ALBERT

Description: In general, the complex electrical resistivity in the subsurface is anisotropic. Despite this, algorithms for the tomographic inversion of complex resistivity data commonly assume isotropy, mainly due to the lack of anisotropic modelling and inversion schemes, potentially leading to artefacts in the inversion results in the presence of anisotropy. The development of an effective anisotropic complex resistivity inversion algorithm which utilizes the gradient information of some cost function benefits from understanding the characteristics of the problem's sensitivities, that is, the partial derivative of the impedance forward response with respect to the complex conductivities in the different spatial directions, as well as with respect to the different ratios of complex conductivities, that is, the different anisotropy ratios. We here derive expressions for these sensitivities and, based on a 2.5-D finite-element modelling algorithm, we compute and discuss sensitivity distributions as well as measurement response curves of typical surface and cross-borehole measurement configurations for 2-D subsurface anisotropic complex resistivity distributions. Depending on the electrode layout and measurement configuration, the sensitivity with respect to the conductivity in a particular direction shows a unique pattern, while for other directions sensitivity patterns are qualitatively similar. These sensitivity characteristics translate into important equivalences between impedance responses of local anisotropic and isotropic anomalies, for both magnitude and phase. Accordingly, with collinear surface arrays only the complex conductivity in the direction of the electrode layout can be unambiguously resolved, and with cross-borehole arrays only the conductivity in the vertical direction, provided an in-hole current injection is used. Nevertheless, anisotropy ratios involving these resolvable conductivity components are likewise detectable. The distinct shape of the measurement response curves, reflecting the distinct spatial patterns of the corresponding sensitivity distributions, suggest that optimized measurement configurations can be inferred for specific exploration questions involving electrical anisotropy and given electrode layouts. The gained insight into the characteristics of the sensitivity distributions of complex resistivity measurements in case of subsurface anisotropy should guide the implementation of effective anisotropic complex resistivity inversion schemes and lead to a routine use of such schemes in any resistivity and induced polarization surveys whenever subsurface electrical anisotropy could be encountered.

Description: The streaming potential phenomenon is an electrokinetic effect that occurs in porous media. It is characterized by an electrokinetic (EK) coefficient. The aim of this paper is to simulate the EK coefficient in unsaturated conditions using the Lattice Boltzmann method in a 2-D capillary channel. The multiphase flow is simulated with the model of Shan & Chen. The Poisson–Boltzmann equation is solved by implementing the model of Chai & Shi. The streaming potential response shows a non-monotonous behaviour due to the combination of the increase of charge density and decrease of flow velocity with decreasing water saturation. Using a potential of –20 mV at the air–water interface, an enhancement of a factor 5–30 of the EK coefficient, compared to the saturated state, can be observed due to the positive charge excess at this interface which is magnified by the fluid velocity away from the rock surface. This enhancement is correlated to the fractioning of the bubbles, and to the dynamic state of these bubbles, moving or entrapped in the crevices of the channel.

Description: Earlier, vitamin C was demonstrated to sterilize Mycobacterium tuberculosis culture via Fenton's reaction at high concentration. It alters the regulatory pathways associated with stress response and dormancy. Since (p)ppGpp is considered to be the master regulator of stress response and is responsible for bacterial survival under stress, we tested the effect of vitamin C on the formation of (p)ppGpp. In vivo estimation of (p)ppGpp showed a decrease in (p)ppGpp levels in vitamin C-treated M. smegmatis cells in comparison to the untreated cells. Furthermore, in vitro (p)ppGpp synthesis using Rel MSM enzyme was conducted in order to confirm the specificity of the inhibition in the presence of variable concentrations of vitamin C. We observed that vitamin C at high concentration can inhibit the synthesis of (p)ppGpp. We illustrated binding of vitamin C to Rel MSM by isothermal titration calorimetry. Enzyme kinetics was followed where K 0.5 was found to be increased with the concomitant reduction of V max value suggesting mixed inhibition. Both long-term survival and biofilm formation were inhibited by vitamin C. The experiments suggest that vitamin C has the potential to be developed as the inhibitor of (p)ppGpp synthesis and stress response, at least in the concentration range used here.

Description: Although nitric oxide (NO) is an important signaling molecule in bacteria and higher organisms, excessive intracellular NO is highly reactive and dangerous. Therefore, living cells need strict regulation systems for cellular NO homeostasis. Recently, we discovered that Streptomyces coelicolor A3(2) retains the nitrogen oxide cycle (NO 3 – -〉NO 2 – -〉NO-〉NO 3 – ) and nitrite removal system. The nitrogen oxide cycle regulates cellular NO levels, thereby controlling secondary metabolism initiation (red-pigmented antibiotic, RED production) and morphological differentiation. Nitrite induces gene expression in neighboring cells, suggesting another role for this cycle as a producer of transmittable intercellular communication molecules. Here, we demonstrated that ammonium-producing nitrite reductase (NirBD) is involved in regulating NO homeostasis in S. coelicolor A3(2). NirBD was constitutively produced in culture independently of GlnR, a known transcriptional factor. NirBD cleared the accumulated nitrite from the medium. Nir deletion mutants showed increased NO-dependent gene expression at later culture stages, whereas the wild-type M145 showed decreased expression, suggesting that high NO concentration was maintained in the mutant. Moreover, the nir deletion mutant produced more RED than that produced by the wild-type M145. These results suggest that NO 2 – removal by NirBD is important to regulate NO homeostasis and to complete NO signaling in S. coelicolor .

Description: Joint inversion that simultaneously inverts multiple geophysical data sets to recover a common Earth model is increasingly being applied to exploration problems. Petrophysical data can serve as an effective constraint to link different physical property models in such inversions. There are two challenges, among others, associated with the petrophysical approach to joint inversion. One is related to the multimodality of petrophysical data because there often exist more than one relationship between different physical properties in a region of study. The other challenge arises from the fact that petrophysical relationships have different characteristics and can exhibit point, linear, quadratic, or exponential forms in a crossplot. The fuzzy c-means (FCM) clustering technique is effective in tackling the first challenge and has been applied successfully. We focus on the second challenge in this paper and develop a joint inversion method based on variations of the FCM clustering technique. To account for the specific shapes of petrophysical relationships, we introduce several different fuzzy clustering algorithms that are capable of handling different shapes of petrophysical relationships. We present two synthetic and one field data examples and demonstrate that, by choosing appropriate distance measures for the clustering component in the joint inversion algorithm, the proposed joint inversion method provides an effective means of handling common petrophysical situations we encounter in practice. The jointly inverted models have both enhanced structural similarity and increased petrophysical correlation, and better represent the subsurface in the spatial domain and the parameter domain of physical properties.

Description: Coal seam fires are a worldwide disaster of both ecological and economic importance. Their remote detection from the ground surface or using airborne techniques is required for developing efficient strategies to extinguish them. We investigate here the use of time-domain-induced polarization to localize coal seam fires. For laboratory experiments, we first introduce a modified time-domain-induced polarization methodology to quickly acquire and invert the secondary voltage distribution mapped after the shutdown of the primary current. A set of sandbox experiments is conducted in which coal is embedded into humidified sand. Raw coal alone generates significant induced polarization anomalies, above those shown by the sand. Even higher induced polarization anomalies are detected in presence of a coal seam fire. We postulate that the higher chargeability is due to the pyrolysis, which may enhance electronic polarization or the polarization associated with the cation exchange capacity (CEC) of the material. The position of the coal seam fire is well recovered inside the tank by inverting the secondary voltages in term of a source current density distribution. We also collected field data over a recognized coal seam fire in Colorado, USA. A chargeability anomaly (~800 mV V –1 ) and a resistivity anomaly (~1 Ohm m) are observed at the position of the coal seam fire. We propose a normalized burning front index (a scaled normalized chargeability) to image and localize, without ambiguity, the position of the coal seam fire in the subsurface. The 3-D reconstructed target is located below a negative self-potential anomaly (similarly to what is observed in laboratory experiments) and a temperature anomaly recorded at a depth of 30 cm.

Description: Rock permeability is an important parameter for the formation evaluation. In this paper, a new method with streaming current is proposed to determine the sample permeability based on the electrokinetic effects, and is proved by the experimental measurements. Corresponding to this method, we have designed an experimental setup and a test system, then performed the streaming current (potential) and electro-osmosis pressure experiments with 23 sandstone samples at 0.05 mol l –1 NaCl solution. The streaming current (potential) coefficient and electro-osmosis pressure coefficient are obtained, respectively, with the experimental data at low frequencies with AC lock-in technique. The electrokinetic permeabilities are further calculated with these coefficients. The results are consistent well with the gas permeability measured with Darcy's law, which verifies the current method for estimating rock permeability. Our measurements are also analysed and compared with previous measurements. The results indicate that our method can reflect the essence of electrokinetic effects better and simplify the electrokinetic measurements as well. In addition, we discuss the influences of experimental artefacts (core holder and confining pressure installation) on the electrokinetic data. The results show that the trough phenomenon, appeared in frequency curves of streaming current (potential) coefficients, is induced by the resonance of the core-holder/vibrator system. This is important for the design of electrokinetic setup and the analysis of low-frequency response of the electrokinetic coupling coefficients.

Description: Multiparameter full waveform inversion (FWI) applied to an elastic orthorhombic model description of the subsurface requires in theory a nine-parameter representation of each pixel of the model. Even with optimal acquisition on the Earth surface that includes large offsets, full azimuth, and multicomponent sensors, the potential for trade-off between the elastic orthorhombic parameters are large. The first step to understanding such trade-off is analysing the scattering potential of each parameter, and specifically, its scattering radiation patterns. We investigate such radiation patterns for diffraction and for scattering from a horizontal reflector considering a background isotropic model. The radiation patterns show considerable potential for trade-off between the parameters and the potentially limited resolution in their recovery. The radiation patterns of C 11 , C 22 , and C 33 are well separated so that we expect to recover these parameters with limited trade-offs. However, the resolution of their recovery represented by recovered range of model wavenumbers varies between these parameters. We can only invert for the short wavelength components (reflection) of C 33 while we can mainly invert for the long wavelength components (transmission) of the elastic coefficients C 11 and C 22 if we have large enough offsets. The elastic coefficients C 13 , C 23 , and C 12 suffer from strong trade-offs with C 55 , C 44 , and C 66 , respectively. The trade-offs between C 13 and C 55 , as well as C 23 and C 44 , can be partially mitigated if we acquire P – SV and SV – SV waves. However, to reduce the trade-offs between C 12 and C 66 , we require credible SH – SH waves. The analytical radiation patterns of the elastic constants are supported by numerical gradients of these parameters.

Description: The 16S rRNA gene (16S rDNA) codes for RNA that plays a fundamental role during translation in the ribosome and is used extensively as a marker gene to establish relationships among bacteria. However, the complementary non-coding 16S rDNA (nc16S rDNA) has been ignored. An idea emerged in the course of analyzing bacterial 16S rDNA sequences in search for nucleotide composition and substitution patterns: Does the nc16S rDNA code? If so, what does it code for? More importantly: Does 16S rDNA evolution reflect its own evolution or the evolution of its counterpart nc16S rDNA? The objective of this minireview is to discuss these thoughts. nc strands often encode small RNAs (sRNAs), ancient components of gene regulation. nc16S rDNA sequences from different bacterial groups were used to search for possible matches in the Bacterial Small Regulatory RNA Database. Intriguingly, the sequence of one published sRNA obtained from Legionella pneumophila (GenBank: AE017354.1) showed high non-random similarity with nc16S rDNA corresponding in part to the V5 region especially from Legionella and relatives. While the target(s) of this sRNA is unclear at the moment, its mere existence might open up a new chapter in the use of the 16S rDNA to study relationships among bacteria.

Description: Thermotolerance of the fungus Fomes sp. EUM1 was evaluated in solid state fermentation (SSF). This thermotolerant strain improved both hyphal invasiveness (38%) and length (17%) in adverse thermal conditions exceeding 30°C and to a maximum of 40°C. In contrast, hyphal branching decreased by 46% at 45°C. The production of cellulases over corn stover increased 1.6-fold in 30°C culture conditions, xylanases increased 2.8-fold at 40°C, while laccase production improved 2.7-fold at 35°C. Maximum production of lignocellulolytic enzymes was obtained at elevated temperatures in shorter fermentation times (8–6 days), although the proteases appeared as a thermal stress response associated with a drop in lignocellulolytic activities. Novel and multiple isoenzymes of xylanase (four bands) and cellulase (six bands) were secreted in the range of 20–150 kDa during growth in adverse temperature conditions. However, only a single laccase isoenzyme (46 kDa) was detected. This is the first report describing the advantages of a thermotolerant white-rot fungus in SSF. These results have important implications for large-scale SSF, where effects of metabolic heat are detrimental to growth and enzyme production, which are severely affected by the formation of high temperature gradients.

Description: Typically, seismic data are sparsely and irregularly sampled due to limitations in the survey environment and these cause problems for key seismic processing steps such as surface-related multiple elimination or wave-equation-based migration. Various interpolation techniques have been developed to alleviate the problems caused by sparse and irregular sampling. Among many interpolation techniques, matching pursuit interpolation is a robust tool to interpolate the regularly sampled data with large receiver separation such as crossline data in marine seismic acquisition when both pressure and particle velocity data are used. Multicomponent matching pursuit methods generally used the sinusoidal basis function, which have shown to be effective for interpolating multicomponent marine seismic data in the crossline direction. In this paper, we report the use of wavelet basis functions which further enhances the performance of matching pursuit methods for de-aliasing than sinusoidal basis functions. We also found that the range of the peak wavenumber of the wavelet is critical to the stability of the interpolation results and the de-aliasing performance and that the range should be determined based on Nyquist criteria. In addition, we reduced the computational cost by adopting the inner product of the wavelet and the input data to find the parameters of the wavelet basis function instead of using L-2 norm minimization. Using synthetic data, we illustrate that for aliased data, wavelet-based matching pursuit interpolation yields more stable results than sinusoidal function-based one when we use not only pressure data only but also both pressure and particle velocity together.

