or source depth, the uniform ocean layer method introduces large biases, depending on the location of the rupture with respect to the continent. The same holds for interpreting shallow slip when horizontal motions are not properly accounted for in the ocean contribution. In both cases the depth at which slip occurs will be underestimated.

Beschreibung: The Tonga-Kermadec forearc is deforming in response to on-going subduction of the Pacific Plate beneath the Indo-Australian Plate. Previous research has focussed on the structural development of the forearc where large bathymetric features such as the Hikurangi Plateau and Louisville Ridge seamount chain are being subducted. Consequently, knowledge of the ‘background’ forearc in regions of normal plate convergence is limited. We report on an ~250-km-long multichannel seismic reflection profile that was shot perpendicular to the Tonga-Kermadec trench at ~28°S to determine the lateral and temporal variations in the structure, stratigraphy and deformation of the Kermadec forearc resulting solely from Pacific Plate subduction. Interpretation of the seismic profile, in conjunction with regional swath bathymetry data, shows that the Pacific Plate exhibits horst and graben structures that accommodate bending-induced extensional stresses, generated as the trenchward dip of the crust increases. Trench infill is also much thicker than expected at 1 km which, we propose, results from increased sediment flux into and along the trench. Pervasive normal faulting of the mid-trench slope most likely accommodates the majority of the observed forearc extension in response to basal subduction erosion, and a structural high is located between the mid- and upper-trench slopes. We interpret this high as representing a dense and most likely structurally robust region of crust lying beneath this region. Sediment of the upper-trench slope documents depositional hiatuses and on-going uplift of the arc. Strong along-arc currents appear to erode the Kermadec volcanic arc and distribute this sediment to the surrounding basins, while currents over the forearc redistribute deposits as sediment waves. Minor uplift of the transitional Kermadec forearc, observed just to the north of the profile, appears to relate to an underlying structural trend as well as subduction of the Louisville Ridge seamount chain 250 km to the north. Relative uplift of the Kermadec arc is observed from changes in the tilt of upper-trench slope deposits and extensional faulting of the basement immediately surrounding the Louisville Ridge.

Beschreibung: While mRNA stability has been demonstrated to control rates of translation, generating both global and local synonymous codon biases in many unicellular organisms, this explanation cannot adequately explain why codon bias strongly tracks neighboring intergene GC content; suggesting that structural dynamics of DNA might also influence codon choice. Because minor groove width is highly governed by 3-base periodicity in GC, the existence of triplet-based codons might imply a functional role for the optimization of local DNA molecular dynamics via GC content at synonymous sites (GC3). We confirm a strong association between GC3-related intrinsic DNA flexibility and codon bias across 24 different prokaryotic multiple whole-genome alignments. We develop a novel test of natural selection targeting synonymous sites and demonstrate that GC3-related DNA backbone dynamics have been subject to moderate selective pressure, perhaps contributing to our observation that many genes possess extreme DNA backbone dynamics for their given protein space. This dual function of codons may impose universal functional constraints affecting the evolution of synonymous and non-synonymous sites. We propose that synonymous sites may have evolved as an ‘accessory’ during an early expansion of a primordial genetic code, allowing for multiplexed protein coding and structural dynamic information within the same molecular context.

Beschreibung: Long-term volcanic subsidence provides insight into intereruptive processes, which comprise the longest portion of the eruptive cycle. Ground-based geodetic surveys of Medicine Lake Volcano (MLV), northern CA, document subsidence at rates of ~–10 mm yr –1 between 1954 and 2004. The long observation period plus the duration and stable magnitude of this signal presents an ideal opportunity to study long-term volcanic deformation, but this first requires accurate knowledge of the geometry and magnitude of the source. Best-fitting analytical source models to past levelling and GPS data sets show conflicting source parameters—primarily the model depth. To overcome this, we combine multiple tracks of InSAR data, each with a different look angle, to improve upon the spatial resolution of ground-based measurements. We compare the results from InSAR to those of past geodetic studies, extending the geodetic record to 2011 and demonstrating that subsidence at MLV continues at ~–10 mm yr –1 . Using geophysical inversions, we obtain the best-fitting analytical source model—a sill located at 9–10 km depth beneath the caldera. This model geometry is similar to those of past studies, providing a good fit to the high spatial density of InSAR measurements, while accounting for the high ratio of vertical to horizontal deformation derived from InSAR and recorded by existing levelling and GPS data sets. We discuss possible causes of subsidence and show that this model supports the hypothesis that deformation at MLV is driven by tectonic extension, gravitational loading, plus a component of volume loss at depth, most likely due to cooling and crystallization within the intrusive complex that underlies the edifice. Past InSAR surveys at MLV, and throughout the Cascades, are of variable success due to dense vegetation, snow cover and atmospheric artefacts. In this study, we demonstrate how InSAR may be successfully used in this setting by applying a suite of multitemporal analysis methods that account for atmospheric and orbital noise sources. These methods include: a stacking strategy based upon the noise characteristics of each data set; pixelwise rate-map formation (-RATE) and persistent scatterer InSAR (StaMPS).

Beschreibung: In the literature, the inverted coseismic slip models from seismological and geodetic data for the 2011 Tohoku-Oki earthquake portray significant discrepancies, in particular regarding the intensity and the distribution of the rupture near the trench. For a megathrust earthquake, it is difficult to discern the slip along the shallow part of the fault from the geodetic data, which are often acquired on land. In this paper, we discuss the uncertainties in the slip distribution inversion using the geodetic data for the 2011 Tohoku earthquake and the Fully Bayesian Inversion method. These uncertainties are due to the prior information regarding the boundary conditions at the edges of the fault, the dip subduction angle and the smoothing operator. Using continuous GPS data from the Japan Island, the results for the rigid and free boundary conditions show that they produce remarkably different slip distributions at shallow depths, with the latter producing a large slip exceeding 30 m near the surface. These results indicate that the smoothing operator (gradient or Laplacian schemes) does not severely affect the slip pattern. To better invert the coseismic slip, we then introduce the ocean bottom GPS (OB-GPS) data, which improve the resolution of the shallow part of the fault. We obtain a near-trench slip greater than 40 m that reaches the Earth's surface, regardless of which boundary condition is used. Additionally, we show that using a mean dip angle for the fault as derived from subduction models is adequate if the goal is to invert for the general features of the slip pattern of this megathrust event.

Beschreibung: The thermophilic fungus Chaetomium thermophilum holds great promise for structural biology. To increase the efficiency of its biochemical and structural characterization and to explore its thermophilic properties beyond those of individual proteins, we obtained transcriptomics and proteomics data, and integrated them with computational annotation methods and a multitude of biochemical experiments conducted by the structural biology community. We considerably improved the genome annotation of Chaetomium thermophilum and characterized the transcripts and expression of thousands of genes. We furthermore show that the composition and structure of the expressed proteome of Chaetomium thermophilum is similar to its mesophilic relatives. Data were deposited in a publicly available repository and provide a rich source to the structural biology community.

Beschreibung: New methods are required to combine the information contained in the passive electrical and seismic signals to detect, localize and monitor hydromechanical disturbances in porous media. We propose a field experiment showing how passive seismic and electrical data can be combined together to detect a preferential flow path associated with internal erosion in a Earth dam. Continuous passive seismic and electrical (self-potential) monitoring data were recorded during a 7-d full-scale levee (earthen embankment) failure test, conducted in Booneschans, Netherlands in 2012. Spatially coherent acoustic emissions events and the development of a self-potential anomaly, associated with induced concentrated seepage and internal erosion phenomena, were identified and imaged near the downstream toe of the embankment, in an area that subsequently developed a series of concentrated water flows and sand boils, and where liquefaction of the embankment toe eventually developed. We present a new 4-D grid-search algorithm for acoustic emissions localization in both time and space, and the application of the localization results to add spatially varying constraints to time-lapse 3-D modelling of self-potential data in the terms of source current localization. Seismic signal localization results are utilized to build a set of time-invariant yet spatially varying model weights used for the inversion of the self-potential data. Results from the combination of these two passive techniques show results that are more consistent in terms of focused ground water flow with respect to visual observation on the embankment. This approach to geophysical monitoring of earthen embankments provides an improved approach for early detection and imaging of the development of embankment defects associated with concentrated seepage and internal erosion phenomena. The same approach can be used to detect various types of hydromechanical disturbances at larger scales.

Beschreibung: We propose a two-phase damage theory in a viscoelastic medium to study the pressure and porosity diffusion in fractured near-surface porous rocks. The key ingredient in the viscoelastic theory is that the pressure difference between solid and fluid is divided into three parts, which contribute to reversible elastic potential energy, irreversible viscous entropy production and surface energy stored during deformation. The resulting continuum description of weakening and failure (distributed void generation and microcracking) in a linear Kelvin body accounts for surface energy being created by both viscous and elastic deformational work. The model shows that while non-linear permeability models leads to an enhanced diffusivity, damage makes the matrix more compressible if we assume the geometry/size of cracks remain unchanged. The net effect is that the porosity diffusivity is reduced causing fluid infiltration to accumulate closer to the injection source, leading to a slower fluid diffusion during hydraulic fracturing with a fixed porosity boundary condition. However if a constant overpressure boundary condition is applied, a weakened matrix with damage leads to greater pressure diffusivity than for porosity.

Beschreibung: Complications arise in the interpretation of gravity fields because of interference from systematic degradations, such as boundary blurring and distortion. The major sources of these degradations are the various systematic errors that inevitably occur during gravity field data acquisition, discretization and geophysical forward modelling. To address this problem, we evaluate deconvolution method that aim to detect the clear horizontal boundaries of anomalous sources by the suppression of systematic errors. A convolution-based multilayer projection model, based on the classical 3-D gravity field forward model, is innovatively derived to model the systematic error degradation. Our deconvolution algorithm is specifically designed based on this multilayer projection model, in which three types of systematic error are defined. The degradations of the different systematic errors are considered in the deconvolution algorithm. As the primary source of degradation, the convolution-based systematic error is the main object of the multilayer projection model. Both the random systematic error and the projection systematic error are shown to form an integral part of the multilayer projection model, and the mixed norm regularization method and the primal-dual optimization method are therefore employed to control these errors and stabilize the deconvolution solution. We herein analyse the parameter identification and convergence of the proposed algorithms, and synthetic and field data sets are both used to illustrate their effectiveness. Additional synthetic examples are specifically designed to analyse the effects of the projection systematic error, which is caused by the uncertainty associated with the estimation of the impulse response function.

Beschreibung: We define an algorithm in the time domain for computing the unwrapped instantaneous phase and its derivative, the instantaneous frequency, using only derivatives and integrals. It does not require user-defined parameters, like most algorithms proposed so far. We validate and compare its performance with respect to open-source and commercial software by synthetic and real data examples.