Description: The rhizobacterium Serratia plymuthica 4Rx13 emits the novel and unique volatile sodorifen (C 16 H 26 ), which has a polymethylated bicyclic structure. Transcriptome analysis revealed that gene SOD_c20750 (annotated as terpene cyclase) is involved in the biosynthesis of sodorifen. Here we show that this gene is located in a small cluster of four genes ( SOD_c20750 – SOD_c20780 ), and the analysis of the knockout mutants demonstrated that SOD_c20760 (annotated as methyltransferase) and SOD_c20780 (annotated as isopentenyl pyrophosphate (IPP) isomerase) are needed for the biosynthesis of sodorifen, while a sodorifen-negative phenotype was not achieved with the SOD_c20770 (annotated as deoxy-xylulose-5-phosphate (DOXP) synthase) mutant. Altogether, the function of this new gene cluster was assigned to the biosynthesis of this structurally unusual volatile compound sodorifen.

Description: Among staphylococci Staphylococcus saprophyticus is the only species that is typically uropathogenic and an important cause of urinary tract infections in young women. The amino acid D-serine occurs in relatively high concentrations in human urine and has a bacteriostatic or toxic effect on many bacteria. In uropathogenic Escherichia coli and S. saprophyticus , the amino acid regulates the expression of virulence factors and can be used as a nutrient. The ability of uropathogens to respond to or to metabolize D-serine has been suggested as a factor that enables colonization of the urinary tract. Until now nothing is known about D-serine transport in S.   saprophyticus . We generated mutants of putative transporter genes in S.   saprophyticus 7108 that show homology to the D-serine transporter cyc A of E. coli and tested them in a D-serine depletion assay to analyze the D-serine uptake rate of the cells. The mutant of SPP1070 showed a strong decrease in D-serine uptake. Therefore, SSP1070 was identified as a major D-serine transporter in S. saprophyticus 7108 and was named D-serine transporter A (DstA). D-serine caused a prolonged lag phase of S. saprophyticus in a chemically defined medium. This negative effect was dependent on the presence of DstA.

Description: Deep sea pockmarks underlain by chimney-like or pipe structures that contain methane hydrate are abundant along the Norwegian continental margin. In such hydrate provinces the interaction between hydrate formation and fluid flow has significance for benthic ecosystems and possibly climate change. The Nyegga region, situated on the western Norwegian continental slope, is characterized by an extensive pockmark field known to accommodate substantial methane gas hydrate deposits. The aim of this study is to detect and delineate both the gas hydrate and free gas reservoirs at one of Nyegga's pockmarks. In 2012, a marine controlled-source electromagnetic (CSEM) survey was performed at a pockmark in this region, where high-resolution 3-D seismic data were previously collected in 2006. 2-D CSEM inversions were computed using the data acquired by ocean bottom electrical field receivers. Our results, derived from unconstrained and seismically constrained CSEM inversions, suggest the presence of two distinctive resistivity anomalies beneath the pockmark: a shallow vertical anomaly at the underlying pipe structure, likely due to gas hydrate accumulation, and a laterally extensive anomaly attributed to a free gas zone below the base of the gas hydrate stability zone. This work contributes to a robust characterization of gas hydrate deposits within subseafloor fluid flow pipe structures.

Description: One of the main applications of magnetic field measurements in boreholes is the detection of unexploded ordnance or buried utility structures like pipes or tiebacks. Even though the advantage of fully oriented magnetic vector measurements have long been recognized and could significantly reduce costs and risks, the tools used for those purposes typically measure only the total magnetic field, the vertical and horizontal components or gradients thereof. The Göttingen Bohrloch Magnetometer uses three fibre optic gyros to record its orientation and thus enables us to compute high-quality three-component magnetic vector data regardless of borehole orientation. The measurements described in this paper were run in the scientific borehole Cuxhaven Lüdingworth 1/1A, which was drilled as a part of the ‘Coastal Aquifer Test field’ project to study the dynamics of the saltwater/freshwater interface. As the drill string got stuck during drilling of the first borehole, a second hole was drilled in the immediate vicinity. The drill string lies at a depth between 80 and 114 m at a distance of only 2.5 m southeast of the borehole used for the measurements, making it an ideal target to demonstrate the benefits of vector magnetic surveys. Although the theories to calculate magnetic fields of objects with different shapes is well established and do not need to be tested, they almost exclusively include approximations of the geometry. It is not obvious whether these approximations are suited to describe real data, or whether additional effects or refinements have to be considered. We use both a simplified monopole model and a cylinder model to fit the data and are able to determine the position of the drill string within a statistical error of approximately 10 cm. Additionally, we show that the location of the drill string could not have been determined by measurements of the total field or its horizontal and vertical component alone and that those methods would require the drilling of additional boreholes to obtain an unambiguous result.

Description: Genes encoding fluoride transporters have been identified in bacterial and archaeal species. The genome sequence of the cariogenic  Streptococcus mutans  bacteria suggests the presence of a putative fluoride transporter, which is referred to as a chloride channel permease. Two homologues of this gene (GenBank locus tags SMU_1290c and SMU_1289c) reside in tandem in the genome of  S. mutans . The aim of this study was to determine whether the chloride channel permeases contribute to fluoride resistance. We constructed SMU_1290c- and SMU_1289c-knockout  S. mutans  UA159 strains. We also constructed a double-knockout strain lacking both genes. SMU_1290c or SMU_1289c was transformed into a fluoride transporter- disrupted  Escherichia coli strain. All bacterial strains were cultured under appropriate conditions with or without sodium fluoride, and fluoride resistance was evaluated. All three gene-knockout  S. mutans  strains showed lower resistance to sodium fluoride than did the wild-type strain. No significant changes in resistance to other sodium halides were recognized between the wild-type and double-knockout strains. Both SMU_1290c and SMU_1289c transformation rescued fluoride transporter-disrupted  E. coli  cell from fluoride toxicity. We conclude that the chloride channel permeases contribute to fluoride resistance in  S. mutans .

Description: Clostridium thermocellum and Thermoanaerobacterium saccharolyticum are bacteria under investigation for production of biofuels from plant biomass. Thermoanaerobacterium saccharolyticum has been engineered to produce ethanol at high yield (〉90% of theoretical) and titer (〉70 g/l). Efforts to engineer C. thermocellum have not, to date, been as successful, and efforts are underway to transfer the ethanol production pathway from T. saccharolyticum to C. thermocellum . One potential challenge in transferring metabolic pathways is the possibility of incompatible levels of nicotinamide cofactors. These cofactors (NAD + , NADH, NADP + and NADPH) and their oxidation state are important in the context of microbial redox metabolism. In this study we directly measured the concentrations and reduced oxidized ratios of these cofactors in a number of strains of C. thermocellum and T. saccharolyticum by using acid/base extraction and enzymatic assays. We found that cofactor ratios are maintained in a fairly narrow range, regardless of the metabolic network modifications considered. We have found that the ratios are similar in both organisms, which is a relevant observation in the context of transferring the T. saccharolyticum ethanol production pathway to C. thermocellum .

Description: Chromosome-long haplotyping of human genomes is important to identify genetic variants with differing gene expression, in human evolution studies, clinical diagnosis, and other biological and medical fields. Although several methods have realized haplotyping based on sequencing technologies or population statistics, accuracy and cost are factors that prohibit their wide use. Borrowing ideas from group testing theories, we proposed a clone-based haplotyping method by overlapping pool sequencing. The clones from a single individual were pooled combinatorially and then sequenced. According to the distinct pooling pattern for each clone in the overlapping pool sequencing, alleles for the recovered variants could be assigned to their original clones precisely. Subsequently, the clone sequences could be reconstructed by linking these alleles accordingly and assembling them into haplotypes with high accuracy. To verify the utility of our method, we constructed 130 110 clones in silico for the individual NA12878 and simulated the pooling and sequencing process. Ultimately, 99.9% of variants on chromosome 1 that were covered by clones from both parental chromosomes were recovered correctly, and 112 haplotype contigs were assembled with an N50 length of 3.4 Mb and no switch errors. A comparison with current clone-based haplotyping methods indicated our method was more accurate.

Description: Sexual differentiation of malaria parasites into gametocytes in the vertebrate host and subsequent gamete fertilization in mosquitoes is essential for the spreading of the disease. The molecular processes orchestrating these transitions are far from fully understood. Here, we report the first transcriptome analysis of male and female Plasmodium falciparum gametocytes coupled with a comprehensive proteome analysis. In male gametocytes there is an enrichment of proteins involved in the formation of flagellated gametes; proteins involved in DNA replication, chromatin organization and axoneme formation. On the other hand, female gametocytes are enriched in proteins required for zygote formation and functions after fertilization; protein-, lipid- and energy-metabolism. Integration of transcriptome and proteome data revealed 512 highly expressed maternal transcripts without corresponding protein expression indicating large scale translational repression in P. falciparum female gametocytes for the first time. Despite a high degree of conservation between Plasmodium species, 260 of these ‘repressed transcripts’ have not been previously described. Moreover, for some of these genes, protein expression is only reported in oocysts and sporozoites indicating that repressed transcripts can be partitioned into short- and long-term storage. Finally, these data sets provide an essential resource for identification of vaccine/drug targets and for further mechanistic studies.

Description: Identifying large-scale structural variation in cancer genomes continues to be a challenge to researchers. Current methods rely on genome alignments based on a reference that can be a poor fit to highly variant and complex tumor genomes. To address this challenge we developed a method that uses available breakpoint information to generate models of structural variations. We use these models as references to align previously unmapped and discordant reads from a genome. By using these models to align unmapped reads, we show that our method can help to identify large-scale variations that have been previously missed.

Description: Cadmium is a widespread environmental pollutant and poses some potential risks to human health. However, the signaling events controlling cadmium toxicity are not fully understood. In this study, we examined the effect of cadmium chloride on cell viability and the intracellular nitric oxide (NO) level in yeast cells. The results showed that exposure of yeast cells to cadmium (0–100 μM) could induce cell killing with significantly increased intracellular NO levels. Morphological analysis of the nuclei with 4 ' ,6-diamidino-2-phenylindole staining and DNA strand breaks analysis showed that cadmium at 50 μM can induce cell apoptosis in yeast cells. Treatment of yeast cells with cadmium (50 μM) and the nitric oxide scavenger c-PTIO [2-(4-carboxyphenyl)-4,4,5,5-teramethylimidazoline-1-oxyl-3-oxide; 0.2 mM] showed that c-PTIO attenuated the cadmium-induced cell killing. Our findings indicated that cadmium-induced yeast cell killing is mediated by a directly increased intracellular NO level.

Description: Although seismic attenuation measurements have great potential to enhance our knowledge of physical conditions and rock properties, their application is limited because robust methods for improving both the resolution and accuracy of attenuation estimates have not yet been established. We propose attenuation estimation methods for zero-offset vertical seismic profile (VSP) data by combining seismic interferometry (SI) and the modified median frequency shift (MMFS) method developed for attenuation estimation using sonic waveform data. The configuration of zero-offset VSP data is redatumed to that of the sonic logging measurement by adopting two types of SI: deconvolution interferometry and crosscorrelation interferometry (CCI). Then, we can apply the MMFS method to the redatumed VSP data. Although the amplitude information estimated from CCI is biased, we propose a correction method for this bias to correctly estimate attenuation. First, to investigate the performance both in resolution and accuracy, we apply different trace separations to synthetic data with random noise at different signal-to-noise ratio levels. Second, we estimate the influence of residual reflection events after wavefield separation on attenuation estimation. The proposed methods provide more stable attenuation estimates in comparison with the spectral ratio method because the mean-median procedure suppresses random events and characteristic features caused by residual reflection events in spectral domain. Our numerical experiments also imply that the proposed methods can estimate scattering attenuation values even if frequency components are not changed between the two receiver depths. Finally, by preliminarily applying the proposed methods to field VSP data, we demonstrate the advantages of the proposed method in the resolution and stability of attenuation estimates and these observations correlate with those of numerical tests.

Description: Effective uptake of fermentable substrates is a fundamentally important aspect of any fermentation process. The solventogenic bacterium Clostridium beijerinckii is noted for its ability to ferment a wide range of carbohydrates, yet few of its sugar transport systems have been characterized. In common with other anaerobes, C. beijerinckii shows a marked dependence on the PEP-dependent phosphotransferase system (PTS) for sugar accumulation. In this study, the gene cbe0751 encoding the sugar-specific domains of a phosphotransferase belonging to the glucose family was cloned into an Escherichia coli strain lacking the ability to take up and phosphorylate glucose. Transformants gained ability to ferment glucose, and also mannose, and further analysis of a selected transformant demonstrated that it could take up and phosphorylate glucose, confirming that cbe0751 encodes a glucose PTS which also recognizes mannose as a substrate. RT-PCR analysis showed that cbe0751 was expressed in cultures grown on both substrates, but also to varying extents during growth on some other carbon sources. Although analogue inhibition studies suggested that Cbe0751 is not the only glucose PTS in C. beijerinckii , this system should nevertheless be regarded as a potential target for metabolic engineering to generate a strain showing improved sugar fermentation properties.

Description: Heavy metals are dense chemicals with dual biological role as micronutrients and intoxicants. A few hypersaline environmental systems are naturally enriched with heavy metals, while most metal-contaminated sites are a consequence of human activities. Numerous halotolerant and moderately halophilic Bacteria possess metal tolerance, whereas a few archaeal counterparts share similar features. The main mechanisms underlying heavy metal resistance in halophilic Bacteria and Archaea include extracellular metal sequestration by biopolymers, metal efflux mediated by specific transporters and enzymatic detoxification. Biotransformation of metals by halophiles has implications both for trace metal turnover in natural saline ecosystems and for development of novel bioremediation strategies.

Description: We screened for a gene that inhibits streptomycin production in Streptomyces griseus when it is introduced on a high-copy-number plasmid pIJ702, and obtained a plasmid pKM545. The introduction of pKM545 abolished streptomycin production on all media tested including YMP-sugar and Nutrient broth. S1 protection analysis demonstrated that the introduction of this plasmid downregulated the transcriptional activity of the promoter preceding strR , the pathway-specific transcriptional regulator for streptomycin biosynthesis. The 2.8-kb Bam HI fragment cloned onto pKM545 contained two coding sequences SGR_5442 and 5443. These coding sequences and the two downstream ones (SGR_5444 and 5445) constituted a possible operon structure designated to be rspABCD (regulation of streptomycin production). RspB and RspC exhibited a marked similarity with an ATP-binding domain and a membrane-associating domain of an ABC-2 type transporter, respectively, suggesting that the Rsp proteins comprise a membrane exporter. The gene cluster consisting of the rsp operon and the upstream divergent small coding sequence (SGR_5441) was widely distributed to Streptomyces genome. An rspB mutant of S. griseus produced 3-fold streptomycin of the parental strain in YMP liquid medium. The evidence implies that the Rsp translocator is involved in the export of a substance that specifies the expression level of streptomycin biosynthesis genes in S. griseus .

Description: We previously demonstrated that in Streptomyces coelicolor two-component system AfsQ1/Q2 activates the production of the yellow-colored coelimycin P2 (also named as yCPK) on glutamate-supplemented minimal medium, and the response regulator AfsQ1 could specifically bind to the intergenic region between two structural genes, cpkA and cpkD . Here, a more in-depth investigation was performed to elucidate the mechanism underlying the role of AfsQ1/Q2 in regulating coelimycin P2 biosynthesis. Deletion of afsQ1/Q2 resulted in markedly decreased expression of the whole coelimycin P2 biosynthetic gene cluster. Electrophoretic mobility shift assays revealed that AfsQ1 bound only to the target site identified previously, but not to any other promoters in the gene cluster. Mutations of AfsQ1-binding motif only resulted in drastically reduced transcription of the cpkA/B/C operon (encoding three type I polyketide synthases) and intriguingly, led to enhanced expression of some coelimcyin P2 genes, particularly accA1 and scF . These results suggested the direct role of AfsQ1/Q2 in regulating coelimycin production, which is directly mediated by the structural genes, but not the cluster-situated regulatory genes, and also implied that other unknown mechanisms may be involved in AfsQ1/Q2-mediated regulation of coelimycin P2 biosynthesis.

Description: Sulfate-reducing bacteria (SRBs) gain their energy by coupling the oxidation of organic substrate to the reduction of sulfate to sulfide. Several SRBs are able to use alternative terminal electron acceptors to sulfate such as nitrate. Nitrate-reducing SRBs have been isolated from a diverse range of environments. In order to be able to understand the significance of nitrate reduction in SRBs, we need to examine the ecology and physiology of the nitrate-reducing SRB isolates.

Description: Capacitively Coupled Resistivity (CCR) is conventionally used to emulate DC resistivity measurements and may provide important information about the ice content of material in periglacial areas. The application of CCR theoretically enables the determination of both electrical parameters, that is, the resistivity and the electrical permittivity, by analysing magnitude and phase shift spectra. The electrical permittivity may dominate the impedance, especially in periglacial areas or regions of hydrogeological interest. However, previous theoretical work suggested that the phase shift may strongly depend on electrode height above ground, implying that electrode height must be known with great accuracy to determine electrical permittivity. Here, we demonstrate with laboratory test measurements, theoretical modelling and by analysing the Jacobian matrix of the inversion that the sensitivity towards electrode height is drastically reduced if the electrical permittivity is frequency dependent in a way that is typical for ice. For the first time, we used a novel broad-band CCR device ‘Chameleon’ for a field test located in one of the ridge galleries beneath the crest of Mount Zugspitze. A permanently ice covered bottom of a tunnel was examined. For the inversion of the measured spectra, the frequency dependence of the electrical parameters was parametrized in three different ways: A Debye Model for pure ices, a Cole–Cole Model for pure ices and a dual Cole–Cole Model including interfacial water additionally. The frequency-dependent resistivity and permittivity spectra obtained from the inversion, including low- and high-frequency limits, agree reasonably well with laboratory and field measurements reported in the literature.

Description: We evaluate the applicability and the effectiveness of GPR attribute analysis for high-resolution glacier imaging and characterization, testing this approach on 4-D GPR multifrequency data collected in a small glacier in the Eastern Alps, by repeating the acquisition along the same profiles in four different periods of the year 2013. The main objectives are to image and characterize the glacier's inner structure and to quantitatively monitor the seasonal thawing of near-surface frozen materials (snow/firn). A multiattribute approach is used to characterize the subsurface through different attribute categories, including instantaneous and textural attributes considering not only amplitude-, phase- and frequency-related attributes, but also other more complex and integrated parameters. We combine information from more than one attribute into a single image with composite displays, using overlays or mixed displays. The results demonstrate that the developed GPR attribute analysis can provide significant improvements in the discrimination of GPR signals, and obtain enhanced and more constrained data interpretations.

Description: The ADP-ribosylating enzymes are encoded in many pathogenic bacteria in order to affect essential functions of the host. In this study, we show that Neisseria gonorrhoeae possess a locus that corresponds to the ADP-ribosyltransferase NarE, a previously characterized enzyme in N. meningitidis . The 291 bp coding sequence of gonococcal narE shares 100% identity with part of the coding sequence of the meningococcal narE gene due to a frameshift previously described, thus leading to a 49-amino-acid deletion at the N-terminus of gonococcal NarE protein. However, we found a promoter region and a GTG start codon, which allowed expression of the protein as demonstrated by RT-PCR and western blot analyses. Using a gonococcal NarE–6xHis fusion protein, we demonstrated that the gonococcal enzyme underwent auto-ADP-ribosylation but to a lower extent than meningococcal NarE. We also observed that gonoccocal NarE exhibited ADP-ribosyltransferase activity using agmatine and cell-free host proteins as ADP-ribose acceptors, but its activity was inhibited by human β-defensins. Taken together, our results showed that NarE of Neisseria gonorrhoeae is a functional enzyme that possesses key features of bacterial ADP-ribosylating enzymes.

Description: The effect of fructose 1,6-bisphosphate (Fru 1,6-P 2 ) on the regulatory enzymes of pentose phosphate pathway of Escherichia coli was examined. Fru 1,6-P 2 inhibited E. coli transaldolase (EC 2.2.1.2) competitively against fructose 6-phosphate and uncompetitively against erythrose 4-phosphate, whereas Fru 1,6-P 2 did not affect glucose 6-phosphate dehydrogenase (EC 1.1.1.49) and 6-phosphogluconate dehydrogenase (EC 1.1.1.44). Kinetic results can be explained by assuming that transaldolase has two kinds of binding sites for Fru 1,6-P 2 : a competitive binding site for fructose 6-phosphate and a second binding site on the enzyme-erythrose 4-phosphate complex. Fru 1,6-P 2 increased resulting from the stimulation of glycolysis, can inhibit transaldolase and further participates in the elevation of the concentration of ribose 5-phosphate that can be preferentially utilized for anabolic reaction in exponential phase of E. coli .

Description: Migration velocity analysis aims at automatically updating the large-scale components of the velocity model, called macromodel. Extended Common Image Gathers are panels used to evaluate focusing after imaging and are constructed as a function of a spatial shift introduced in the imaging condition. We investigate how transmitted waves can also be used in migration velocity analysis: instead of back-propagating the residuals associated with reflected waves, we propose to back-propagate the full wavefield. The image function, equivalent to the migrated section for reflected data, does not exhibit localized events in space along horizons but is still sensitive to the choice of the background velocity model and can thus be coupled to the same objective function defined in the image domain. In order to enhance the benefits of direct waves, we consider a cross-well configuration. Direct waves provide a large illumination between two vertical wells. Associated Common Image Gathers present different characteristics than the ones associated with reflected waves in surface acquisition. In particular, energy is spread over up to the maximum penetration depth. We invert cross-well seismic data along two lines. In the first case, the input data contain the full wavefield dominated by transmitted waves. It demonstrates the possibility to handle transmitted waves to determine the velocity model. It appears that the misfit in the data domain is largely reduced after inversion. In the second case, we use the same algorithm, but with reflected observed data only, as in a classical approach. Most of velocity updates are localized around the reflectivity, leading to an incorrect final model. This demonstrates the benefit of transmitted waves for migration velocity analysis in a cross-well configuration.

Description: An integrated approach to interpret Self-Potential (SP) anomalies based on spectral analysis and tomographic methods is presented. The Maximum Entropy Method (MEM) is used for providing accurate estimates of the depth of the anomaly source. The 2-D tomographic inversion technique, based on the underground charge occurrence probability (COP) function, is, then, used to fully characterize the anomalous body, as the MEM is not helpful in delineating the shape of the anomaly source. The proposed integrated approach is applied for the inversion of synthetic SP data generated by geometrically simple anomalous bodies, such as cylinders and inclined sheets. This numerical study has allowed the determination of mathematical relationships between zero lines of the COP distributions, the polarization angles and the positions along the profile of the causative sources, which have been of great help for interpreting the related SP anomalies. Finally, the analysis of field examples shows the high potential applicability of the proposed integrated approach for SP data inversion.

Description: Volcanic and tectonic events are the main processes involved in the generation of the oceanic crust and responsible for the seismicity associated with seafloor spreading. To monitor this activity, usually not or poorly detected by land-based seismological stations, we deployed from February 2012 to February 2013 a network of autonomous hydrophones to compare the behaviour of the ultraslow-spreading Southwest Indian ridge (SWIR) with that of the intermediate-spreading Southeast Indian ridge (SEIR). The rate of seismicity is similar for both ridges, suggesting that there is no systematic relationship between seismicity and spreading rates. The along-axis distribution of the seismic events, however, does differ, reflecting the rate dependence of accretion modes. Earthquakes are sparse and regularly spaced and scattered along the SWIR, reflecting prevailing tectonic processes. By contrast, along the SEIR, events are irregularly distributed and focus at ridge-segment ends and transforms faults, reflecting the ridge segmentation; only two swarms occurred at a segment centre and are probably caused by a magmatic event. This seismicity distribution thus looks controlled by segment-scale crustal heterogeneities along the SEIR and by regional-scale contrasting accretion processes along the SWIR, probably driven by different lithospheric and asthenospheric dynamics on either side of the Melville fracture zone. The comparison of hydroacoustic and teleseismic catalogues shows that, along these spreading ridges, the background seismicity observed in 1 yr by a hydroacoustic network is representative of the seismicity observed over two decades by land-based networks.

Description: The Hdr (heterodisulfide reductase)-like enzyme is predicted, from gene transcript profiling experiments previously published, to be essential in oxidative sulfur metabolism in a number of bacteria and archaea. Nevertheless, no biochemical and physicochemical data are available so far about this enzyme. Genes coding for it were identified in Aquifex aeolicus , a Gram-negative, hyperthermophilic, chemolithoautotrophic and microaerophilic bacterium that uses inorganic sulfur compounds as electron donor to grow. We provide biochemical evidence that this Hdr-like enzyme is present in this sulfur-oxidizing prokaryote (cultivated with thiosulfate or elemental sulfur). We demonstrate, by immunolocalization and cell fractionation, that Hdr-like enzyme is associated, presumably monotopically, with the membrane fraction. We show by co-immunoprecipitation assay or partial purification, that the Hdr proteins form a stable complex composed of at least five subunits, HdrA, HdrB1, HdrB2, HdrC1 and HdrC2, present in two forms of high molecular mass on native gel (~240 and 450 kDa). These studies allow us to propose a revised model for dissimilatory sulfur oxidation pathways in A. aeolicus , with Hdr predicted to generate sulfite.

Description: Geobacter sulfurreducens is an anaerobic soil bacterium that is involved in biogeochemical cycles of elements such as Fe and Mn. Although significant progress has been made in the understanding of the electron transfer processes in G. sulfurreducens , little is known about the regulatory mechanisms involved in their control. To expand the study of gene regulation in G. sulfurreducens , we carried out a genome-wide identification of transcription start sites (TSS) by 5'RACE and by deep RNA sequencing of primary mRNAs in two growth conditions. TSSs were identified along G. sulfurreducens genome and over 50% of them were located in the upstream region of the associated gene, and in some cases we detected genes with more than one TSS. Our global mapping of TSSs contributes with valuable information, which is needed for the study of transcript structure and transcription regulation signals and can ultimately contribute to the understanding of transcription initiation phenomena in G. sulfurreducens .

Description: Advanced sequencing technologies have generated a plethora of data for many chromatin marks in multiple tissues and cell types, yet there is lack of a generalized tool for optimal utility of those data. A major challenge is to quantitatively model the epigenetic dynamics across both the genome and many cell types for understanding their impacts on differential gene regulation and disease. We introduce IDEAS, an i ntegrative and d iscriminative e pigenome a nnotation s ystem, for jointly characterizing epigenetic landscapes in many cell types and detecting differential regulatory regions. A key distinction between our method and existing state-of-the-art algorithms is that IDEAS integrates epigenomes of many cell types simultaneously in a way that preserves the position-dependent and cell type-specific information at fine scales, thereby greatly improving segmentation accuracy and producing comparable annotations across cell types.

Description: DnpA, a putative de- N -acetylase of the PIG-L superfamily, is required for antibiotic tolerance in Pseudomonas aeruginosa . Exactly how dnpA (gene locus PA5002) directs the formation of antibiotic-tolerant persister cells is currently unknown. Previous research provided evidence for a role in surface-associated process(es), possibly in lipopolysaccharide biosynthesis. In silico sequence analysis of DnpA predicts a single transmembrane domain and N in /C out orientation of DnpA. In contrast, we here show that DnpA is an integral inner membrane protein containing two transmembrane domains, with the major C-terminal part located at the cytoplasmic face. Correct insertion into the inner membrane is necessary for DnpA to promote fluoroquinolone tolerance. The membrane localization of DnpA further supports its role in cell envelope-associated process(es). In addition to shedding light on the biological role of DnpA, this study highlights the risks of overreliance on the predictive value of bioinformatics tools and the importance of rigorous experimental validation of in silico predictions.