Beschreibung: Simulating electromagnetic fields in the quasi-static regime by solving Maxwell's equations is a central task in many geophysical applications. In most cases, geophysical targets of interest exhibit complex topography and bathymetry as well as layers and faults. Capturing these effects with a sufficient level of detail is a huge challenge for numerical simulations. Standard techniques require a very fine discretization that can result in an impracticably large linear system to be solved. A remedy is to use locally refined and adaptive meshes, however, the potential coarsening is limited in the presence of highly heterogeneous and anisotropic conductivities. In this paper, we discuss the application of multiscale finite volume (MSFV) methods to Maxwell's equations in frequency domain. Given a partition of the fine mesh into a coarse mesh the idea is to obtain coarse-to-fine interpolation by solving local versions of Maxwell's equations on each coarsened grid cell. By construction, the interpolation accounts for fine scale conductivity changes, yields a natural homogenization, and reduces the fine mesh problem dramatically in size. To improve the accuracy for singular sources, we use an irregular coarsening strategy. We show that using MSFV methods we can simulate electromagnetic fields with reasonable accuracy in a fraction of the time as compared to state-of-the-art solvers for the fine mesh problem, especially when considering parallel platforms.

Beschreibung: Wave-equation tomography (WT) and full waveform inversion (FWI) are combined through a hybrid misfit function to estimate high-resolution subsurface structures starting from a poorly constrained initial velocity model. Both methods share the same wavefield forward modelling and inversion schemes, while they differ only on the ways to calculate misfit functions and hence the ways to sample in the model space. Aiming at minimizing the cross-correlation phase delay between synthetic and real data, WT can be used to retrieve the long- and middle-wavelength model components, which are essential to FWI. Compared to ray-based traveltime tomography that is based on asymptotic high-frequency approximation, WT provides a better resolution by exploring the band-limited feature of seismic wavefield. On the other hand, FWI is capable of resolving the short-wavelength model component, complementing the WT. In this study, we apply WT to surface first-arrival refraction data, and apply FWI to both refraction and reflection data. We assign adaptive weights to the two different misfit measurements and build a progressive inversion strategy. To illustrate the advantage of our strategy over conventional ‘ray tomography + FWI’ approach, we show in a synthetic lens test that WT can provide extra subsurface information that is critical for a successful FWI application. To further show the efficiency, we test our strategy on the 2-D Marmousi model where satisfactory inversion results are achieved without much manual intervention. Finally, we apply the inversion strategy to a deep-water seismic data set acquired offshore Sumatra with a 12-km-long streamer. In order to alleviate several practical problems posed by the deep-water setting, we apply downward continuation (DC) to generate a virtual ocean bottom experiment data set prior to inversion. The new geometry after DC boosts up the shallow refractions, as well as avoiding cumbersome modelling through the thick water column, thus reducing the computation cost by 85 per cent. The inversion result from the new data set shows high-resolution shallow sediment structures and the migration images prove the superiority of the inverted model over a conventional tomography model.

Beschreibung: The activation of Late Quaternary faults in the Central Apennines (Italy) could generate earthquakes with magnitude of about 6.5, and the Monte Vettore fault system probably belongs to the same category of seismogenetic faults. Such structure has been defined ‘silent’, because of its geological and geomorphological evidences of past activation, but the absence of historical records in the seismic catalogues to be associated with its activation. The ‘Piano di Castelluccio’ intramountain basin, resulting from the Quaternary activity of normal faults, is characterized by a secondary fault strand highlighted by a NW–SE fault scarp: it has been already studied through palaeoseismological trenches, which highlighted evidences of Quaternary shallow faulting due to strong earthquakes, and through a 2-D ground penetrating radar (GPR) survey, showing the first geophysical signature of faulting for this site. Within the same place, a 3-D GPR volume over a 20 20 m area has been collected. The collection of radar echoes in three dimensions allows to map both the vertical and lateral continuity of shallow geometries of the fault zone (Fz), imaging features with high resolution, ranging from few metres to centimetres and therefore imaging also local variations at the microscale. Several geophysical markers of faulting, already highlighted on this site, have been taken as reference to plan the 3-D survey. In this paper, we provide the first 3-D subsurface imaging of an active shallow fault belonging to the Umbria-Marche Apennine highlighting the subsurface fault geometry and the stratigraphic sequence up to a depth of about 5 m. From our data, geophysical faulting signatures are clearly visible in three dimensions: diffraction hyperbolas, truncations of layers, local attenuated zones and varying dip of the layers have been detected within the Fz. The interpretation of the 3-D data set provided qualitative and quantitative geological information in addition to the fault location, like its geometry, boundaries and an estimation of the fault throw.

Beschreibung: Shotgun metagenome sequencing has become a fast, cheap and high-throughput technology for characterizing microbial communities in complex environments and human body sites. However, accurate identification of microorganisms at the strain/species level remains extremely challenging. We present a novel k -mer-based approach, termed GSMer, that identifies genome-specific markers (GSMs) from currently sequenced microbial genomes, which were then used for strain/species-level identification in metagenomes. Using 5390 sequenced microbial genomes, 8 770 321 50-mer strain-specific and 11 736 360 species-specific GSMs were identified for 4088 strains and 2005 species (4933 strains), respectively. The GSMs were first evaluated against mock community metagenomes, recently sequenced genomes and real metagenomes from different body sites, suggesting that the identified GSMs were specific to their targeting genomes. Sensitivity evaluation against synthetic metagenomes with different coverage suggested that 50 GSMs per strain were sufficient to identify most microbial strains with ≥0.25 x coverage, and 10% of selected GSMs in a database should be detected for confident positive callings. Application of GSMs identified 45 and 74 microbial strains/species significantly associated with type 2 diabetes patients and obese/lean individuals from corresponding gastrointestinal tract metagenomes, respectively. Our result agreed with previous studies but provided strain-level information. The approach can be directly applied to identify microbial strains/species from raw metagenomes, without the effort of complex data pre-processing.

Beschreibung: To estimate the seismic hazard, the geometry (dip, length and orientation) and the dynamics (type of displacements and amplitude) of the faults in the area of interest need to be understood. In this paper, in addition to geomorphologic observations, we present the results of two ground penetrating radar (GPR) campaigns conducted in 2010 and 2011 along the Emeelt fault in the vicinity of Ulaanbaatar, capital of Mongolia, located in an intracontinental region with low deformation rate that induces long recurrence time between large earthquakes. As the geomorphology induced by the fault activity has been highly smoothed by erosion processes since the last event, the fault location and geometry is difficult to determine precisely. However, by using GPR first, a non-destructive and fast investigation, the fault and the sedimentary deposits near the surface can be characterized and the results can be used for the choice of trench location. GPR was performed with a 50 MHz antenna over 2-D lines and with a 500 MHz antenna for pseudo-3-D surveys. The 500 MHz GPR profiles show a good consistency with the trench observations, dug next to the pseudo-3-D surveys. The 3-D 500 MHz GPR imaging of a palaeochannel crossed by the fault allowed us to estimate its lateral displacement to be about 2 m. This is consistent with a right lateral strike-slip displacement induced by an earthquake around magnitude 7 or several around magnitude 6. The 2-D 50 MHz profiles, recorded perpendicular to the fault, show a strong reflection dipping to the NE, which corresponds to the fault plane. Those profiles provided complementary information on the fault such as its location at shallow depth, its dip angle (from 23° to 35°) and define its lateral extension.

Beschreibung: We propose to test if gravimetry can prove useful in discriminating different models of long-term deep crustal processes in the case of the Taiwan mountain belt. We discuss two existing tectonic models that differ in the deep processes proposed to sustain the long-term growth of the orogen. One model assumes underplating of the uppermost Eurasian crust with subduction of the deeper part of the crust into the mantle. The other one suggests the accretion of the whole Eurasian crust above crustal-scale ramps, the lower crust being accreted into the collisional orogen. We compute the temporal gravity changes caused only by long-term rock mass transfers at depth for each of them. We show that the underplating model implies a rate of gravity change of –6 x 10 –2 μGal yr –1 , a value that increases to 2 x 10 –2 μGal yr –1 if crustal subduction is neglected. If the accretion of the whole Eurasian crust occurs, a rate of 7 x 10 –2 μGal yr –1 is obtained. The two models tested differ both in signal amplitude and spatial distribution. The yearly gravity changes expected by long-term deep crustal mass processes in Taiwan are two orders of magnitude below the present-day uncertainty of land-based gravity measurements. Assuming that these annually averaged long-term gravity changes will linearly accumulate with ongoing mountain building, multidecadal time-series are needed to identify comparable rates of gravity change. However, as gravity is sensitive to any mass redistribution, effects of short-term processes such as seismicity and surface mass transfers (erosion, sedimentation, ground-water) may prevent from detecting any long-term deep signal. This study indicates that temporal gravity is not appropriate for deciphering the long-term deep crustal processes involved in the Taiwan mountain belt.

Beschreibung: A new method to obtain the statistics of a geostatistical model is introduced. The method elicits the statistical information from a geological expert directly, by iteratively updating a population of vectors of statistics, based on the expert's subjective opinion of the corresponding geological simulations. Thus, it does not require the expert to have knowledge of the mathematical and statistical details of the model. The process uses a genetic algorithm to generate new vectors. We demonstrate the methodology for a particular geostatistical model used to model rock pore-space, which simulates the spatial distribution of matrix and pores over a 2-D grid, using multipoint statistics specified by conditional probabilities. Experts were asked to use the algorithm to estimate the statistics of a given target pore-space image with known statistics; thus, their numerical rates of convergence could be calculated. Convergence was measured for all experts, showing that the algorithm can be used to find appropriate probabilities given the expert's subjective input. However, considerable and apparently irreducible residual misfit was found between the true statistics and the estimates of statistics obtained by the experts, with the root-mean-square error on the conditional probabilities typically 〉0.1. This is interpreted as the limit of the experts’ abilities to distinguish between realizations of different spatial statistics using the algorithm. More accurate discrimination is therefore likely to require complementary elicitation techniques or sources of information independent of expert opinion.

Beschreibung: The computation of quasi-static deformation for axisymmetric viscoelastic structures on a gravitating spherical earth is addressed using the spectral element method (SEM). A 2-D spectral element domain is defined with respect to spherical coordinates of radius and angular distance from a pole of symmetry, and 3-D viscoelastic structure is assumed to be azimuthally symmetric with respect to this pole. A point dislocation source that is periodic in azimuth is implemented with a truncated sequence of azimuthal order numbers. Viscoelasticity is limited to linear rheologies and is implemented with the correspondence principle in the Laplace transform domain. This leads to a series of decoupled 2-D problems which are solved with the SEM. Inverse Laplace transform of the independent 2-D solutions leads to the time-domain solution of the 3-D equations of quasi-static equilibrium imposed on a 2-D structure. The numerical procedure is verified through comparison with analytic solutions for finite faults embedded in a laterally homogeneous viscoelastic structure. This methodology is applicable to situations where the predominant structure varies in one horizontal direction, such as a structural contrast across (or parallel to) a long strike-slip fault.