Description: Cell wall impermeability and active efflux of drugs are among the primary reasons for drug resistance in mycobacteria. Efflux pumps are tripartite membrane localized transport proteins that expel drug molecules outside the cells. Several of such efflux pumps are annotated in mycobacteria, but few have been characterized, like MSMEG_2991, a putative efflux pump permease of Mycobacterium smegmatis . To substantiate this, we overexpressed MSMEG_2991 protein in Escherichia coli 2443. Expression of MSMEG_2991 elevated the resistance towards structurally unrelated groups of antibiotics. An active antibiotic efflux pump nature of MSMEG_2991 was revealed by assessing the acquisition of ciprofloxacin in the absence and presence of the efflux pump inhibitor, carbonyl cyanide m-chlorophenyl hydrazone, indicating the involvement of proton-motive force (pmf) during the efflux activity. MSMEG_2991 expression elevated biofilm formation in E. coli by 4-fold, keeping parity to some of the earlier reported efflux pumps. In silico analysis suggested the presence of 12 transmembrane helices in MSMEG_2991 resembling EmrD efflux pump of E. coli . Based on in vivo and in silico analyses, MSMEG_2991 may be designated as a pmf-mediated multidrug efflux pump protein that expels diverse groups of antibiotics and might as well be involved in the biofilm enhancement.

Description: Bacterial small RNAs (sRNAs) play essential roles in the post-transcriptional control of gene expression. To improve their detection by conventional microarrays, we designed a custom microarray containing a group of probes targeting known and some putative Escherichia coli sRNAs. To assess its potential in detection of sRNAs, RNA profiling experiments were performed with total RNA extracted from E. coli MG1655 cells exponentially grown in rich (Luria–Bertani) and minimal (M9/glucose) media. We found that many sRNAs could yield reasonably strong and statistically significant signals corresponding to nearly all sRNAs annotated in the EcoCyc database. Besides differential expression of two sRNAs (GcvB and RydB), expression of other sRNAs was less affected by the composition of the growth media. Other examples of the differentially expressed sRNAs were revealed by comparing gene expression of the wild-type strain and its isogenic mutant lacking functional poly(A) polymerase I ( pcnB ). Further, northern blot analysis was employed to validate these data and to assess the existence of new putative sRNAs. Our results suggest that the use of custom microarrays with improved capacities for detection of sRNAs can offer an attractive opportunity for efficient gene expression profiling of sRNAs and their target mRNAs at the whole transcriptome level.

Description: Escherichia coli DedA/Tvp38 family proteins YghB and YqjA are putative membrane transporters with 62% amino acid identity and overlapping functions. An E. coli strain (BC202) with nonpolar yghB and yqjA mutations displays cell-division defects and temperature sensitivity and is sensitive to antibiotics and alkaline pH. In this study, we performed site-directed mutagenesis on conserved, charged amino acids of YqjA and YghB. We discovered two conserved predicted membrane-embedded arginines (R130 and R136) that are critical for function in both proteins as defined by their ability to complement BC202 phenotypes, when expressed from a plasmid. Lysine can substitute for arginine at position R130 indicating a charge dependence at this position, but could not substitute at R136. In light of the established role that arginine plays in the translocation mechanism of numerous membrane transporters, we hypothesize that these amino acids play a role in the transport mechanism of these DedA/Tvp38 family proteins.

Description: Candida albicans is a dimorphic yeast commonly found on human mucosal membranes that switches from yeast to hyphal morphology in response to environmental factors. The change to hyphal growth requires histone H3 modifications by the yeast-specific histone acetyltransferase Rtt109. In addition to its role in morphogenesis, Rtt109-dependent acetylation of histone H3 lysine residues 9 and 56 has regulatory functions during DNA replication and repair. Boric acid (BA) is a broad-spectrum agent that specifically inhibits C. albicans hyphal growth, locking the fungus in its harmless commensal yeast state. The present study characterizes the effect of BA on C. albicans histone acetylation in respect to specificity, time-course and significance. We demonstrate that sublethal concentrations of BA reduce H3K9/H3K56 acetylation, both on a basal level and in response to genotoxic stress. Acetylation at other selected histone sites were not affected by BA. qRT-PCR expression analysis of the DNA repair gene Rad51 indicated no elevated level of genotoxic stress during BA exposure. A forward-mutation analysis demonstrated the BA does not increase spontaneous or induced mutations . The findings suggest that DNA repair remains effective even when histone H3 acetylation decreases and dispels the notion that BA treatment impairs genome integrity in yeast.

Description: The impact of gene mutations on the growth of the cells can be studied using pure cultures. However, the importance of certain proteins and pathways can be also examined via co-culturing wild type and its mutant derivative. Here, the relative fitness of a mutant strain that lacks the global nitrogen regulator, CodY, was examined in Bacillus cereus , a food poisoning Gram-positive bacterium. Fitness measurements revealed that the codY strain was outcompeted when cocultured with the wild-type ATCC 14579 under various rich laboratory medium, and also when inoculated in certain beverages. In nutrient-poor minimal medium, the codY mutant had comparable fitness to the wild-type strain. Interestingly, the relative fitness of the codY strain was antagonistic when it was cultivated in apple or orange juices due to unknown properties of these beverages, highlighting the importance of chemical composition of the test medium during the bacterial fitness measurements.

Description: Out of 8000 candidates from a genetic screening for Pseudomonas putida KT2442 mutants showing defects in biofilm formation, 40 independent mutants with diminished levels of biofilm were analyzed. Most of these mutants carried insertions in genes of the lap cluster, whose products are responsible for synthesis, export and degradation of the adhesin LapA. All mutants in this class were strongly defective in biofilm formation. Mutants in the flagellar regulatory genes fleQ and flhF showed similar defects to that of the lap mutants . On the contrary, transposon insertions in the flagellar structural genes fliP and flgG , that also impair flagellar motility, had a modest defect in biofilm formation. A mutation in gacS , encoding the sensor element of the GacS/GacA two-component system, also had a moderate effect on biofilm formation. Additional insertions targeted genes involved in cell envelope function: PP3222, encoding the permease element of an ABC-type transporter and tolB , encoding the periplasmic component of the Tol-OprL system required for outer membrane stability. Our results underscore the central role of LapA, suggest cross-regulation between motility and adhesion functions and provide insights on the role of cell envelope trafficking and maintenance for biofilm development in P. putida .

Description: Horizontal gene transfer plays a significant role in bacterial evolution and has major clinical importance. Thus, it is vital to understand the mechanisms and kinetics of genetic transformations. Natural transformation is the driving mechanism for horizontal gene transfer in diverse genera of bacteria. Our study introduces a simple and rapid method for the investigation of natural transformation. This highly sensitive system allows the detection of a transformation event directly from a bacterial population without any separation step or selection of cells. The system is based on the bacterial luciferase operon from Photorhabdus luminescens . The studied molecular tools consist of the functional modules luxCDE and luxAB , which involve a replicative plasmid and an integrative gene cassette. A well-established host for bacterial genetic investigations, Acinetobacter baylyi ADP1, is used as the model bacterium. We show that natural transformation followed by homologous recombination or plasmid recircularization can be readily detected in both actively growing and static biofilm-like cultures, including very rare transformation events. The system allows the detection of natural transformation within 1 h of introducing sample DNA into the culture. The introduced method provides a convenient means to study the kinetics of natural transformation under variable conditions and perturbations.

Description: The legume–rhizobial interaction results in the formation of symbiotic nodules in which rhizobia fix nitrogen. During the process of symbiosis, reactive oxygen species (ROS) are generated. Thus, the response of rhizobia to ROS is important for successful nodulation and nitrogen fixation. In this study, we investigated how Azorhizobium caulinodans , a rhizobium that forms both root and stem nodules on its host plant, regulates ROS resistance. We found that in-frame deletions of a gene encoding the putative catalase-peroxidase katG or a gene encoding a LysR-family regulatory protein, oxyR , exhibited increased sensitivity to H 2 O 2 . We then showed that OxyR positively regulated katG expression in an H 2 O 2 -independent fashion. Furthermore, we found that deletion of katG or oxyR led to significant reduction in the number of stem nodules and decrease of nitrogen fixation capacities in symbiosis. Our results revealed that KatG and OxyR are not only critical for antioxidant defense in vitro , but also important for nodule formation and nitrogen fixation during interaction with plant hosts.

Description: Geothermal well GRT-1 (Rittershoffen, Alsace) was drilled in 2012. Its open-hole section (extending down to a depth of 2.6 km) penetrated fractured sandstones and granite. In 2013, the well was subjected to Thermal, Chemical and Hydraulic (TCH) stimulation, which improved the injectivity index fivefold. The goal of the study was to assess the impact of the stimulation by comparing pre- and post-stimulation well-logging (acoustic and temperature [ T ] logs) and mud-logging data. This comparison revealed modifications of almost all the natural fractures. However, not all of these fractures are associated with permeability enhancement, and the post-stimulation T logs are important for characterizing this enhancement. Chemical alteration due to mechanical erosion at the tops and bottoms of the fractures was observed in the sandstones. These zones display indications of very small new permeability after the TCH stimulation. Because a major fault zone caved extensively where it crosses the borehole, it was not imaged in the acoustic logs. However, this originally permeable zone was enhanced as demonstrated by the T logs. Based on the natural injectivity of this fault zone, hydraulic erosion and thermal microcracking of its internal quartz veins are associated with this permeability enhancement. Although local changes in the borehole wall observed in the acoustic images cannot be directly linked to the improved injectivity index, the comparison of the acoustic image logs allows for identification of fracture zones impacted by the TCH stimulation.

Description: We investigate the spatial coherence of underwater ambient noise using a yearlong time-series measured off Ascension Island. Qualitative agreement with observed cross-correlations is achieved using a simple range-dependent model, constrained by earlier, active tomographic studies in the area. In particular, the model correctly predicts the existence of two weakly dispersive normal modes in the microseism frequency range, with the group speed of one of the normal modes being smaller than the sound speed in water. The agreement justifies our interpretation of the peaks of the measured cross-correlation function of ambient noise as modal arrivals, with dispersion that is sensitive to crustal velocity structure. Our observations are consistent with Scholte to Moho head wave coupled propagation, with double mode conversion occurring due to the bathymetric variations between receivers. We thus demonstrate the feasibility of interrogating crustal properties using noise interferometry of moored hydrophone data at ranges in excess of 120 km.

Description: Recent tectonic reconstructions of the South Atlantic have partitioned the ocean basin into several segments based upon one or more proposed intraplate South American deformation zones. In several of these reconstructions, opening of the southern segment(s) by seafloor spreading prior to Aptian-Albian time is accompanied by contemporaneous strike-slip motion along an intraplate boundary extending southeastward from the Andean Cochabamba—Santa Cruz bend to the Rio Grande Fracture Zone (RGFZ). We have examined new magnetic data over the Pelotas, Santos and Campos Basins, offshore Argentina and Brazil, acquired by ION-GXT in tandem with long-offset, long record seismic reflection data, and identified seafloor spreading anomalies M4, M3, M2 and M0 (~131, ~129, ~128 and ~125 Ma). Integrating these results with our earlier work, we have been able to correlate magnetochrons M4, M3, M2 and M0 north and south of the RGFZ on the South American margin, and north and south of the Walvis Ridge on the African side. Our results are therefore inconsistent with diachronous opening models that involve substantial continental strike-slip motion north of RGFZ during M4 to M0 time. Although the ocean basin may have opened from south to north, our results indicate that seafloor spreading began north of the RGFZ earlier than previously proposed.

Description: Peptidoglycan (also known as murein) is an important envelope component of bacteria, and its turnover usually takes place at considerable levels during normal growth. Amino sugars and murein tripeptide resulting from murein degradation are used for resynthesis of peptidoglycan or as self-generated nutrients or energy sources for cell growth. PgrR (regulator of peptide glycan recycling; formerly YcjZ) was recently identified as a repressor of several genes participating in uptake and degradation of murein tripeptide. In this study, we identified the ycjG gene involved in murein tripeptide degradation as a new direct target of PgrR. The expression of PgrR-regulated genes including ycjY , mppA , mpaA and ycjG was repressed in the presence of a good nitrogen source, but their expression increased under poor nitrogen conditions. Under nitrogen starvation, the pgrR mutant cells exhibited faster growth than wild-type cells, implying that derepression of genes under the control of PgrR may help cells overcome nitrogen limitation. Therefore, these results suggest that nitrogen starvation induces derepression of PgrR-controlled genes involved in uptake and degradation of murein tripeptide, and this may stimulate the utilization of murein tripeptide as a nitrogen source.

Description: A majority of large-scale bacterial genome rearrangements involve mobile genetic elements such as insertion sequence (IS) elements. Here we report novel insertions and excisions of IS elements and recombination between homologous IS elements identified in a large collection of Escherichia coli mutation accumulation lines by analysis of whole genome shotgun sequencing data. Based on 857 identified events (758 IS insertions, 98 recombinations and 1 excision), we estimate that the rate of IS insertion is 3.5 x 10 –4 insertions per genome per generation and the rate of IS homologous recombination is 4.5 x 10 –5 recombinations per genome per generation. These events are mostly contributed by the IS elements IS 1 , IS 2 , IS 5 and IS 186 . Spatial analysis of new insertions suggest that transposition is biased to proximal insertions, and the length spectrum of IS-caused deletions is largely explained by local hopping. For any of the ISs studied there is no region of the circular genome that is favored or disfavored for new insertions but there are notable hotspots for deletions. Some elements have preferences for non-coding sequence or for the beginning and end of coding regions, largely explained by target site motifs. Interestingly, transposition and deletion rates remain constant across the wild-type and 12 mutant E. coli lines, each deficient in a distinct DNA repair pathway. Finally, we characterized the target sites of four IS families, confirming previous results and characterizing a highly specific pattern at IS 186 target-sites, 5'-GGGG(N6/N7)CCCC-3'. We also detected 48 long deletions not involving IS elements.

Description: Determining thin layer thickness is very important for reservoir characterization and CO 2 quantification. Given its high time–frequency resolution and robustness, the complex spectral decomposition method was applied on time-lapse 3-D seismic data from the Ketzin pilot site for CO 2 storage to evaluate the frequency-dependent characteristics of thin layers at the injection level. Higher temporal resolution and more stratigraphic details are seen in the all-frequency and monochromatic reflectivity amplitude sections obtained by complex spectral decomposition compared to the stacked sections. The mapped geologic discontinuities within the reservoir are consistent with the preferred orientation of CO 2 propagation. Tuning frequency mapping shows the thicknesses of the reservoir sandstone and gaseous CO 2 is consistent with the measured thickness of the sandstone unit from well logging. An attempt to discriminate between pressure effects and CO 2 saturation using the extracted tuning frequency indicates that CO 2 saturation is the main contributor to the amplitude anomaly at the Ketzin site. On the basis of determined thickness of gaseous CO 2 in the reservoir, quantitative analysis of the amount of CO 2 was performed and shows a discrepancy between the injected and calculated CO 2 mass. This may be explained by several uncertainties, like structural reservoir heterogeneity, a limited understanding of the complex subsurface conditions, error of determined tuning frequency, the presence of ambient noise and ongoing CO 2 dissolution.

Description: Comprehensive analytical solutions to 3-D axisymmetric problems for static response of multilayered thermoelastic media subjected to surface loads and containing sources are presented in this study. The solution strategy employs Laplace and Hankel transforms to the field variables. The problem is formulated in cylindrical coordinate system and in this coordinate system vector surface harmonics and generalized propagator matrix are introduced to find the solution for the problem for the behaviour of thermoelastic multilayered media subject to surface loads and containing heat sources. A high-order adaptive Gaussian quadrature method with continued fraction expansions is employed to approximate the integral solutions expressed in terms of semi-infinite Hankel-type integrals. It is the first time to apply the proposed solution method to investigate the behaviour of repository for heat-emitting high-level nuclear waste (HLW) in a geological formation where the HLW can be regarded as a decaying with time point heat source.

Description: We present a series of processes for understanding and analysing controlled-source electromagnetic (CSEM) responses for a conductive and permeable earth. To realize the CSEM response, a new 3-D CSEM forward modelling algorithm based on an edge finite element method for both electrically conductive and magnetically permeable heterogeneities is developed. The algorithm shows highly accurate results in validation tests against a semi-analytic solution for stratified earth and an integral form of the scattered field. We describe the vector behaviour of an anomalous magnetic field originating from a conductive and permeable anomaly when the loop sources are deployed over a conductive half-space. The CSEM response of the conductive and permeable anomaly is classified into three effects originating from: conductivity perturbations, permeability perturbations and the coupling of these two effects. The separated individual results and the corresponding integral equation form of the anomalous field help to better understand the physical behaviour. We confirm the characteristic features of the CSEM response from the conductive and permeable anomaly, for example, (1) the general dominance of the induction effect in the out-of-phase response accompanied by a non-negligible magnetization effect from the magnetic anomaly in a conductive half-space and (2) the dominance of near frequency-independent magnetization effects in the in-phase response at relatively low frequencies and change in ruling part of the in-phase response into the induction effect as the frequency increases. We also demonstrate the effect of coupling mode and show that its maximum contribution is limited to a few per cent level of other two modes, induction and magnetization mode, even when the heterogeneity of our model is strong. In our synthetic survey, using examples of land-based profiling surveys of low induction number and intermediate regime, we find that the effect of magnetization can be used as an indicator of the existence of magnetic material. One important point to note from this study is the importance of accurate cancelling-out or estimation of free-space responses, which can mask the magnetic responses to imaging the varying magnetic property.

Description: Extraction of fluids from subsurface reservoirs induces changes in pore pressure, leading not only to geomechanical changes, but also perturbations in seismic velocities and hence observable seismic attributes. Time-lapse seismic analysis can be used to estimate changes in subsurface hydromechanical properties and thus act as a monitoring tool for geological reservoirs. The ability to observe and quantify changes in fluid, stress and strain using seismic techniques has important implications for monitoring risk not only for petroleum applications but also for geological storage of CO 2 and nuclear waste scenarios. In this paper, we integrate hydromechanical simulation results with rock physics models and full-waveform seismic modelling to assess time-lapse seismic attribute resolution for dynamic reservoir characterization and hydromechanical model calibration. The time-lapse seismic simulations use a dynamic elastic reservoir model based on a North Sea deep reservoir undergoing large pressure changes. The time-lapse seismic traveltime shifts and time strains calculated from the modelled and processed synthetic data sets (i.e. pre-stack and post-stack data) are in a reasonable agreement with the true earth models, indicating the feasibility of using 1-D strain rock physics transform and time-lapse seismic processing methodology. Estimated vertical traveltime shifts for the overburden and the majority of the reservoir are within ±1 ms of the true earth model values, indicating that the time-lapse technique is sufficiently accurate for predicting overburden velocity changes and hence geomechanical effects. Characterization of deeper structure below the overburden becomes less accurate, where more advanced time-lapse seismic processing and migration is needed to handle the complex geometry and strong lateral induced velocity changes. Nevertheless, both migrated full-offset pre-stack and near-offset post-stack data image the general features of both the overburden and reservoir units. More importantly, the results from this study indicate that integrated seismic and hydromechanical modelling can help constrain time-lapse uncertainty and hence reduce risk due to fluid extraction and injection.

Description: A new plate motions model for the northwest Africa–North America Plate pair during the Oligocene and early Miocene is presented. The model is accompanied by a high-resolution isochron map for the central Atlantic region, resulting from a re-examination of 423 ship tracks from the NGDC data base for the area between the 15°20' FZ and the Azores triple junction. A new digital model of fracture zones for this region and a set of 309 magnetic profiles crossing the Oligocene to recent oceanic crust within the study area allowed to determine accurate finite reconstruction poles for the North America–northwest Africa conjugate plate pair between the early Miocene (Chron 6) and the early Oligocene (Chron 13). For times older than Chron 7 (~25 Ma), the finite reconstruction poles were calculated using a reliable data set coming exclusively from the region south of the Canary Islands FZ (~32°N), which allowed to test the rigidity of the northwest African oceanic lithosphere during the Oligocene–early Miocene phase of Atlas orogeny. A comparison of theoretical magnetic isochrons with observed magnetic lineations systematically shows that anomalously high spreading rates occurred in the area north of the Canary Islands FZ before Chron 7, thereby suggesting that the formation of the Atlas mountain, rather than being a localized intracontinental process, was logically linked to the central Atlantic spreading history. Thus, an independent Moroccan Plate could have existed during the Oligocene–early Miocene time interval, which included both the oceanic lithosphere north of the Canary Islands FZ and the northern Maghrebian areas of Morocco, Algeria and Tunisia. In this eventuality, the Atlas mountain belt should be reinterpreted as a giant flower structure associated with dextral transpression.

Description: Spectral induced polarization measurements are affected by temperature variations due to a variety of temperature-dependent parameters that control the complex electrical conductivity. Most important is the influence of the ion mobility, which increases with increasing temperature. It is responsible for the increase of the conductivity of the fluid in the pores with temperature and influences the electrical double layer on the mineral surface. This work is based on laboratory measurements of 13 sandstone samples from different sources with different geological and petrophysical characteristics. We measured the complex impedance in a frequency range from 0.01 to 100 Hz and a temperature range from 0 to 40 °C. The main observation is a decrease of the characteristic time (defined by the inverse of the frequency, at which the phase shift is maximum) with increasing temperature. The strength of this decrease differs from one sample to another. The temperature dependence of the phase shift magnitude cannot easily be generalized, as it depends on the particular sample. The experimental findings suggest that neglecting the influence of temperature on complex conductivity may lead to significant errors when estimating hydraulic conductivity from relaxation time. We also simulate the temperature dependence with a theoretical model of membrane polarization and review some of the model properties, with an emphasis on the temperature dependence of the parameters. The model reproduces several features characterizing the measured data, including the temperature dependence of the characteristic times. Computed tomography and microscope images of the pore structure of three samples also allow us to associate differences in the geometrical parameters used in the modelling with pore scale parameters of the actual samples.

Description: Knowledge about the pressure dependency of elastic and electrical properties is important for a variety of geophysical applications. We present a technique to invert for the stiff and compliant porosity from velocity measurements made as a function of differential pressure on saturated sandstones. A dual porosity concept is used for dry rock compressibility and a squirt model is employed for the pressure and frequency dependent elastic properties of the rocks when saturated. The total porosity obtained from inversion shows satisfactory agreement with experimental results. The electrical cementation factor was determined using the inverted porosity in combination with measured electrical conductivity. It was found that cementation factor increased exponentially with increasing differential pressure during isostatic loading. Elastic compressibility, electrical cementation factor and electrical conductivity of the saturated rocks correlate linearly with compliant porosity, and electrical cementation factor and electrical conductivity exhibit linear correlations with elastic compressibility of the saturated rocks under loading. The results show that the dual porosity concept is sufficient to explain the pressure dependency of elastic, electrical and joint elastic-electrical properties of saturated porous sandstones.

Description: A new plate motions model for the northwest Africa–North America Plate pair during the Oligocene and early Miocene is presented. The model is accompanied by a high-resolution isochron map for the central Atlantic region, resulting from a re-examination of 423 ship tracks from the NGDC data base for the area between the 15°20' FZ and the Azores triple junction. A new digital model of fracture zones for this region and a set of 309 magnetic profiles crossing the Oligocene to recent oceanic crust within the study area allowed to determine accurate finite reconstruction poles for the North America–northwest Africa conjugate plate pair between the early Miocene (Chron 6) and the early Oligocene (Chron 13). For times older than Chron 7 (~25 Ma), the finite reconstruction poles were calculated using a reliable data set coming exclusively from the region south of the Canary Islands FZ (~32°N), which allowed to test the rigidity of the northwest African oceanic lithosphere during the Oligocene–early Miocene phase of Atlas orogeny. A comparison of theoretical magnetic isochrons with observed magnetic lineations systematically shows that anomalously high spreading rates occurred in the area north of the Canary Islands FZ before Chron 7, thereby suggesting that the formation of the Atlas mountain, rather than being a localized intracontinental process, was logically linked to the central Atlantic spreading history. Thus, an independent Moroccan Plate could have existed during the Oligocene–early Miocene time interval, which included both the oceanic lithosphere north of the Canary Islands FZ and the northern Maghrebian areas of Morocco, Algeria and Tunisia. In this eventuality, the Atlas mountain belt should be reinterpreted as a giant flower structure associated with dextral transpression.

Description: Genetic variants in or near miRNA genes can have profound effects on miRNA expression and targeting. As user-friendly software for the impact prediction of miRNA variants on a large scale is still lacking, we created a tool called miRVaS. miRVaS automates this prediction by annotating the location of the variant relative to functional regions within the miRNA hairpin (seed, mature, loop, hairpin arm, flanks) and by annotating all predicted structural changes within the miRNA due to the variant. In addition, the tool defines the most important region that is predicted to have structural changes and calculates a conservation score that is indicative of the reliability of the structure prediction. The output is presented in a tab-separated file, which enables fast screening, and in an html file, which allows visual comparison between wild-type and variant structures. All separate images are provided for downstream use. Finally, we tested two different approaches on a small test set of published functionally validated genetic variants for their capacity to predict the impact of variants on miRNA expression.

Description: Analysis of RNA-seq data often detects numerous ‘non-co-linear’ (NCL) transcripts, which comprised sequence segments that are topologically inconsistent with their corresponding DNA sequences in the reference genome. However, detection of NCL transcripts involves two major challenges: removal of false positives arising from alignment artifacts and discrimination between different types of NCL transcripts ( trans -spliced, circular or fusion transcripts). Here, we developed a new NCL-transcript-detecting method (‘NCLscan’), which utilized a stepwise alignment strategy to almost completely eliminate false calls (〉98% precision) without sacrificing true positives, enabling NCLscan outperform 18 other publicly-available tools (including fusion- and circular-RNA-detecting tools) in terms of sensitivity and precision, regardless of the generation strategy of simulated dataset, type of intragenic or intergenic NCL event, read depth of coverage, read length or expression level of NCL transcript. With the high accuracy, NCLscan was applied to distinguishing between trans -spliced, circular and fusion transcripts on the basis of poly(A)- and nonpoly(A)-selected RNA-seq data. We showed that circular RNAs were expressed more ubiquitously, more abundantly and less cell type-specifically than trans -spliced and fusion transcripts. Our study thus describes a robust pipeline for the discovery of NCL transcripts, and sheds light on the fundamental biology of these non-canonical RNA events in human transcriptome.

Description: The central metabolite acetyl phosphate (acP) has long been proposed to influence transcription regulation by directly transferring its phosphoryl group to a number of response regulators in many bacterial species. Here, we provide in vitro evidence for this proposition and demonstrate, using an in vitro transcription system, that acP-dependent phosphorylation of aspartate 51 of CpxR induces transcription of one of its regulon members in E. coli , cpxP . We also used this in vitro transcription system to extend our previously reported in vivo data that hypothesized that acetylation of RNA polymerase (RNAP) influences acP-dependent cpxP transcription, using glutamine as a genetic mimic for acetylated arginine 291 of the carboxy-terminal domain of RNAP α subunit. The data we present here lend strong support to the hypothesis that acP has a direct effect on transcription regulation in E. coli via phosphorylation of CpxR, and that RNAP acetylation can modulate this response.