Beschreibung: The geodetic rates for the gravity variation and vertical uplift in polar regions subject to past and present-day ice-mass changes (PDIMCs) provide important insight into the rheological structure of the Earth. We provide an update of the rates observed at Ny-Ålesund, Svalbard. To do so, we extract and remove the significant seasonal content from the observations. The rate of gravity variations, derived from absolute and relative gravity measurements, is –1.39 ± 0.11 μGal yr –1 . The rate of vertical displacements is estimated using GPS and tide gauge measurements. We obtain 7.94 ± 0.21 and 8.29 ± 1.60 mm yr –1 , respectively. We compare the extracted signal with that predicted by GLDAS/Noah and ERA-interim hydrology models. We find that the seasonal gravity variations are well-represented by local hydrology changes contained in the ERA-interim model. The phase of seasonal vertical displacements are due to non-local continental hydrology and non-tidal ocean loading. However, a large part of the amplitude of the seasonal vertical displacements remains unexplained. The geodetic rates are used to investigate the asthenosphere viscosity and lithosphere/asthenosphere thicknesses. We first correct the updated geodetic rates for those induced by PDIMCs in Svalbard, using published results, and the sea level change due to the melting of the major ice reservoirs. We show that the latter are at the level of the geodetic rate uncertainties and are responsible for rates of gravity variations and vertical displacements of –0.29 ± 0.03 μGal yr –1 and 1.11 ± 0.10 mm yr –1 , respectively. To account for the late Pleistocene deglaciation, we use the global ice evolution model ICE-3G. The Little Ice Age (LIA) deglaciation in Svalbard is modelled using a disc load model with a simple linear temporal evolution. The geodetic rates at Ny-Ålesund induced by the past deglaciations depend on the viscosity structure of the Earth. We find that viscous relaxation time due to the LIA deglaciation in Svalbard is more than 60 times shorter than that due to the Pleistocene deglaciation. We also find that the response to past and PDIMCs of an Earth model with asthenosphere viscosities ranging between 1.0 and 5.5 x 10 18 Pa s and lithosphere (resp. asthenosphere) thicknesses ranging between 50 and 100 km (resp. 120 and 170 km) can explain the rates derived from geodetic observations.

Beschreibung: We show analytically that a well-known transfer function previously derived for the scalar acoustic problem that converts measurements from a 3-D (real-world) setting to a 2-D equivalent is directly applicable to the vector electromagnetic borehole ground penetrating radar problem. We also show that the transfer function's precision is improved for the low-loss case through the use of complex velocity. The transfer function has a strong effect on amplitude, and is therefore a critical preprocessing step for 2-D full-wave inversion when finding conductivity is of concern. We demonstrate the effectiveness of the transfer function through various numerical experiments and a synthetic frequency-domain full-wave inversion. We also compare the effectiveness of this curved-ray transfer function to a quasi-straight-ray transfer function. The inversion demonstrates the positive effect the transfer functions have on recovering conductivity and also that they are effective even when there are sharp velocity contrasts.

Beschreibung: Understanding the role of a given transcription factor (TF) in regulating gene expression requires precise mapping of its binding sites in the genome. Chromatin immunoprecipitation-exo, an emerging technique using exonuclease to digest TF unbound DNA after ChIP, is designed to reveal transcription factor binding site (TFBS) boundaries with near-single nucleotide resolution. Although ChIP-exo promises deeper insights into transcription regulation, no dedicated bioinformatics tool exists to leverage its advantages. Most ChIP-seq and ChIP-chip analytic methods are not tailored for ChIP-exo, and thus cannot take full advantage of high-resolution ChIP-exo data. Here we describe a novel analysis framework, termed MACE (model-based analysis of ChIP-exo) dedicated to ChIP-exo data analysis. The MACE workflow consists of four steps: (i) sequencing data normalization and bias correction; (ii) signal consolidation and noise reduction; (iii) single-nucleotide resolution border peak detection using the Chebyshev Inequality and (iv) border matching using the Gale-Shapley stable matching algorithm. When applied to published human CTCF, yeast Reb1 and our own mouse ONECUT1/HNF6 ChIP-exo data, MACE is able to define TFBSs with high sensitivity, specificity and spatial resolution, as evidenced by multiple criteria including motif enrichment, sequence conservation, direct sequence pileup, nucleosome positioning and open chromatin states. In addition, we show that the fundamental advance of MACE is the identification of two boundaries of a TFBS with high resolution, whereas other methods only report a single location of the same event. The two boundaries help elucidate the in vivo binding structure of a given TF, e.g. whether the TF may bind as dimers or in a complex with other co-factors.

Beschreibung: The 2 principle and the unbiased predictive risk estimator are used to determine optimal regularization parameters in the context of 3-D focusing gravity inversion with the minimum support stabilizer. At each iteration of the focusing inversion the minimum support stabilizer is determined and then the fidelity term is updated using the standard form transformation. Solution of the resulting Tikhonov functional is found efficiently using the singular value decomposition of the transformed model matrix, which also provides for efficient determination of the updated regularization parameter each step. Experimental 3-D simulations using synthetic data of a dipping dike and a cube anomaly demonstrate that both parameter estimation techniques outperform the Morozov discrepancy principle for determining the regularization parameter. Smaller relative errors of the reconstructed models are obtained with fewer iterations. Data acquired over the Gotvand dam site in the south-west of Iran are used to validate use of the methods for inversion of practical data and provide good estimates of anomalous structures within the subsurface.

Beschreibung: Global navigation satellite systems (GNSSs) have revealed that a mega-thrust earthquake that occurs in an island-arc trench system causes post-seismic crustal deformation. Such crustal deformation data have been interpreted by combining three mechanisms: afterslip, poroelastic rebound and viscoelastic relaxation. It is seismologically important to determine the contribution of each mechanism because it provides frictional properties between the plate boundaries and viscosity estimates in the asthenosphere which are necessary to evaluate the stress behaviour during earthquake cycles. However, the observation sites of GNSS are mostly deployed over land and can detect only a small part of the large-scale deformation, which precludes a clear separation of the mechanisms. To extend the spatial coverage of the deformation area, recent studies started to use satellite gravity data that can detect long-wavelength deformations over the ocean. To date, compared with theoretical models for calculating the post-seismic crustal deformation, a few models have been proposed to interpret the corresponding gravity variations. Previous approaches have adopted approximations for the effects of compressibility, sphericity and self-gravitation when computing gravity changes. In this study, a new spectral-finite element approach is presented to consider the effects of material compressibility for Burgers viscoelastic earth model with a laterally heterogeneous viscosity distribution. After the basic principles are explained, it is applied to the 2004 Sumatra–Andaman earthquake. For this event, post-seismic deformation mechanisms are still a controversial topic. Using the developed approach, it is shown that the spatial patterns of gravity change generated by the above three mechanisms clearly differ from one another. A comparison of the theoretical simulation results with the satellite gravity data obtained from the Gravity Recovery and Climate Experiment reveals that both afterslip and viscoelastic relaxation are occurring. Considering the spatial patterns in satellite gravity fields is an effective method for investigating post-seismic deformation mechanisms.

Beschreibung: Functional mechanisms of biomolecules often manifest themselves precisely in transient conformational substates. Researchers have long sought to structurally characterize dynamic processes in non-coding RNA, combining experimental data with computer algorithms. However, adequate exploration of conformational space for these highly dynamic molecules, starting from static crystal structures, remains challenging. Here, we report a new conformational sampling procedure, KGSrna, which can efficiently probe the native ensemble of RNA molecules in solution. We found that KGSrna ensembles accurately represent the conformational landscapes of 3D RNA encoded by NMR proton chemical shifts. KGSrna resolves motionally averaged NMR data into structural contributions; when coupled with residual dipolar coupling data, a KGSrna ensemble revealed a previously uncharacterized transient excited state of the HIV-1 trans-activation response element stem–loop. Ensemble-based interpretations of averaged data can aid in formulating and testing dynamic, motion-based hypotheses of functional mechanisms in RNAs with broad implications for RNA engineering and therapeutic intervention.

Beschreibung: Binding of transcription factors to their binding sites in promoter regions is the fundamental event in transcriptional gene regulation. When a transcription factor binding site is located within a nucleosome, the DNA has to partially unwrap from the nucleosome to allow transcription factor binding. This reduces the rate of transcription factor binding and is a known mechanism for regulation of gene expression via chromatin structure. Recently a second mechanism has been reported where transcription factor off-rates are dramatically increased when binding to target sites within the nucleosome. There are two possible explanations for such an increase in off-rate short of an active role of the nucleosome in pushing the transcription factor off the DNA: (i) for dimeric transcription factors the nucleosome can change the equilibrium between monomeric and dimeric binding or (ii) the nucleosome can change the equilibrium between specific and non-specific binding to the DNA. We explicitly model both scenarios and find that dimeric binding can explain a large increase in off-rate while the non-specific binding model cannot be reconciled with the large, experimentally observed increase. Our results suggest a general mechanism how nucleosomes increase transcription factor dissociation to promote exchange of transcription factors and regulate gene expression.

Beschreibung: We report and analyse the tsunami recorded in the northwestern Indian Ocean at the Makran region following the M w 7.7 Pakistan inland strike-slip earthquake on 2013 September 24. We analyse eleven tide gauge records as well as one DART record of this tsunami and perform numerical modelling of the tsunami that would be triggered by a range of possible sources. The tsunami registered a maximum wave height of 109 cm at the Qurayat tide gauge station (Oman). The dominant period of the tsunami was around 12 min, although wavelet analysis showed that parts of the tsunami energy were partitioned into a slightly wider period range of 7 and 16 min. Tsunami backward ray tracing showed that the tsunami source was possibly located offshore Jiwani (Pakistan) and that the tsunami was most likely triggered by the main shock. The aftershocks are distributed in the inland region and the coseismic vertical and horizontal displacements are also limited inland implying that the tsunami was generated by secondary sources triggered by the earthquake. Different possible tsunami sources including a mud volcano at the location of the newly generated island, and a mud volcano or diapir at offshore deep water were examined through numerical modelling and all failed to reproduce the observed waveforms. Numerical modelling showed that a submarine slump with a source dimension of about 10–15 km and a thickness of about 100 m located at 61.49°E and 24.62°N, that is, about 60–70 km off the Jiwani coast (Pakistan), seems capable of reasonably reproducing the wave amplitudes and periods of the observed tsunami waveforms. This event was the second instrumentally recorded tsunami in the region, after the Makran tsunami of 1945 November, and provides evidence for a hazard from landslide/slump-generated waves following seismic activity in the area.