Description: Maximizing vertical resolution is a key objective in seismic data processing. Early deconvolution and spectral balancing algorithms assumed that the seismic source wavelet was temporally invariant, or stationary. In practice, seismic scattering and attenuation give rise to non-stationary seismic source wavelets. To address this issue, most conventional time-varying deconvolution wavelet shaping and spectral modelling techniques using the stationary polynomial fitting assume the wavelet to be locally stationary within a small number of overlapping analysis windows while the fitting coefficients are invariant with all the frequencies. In this paper, we show an improvement obtained by modelling smoothly varying spectra of the seismic wavelet using non-stationary polynomial fitting in the time–frequency domain. We first decompose each seismic trace using a generalized S-transform that provides a good time–frequency distribution for the estimation of the time-varying wavelet spectra. We then model the slowly varying source wavelet spectrum at each time sample by a smooth low-order polynomial. Finally, we spectrally balance the modelled wavelet to flatten the seismic response, thereby increasing vertical resolution. We calibrate the algorithm on a simple synthetic and then apply it to a 3-D land survey acquired in western China, showing the value on both vertical slices through seismic amplitude and attribute time slices. Our new algorithm significantly improves the vertical resolution of the seismic signal, while not increasing the noise.

Description: Knowledge about the pressure dependency of elastic and electrical properties is important for a variety of geophysical applications. We present a technique to invert for the stiff and compliant porosity from velocity measurements made as a function of differential pressure on saturated sandstones. A dual porosity concept is used for dry rock compressibility and a squirt model is employed for the pressure and frequency dependent elastic properties of the rocks when saturated. The total porosity obtained from inversion shows satisfactory agreement with experimental results. The electrical cementation factor was determined using the inverted porosity in combination with measured electrical conductivity. It was found that cementation factor increased exponentially with increasing differential pressure during isostatic loading. Elastic compressibility, electrical cementation factor and electrical conductivity of the saturated rocks correlate linearly with compliant porosity, and electrical cementation factor and electrical conductivity exhibit linear correlations with elastic compressibility of the saturated rocks under loading. The results show that the dual porosity concept is sufficient to explain the pressure dependency of elastic, electrical and joint elastic-electrical properties of saturated porous sandstones.

Description: Chaperones are a diverse class of ubiquitous proteins that assist other cellular proteins in folding correctly and maintaining their native structure. Many different chaperones cooperate to constitute the ‘proteostasis’ machinery in the cells. It has been proposed earlier that archaeal organisms could be ideal model systems for deciphering the basic functioning of the ‘protein folding machinery’ in higher eukaryotes. Several chaperone families have been characterized in archaea over the years but mostly one protein at a time, making it difficult to decipher the composition and mechanistics of the protein folding system as a whole. In order to deal with these lacunae, we have developed a database of all archaeal chaperone proteins, CrAgDb ( C haperone r epertoire in A rchaeal g enomes). The data have been presented in a systematic way with intuitive browse and search facilities for easy retrieval of information. Access to these curated datasets should expedite large-scale analysis of archaeal chaperone networks and significantly advance our understanding of operation and regulation of the protein folding machinery in archaea. Researchers could then translate this knowledge to comprehend the more complex protein folding pathways in eukaryotic systems. The database is freely available at http://14.139.227.92/mkumar/cragdb/ .

Description: In order to perform gas exploration and determine the distribution pattern of gas in the Yanchang Oil Field in the eastern part of the North Shaanxi Slope, Ordos Basin, China, gravity and magnetic survey data were systemically collated, processed and interpreted in combination with the drilling data and recent seismic data. The genesis of gravity and magnetic anomalies and the relationship between the characteristics of the gravity and magnetic fields and known gas distribution were explored in order to predict the favourable exploration targets for gas. Gravity anomalies resulted both from the lateral variation in density of the basement rock and lateral lithologic transformation in the sedimentary cover. The regional magnetic anomalies were mainly caused by the basement metamorphic rocks and the residual magnetic anomalies may reflect the amount and general location of the volcanic materials in the overlying strata. The residual gravity and magnetic anomalies generated by high-density sandstone and high content of volcanics in the gas reservoir of the upper Paleozoic distorted and deformed the anomaly curves when they were stacked onto the primary background anomaly. The gas wells were generally found to be located in the anomaly gradient zones, or the distorted part of contour lines, and the flanks of high and low anomalies, or the transitional zones between anomaly highs and lows. The characteristics of gravity and magnetic fields provide significant information that can be used for guidance when exploring the distribution of gas. Based on these characteristics, five favourable areas for gas exploration were identified; these are quasi-equally spaced like a strip extending from the southeast to the northwest.

Description: In bacteria, cysteine can be synthesized from serine by two steps involving an L-serine O-acetyltransferase (SAT) and a cysteine synthase (CysK). While CysK is found in the publicly available annotated genome from Lactobacillus casei ATCC 334, a gene encoding SAT ( cysE ) is missing. In this study, we found that various strains of L. casei grew in a chemically defined medium containing sulfide as the sole sulfur source, indicating the presence of a serine O-acetyltransferase. The gene lying upstream of cysK is predicted to encode a homoserine trans-succinylase ( metA ). To study the function of this gene, it was cloned from L. casei FAM18110. The purified, recombinant protein did not acylate L-homoserine in vitro . Instead, it catalyzed the formation of O-acetyl serine from L-serine and acetyl-CoA. Furthermore, the plasmid expressing the L. casei gene complemented an Escherichia coli cysE mutant strain but not an E. coli metA mutant. This clearly demonstrated that the gene annotated as metA in fact encodes the SAT function and should be annotated as cysE .

Description: Spectral induced polarization measurements are affected by temperature variations due to a variety of temperature-dependent parameters that control the complex electrical conductivity. Most important is the influence of the ion mobility, which increases with increasing temperature. It is responsible for the increase of the conductivity of the fluid in the pores with temperature and influences the electrical double layer on the mineral surface. This work is based on laboratory measurements of 13 sandstone samples from different sources with different geological and petrophysical characteristics. We measured the complex impedance in a frequency range from 0.01 to 100 Hz and a temperature range from 0 to 40 °C. The main observation is a decrease of the characteristic time (defined by the inverse of the frequency, at which the phase shift is maximum) with increasing temperature. The strength of this decrease differs from one sample to another. The temperature dependence of the phase shift magnitude cannot easily be generalized, as it depends on the particular sample. The experimental findings suggest that neglecting the influence of temperature on complex conductivity may lead to significant errors when estimating hydraulic conductivity from relaxation time. We also simulate the temperature dependence with a theoretical model of membrane polarization and review some of the model properties, with an emphasis on the temperature dependence of the parameters. The model reproduces several features characterizing the measured data, including the temperature dependence of the characteristic times. Computed tomography and microscope images of the pore structure of three samples also allow us to associate differences in the geometrical parameters used in the modelling with pore scale parameters of the actual samples.

Description: The streaming potential phenomenon is produced by the flow of an electrolyte in a porous medium and is used for geophysical prospecting. It is quantified through an electrokinetic (EK) coefficient. The dependence of the EK coefficient on the conductivity of the electrolyte is described by the Helmholtz–Smoluchowski (HS) equation. This equation provides successful forecasts of the EK coefficient in the standard range of concentration. However, experimental measurements show deviations to this equation at extreme low and extreme high salinities. The aim of this study is to model the EK coefficient using Lattice Boltzmann simulations in a 2-D capillary channel, with a view to understanding these deviations. The effect of the constitutive parameters of the HS equation such as the permittivity and the viscosity is discussed. The validity of the HS equation using strong potentials is assessed. Finally, a model of bulk fluid conductivity is derived. This model allows to take into account the change of local ionic distribution in the vicinity of the mineral. It appears to have a significant impact on the derivation of potentials at low salinities and in the presence of polyvalent counterions.

Description: Extraction of fluids from subsurface reservoirs induces changes in pore pressure, leading not only to geomechanical changes, but also perturbations in seismic velocities and hence observable seismic attributes. Time-lapse seismic analysis can be used to estimate changes in subsurface hydromechanical properties and thus act as a monitoring tool for geological reservoirs. The ability to observe and quantify changes in fluid, stress and strain using seismic techniques has important implications for monitoring risk not only for petroleum applications but also for geological storage of CO 2 and nuclear waste scenarios. In this paper, we integrate hydromechanical simulation results with rock physics models and full-waveform seismic modelling to assess time-lapse seismic attribute resolution for dynamic reservoir characterization and hydromechanical model calibration. The time-lapse seismic simulations use a dynamic elastic reservoir model based on a North Sea deep reservoir undergoing large pressure changes. The time-lapse seismic traveltime shifts and time strains calculated from the modelled and processed synthetic data sets (i.e. pre-stack and post-stack data) are in a reasonable agreement with the true earth models, indicating the feasibility of using 1-D strain rock physics transform and time-lapse seismic processing methodology. Estimated vertical traveltime shifts for the overburden and the majority of the reservoir are within ±1 ms of the true earth model values, indicating that the time-lapse technique is sufficiently accurate for predicting overburden velocity changes and hence geomechanical effects. Characterization of deeper structure below the overburden becomes less accurate, where more advanced time-lapse seismic processing and migration is needed to handle the complex geometry and strong lateral induced velocity changes. Nevertheless, both migrated full-offset pre-stack and near-offset post-stack data image the general features of both the overburden and reservoir units. More importantly, the results from this study indicate that integrated seismic and hydromechanical modelling can help constrain time-lapse uncertainty and hence reduce risk due to fluid extraction and injection.

Description: We study the accuracy and numerical stability of three eigenvector sets for modelling the coupled poroelastic and electromagnetic layered-Earth response. We use a known eigenvector set, its flux-normalized version and a newly derived flux-normalized set. The new set is chosen such that the system is properly uncoupled when the coupling between the poroelastic and electromagnetic fields vanishes. We carry out two different numerical stability tests: the first test focuses on the internal system, eigenvector and eigenvalue consistency; the second test investigates the stability and preciseness of the flux-normalized systems by looking at identity relations. We find that the known set shows the largest deviation for both tests, whereas the new set performs best. In two additional numerical modelling experiments, these numerical inaccuracies are shown to generate numerical noise levels comparable to small signals, such as signals coming from the important interface conversion responses, especially when the coupling coefficient is small. When coupling vanishes completely, the known set does not produce proper results. The new set produces numerically stable and accurate results in all situations. We therefore strongly recommend to use this newly derived set for future layered-Earth seismo-electromagnetic modelling experiments.

Description: Visualization of chromosomal dynamics is important for understanding many fundamental intra-nuclear processes. Efficient and reliable live-cell multicolor labeling of chromosomal loci can realize this goal. However, the current methods are constrained mainly by insufficient labeling throughput, efficiency, flexibility as well as photostability. Here we have developed a new approach to realize dual-color chromosomal loci imaging based on a modified single-guide RNA (sgRNA) of the CRISPR/Cas9 system. The modification of sgRNA was optimized by structure-guided engineering of the original sgRNA, consisting of RNA aptamer insertions that bind fluorescent protein-tagged effectors. By labeling and tracking telomeres, centromeres and genomic loci, we demonstrate that the new approach is easy to implement and enables robust dual-color imaging of genomic elements. Importantly, our data also indicate that the fast exchange rate of RNA aptamer binding effectors makes our sgRNA-based labeling method much more tolerant to photobleaching than the Cas9-based labeling method. This is crucial for continuous, long-term tracking of chromosomal dynamics. Lastly, as our method is complementary to other live-cell genomic labeling systems, it is therefore possible to combine them into a plentiful palette for the study of native chromatin organization and genome ultrastructure dynamics in living cells.

Description: The Quaternary Añavieja-Dévanos tufa system is located in the northern sector of the Iberian Chain. It has been previously tackled by means sedimentological studies focused on the available outcrops and some boreholes. They have permitted the proposal of a sedimentary scenario that fits with a pool-barrage fluvial tufa model. However a better knowledge of the characteristics and internal distribution of the usually non-outcropping pool deposits as well as of its relationship with barrage deposits has not been evaluated in detail yet. Palaeoenvironmental studies on tufas are usually biased because tufas are commonly delicate facies exposed to intense erosion during water level fall stages; for this reason outcrops are usually scarce and very often coincide with the most cemented barrage deposits. In order to analyse the internal characteristics of the tufa deposits under study, but also the lateral correlation among different facies, ground penetrating radar (GPR) has been employed both for the evaluation of its applicability in such kind of environments and to improve, if possible, the sedimentary model using geophysical data in sectors without outcrops. A GPR survey including different antennas ranging from 50 to 500 MHz along different sectors and its comparison with natural outcrops has been carried out. GPR results have permitted to deduce clear differences between pool and barrage deposits and to recognise its internal structure and geometrical relationships. The survey also permitted an approach to different scales of heterogeneities in the radarfacies evaluation by using distinct antennas and therefore, reaching different resolutions and penetrations. The resulting integration from different antennas allows three different attenuant and eight reflective radarfacies to be defined permitting a better approach to the real extension of the pool areas. These results have permitted to decipher the horizontal and vertical facies changes and the identification of a scarcer development of pool deposits than expected in the studied system.

Description: Two novel transient controlled source electromagnetic methods called circular electrical dipole (CED) and differential electrical dipole (DED) are theoretically analysed for applications in shallow marine environments. 1-D and 3-D time-domain modelling studies are used to investigate the detectability and applicability of the methods when investigating resistive layers/targets representing hydrocarbon-saturated formations. The results are compared to the conventional time-domain horizontal electrical dipole (HED) and vertical electrical dipole (VED) sources. The applied theoretical modelling studies demonstrate that CED and DED have higher signal detectability towards resistive targets compared to TD-CSEM, but demonstrate significantly poorer signal amplitudes. Future CED/DED applications will have to solve this issue prior to measuring. Furthermore, the two novel methods have very similar detectability characteristics towards 3-D resistive targets embedded in marine sediments as VED while being less susceptible towards non-verticality. Due to the complex transmitter design of CED/DED the systems are prone to geometrical errors. Modelling studies show that even small transmitter inaccuracies have strong effects on the signal characteristics of CED making an actual marine application difficult at the present time. In contrast, the DED signal is less affected by geometrical errors in comparison to CED and may therefore be more adequate for marine applications.