Beschreibung: Joint inversion of different geophysical data sets is becoming a more popular and powerful tool, and it has been performed on data sensitive both to the same physical parameter and to different physical parameters. Joint inversion is undertaken to reduce acceptable model space and to increase sensitivity to model parameters that one method alone is unable to resolve adequately. We examine and implement a novel hybrid joint inversion approach. In our inversion scheme a model—the reference model—is fixed, and the information shared with the subsurface structure obtained from another method will be maximized; in our case conductivity structures from magnetotelluric (MT) inversion. During inversion, the joint probability distribution of the MT and the specified reference model is estimated and its entropy minimized in order to guide the inversion result towards a solution that is statistically compatible with the reference model. The powerful feature of this technique is that no explicit relationships between estimated model parameters and reference model ones are presumed: if a link exists in data then it is highlighted in the estimation of the joint probability distribution, if no link is required, then none is enforced. Tests performed verify the robustness of this method and the advantages of it in a 1-D anisotropic scenario are demonstrated. A case study was performed with data from Central Germany, effectively fitting an MT data set from a single station within as minimal an amount of anisotropy as required.

Beschreibung: Some of the major geothermal anomalies in central Europe are linked to tectonic structures within the top of crystalline basement, which modify strongly the top of this basement. Their assessment is a major challenge in exploration geophysics. Gravity has been proven to be suitable for the detection of mainly large scale lithological and structural inhomogeneities. Indeed, it is well known and proven by different wells that, for example, in northern Switzerland extended negative anomalies are linked to such structures. Due to depth limitation of wells, there vertical extension is often unknown. In this study, we have investigated the potential of gravity for the geometrical characterization of such basement structures. Our approach consists in the combination of the series of Butterworth filters, geological modelling and best-fitting between observed and computed residual anomalies. In this respect, filters of variable wavelength are applied to observed and computed gravity data. The geological model is discretized into a finite element mesh. Near-surface anomalies and the effect of the sedimentary cover were eliminated using cut-off wavelength of 10 km and geological and seismic information. We analysed the potential of preferential Butterworth filtering in a sensitivity study and applied the above mentioned approach to part of the Swiss molasses basin. Sensitivity analyses reveal that such sets of residual anomalies represents a pseudo-tomography revealing the distribution of different structures with depth. This finding allows for interpreting negative anomalies in terms of 3-D volumes. Best-fitting then permits determination of the most likely 3-D geometries of such basement structures. Our model fits both, geological observations and gravity: among 10 deep boreholes in the studied area, six reach the respective units and confirm our distribution of the negative (and positive) anomalies.

Beschreibung: Several slope failures are observed near the deformation front on the frontal ridges of the northern Cascadia accretionary margin off Vancouver Island. The cause for these events is not clear, although several lines of evidence indicate a possible connection between the occurrence of gas hydrate and submarine landslide features. The presence of gas hydrate is indicated by a prominent bottom-simulating reflector (BSR), at a depth of ~265–275 m beneath the seafloor (mbsf), as interpreted from vertical-incidence and wide-angle seismic data beneath the ridge crests of the frontal ridges. For one slide, informally called Slipstream Slide, the velocity structure inferred from tomography analyses shows anomalous high velocities values of about 2.0 km s –1 at shallow depths of 100 mbsf. The estimated depth of the glide plane (100 ± 10 m) closely matches the depth of these shallow high velocities. In contrast, at a frontal ridge slide just to the northwest (informally called Orca Slide), the glide plane occurs at the same depth as the current BSR. Our new results indicate that the glide plane of the Slipstream slope failure is associated with the contrast between sediments strengthened by gas hydrate and overlying sediments where little or no hydrate is present. In contrast, the glide plane of Orca Slide is between sediment strengthened by hydrate underlain by sediments beneath the gas hydrate stability zone, possibly containing free gas. Additionally, a set of margin perpendicular normal faults are imaged from seafloor down to BSR depth at both frontal ridges. As inferred from the multibeam bathymetry, the estimated volume of the material lost during the slope failure at Slipstream Slide is about 0.33 km 3 , and ~0.24 km 3 of this volume is present as debris material on the ocean basin floor. The 20 per cent difference is likely due to more widely distributed fine sediments not easily detectable as bathymetric anomalies. These volume estimates on the Cascadia margin are approaching the mass failure volume for other slides that have generated large tsunamis—for example 1–3 km 3 for a 1998 Papua New Guinea slide.

Beschreibung: The method presented here aims to assess the tsunami threat very rapidly after the occurrence of a large earthquake, using as input the parameters of the seismic source, and an approach based on Green's summation. We show that the main weakness of the approach (the need to consider only linear shallow water propagation) is largely compensated by the advantages in terms of computing performance and independence with respect to pre-computed scenarios. To test the approach and to illustrate its implementation in a real environment, we focus on the Sea of Oman, a tsunamigenic area characterized by Makran subduction zone which detailed structure is partially unknown and where secondary tsunami sources must also be taken into account, both for hazard studies and warning purposes. The potential source area is partitioned into a grid of unity water sources. A shallow water (SW) numerical model is used to pre-compute the corresponding empirical Green's functions on several points of interest located on the coasts of Iran, Pakistan and Oman. The comparison between Green's summation and the direct SW computation using the full resolution of the bathymetric grid shows that the accuracy is good enough for practical applications.

Beschreibung: Clay minerals as products of hydrothermal alteration significantly influence the hydraulic and mechanical properties of crystalline rock. Therefore, the localization and characterization of alteration zones by downhole measurements is a great challenge for the development of geothermal reservoirs. The magnetite bearing granite of the geothermal site in Soultz-sous-Forêts (France) experienced hydrothermal alteration during several tectonic events and clay mineral formation is especially observed in alteration halos around fracture zones. During the formation of clay minerals, magnetite was oxidized into hematite, which significantly reduces the magnetic susceptibility of the granite from ferrimagnetic to mostly paramagnetic values. The aim of this study was to find out if there exists a correlation between synthetic clay content logs (SCCLs) and measurements of magnetic susceptibility on cuttings in the granite in order to characterize their alteration mineralogy. Such a correlation has been proven for core samples of the EPS1 reference well. SCCLs were created from gamma ray and fracture density logs using a neural network. These logs can localize altered fracture zones in the GPK1-4 wells, where no core material is available. Mass susceptibility from 261 cutting samples of the wells GPK1–GPK4 was compared with the neural network derived synthetic logs. We applied a combination of temperature dependent magnetic susceptibility measurements with optical and electron microscopy, and energy dispersive X-ray spectroscopy to discriminate different stages of alteration. We found, that also in the granite cuttings an increasing alteration grade is characterized by an advancing oxidation of magnetite into hematite and a reduction of magnetic susceptibility. A challenge to face for the interpretation of magnetic susceptibility data from cuttings material is that extreme alteration grades can also display increased susceptibilities due to the formation of secondary magnetite. Low magnetic susceptibility can also be attributed to primary low magnetite content, if the granite facies changes. In order to interpret magnetic susceptibility from cuttings, contaminations with iron from wear debris of the drilling tools must be eliminated. Provided that the magnetic mineralogy of the granite is known in detail, this method in combination with petrographic investigations is suited to indicate and characterize hydrothermal alteration and the appearance of clay.

Beschreibung: Elastic and mechanical weakening from water saturation are widely known to occur in sedimentary rocks, and particularly in carbonate rocks. To improve our understanding of the physics underlying this phenomenon, ultrasonic ( f  ~ 0.5 MHz) elastic properties are measured on a large suite of clean limestones and sandstones at very low saturations from relative humidity (RH) variations at ambient conditions. Measurements clearly highlight an elastic weakening (i.e. decrease in elastic wave velocity) from moisture adsorption. P - and S -wave velocities are similarly affected by adsorption, but in a different way for limestones and sandstone samples. While the elastic properties of limestone samples show almost no RH dependence, a large weakening is observed for samples of Fontainebleau sandstone that increases with the samples’ porosity. The main elastic weakening effect is likely to result from adsorption of fluid at grain contacts. It thus affects particularly granular rocks such as sandstones while well-cemented limestones are not affected. The granular model from Murphy et al. , accounting for surface energy effects, proves to be appropriate. Applying this model, it is shown that (i) P - and S -wave velocities have the same dependence on surface energy, which is consistent with the measurements and (ii) surface energy values obtained from the ultrasonic data using this model correlate with RH, and are consistent with the expected value for quartz crystals at vapour pressure. Yet, porosity, which relates to degree of cementation in the particular case of Fontainebleau sandstone, appears to be an additional parameter. A modified model is thus derived using the cementation model from Digby, accounting for a bonding radius at grain contact. It proves to apply well to the measured data. The fundamental difference between limestones’ and sandstones’ dependence to RH appears to be related to a microstructural difference. Saturation variations from RH increase depend on specific surface area, which is particularly low in Fontainebleau sandstones and large in microporous limestones. However elastic weakening from RH is more important in sandstones owing to their granular microstructure.

Beschreibung: High-throughput sequencing technologies, including RNA-seq, have made it possible to move beyond gene expression analysis to study transcriptional events including alternative splicing and gene fusions. Furthermore, recent studies in cancer have suggested the importance of identifying transcriptionally altered loci as biomarkers for improved prognosis and therapy. While many statistical methods have been proposed for identifying novel transcriptional events with RNA-seq, nearly all rely on contrasting known classes of samples, such as tumor and normal. Few tools exist for the unsupervised discovery of such events without class labels. In this paper, we present SigFuge for identifying genomic loci exhibiting differential transcription patterns across many RNA-seq samples. SigFuge combines clustering with hypothesis testing to identify genes exhibiting alternative splicing, or differences in isoform expression. We apply SigFuge to RNA-seq cohorts of 177 lung and 279 head and neck squamous cell carcinoma samples from the Cancer Genome Atlas, and identify several cases of differential isoform usage including CDKN2A , a tumor suppressor gene known to be inactivated in a majority of lung squamous cell tumors. By not restricting attention to known sample stratifications, SigFuge offers a novel approach to unsupervised screening of genetic loci across RNA-seq cohorts. SigFuge is available as an R package through Bioconductor.

Beschreibung: A novel experimental method is introduced to estimate the Thomsen's elastic anisotropy parameters and of a transversely isotropic shale under variable stress and saturation conditions. The method consists in recording P -wave velocities along numerous paths on a cylindrical specimen using miniature ultrasonic transducers. Such an overdetermined set of measurements is specifically designed to reduce the uncertainty associated with the determination of Thomsen's parameter compared to the classical method for which a single off-axis measurement is used (usually at 45° to the specimen's axis). This method is applied to a specimen of Opalinus Clay recovered from the Mont-Terri Underground Research Laboratory in Switzerland. The specimen is first saturated with brine at low effective pressure and then subjected to an effective pressure cycle up to 40 MPa, followed by a triaxial loading up to failure. During saturation and deformation, the evolution of P -wave velocities along a maximum of 240 ray paths is monitored and Thomsen's parameters α , and are computed by fitting Thomsen's weak anisotropy model to the data. The values of and obtained at the highest confining pressures reached during the experiment are comparable with those predicted from X-ray diffraction texture analysis and modelling for Opalinus Clay reported in the literature. These models neglect the effect of soft-porosity on elastic properties, but become relevant when soft porosity is closed at high effective pressure.