Description: We purified a chitinase from pilei extractions of Coprinopsis cinerea fruiting bodies by ammonium sulfate precipitation and CM Sepharose cation exchange chromatography. MALDI-TOF/TOF MS analysis characterized this purified chitinase as a putative class V chitinase, ChiB1. ChiB1 hydrolyzed colloidal chitin and chitosan, whereas it did not hydrolyze chitin powder. ChiB1 cleaved only pNP-(GlcNAc) 2 , rather than pNP-GlcNAc or pNP-(Glc-NAc) 3 , to release nitrophenol. ChiB1 preferably and progressively released (GlcNAc)2 from (GlcNAc)6 and digested (GlcNAc)6 to two molecules of (GlcNAc)3 in a small proportion, but did not split (GlcNAc) 2 , so it is an exochitinase. ChiB1 has an optimum temperature range of 35°C to 40°C and an optimum pH of 5.0. ChiB1 exhibited Km and Vmax values of 2.63 mg ml –1 and 2.31 μmol min –1  mg protein –1 for colloidal chitin, respectively. The ChiB1 gene, along with another putative endochitinase (class III chitinase gene), was expressed dominantly among eight predicted chitinase genes in the genome, and its expression level increased with the maturation of fruiting bodies. ChiB1 incubation released a large amount of soluble β-glucan fractions from alkali-insoluble cell wall fractions of C. cinerea fruiting bodies, thereby it may promote the degradation of cell walls in synergy with the β-1,3-glucanases during pileus autolysis.

Description: Acidithiobacillus ferrivorans is an acidophile implicated in low-temperature biomining for the recovery of metals from sulfide minerals. Acidithiobacillus ferrivorans obtains its energy from the oxidation of inorganic sulfur compounds, and genes encoding several alternative pathways have been identified. Next-generation sequencing of At. ferrivorans RNA transcripts identified the genes coding for metabolic and electron transport proteins for energy conservation from tetrathionate as electron donor. RNA transcripts suggested that tetrathionate was hydrolyzed by the tetH1 gene product to form thiosulfate, elemental sulfur and sulfate. Despite two of the genes being truncated, RNA transcripts for the SoxXYZAB complex had higher levels than for thiosulfate quinone oxidoreductase ( doxDA genes). However, a lack of heme-binding sites in soxX suggested that DoxDA was responsible for thiosulfate metabolism. Higher RNA transcript counts also suggested that elemental sulfur was metabolized by heterodisulfide reductase ( hdr genes) rather than sulfur oxygenase reductase ( sor ). The sulfite produced as a product of heterodisulfide reductase was suggested to be oxidized by a pathway involving the sat gene product or abiotically react with elemental sulfur to form thiosulfate. Finally, several electron transport complexes were involved in energy conservation. This study has elucidated the previously unknown At. ferrivorans tetrathionate metabolic pathway that is important in biomining.

Description: TRP1 is a frequently used auxotrophic marker for genetic modifications and selections in trp – budding yeast strains, including the commonly used wild-type strain W303a. However, we found that introduction of the TRP1 gene into a trp – strain significantly affected vegetative growth at low and high temperatures. Therefore, caution should be needed when working in a trp – background strain and using the TRP1 marker to study stress response phenotypes, particularly when analyzing temperature sensitivities.

Description: An L-configured, three-component short period seismic array was deployed on the Ross Ice Shelf, Antarctica during November 2014. Polarization analysis of ambient noise data from these stations shows linearly polarized waves for frequency bands between 0.2 and 2 Hz. A spectral peak at about 1.6 Hz is interpreted as the resonance frequency of the water column and is used to estimate the water layer thickness below the ice shelf. The frequency band from 4 to 18 Hz is dominated by Rayleigh and Love waves propagating from the north that, based on daily temporal variations, we conclude were generated by field camp activity. Frequency–slowness plots were calculated using beamforming. Resulting Love and Rayleigh wave dispersion curves were inverted for the shear wave velocity profile within the firn and ice to ~150 m depth. The derived density profile allows estimation of the pore close-off depth and the firn–air content thickness. Separate inversions of Rayleigh and Love wave dispersion curves give different shear wave velocity profiles within the firn. We attribute this difference to an effective anisotropy due to fine layering. The layered structure of firn, ice, water and the seafloor results in a characteristic dispersion curve below 7 Hz. Forward modelling the observed Rayleigh wave dispersion curves using representative firn, ice, water and sediment structures indicates that Rayleigh waves are observed when wavelengths are long enough to span the distance from the ice shelf surface to the seafloor. The forward modelling shows that analysis of seismic data from an ice shelf provides the possibility of resolving ice shelf thickness, water column thickness and the physical properties of the ice shelf and underlying seafloor using passive-source seismic data.

Description: The factors driving the dominance of the Calvin–Benson–Bassham cycle (CBB) or reductive citric acid cycle (rCAC) in autotrophic microorganisms in different habitats are debated. Based on costs for synthesizing a few metabolic intermediates, it has been suggested that the CBB poses a disadvantage due to higher metabolic cost. The purpose of this study was to extend this estimate of cost from metabolite synthesis to biomass synthesis. For 12 gammaproteobacteria (CBB) and five epsilonproteobacteria (rCAC), the amount of ATP to synthesize a gram of biomass from CO 2 was calculated from genome sequences via metabolic maps. The eleven central carbon metabolites needed to synthesize biomass were all less expensive to synthesize via the rCAC (66%–89% of the ATP needed to synthesize them via CBB). Differences in cell compositions did result in differing demands for metabolites among the organisms, but the differences in cost to synthesize biomass were small among organisms that used a particular pathway (e.g. rCAC), compared to the difference between pathways (rCAC versus CBB). The rCAC autotrophs averaged 0.195 moles ATP per g biomass, while their CBB counterparts averaged 0.238. This is the first in silico estimate of the relative expense of both pathways to generate biomass.

Description: We present an algorithm to recover the Bayesian posterior model probability density function of subsurface elastic parameters, as required by the full pressure field recorded at an ocean bottom cable due to an impulsive seismic source. Both the data noise and source wavelet are estimated by our algorithm, resulting in robust estimates of subsurface velocity and density. In contrast to purely gradient based approaches, our method avoids model regularization entirely and produces 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. Our algorithm is trans-dimensional and performs model selection, sampling over a wide range of model parametrizations. We follow a frequency domain approach and derive the corresponding likelihood in the frequency domain. We present first a synthetic example of a reservoir at 2 km depth with minimal acoustic impedance contrast, which is difficult to study with conventional seismic amplitude versus offset changes. Finally, we apply our methodology to survey data collected over the Alba field in the North Sea, an area which is known to show very little lateral heterogeneity but nevertheless presents challenges for conventional post migration seismic amplitude versus offset analysis.

Description: A novel application of seismic interferometry (SI) and Marchenko imaging using both surface and borehole data is presented. A series of redatuming schemes is proposed to combine both data sets for robust deep local imaging in the presence of velocity uncertainties. The redatuming schemes create a virtual acquisition geometry where both sources and receivers lie at the horizontal borehole level, thus only a local velocity model near the borehole is needed for imaging, and erroneous velocities in the shallow area have no effect on imaging around the borehole level. By joining the advantages of SI and Marchenko imaging, a macrovelocity model is no longer required and the proposed schemes use only single-component data. Furthermore, the schemes result in a set of virtual data that have fewer spurious events and internal multiples than previous virtual source redatuming methods. Two numerical examples are shown to illustrate the workflow and to demonstrate the benefits of the method. One is a synthetic model and the other is a realistic model of a field in the North Sea. In both tests, improved local images near the boreholes are obtained using the redatumed data without accurate velocities, because the redatumed data are close to the target.

Description: Bacteria carry a number of genes that cause cell growth arrest or cell lysis upon expression. Notably, defective prophages retain many lysis proteins. Here, we identified a novel lytic gene, ydfD , on the Qin prophage segment of the Escherichia coli genome. YdfD lyses 99.9% of cells within 2 h of its induction. The co-expression of the upstream gene, dicB , encoding a cell division inhibitor, as well as sulA , encoding another cell division inhibitor, abolished YdfD-induced cell lysis. These results imply that YdfD-induced lysis is a cell division-dependent event. We further found that by deleting the hydrophobic 22-residue N-terminal domain, the resulting 42-residue C-terminal domain was still toxic to cause cell lysis. We propose that YdfD, associated with the cytoplasmic membrane, inhibits an essential cellular process(s).

Description: Many bacteria are capable of accumulating intracellular granules of polyhydroxyalkanoates (PHA). In this work, we developed confocal microscopy analysis of bacterial cells to study changes in the diameters of cells as well as PHA granules during growth and PHA accumulation in the bacterium Cupriavidus necator H16 (formerly Ralstonia eutropha ). The cell envelope was stained by DiD ® fluorescent probe and PHA granules by Nile Red. Signals from both probes were separated based on their spectral and fluorescence life-time properties. During growth and PHA accumulation, bacterial cells increased their length but the width of the cells remained constant. The volume fraction of PHA granules in cells increased during PHA accumulation, nevertheless, its value did not exceed 40 vol. % regardless of the PHA weight content. It seems that bacterial cultures lengthen the cells in order to control the PHA volume portion. However, since similar changes in cell length were also observed in a PHA non-accumulating mutant, it seems that there is no direct control mechanism, which regulates the prolongation of the cells with respect to PHA granules volume. It is more likely that PHA biosynthesis and the length of cells are influenced by the same external stimuli such as nutrient limitation.

Description: The effect of the selection of different nuclear magnetic resonance (NMR) relaxation times for permeability estimation is investigated for a set of fully brine-saturated rocks acquired from Cretaceous carbonate reservoirs in the North Sea and Middle East. Estimators that are obtained from the relaxation times based on the Pythagorean means are compared with estimators that are obtained from the relaxation times based on the concept of a cumulative saturation cut-off. Select portions of the longitudinal ( T 1 ) and transverse ( T 2 ) relaxation-time distributions are systematically evaluated by applying various cut-offs, analogous to the Winland-Pittman approach for mercury injection capillary pressure (MICP) curves. Finally, different approaches to matching the NMR and MICP distributions using different mean-based scaling factors are validated based on the performance of the related size-scaled estimators. The good results that were obtained demonstrate possible alternatives to the commonly adopted logarithmic mean estimator and reinforce the importance of NMR-MICP integration to improving carbonate permeability estimates.

Description: Three putative regulatory genes, namely ttmRI, ttmRII and ttmRIII , which are present in the tetramycin ( ttm ) biosynthetic gene cluster, were found in Streptomyces ahygroscopicus . Disruption of ttmRI, ttmRII or ttmRIII reduced tetramycin production, and their complementation restored production to varying degrees. Gene expression analysis of the wild-type (WT) and mutant strains through reverse transcriptase–polymerase chain reaction (RT-PCR) of the ttm gene cluster showed that the expression levels of most of the biosynthetic genes were reduced in ttmRI , ttmRII and ttmRIII . Electrophoretic mobility shift assays demonstrated that TtmRI, TtmRII and TtmRIII bound the promoters of several genes in the ttm gene cluster. This study found that these three proteins are a group of positive regulators that activate the transcription of the ttm gene cluster in S. ahygroscopicus . In addition, ttmRII had a reduced degree of grey pigmentation. Thus, TtmRII has a pleiotropic regulatory function in the tetramycin biosynthetic pathway and in development.

Description: Sulfide:quinone oxidoreductase (SQR) is the primary sulfide-oxidizing enzyme found in all three domains of life. Of the six phylogenetically distinct types of SQR, four have representatives that have been biochemically characterized. The genome of Chlorobaculum tepidum encodes three SQR homologs. One of these, encoded by CT1087, is a type VI SQR that has been previously shown to be required for growth at high sulfide concentrations and to be expressed in sulfide-dependent manner. Therefore, CT1087 was hypothesized to be a high sulfide adapted SQR. CT1087 was expressed in Escherichia coli with an N-terminal His-tag (CT1087NHis 6 ) and purified by Ni-NTA chromatography. CT1087NHis 6 was active and contained FAD as a strongly bound cofactor. The measured kinetic parameters for CT1087NHis 6 indicate a low affinity for sulfide and a high enzymatic turnover rate consistent with the hypothesis for its function inferred from genetic and expression data. These are the first kinetic data for a type VI SQR and have implications for structure-function analyses of all SQR's.

Description: Static time-delay corrections are frequency independent and ignore velocity variations away from the assumed vertical ray path through the subsurface. There is therefore a clear potential for improvement if the finite frequency nature of wave propagation can be properly accounted for. Such a method is presented here based on the Born approximation, the assumption of surface consistency and the misfit of instantaneous phase. The concept of instantaneous phase lends itself very well for sweep-like signals, hence these are the focus of this study. Analytical sensitivity kernels are derived that accurately predict frequency-dependent phase shifts due to P -wave anomalies in the near surface. They are quick to compute and robust near the source and receivers. An additional correction is presented that re-introduces the nonlinear relation between model perturbation and phase delay, which becomes relevant for stronger velocity anomalies. The phase shift as function of frequency is a slowly varying signal, its computation therefore does not require fine sampling even for broad-band sweeps. The kernels reveal interesting features of the sensitivity of seismic arrivals to the near surface: small anomalies can have a relative large impact resulting from the medium field term that is dominant near the source and receivers. Furthermore, even simple velocity anomalies can produce a distinct frequency-dependent phase behaviour. Unlike statics, the predicted phase corrections are smooth in space. Verification with spectral element simulations shows an excellent match for the predicted phase shifts over the entire seismic frequency band. Applying the phase shift to the reference sweep corrects for wavelet distortion, making the technique akin to surface consistent deconvolution, even though no division in the spectral domain is involved. As long as multiple scattering is mild, surface consistent finite frequency phase corrections outperform traditional statics for moderately large velocity contrasts, also in the case of a free surface with topography. The kernels could also be used to update the velocity model, if one has knowledge of (wave propagation related only) surface consistent phase operators.