Beschreibung: We present a novel accurate and efficient goal-oriented adaptive finite-element method solution for complex multi-electrodes resistivity system with arbitrary smooth surface topographies. A simple Green's function of a half-space model is adopted to eliminate the singularity. A unified boundary value problem for the regular potential is formulated for a multi-electrodes system so that it shares a common system matrix. In addition, a goal-oriented error estimation technique is developed to generate an optimal common grid so that highly accurate solutions are obtained with minimum computation cost. Synthetic models are used to verify our algorithm and excellent agreements are obtained by comparing with other methods.

Beschreibung: On 2008 October 5, a magnitude 6.6 earthquake struck the eastern termination of the intermontane Alai valley between the southern Tien Shan and the northern Pamir of Kyrgyzstan. The shallow thrust earthquake occurred in the footwall of the Main Pamir thrust, where the Pamir orogen is colliding with the southern Tien Shan mountains. We measure the coseismic surface displacements using SAR (Synthetic Aperture RADAR) data; the results show clear gradients in the vertical and horizontal directions along a complex pattern of surface ruptures and active faults. To integrate and to interpret these observations in the context of the regional tectonics, we complement the SAR data analysis with seismological data and geological field observations. While the main moment release of the Nura earthquake appears to be on the Pamir Frontal thrust, the main surface displacements and surface rupture occurred in the footwall along the NE–SW striking Irkeshtam fault. With InSAR data from ascending and descending tracks along with pixel offset measurements, we model the Nura earthquake source as a segmented rupture. One fault segment corresponds to high-angle brittle faulting at the Pamir Frontal thrust and two more fault segments show moderate-angle and low-friction thrusting at the Irkeshtam fault. Our integrated analysis of the coseismic deformation argues for rupture segmentation and strain partitioning associated to the earthquake. It possibly activated an orogenic wedge in the easternmost segment of the Pamir-Alai collision zone. Further, the style of the segmentation may be associated with the presence of Palaeogene evaporites.

Beschreibung: The Geological Survey of Sweden has been collecting airborne tensor very low frequency data (VLF) over several decades, covering large parts of the country. The data has been an invaluable source of information for identifying conductive structures that can among other things be related to water-filled fault zones, wet sediments that fill valleys or ore mineralizations. Because the method only uses two differently polarized plane waves of very similar frequency, vertical resolution is low and interpretation is in most cases limited to maps that are directly derived from the data. Occasionally, 2-D inversion is carried out along selected profiles. In this paper, we present for the first time a 3-D inversion for tensor VLF data in order to further increase the usefulness of the data set. The inversion is performed using a non-linear conjugate gradient scheme (Polak-Ribière) with an inexact line-search. The gradient is obtained by an algebraic adjoint method that requires one additional forward calculation involving the adjoint system matrix. The forward modelling is based on integral equations with an analytic formulation of the half-space Green's tensor. It avoids typically required Hankel transforms and is particularly amenable to singularity removal prior to the numerical integration over the volume elements. The system is solved iteratively, thus avoiding construction and storage of the dense system matrix. By using fast 3-D Fourier transforms on nested grids, subsequently farther away interactions are represented with less detail and therefore with less computational effort, enabling us to bridge the gap between the relatively short wavelengths of the fields (tens of metres) and the large model dimensions (several square kilometres). We find that the approximation of the fields can be off by several per cent, yet the transfer functions in the air are practically unaffected. We verify our code using synthetic calculations from well-established 2-D methods, and trade modelling accuracy off against computational effort in order to keep the inversion feasible in both respects. Our compromise is to limit the permissible resistivity to not fall below 100 m to maintain computational domains as large as 10 10 km 2 and computation times on the order of a few hours on standard PCs. We investigate the effect of possible local violations of these limits. Even though the conductivity magnitude can then not be recovered correctly, we do not observe any structural artefacts related to this in our tests. We invert a data set from northern Sweden, where we find an excellent agreement of known geological features, such as contacts or fault zones, with elongated conductive structures, while high resistivity is encountered in probably less disturbed geology, often related to topographic highs, which have survived predominantly glacial erosion processes. As expected from synthetic studies, the resolution is laterally high, but vertically limited down to the top of conductive structures.

Beschreibung: Induced polarization is a geophysical method looking to image and interpret low-frequency polarization mechanisms occurring in porous media. Below 10 kHz, the quadrature conductivity of metal-free sandy and clayey materials exhibits a distribution of relaxation times, which can be related to the pore size distribution of these porous materials. When the polarization spectra are fitted with a Cole–Cole model, we first observe that the main relaxation time is controlled by the main pore size of the material and that the Cole–Cole exponent c is never much above 0.5, a value corresponding to a Warburg function. The complex conductivity is then obtained through a convolution product between the pore size distribution and such Warburg function. We also provide a way to recover the pore size distribution by performing a deconvolution of measured spectra using the Warburg function. A new dataset of mercury porosimetry and induced polarization data of six siliciclastic materials supports the hypothesis that the Cole–Cole relaxation time is strongly controlled by the pore size, and especially the characteristic pore size corresponding to the peak of the pore size distribution from mercury porosimetry. The distribution of the pore throat sizes of these materials seems fairly well recovered using the Warburg decomposition of the spectral induced polarization spectra but additional data will be needed to confirm this finding.

Beschreibung: We propose a new, simple and efficient method to image electrical resistivity between a set of wells. Our procedure consists of two steps: first, we map the interfaces between various subsurface formations using seismoelectric conversions; second, we derive the formation resistivity using image-guided cross-well electric tomography. In the first step, we focus seismic energy at a set of points located on a regular grid between wells, which enables us to map the geological formations in terms of heterogeneities in electrical, hydraulic and/or seismic properties. The density of the scanning points (i.e. the seismoelectric image resolution) is related to the wavelength of the seismic impulse used to scan the formations. Each time the seismic energy is focused at a point, the resulting electrical potential burst (equivalent to the one generated by a volumetric seismic source) is recorded remotely at a set of electrodes positioned in wells (the reference electrode can be located on the ground surface or far enough to be considered at infinity). We construct a high-resolution ‘seismoelectric’ image by assigning the electrical potential simulated at these fixed electrodes to the location of the seismic focus. In a follow-up step, the structure of this image is used in image-guided inversion to improve electrical resistivity tomography between the two wells. The structural information from the seismoelectric image is used to impose constraints on the model covariance matrix used in the inversion of the electrical resistivity data. This approach offers new perspectives in recovering fine structure of resistivity (high definition resistivity tomography) between the wells, which cannot be resolved through conventional cross-well resistivity or from seismic tomography alone.

Beschreibung: The terrestrial reference frame is a cornerstone for modern geodesy and its applications for a wide range of Earth sciences. The underlying assumption for establishing a terrestrial reference frame is that the motion of the solid Earth's figure centre relative to the mass centre of the Earth system on a multidecadal timescale is linear. However, past international terrestrial reference frames (ITRFs) showed unexpected accelerated motion in their translation parameters. Based on this underlying assumption, the inconsistency of relative origin motions of the ITRFs has been attributed to data reduction imperfection. We investigated the impact of surface mass loading from atmosphere, ocean, snow, soil moisture, ice sheet, glacier and sea level from 1983 to 2008 on the geocentre variations. The resultant geocentre time-series display notable trend acceleration from 1998 onward, in particular in the z -component. This effect is primarily driven by the hydrological mass redistribution in the continents (soil moisture, snow, ice sheet and glacier). The acceleration is statistically significant at the 99 per cent confidence level as determined using the Mann–Kendall test, and it is highly correlated with the satellite laser ranging determined translation series. Our study, based on independent geophysical and hydrological models, demonstrates that, in addition to systematic errors from analysis procedures, the observed non-linearity of the Earth-system behaviour at interannual timescales is physically driven and is able to explain 42 per cent of the disparity between the origins of ITRF2000 and ITRF2005, as well as the high level of consistency between the ITRF2005 and ITRF2008 origins.

Beschreibung: This paper presents a novel mathematical reformulation of the theory of the free wobble/nutation of an axisymmetric reference earth model in hydrostatic equilibrium, using the linear momentum description. The new features of this work consist in the use of (i) Clairaut coordinates (rather than spherical polars), (ii) standard spherical harmonics (rather than generalized spherical surface harmonics), (iii) linear operators (rather than J-square symbols) to represent the effects of rotational and ellipticity coupling between dependent variables of different harmonic degree and (iv) a set of dependent variables all of which are continuous across material boundaries. The resulting infinite system of coupled ordinary differential equations is given explicitly, for an elastic solid mantle and inner core, an inviscid outer core and no magnetic field. The formulation is done to second order in the Earth's ellipticity. To this order it is shown that for wobble modes (in which the lowest harmonic in the displacement field is degree 1 toroidal, with azimuthal order m  = ±1), it is sufficient to truncate the chain of coupled displacement fields at the toroidal harmonic of degree 5 in the solid parts of the earth model. In the liquid core, however, the harmonic expansion of displacement can in principle continue to indefinitely high degree at this order of accuracy. The full equations are shown to yield correct results in three simple cases amenable to analytic solution: a general earth model in rigid rotation, the tiltover mode in a homogeneous solid earth model and the tiltover and Chandler periods for an incompressible homogeneous solid earth model. Numerical results, from programmes based on this formulation, are presented in part II of this paper.

Beschreibung: Numerical solutions are presented for the formulation of the linear momentum description of Earth's dynamics using Clairaut coordinates. We have developed a number of methods to integrate the equations of motion, including starting at the Earth's centre of mass, starting at finite radius and separating the displacement associated with the primary rigid rotation. We include rotation and ellipticity to second order up to spherical harmonic T $_5^m$ , starting with the primary displacement T $_1^m$ with m  = ±1. We are able to confirm many of the previous results for models PREM (with no surface ocean) and 1066A, both in their original form and with neutrally stratified liquid cores. Our period search ranges from the near-seismic band [0.1 sidereal days (sd)] to 3500 sd, within which we have identified the four well-known wobble-nutation modes: the Free Core Nutation (retrograde) at –456 sd, the Free Inner Core Nutation (FICN, prograde) at 468 sd, the Chandler Wobble (prograde) at 402 sd, and the Inner Core Wobble (ICW, prograde) at about 2842 sd (7.8 yr) for neutral PREM. The latter value varies significantly with earth model and integration method. In addition we have verified to high accuracy the tilt-over mode at 1 sd within a factor 10 –6 . In an exhaustive search we found no additional near-diurnal wobble modes that could be identified as nutations. We show that the eigenfunctions for the as-yet-unidentified ICW are extremely sensitive to the details of the earth model, especially the core stability profile and there is no well-defined sense of its wobble relative to the mantle. Calculations are also done for a range of models derived from PREM with homogeneous layers, as well as with incompressible cores. For this kind of model the ICW ceases to have just a simple IC rigid motion when the fluid compressibility is either unchanged or multiplied by a factor 10; in this case the outer core exhibits oscillations that arise from an unstable fluid density stratification. For the FICN our results for the truncation at harmonic T 5 show less change from the T 3 truncation than a similar result reported elsewhere. Finally, we give a thorough discussion of the complete spectrum of the characteristic determinant including the location of poles and non-wobble gravity modes, and discuss in general the dynamics of the inviscid core at periods short compared to those involved in the geodynamo.

Beschreibung: Considering the drawback of existing global weighted mean temperature model, this paper uses 2006–2012 NCEP reanalysis data to establish global empirical model for mapping zenith wet delays onto precipitable water—GTm_N, takes the influence of half-year periodicity of Tm into account when modelling and estimate the initial phase of each cycle. In order to evaluate the precision of GTm_N, we use three different Tm data sets from the NCEP during 2013, 650 radiosonde stations and COSMIC occultation in 2011 to test this model. The results show that GTm_N has higher precision in both ocean and continental area in every moment of every day. The accuracy of GTm_N is higher than Bevis formulas and GTm_II models. In addition, the actual surface temperature is not required in GTm_N model, and it will have wide application in GPS meteorology.

Beschreibung: In subduction zones, shallow subsurface structures are the manifestation of the plate interactions at depth. However, significant water depths, rough bathymetry and presence of heavily deformed accretionary wedge materials hamper imaging of the near-surface features to a great extent using conventional imaging techniques. In this study, we show results using an integrated processing technique to a multichannel seismic data set acquired in 2006 from the northwestern offshore Sumatra. We start with first downward continuing the 12-km-long surface streamer data to the seafloor, followed by a high-resolution traveltime tomography of refracted phases to determine a detailed velocity–depth model of subsurface, which in turns, is used for pre-stack depth migration in order to delineate the shallow subsurface structures beneath the trench, subduction front and outer accretionary wedge. Our velocity–depth model and the depth migrated image depict variation of sediment properties across the front and structures of uppermost sedimentary sequence with an unprecedented high resolution providing the precise location of the frontal and conjugate thrusts, highly folded sedimentary sequences, which in turns describe their relationship with the top of the subducting plate and factors that control rupture propagation to the trench. Furthermore, we estimate the porosity distribution across the front, where we find a 12 and 18 per cent decrease in porosity beneath the deformation front and the inner accretionary plateau at 500 m below the seafloor, respectively, which we interpret to be due to the compaction. A significant decrease in porosity at the plate interface below 5–6-km thick sediments near the deformation front would increase the coupling, leading to the rupture propagation up to the trench, uplifting 4.5 km water and producing large tsunami.

Beschreibung: We present a scale- and parameter-adaptive method to pre-condition the gradient of the parameters to be inverted in time-domain 2-D elastic full-waveform inversion (FWI). The proposed technique, which relies on a change of variables of the model parameters, allows to balance the value of the gradient of the Lamé parameters and density throughout the model in each step of the multiscale inversion. The main difference compared to existing gradient pre-conditioners is that the variables are automatically selected based on a least-squares minimization criteria of the gradient weight, which corresponds to the product of the gradient by a power of the parameter to be inverted. Based on numerical tests made with (1) a modified version of the Marmousi-2 model, and (2) a high-velocity and density local anomaly model, we illustrate that the value of the power helps to balance the gradient throughout the model. In addition, we show that a particular value exists for each parameter that optimizes the inversion results in terms of accuracy and efficiency. For the two models, the optimal power is ~2.0–2.5 and ~1.5 for the first and second Lamé parameters, respectively; and between 3 and 6, depending on the inverted frequency, for density. These power values provide the fastest and most accurate inversion results for the three parameters in the framework of multiscale and multishooting FWI using three different optimization schemes.

Beschreibung: Revealing the clonal composition of a single tumor is essential for identifying cell subpopulations with metastatic potential in primary tumors or with resistance to therapies in metastatic tumors. Sequencing technologies provide only an overview of the aggregate of numerous cells. Computational approaches to de-mix a collective signal composed of the aberrations of a mixed cell population of a tumor sample into its individual components are not available. We propose an evolutionary framework for deconvolving data from a single genome-wide experiment to infer the composition, abundance and evolutionary paths of the underlying cell subpopulations of a tumor. We have developed an algorithm (TrAp) for solving this mixture problem. In silico analyses show that TrAp correctly deconvolves mixed subpopulations when the number of subpopulations and the measurement errors are moderate. We demonstrate the applicability of the method using tumor karyotypes and somatic hypermutation data sets. We applied TrAp to Exome-Seq experiment of a renal cell carcinoma tumor sample and compared the mutational profile of the inferred subpopulations to the mutational profiles of single cells of the same tumor. Finally, we deconvolve sequencing data from eight acute myeloid leukemia patients and three distinct metastases of one melanoma patient to exhibit the evolutionary relationships of their subpopulations.

Beschreibung: MicroRNAs (miRNAs) constitute an important class of small regulatory RNAs that are derived from distinct hairpin precursors (pre-miRNAs). In contrast to mature miRNAs, which have been characterized in numerous genome-wide studies of different organisms, research on global profiling of pre-miRNAs is limited. Here, using massive parallel sequencing, we have performed global characterization of both mouse mature and precursor miRNAs. In total, 87 369 704 and 252 003 sequencing reads derived from 887 mature and 281 precursor miRNAs were obtained, respectively. Our analysis revealed new aspects of miRNA/pre-miRNA processing and modification, including eight Ago2-cleaved pre-miRNAs, eight new instances of miRNA editing and exclusively 5' tailed mirtrons. Furthermore, based on the sequences of both mature and precursor miRNAs, we developed a miRNA discovery pipeline, miRGrep, which does not rely on the availability of genome reference sequences. In addition to 239 known mouse pre-miRNAs, miRGrep predicted 41 novel ones with high confidence. Similar as known ones, the mature miRNAs derived from most of these novel loci showed both reduced abundance following Dicer knockdown and the binding with Argonaute2. Evaluation on data sets obtained from Caenorhabditis elegans and Caenorhabditis sp.11 demonstrated that miRGrep could be widely used for miRNA discovery in metazoans, especially in those without genome reference sequences.

Beschreibung: DNA methylation is one of the most important epigenetic alterations involved in the control of gene expression. Bisulfite sequencing of genomic DNA is currently the only method to study DNA methylation patterns at single-nucleotide resolution. Hence, next-generation sequencing of bisulfite-converted DNA is the method of choice to investigate DNA methylation profiles at the genome-wide scale. Nevertheless, whole genome sequencing for analysis of human methylomes is expensive, and a method for targeted gene analysis would provide a good alternative in many cases where the primary interest is restricted to a set of genes. Here, we report the successful use of a custom Agilent SureSelect Target Enrichment system for the hybrid capture of bisulfite-converted DNA. We prepared bisulfite-converted next-generation sequencing libraries, which are enriched for the coding and regulatory regions of 174 ADME genes (i.e. genes involved in the metabolism and distribution of drugs). Sequencing of these libraries on Illumina’s HiSeq2000 revealed that the method allows a reliable quantification of methylation levels of CpG sites in the selected genes, and validation of the method using pyrosequencing and the Illumina 450K methylation BeadChips revealed good concordance.

Beschreibung: It is a challenge to classify protein-coding or non-coding transcripts, especially those re-constructed from high-throughput sequencing data of poorly annotated species. This study developed and evaluated a powerful signature tool, Coding-Non-Coding Index (CNCI), by profiling adjoining nucleotide triplets to effectively distinguish protein-coding and non-coding sequences independent of known annotations. CNCI is effective for classifying incomplete transcripts and sense–antisense pairs. The implementation of CNCI offered highly accurate classification of transcripts assembled from whole-transcriptome sequencing data in a cross-species manner, that demonstrated gene evolutionary divergence between vertebrates, and invertebrates, or between plants, and provided a long non-coding RNA catalog of orangutan. CNCI software is available at http://www.bioinfo.org/software/cnci .

Beschreibung: The epigenetic modification of 5-hydroxymethylcytosine (5hmC) is receiving great attention due to its potential role in DNA methylation reprogramming and as a cell state identifier. Given this interest, it is important to identify reliable and cost-effective methods for the enrichment of 5hmC marked DNA for downstream analysis. We tested three commonly used affinity-based enrichment techniques; (i) antibody, (ii) chemical capture and (iii) protein affinity enrichment and assessed their ability to accurately and reproducibly report 5hmC profiles in mouse tissues containing high (brain) and lower (liver) levels of 5hmC. The protein-affinity technique is a poor reporter of 5hmC profiles, delivering 5hmC patterns that are incompatible with other methods. Both antibody and chemical capture-based techniques generate highly similar genome-wide patterns for 5hmC, which are independently validated by standard quantitative PCR (qPCR) and glucosyl-sensitive restriction enzyme digestion (gRES-qPCR). Both antibody and chemical capture generated profiles reproducibly link to unique chromatin modification profiles associated with 5hmC. However, there appears to be a slight bias of the antibody to bind to regions of DNA rich in simple repeats. Ultimately, the increased specificity observed with chemical capture-based approaches makes this an attractive method for the analysis of locus-specific or genome-wide patterns of 5hmC.

Beschreibung: Methylation-specific fluorescence in situ hybridization (MeFISH) was developed for microscopic visualization of DNA methylation status at specific repeat sequences in individual cells. MeFISH is based on the differential reactivity of 5-methylcytosine and cytosine in target DNA for interstrand complex formation with osmium and bipyridine-containing nucleic acids (ICON). Cell nuclei and chromosomes hybridized with fluorescence-labeled ICON probes for mouse major and minor satellite repeats were treated with osmium for crosslinking. After denaturation, fluorescent signals were retained specifically at satellite repeats in wild-type, but not in DNA methyltransferase triple-knockout (negative control) mouse embryonic stem cells. Moreover, using MeFISH, we successfully detected hypomethylated satellite repeats in cells from patients with immunodeficiency, centromeric instability and facial anomalies syndrome and 5-hydroxymethylated satellite repeats in male germ cells, the latter of which had been considered to be unmethylated based on anti-5-methylcytosine antibody staining. MeFISH will be suitable for a wide range of applications in epigenetics research and medical diagnosis.

Beschreibung: The coupling of chromosome conformation capture (3C) with next-generation sequencing technologies enables the high-throughput detection of long-range genomic interactions, via the generation of ligation products between DNA sequences, which are closely juxtaposed in vivo . These interactions involve promoter regions, enhancers and other regulatory and structural elements of chromosomes and can reveal key details of the regulation of gene expression. 3C-seq is a variant of the method for the detection of interactions between one chosen genomic element (viewpoint) and the rest of the genome. We present r3Cseq , an R/Bioconductor package designed to perform 3C-seq data analysis in a number of different experimental designs. The package reads a common aligned read input format, provides data normalization, allows the visualization of candidate interaction regions and detects statistically significant chromatin interactions, thus greatly facilitating hypothesis generation and the interpretation of experimental results. We further demonstrate its use on a series of real-world applications.

Beschreibung: Genomic deletions induced by imprecise excision of transposons have been used to disrupt gene functions in Drosophila . To determine the excision properties of Tol2 , a popular transposon in zebrafish, we took advantage of two transgenic zebrafish lines Et(gata2a:EGFP)pku684 and Et(gata2a:EGFP)pku760 , and mobilized the transposon by injecting transposase mRNA into homozygous transgenic embryos. Footprint analysis showed that the Tol2 transposons were excised in either a precise or an imprecise manner. Furthermore, we identified 1093-bp and 1253-bp genomic deletions in Et(gata2a:EGFP)pku684 founder embryos flanking the 5' end of the original Tol2 insertion site, and a 1340-bp deletion in the Et(gata2a:EGFP)pku760 founder embryos flanking the 3' end of the insertion site. The mosaic Et(gata2a:EGFP)pku684 embryos were raised to adulthood and screened for germline transmission of Tol2 excision in their F 1 progeny. On average, ~42% of the F 1 embryos displayed loss or altered EGFP patterns, demonstrating that this transposon could be efficiently excised from the zebrafish genome in the germline. Furthermore, from 59 founders, we identified one that transmitted the 1093-bp genomic deletion to its offspring. These results suggest that imprecise Tol2 transposon excision can be used as an alternative strategy to achieve gene targeting in zebrafish.

Beschreibung: Combinations of histone modifications have significant biological roles, such as maintenance of pluripotency and cancer development, but cannot be analyzed at the single cell level. Here, we visualized a combination of histone modifications by applying the in situ proximity ligation assay, which detects two proteins in close vicinity (~30 nm). The specificity of the method [designated as imaging of a combination of histone modifications (iChmo)] was confirmed by positive signals from H3K4me3/acetylated H3K9, H3K4me3/RNA polymerase II and H3K9me3/H4K20me3, and negative signals from H3K4me3/H3K9me3. Bivalent modification was clearly visualized by iChmo in wild-type embryonic stem cells (ESCs) known to have it, whereas rarely in Suz12 knockout ESCs and mouse embryonic fibroblasts known to have little of it. iChmo was applied to analysis of epigenetic and phenotypic changes of heterogeneous cell population, namely, ESCs at an early stage of differentiation, and this revealed that the bivalent modification disappeared in a highly concerted manner, whereas phenotypic differentiation proceeded with large variations among cells. Also, using this method, we were able to visualize a combination of repressive histone marks in tissue samples. The application of iChmo to samples with heterogeneous cell population and tissue samples is expected to clarify unknown biological and pathological significance of various combinations of epigenetic modifications.

Beschreibung: Nucleosome positioning on the chromatin strand plays a critical role in regulating accessibility of DNA to transcription factors and chromatin modifying enzymes. Hence, detailed information on nucleosome depletion or movement at cis -acting regulatory elements has the potential to identify predicted binding sites for trans -acting factors. Using a novel method based on enrichment of mononucleosomal DNA by bacterial artificial chromosome hybridization, we mapped nucleosome positions by deep sequencing across 250 kb, encompassing the cystic fibrosis transmembrane conductance regulator ( CFTR ) gene. CFTR shows tight tissue-specific regulation of expression, which is largely determined by cis -regulatory elements that lie outside the gene promoter. Although multiple elements are known, the repertoire of transcription factors that interact with these sites to activate or repress CFTR expression remains incomplete. Here, we show that specific nucleosome depletion corresponds to well-characterized binding sites for known trans -acting factors, including hepatocyte nuclear factor 1, Forkhead box A1 and CCCTC-binding factor. Moreover, the cell-type selective nucleosome positioning is effective in predicting binding sites for novel interacting factors, such as BAF155. Finally, we identify transcription factor binding sites that are overrepresented in regions where nucleosomes are depleted in a cell-specific manner. This approach recognizes the glucocorticoid receptor as a novel trans -acting factor that regulates CFTR expression in vivo .

Beschreibung: Here, we describe an approach to isolate native chromatin sections without genomic engineering for label-free proteomic identification of associated proteins and histone post-translational modifications. A transcription activator-like (TAL) protein A fusion protein was designed to recognize a unique site in the yeast GAL1 promoter. The TAL-PrA fusion enabled chromatin affinity purification (ChAP) of a small section of native chromatin upstream from the GAL1 locus, permitting mass spectrometric (MS) identification of proteins and histone post-translational modifications regulating galactose-induced transcription. This TAL-ChAP-MS approach allows the biochemical isolation of a specific native genomic locus for proteomic studies and will provide for unprecedented objective insight into protein and epigenetic mechanisms regulating site-specific chromosome metabolism.

Beschreibung: Sequence elements, at all levels—DNA, RNA and protein, play a central role in mediating molecular recognition and thereby molecular regulation and signaling. Studies that focus on measuring and investigating sequence-based recognition make use of statistical and computational tools, including approaches to searching sequence motifs. State-of-the-art motif searching tools are limited in their coverage and ability to address large motif spaces. We develop and present statistical and algorithmic approaches that take as input ranked lists of sequences and return significant motifs. The efficiency of our approach, based on suffix trees, allows searches over motif spaces that are not covered by existing tools. This includes searching variable gap motifs—two half sites with a flexible length gap in between—and searching long motifs over large alphabets. We used our approach to analyze several high-throughput measurement data sets and report some validation results as well as novel suggested motifs and motif refinements. We suggest a refinement of the known estrogen receptor 1 motif in humans, where we observe gaps other than three nucleotides that also serve as significant recognition sites, as well as a variable length motif related to potential tyrosine phosphorylation.

Beschreibung: Cataloging the association of transcripts to genetic variants in recent years holds the promise for functional dissection of regulatory structure of human transcription. Here, we present a novel approach, which aims at elucidating the joint relationships between transcripts and single-nucleotide polymorphisms (SNPs). This entails detection and analysis of modules of transcripts, each weakly associated to a single genetic variant, together exposing a high-confidence association signal between the module and this ‘main’ SNP. To explore how transcripts in a module are related to causative loci for that module, we represent such dependencies by a graphical model. We applied our method to the existing data on genetics of gene expression in the liver. The modules are significantly more, larger and denser than found in permuted data. Quantification of the confidence in a module as a likelihood score, allows us to detect transcripts that do not reach genome-wide significance level. Topological analysis of each module identifies novel insights regarding the flow of causality between the main SNP and transcripts. We observe similar annotations of modules from two sources of information: the enrichment of a module in gene subsets and locus annotation of the genetic variants. This and further phenotypic analysis provide a validation for our methodology.

Beschreibung: Phase variation of surface structures occurs in diverse bacterial species due to stochastic, high frequency, reversible mutations. Multiple genes of Campylobacter jejuni are subject to phase variable gene expression due to mutations in polyC/G tracts. A modal length of nine repeats was detected for polyC/G tracts within C. jejuni genomes. Switching rates for these tracts were measured using chromosomally-located reporter constructs and high rates were observed for cj1139 (G8) and cj0031 (G9). Alteration of the cj1139 tract from G8 to G11 increased mutability 10-fold and changed the mutational pattern from predominantly insertions to mainly deletions. Using a multiplex PCR, major changes were detected in ‘on/off’ status for some phase variable genes during passage of C. jejuni in chickens. Utilization of observed switching rates in a stochastic, theoretical model of phase variation demonstrated links between mutability and genetic diversity but could not replicate observed population diversity. We propose that modal repeat numbers have evolved in C. jejuni genomes due to molecular drivers associated with the mutational patterns of these polyC/G repeats, rather than by selection for particular switching rates, and that factors other than mutational drift are responsible for generating genetic diversity during host colonization by this bacterial pathogen.

Beschreibung: Control of translation in eukaryotes is complex, depending on the binding of various factors to mRNAs. Available data for subsets of mRNAs that are translationally up- and down-regulated in yeast eIF4E-binding protein (4E-BP) deletion mutants are coupled with reported mRNA secondary structure measurements to investigate whether 5'-UTR secondary structure varies between the subsets. Genes with up-regulated translational efficiencies in the caf20 mutant have relatively high averaged 5'-UTR secondary structure. There is no apparent wide-scale correlation of RNA-binding protein preferences with the increased 5'-UTR secondary structure, leading us to speculate that the secondary structure itself may play a role in differential partitioning of mRNAs between eIF4E/4E-BP repression and eIF4E/eIF4G translation initiation. Both Caf20p and Eap1p contain stretches of positive charge in regions of predicted disorder. Such regions are also present in eIF4G and have been reported to associate with mRNA binding. The pattern of these segments, around the canonical eIF4E-binding motif, varies between each 4E-BP and eIF4G. Analysis of gene ontology shows that yeast proteins containing predicted disordered segments, with positive charge runs, are enriched for nucleic acid binding. We propose that the 4E-BPs act, in part, as differential, flexible, polyelectrostatic scaffolds for mRNAs.

Beschreibung: The activation of cryptic 5' splice sites (5' SSs) is often related to human hereditary diseases. The DNA-based mutation screening strategies are commonly used to recognize the cryptic 5' SSs, because features of the local DNA sequence can influence the choice of cryptic 5' SSs. To improve the identification of the cryptic 5' SSs, we developed a structure-based method, named SPO (structure profiles and odds measure), which combines two parameters, the structural feature derived from hydroxyl radical cleavage pattern and odds measure, to assess the likelihood of a cryptic 5' SS activation in competing with its paired authentic 5' SS. Compared to the current tools for identifying activated cryptic 5' SSs, the SPO algorithm achieves higher prediction accuracy than the other methods, including MaxEnt, MDD, Markov model, weight matrix model, Shapiro and Senapathy matrix, R i and G . In addition, the predicted SPO scores from the SPO algorithm exhibited a greater degree of correlation with the strength of cryptic 5' SS activation than that measured from the other seven methods. In conclusion, the SPO algorithm provides an optimal identification of cryptic 5' SSs, can be applied in designing mutagenesis experiments for various splicing events and may be helpful to investigate the relationship between structural variants and human hereditary diseases.

Beschreibung: The Joint BioEnergy Institute Inventory of Composable Elements (JBEI-ICEs) is an open source registry platform for managing information about biological parts. It is capable of recording information about ‘legacy’ parts, such as plasmids, microbial host strains and Arabidopsis seeds, as well as DNA parts in various assembly standards. ICE is built on the idea of a web of registries and thus provides strong support for distributed interconnected use. The information deposited in an ICE installation instance is accessible both via a web browser and through the web application programming interfaces, which allows automated access to parts via third-party programs. JBEI-ICE includes several useful web browser-based graphical applications for sequence annotation, manipulation and analysis that are also open source. As with open source software, users are encouraged to install, use and customize JBEI-ICE and its components for their particular purposes. As a web application programming interface, ICE provides well-developed parts storage functionality for other synthetic biology software projects. A public instance is available at public-registry.jbei.org, where users can try out features, upload parts or simply use it for their projects. The ICE software suite is available via Google Code, a hosting site for community-driven open source projects.

Beschreibung: Bromodeoxyuridine (5-bromo-2'-deoxyuridine, BrdU) is a halogenated nucleotide of low toxicity commonly used to monitor DNA replication. It is considered a valuable tool for in vitro and in vivo studies, including the detection of the small population of neural stem cells (NSC) in the mammalian brain. Here, we show that NSC grown in self-renewing conditions in vitro , when exposed to BrdU, lose the expression of stem cell markers like Nestin, Sox2 and Pax6 and undergo glial differentiation, strongly up-regulating the astrocytic marker GFAP. The onset of GFAP expression in BrdU exposed NSC was paralleled by a reduced expression of key DNA methyltransferases (DNMT) and a rapid loss of global DNA CpG methylation, as we determined by our specially developed analytic assay. Remarkably, a known DNA demethylating compound, 5-aza-2'-deoxycytidine (Decitabine), had similar effect on demethylation and differentiation of NSC. Since our key findings apply also to NSC derived from murine forebrain, our observations strongly suggest more caution in BrdU uses in stem cells research. We also propose that BrdU and its related substances may also open new opportunities for differentiation therapy in oncology.

Beschreibung: In Escherichia coli , the SeqA protein binds specifically to GATC sequences which are methylated on the A of the old strand but not on the new strand. Such hemimethylated DNA is produced by progression of the replication forks and lasts until Dam methyltransferase methylates the new strand. It is therefore believed that a region of hemimethylated DNA covered by SeqA follows the replication fork. We show that this is, indeed, the case by using global ChIP on Chip analysis of SeqA in cells synchronized regarding DNA replication. To assess hemimethylation, we developed the first genome-wide method for methylation analysis in bacteria. Since loss of the SeqA protein affects growth rate only during rapid growth when cells contain multiple replication forks, a comparison of rapid and slow growth was performed. In cells with six replication forks per chromosome, the two old forks were found to bind surprisingly little SeqA protein. Cell cycle analysis showed that loss of SeqA from the old forks did not occur at initiation of the new forks, but instead occurs at a time point coinciding with the end of SeqA-dependent origin sequestration. The finding suggests simultaneous origin de-sequestration and loss of SeqA from old replication forks.

Beschreibung: Live-cell measurement of protein binding to chromatin allows probing cellular biochemistry in physiological conditions, which are difficult to mimic in vitro . However, different studies have yielded widely discrepant predictions, and so it remains uncertain how to make the measurements accurately. To establish a benchmark we measured binding of the transcription factor p53 to chromatin by three approaches: fluorescence recovery after photobleaching (FRAP), fluorescence correlation spectroscopy (FCS) and single-molecule tracking (SMT). Using new procedures to analyze the SMT data and to guide the FRAP and FCS analysis, we show how all three approaches yield similar estimates for both the fraction of p53 molecules bound to chromatin (only about 20%) and the residence time of these bound molecules (~1.8 s). We also apply these procedures to mutants in p53 chromatin binding. Our results support the model that p53 locates specific sites by first binding at sequence-independent sites.

Beschreibung: Understanding the numerous functions that RNAs play in living cells depends critically on knowledge of their three-dimensional structure. Due to the difficulties in experimentally assessing structures of large RNAs, there is currently great demand for new high-resolution structure prediction methods. We present the novel method for the fully automated prediction of RNA 3D structures from a user-defined secondary structure. The concept is founded on the machine translation system. The translation engine operates on the RNA FRABASE database tailored to the dictionary relating the RNA secondary structure and tertiary structure elements. The translation algorithm is very fast. Initial 3D structure is composed in a range of seconds on a single processor. The method assures the prediction of large RNA 3D structures of high quality. Our approach needs neither structural templates nor RNA sequence alignment, required for comparative methods. This enables the building of unresolved yet native and artificial RNA structures. The method is implemented in a publicly available, user-friendly server RNAComposer. It works in an interactive mode and a batch mode. The batch mode is designed for large-scale modelling and accepts atomic distance restraints. Presently, the server is set to build RNA structures of up to 500 residues.

Beschreibung: DNA methylation plays a key role in epigenetic regulation of eukaryotic genomes. Hence the genome-wide distribution of 5-methylcytosine, or the methylome, has been attracting intense attention. In recent years, whole-genome bisulfite sequencing (WGBS) has enabled methylome analysis at single-base resolution. However, WGBS typically requires microgram quantities of DNA as well as global PCR amplification, thereby precluding its application to samples of limited amounts. This is presumably because bisulfite treatment of adaptor-tagged templates, which is inherent to current WGBS methods, leads to substantial DNA fragmentation. To circumvent the bisulfite-induced loss of intact sequencing templates, we conceived an alternative method termed Post-Bisulfite Adaptor Tagging (PBAT) wherein bisulfite treatment precedes adaptor tagging by two rounds of random primer extension. The PBAT method can generate a substantial number of unamplified reads from as little as subnanogram quantities of DNA. It requires only 100 ng of DNA for amplification-free WGBS of mammalian genomes. Thus, the PBAT method will enable various novel applications that would not otherwise be possible, thereby contributing to the rapidly growing field of epigenomics.

Beschreibung: Programmed –1 ribosomal frameshifting is employed in the expression of a number of viral and cellular genes. In this process, the ribosome slips backwards by a single nucleotide and continues translation of an overlapping reading frame, generating a fusion protein. Frameshifting signals comprise a heptanucleotide slippery sequence, where the ribosome changes frame, and a stimulatory RNA structure, a stem–loop or RNA pseudoknot. Antisense oligonucleotides annealed appropriately 3' of a slippery sequence have also shown activity in frameshifting, at least in vitro . Here we examined frameshifting at the U 6 A slippery sequence of the HIV gag/pol signal and found high levels of both –1 and –2 frameshifting with stem–loop, pseudoknot or antisense oligonucleotide stimulators. By examining –1 and –2 frameshifting outcomes on mRNAs with varying slippery sequence-stimulatory RNA spacing distances, we found that –2 frameshifting was optimal at a spacer length 1–2 nucleotides shorter than that optimal for –1 frameshifting with all stimulatory RNAs tested. We propose that the shorter spacer increases the tension on the mRNA such that when the tRNA detaches, it more readily enters the –2 frame on the U 6 A heptamer. We propose that mRNA tension is central to frameshifting, whether promoted by stem–loop, pseudoknot or antisense oligonucleotide stimulator.

Beschreibung: Understanding the categorization of human diseases is critical for reliably identifying disease causal genes. Recently, genome-wide studies of abnormal chromosomal locations related to diseases have mapped 〉2000 phenotype–gene relations, which provide valuable information for classifying diseases and identifying candidate genes as drug targets. In this article, a regularized non-negative matrix tri-factorization (R-NMTF) algorithm is introduced to co-cluster phenotypes and genes, and simultaneously detect associations between the detected phenotype clusters and gene clusters. The R-NMTF algorithm factorizes the phenotype–gene association matrix under the prior knowledge from phenotype similarity network and protein–protein interaction network, supervised by the label information from known disease classes and biological pathways. In the experiments on disease phenotype–gene associations in OMIM and KEGG disease pathways, R-NMTF significantly improved the classification of disease phenotypes and disease pathway genes compared with support vector machines and Label Propagation in cross-validation on the annotated phenotypes and genes. The newly predicted phenotypes in each disease class are highly consistent with human phenotype ontology annotations. The roles of the new member genes in the disease pathways are examined and validated in the protein–protein interaction subnetworks. Extensive literature review also confirmed many new members of the disease classes and pathways as well as the predicted associations between disease phenotype classes and pathways.

Beschreibung: The development of algorithms for designing artificial RNA sequences that fold into specific secondary structures has many potential biomedical and synthetic biology applications. To date, this problem remains computationally difficult, and current strategies to address it resort to heuristics and stochastic search techniques. The most popular methods consist of two steps: First a random seed sequence is generated; next, this seed is progressively modified (i.e. mutated) to adopt the desired folding properties. Although computationally inexpensive, this approach raises several questions such as (i) the influence of the seed; and (ii) the efficiency of single-path directed searches that may be affected by energy barriers in the mutational landscape. In this article, we present RNA-ensign , a novel paradigm for RNA design. Instead of taking a progressive adaptive walk driven by local search criteria, we use an efficient global sampling algorithm to examine large regions of the mutational landscape under structural and thermodynamical constraints until a solution is found. When considering the influence of the seeds and the target secondary structures, our results show that, compared to single-path directed searches, our approach is more robust, succeeds more often and generates more thermodynamically stable sequences. An ensemble approach to RNA design is thus well worth pursuing as a complement to existing approaches. RNA-ensign is available at http://csb.cs.mcgill.ca/RNAensign .

Beschreibung: The mammalian thymine DNA glycosylase (TDG) is implicated in active DNA demethylation via the base excision repair pathway. TDG excises the mismatched base from G:X mismatches, where X is uracil, thymine or 5-hydroxymethyluracil (5hmU). These are, respectively, the deamination products of cytosine, 5-methylcytosine (5mC) and 5-hydroxymethylcytosine (5hmC). In addition, TDG excises the Tet protein products 5-formylcytosine (5fC) and 5-carboxylcytosine (5caC) but not 5hmC and 5mC, when paired with a guanine. Here we present a post-reactive complex structure of the human TDG domain with a 28-base pair DNA containing a G:5hmU mismatch. TDG flips the target nucleotide from the double-stranded DNA, cleaves the N -glycosidic bond and leaves the C1' hydrolyzed abasic sugar in the flipped state. The cleaved 5hmU base remains in a binding pocket of the enzyme. TDG allows hydrogen-bonding interactions to both T/U-based (5hmU) and C-based (5caC) modifications, thus enabling its activity on a wider range of substrates. We further show that the TDG catalytic domain has higher activity for 5caC at a lower pH (5.5) as compared to the activities at higher pH (7.5 and 8.0) and that the structurally related Escherichia coli mismatch uracil glycosylase can excise 5caC as well. We discuss several possible mechanisms, including the amino-imino tautomerization of the substrate base that may explain how TDG discriminates against 5hmC and 5mC.

Beschreibung: Identifying cancer driver genes and pathways among all somatic mutations detected in a cohort of tumors is a key challenge in cancer genomics. Traditionally, this is done by prioritizing genes according to the recurrence of alterations that they bear. However, this approach has some known limitations, such as the difficulty to correctly estimate the background mutation rate, and the fact that it cannot identify lowly recurrently mutated driver genes. Here we present a novel approach, Oncodrive-fm, to detect candidate cancer drivers which does not rely on recurrence. First, we hypothesized that any bias toward the accumulation of variants with high functional impact observed in a gene or group of genes may be an indication of positive selection and can thus be used to detect candidate driver genes or gene modules. Next, we developed a method to measure this bias (FM bias) and applied it to three datasets of tumor somatic variants. As a proof of concept of our hypothesis we show that most of the highly recurrent and well-known cancer genes exhibit a clear FM bias. Moreover, this novel approach avoids some known limitations of recurrence-based approaches, and can successfully identify lowly recurrent candidate cancer drivers.