Description: The seislet transform has been demonstrated to have a better compression performance for seismic data compared with other well-known sparsity promoting transforms, thus it can be used to remove random noise by simply applying a thresholding operator in the seislet domain. Since the seislet transform compresses the seismic data along the local structures, the seislet thresholding can be viewed as a simple structural filtering approach. Because of the dependence on a precise local slope estimation, the seislet transform usually suffers from low compression ratio and high reconstruction error for seismic profiles that have dip conflicts. In order to remove the limitation of seislet thresholding in dealing with conflicting-dip data, I propose a dip-separated filtering strategy. In this method, I first use an adaptive empirical mode decomposition based dip filter to separate the seismic data into several dip bands (5 or 6). Next, I apply seislet thresholding to each separated dip component to remove random noise. Then I combine all the denoised components to form the final denoised data. Compared with other dip filters, the empirical mode decomposition based dip filter is data-adaptive. One only needs to specify the number of dip components to be separated. Both complicated synthetic and field data examples show superior performance of my proposed approach than the traditional alternatives. The dip-separated structural filtering is not limited to seislet thresholding, and can also be extended to all those methods that require slope information.

Description: Noise interferometry is the process by which approximations to acoustic Green's functions, which describe sound propagation between two locations, are estimated by cross-correlating time series of ambient noise measured at those locations. Noise-interferometry-based approximations to Green's functions can be used as the basis for a variety of inversion algorithms, thereby providing a purely passive alternative to active-source ocean acoustic remote sensing. In this paper we give an overview of results from noise interferometry experiments conducted in the Florida Straits at 100 m depth in December 2012, and at 600 m depth in September/October 2013. Under good conditions for noise interferometry, estimates of cross-correlation functions are shown to allow one to perform advanced phase-coherent signal processing techniques to perform waveform inversions, estimate currents by exploiting non-reciprocity, perform time-reversal/back-propagation calculations and investigate modal dispersion using time-warping techniques. Conditions which are favourable for noise interferometry are identified and discussed.

Description: In this paper, we consider a new spectral finite volume method (FVM) for the elastic wave equations. Our new FVM is based on a piecewise constant approximation on a fine mesh and a high-order polynomial reconstruction on a coarser mesh. Our new method is constructed based on two existing techniques, the high-order FVM and the spectral FVM. In fact, we will construct a new method to take advantage of both methods. More precisely, our method has two distinctive features. The first one is that the local polynomial reconstructions are performed on the coarse triangles and the reconstruction matrices for all the coarse triangles are the same. This fact enhances the parallelization of our algorithm. We will present a parallel implementation of our method and show excellent efficiency results. The second one is that, by using a suitable number of finer triangles with a coarse triangle, we obtain an overdetermined reconstruction system, which can enhance the robustness of the reconstruction process. To derive our scheme, standard finite volume technique is applied to each fine triangle, and the high-order reconstructed polynomials, computed on coarse triangles, are used to compute numerical fluxes. We will present numerical results to show the performance of our method. Our method is presented for 2-D problems, but the same methodology can be applied to 3-D.

Description: At the heart of the full waveform inversion (FWI) implementation is wavefield extrapolation, and specifically its accuracy and cost. To obtain accurate, dispersion free wavefields, the extrapolation for modelling is often expensive. Combining an efficient extrapolation with a novel gradient preconditioning can render an FWI implementation that efficiently converges to an accurate model. We, specifically, recast the extrapolation part of the inversion in terms of its spectral components for both data and gradient calculation. This admits dispersion free wavefields even at large extrapolation time steps, which improves the efficiency of the inversion. An alternative spectral representation of the depth axis in terms of sine functions allows us to impose a free surface boundary condition, which reflects our medium boundaries more accurately. Using a newly derived perfectly matched layer formulation for this spectral implementation, we can define a finite model with absorbing boundaries. In order to reduce the nonlinearity in FWI, we propose a multiscale conditioning of the objective function through combining the different directional components of the gradient to optimally update the velocity. Through solving a simple optimization problem, it specifically admits the smoothest approximate update while guaranteeing its ascending direction. An application to the Marmousi model demonstrates the capability of the proposed approach and justifies our assertions with respect to cost and convergence.

Description: Chloroflexus aggregans is an unbranched multicellular filamentous bacterium having the ability of gliding motility. The filament moves straightforward at a constant rate, ~3 μm sec –1 on solid surface and occasionally reverses the moving direction. In this study, we successfully detected movements of glass beads on the cell-surface along long axis of the filament indicating that the cell-surface movement was the direct force for gliding. Microscopic analyses found that the cell-surface movements were confined to a cell of the filament, and each cell independently moved and reversed the direction. To understand how the cellular movements determine the moving direction of the filament, we proposed a discrete-time stochastic model; sum of the directions of the cellular movements determines the moving direction of the filament only when the filament pauses, and after moving, the filament keeps the same directional movement until all the cells pause and/or move in the opposite direction. Monte Carlo simulation of this model showed that reversal frequency of longer filaments was relatively fixed to be low, but the frequency of shorter filaments varied widely. This simulation result appropriately explained the experimental observations. This study proposed the relevant mechanism adequately describing the motility of the multicellular filament in C. aggregans .

Description: Members of the Gram-positive lactic acid bacteria (LAB) are well-known for their beneficial properties as starter cultures and probiotics. Many LAB species produce ribosomally synthesized proteinaceous antibiotics (bacteriocins). Weissella confusa MBF8-1 is a strain isolated from a fermented soybean product that not only produces useful exopolysaccharides but also exhibits bacteriocin activity, which we call weissellicin MBF. Here, we show that bacteriocin production by W. confusa MBF8-1 is specified by a large plasmid, pWcMBF8-1. Plasmid pWcMBF8-1 (GenBank accession number KR350502), which was identified from the W. confusa MBF8-1 draft genome sequence, is 17 643 bp in length with a G + C content of 34.8% and contains 25 open reading frames (ORFs). Six ORFs constitute the weissellicin MBF locus, encoding three putative double-glycine-motif peptides (Bac1, Bac2, Bac3), an ABC transporter complex (BacTE) and a putative immunity protein (BacI). Two ORFs encode plasmid partitioning and mobilization proteins, suggesting that pWcMBF8-1 is transferable to other hosts. To the best of our knowledge, plasmid pWcMBF8-1 not only represents the first large Weissella plasmid to be sequenced but also the first to be associated with bacteriocin production in W. confusa .

Description: Spectral induced polarization (SIP) data are commonly analysed using phenomenological models. Among these models the Cole–Cole (CC) model is the most popular choice to describe the strength and frequency dependence of distinct polarization peaks in the data. More flexibility regarding the shape of the spectrum is provided by decomposition schemes. Here the spectral response is decomposed into individual responses of a chosen elementary relaxation model, mathematically acting as kernel in the involved integral, based on a broad range of relaxation times. A frequently used kernel function is the Debye model, but also the CC model with some other a priorly specified frequency dispersion (e.g. Warburg model) has been proposed as kernel in the decomposition. The different decomposition approaches in use, also including conductivity and resistivity formulations, pose the question to which degree the integral spectral parameters typically derived from the obtained relaxation time distribution are biased by the approach itself. Based on synthetic SIP data sampled from an ideal CC response, we here investigate how the two most important integral output parameters deviate from the corresponding CC input parameters. We find that the total chargeability may be underestimated by up to 80 per cent and the mean relaxation time may be off by up to three orders of magnitude relative to the original values, depending on the frequency dispersion of the analysed spectrum and the proximity of its peak to the frequency range limits considered in the decomposition. We conclude that a quantitative comparison of SIP parameters across different studies, or the adoption of parameter relationships from other studies, for example when transferring laboratory results to the field, is only possible on the basis of a consistent spectral analysis procedure. This is particularly important when comparing effective CC parameters with spectral parameters derived from decomposition results.

Description: Viability for the development of an engineered geothermal system (EGS) in the oilsands region near Fort McMurray, Alberta, is investigated by studying the structure of the Precambrian basement rocks with magnetotellurics (MT). MT data were collected at 94 broad-band stations on two east–west profiles. Apparent resistivity and phase data showed little variation along each profile. The short period MT data detected a 1-D resistivity structure that could be identified as the shallow sedimentary basin underlain by crystalline basement rocks to a depth of 4–5 km. At lower frequencies a strong directional dependence, large phase splits, and regions of out-of-quadrant (OOQ) phase were detected. 2-D isotropic inversions of these data failed to produce a realistic resistivity model. A detailed dimensionality analysis found links between large phase tensor skews (~15°), azimuths, OOQ phases and tensor decomposition strike angles at periods greater than 1 s. Low magnitude induction vectors, as well as uniformity of phase splits and phase tensor character between the northern and southern profiles imply that a 3-D analysis is not necessary or appropriate. Therefore, 2-D anisotropic forward modelling was used to generate a resistivity model to interpret the MT data. The preferred model was based on geological observations of outcropping anisotropic mylonitic basement rocks of the Charles Lake shear zone, 150 km to the north, linked to the study area by aeromagnetic and core sample data. This model fits all four impedance tensor elements with an rms misfit of 2.82 on the southern profile, and 3.3 on the northern. The conductive phase causing the anisotropy is interpreted to be interconnected graphite films within the metamorphic basement rocks. Characterizing the anisotropy is important for understanding how artificial fractures, necessary for EGS development, would form. Features of MT data commonly interpreted to be 3-D (e.g. out of OOQ phase and large phase tensor skew) are shown to be interpretable with this 2-D anisotropic model.

Description: Small non-coding regulatory RNAs (sRNAs) are key players in post-transcriptional regulation of gene expression. Hundreds of sRNAs have been identified in Sinorhizobium meliloti , but their biological function remains unknown for most of them. In this study, we characterized the expression pattern of the gene encoding the 77-nt sRNA MmgR in S. meliloti strain 2011. A chromosomal transcriptional reporter fusion (P mmgR - gfp ) showed that the mmgR promoter is active along different stages of the interaction with alfalfa roots. In pure cultures, P mmgR - gfp activity paralleled the sRNA abundance indicating that mmgR expression is primarily controlled at the level of transcriptional initiation. P mmgR - gfp activity was higher during growth in rhizobial defined medium (RDM) than in TY medium. Furthermore, P mmgR - gfp was induced at 60 min after shifting growing cells from TY to RDM medium, i.e. shorter than the cell doubling time. In defined RDM medium containing NO 3 – , both P mmgR - gfp and MmgR level were repressed by the addition of tryptone or single amino acids, suggesting that mmgR expression depends on the cellular nitrogen (N) status. In silico analysis failed to detect conserved motifs upstream the promoter RNA polymerase binding site, but revealed a strongly conserved motif centered at –28 that may be linked to the observed regulatory pattern by the N source.

Description: A β-glycoside hydrolase was isolated from cell walls material in Coprinopsis cinerea elongating stipes. By analysis of SDS-PAGE, MALDI-TOF/TOF MS and substrate specificity, this enzyme was characterized as an extracellular β-glucosidase which is a trimer consisting of three homosubunits. β-Glucosidase did not degrade β-glucans with modified ends, whereas it hydrolyzed various β-glucans with free ends and related oligosaccharides with β-1,3-, β-1,4- or β-1,6-linkages. Although this β-glucosidase possesses glycosyltransferase activity on laminarioligosaccharides, it did not transfer glucose residues from laminaritriose to β-glucan in stipe cell walls to produce larger β-glucan molecules; instead, it caused a decrease in the molecular size of stipe wall β-glucan by removing glucose. Relatively, the molecular size of wall β-glucans in the elongating apical stipe was less than that found in the non-elongating basal stipes, and this β-glucosidase was more highly expressed in the elongating apical stipe than in non-elongating basal regions. Therefore, we propose that β-glucosidase functions by trimming or cutting the β-glucan side chains on the β-1,3-glucan backbone to prevent them from forming longer branches, keeping the wall plastic to promote diffuse wall growth.

Description: Streptococcus mutans harbours an intracellular, human DPP IV-analogous enzyme Xaa-Pro dipeptidyl-peptidase (EC 3.4.14.11). According to previous reports, an extracellular isozyme in S. gordonii and S. suis has been associated with virulence. Speculating that even an intracellular form may aid in virulence of S. mutans , we have tried to purify, characterize and evaluate enzyme inhibition by specific inhibitors. The native enzyme was partially purified by ion-exchange and gel filtration chromatography. Owing to low yield, the enzyme was overexpressed in Lactococcus lactis and purified by affinity chromatography. The recombinant enzyme (rSm-XPDAP) had a specific activity of 1070 U mg –1 , while the V max and K m were 7 μM min –1 and 89 ± 7 μM ( n = 3), respectively. The serine protease inhibitor phenylmethylsulphonyl fluoride and a DPP IV-specific inhibitor diprotin A proved to be active against rSm-XPDAP. As a novel approach, the evaluation of the effect of anti-human DPP IV (AHD) drugs on rSm-XPDAP activity found saxagliptin to be effective to some extent ( K i = 129 ± 16 μM), which may lead to the synthesis and development of a new class of antimicrobial agents.

Description: Deep geological repositories, isolated from the geosphere by an engineered bentonite barrier, are currently considered the safest solution for high-level radioactive waste (HLRW) disposal. As the physical conditions and properties of the bentonite barrier are anticipated to change with time, seismic tomography was suggested as a viable technique to monitor the physical state and integrity of the barrier and to timely detect any unforeseen failure. To do so, the seismic monitoring system needs to be optimized, and this can be achieved by conducting numerical simulations of wave propagation in the repository geometry. Previous studies treated bentonite as an elastic medium, whereas recent experimental investigations indicate its pronounced viscoelastic behaviour. The aims of this contribution are (i) to numerically estimate the effective attenuation of bentonite as a function of temperature T and water content W c , so that synthetic data can accurately reproduce experimental traces and (ii) assess the feasibility and limitation of the HLRW repository monitoring by simulating the propagation of sonic waves in a realistic repository geometry. A finite difference method was utilized to simulate the wave propagation in experimental and repository setups. First, the input of the viscoelastic model was varied to achieve a match between experimental and numerical traces. The routine was repeated for several values of W c and T , so that quality factors Q p ( W c , T ) and Q s ( W c , T ) were obtained. Then, the full-scale monitoring procedure was simulated for six scenarios, representing the evolution of bentonite's physical state. The estimated Q p and Q s exhibited a minimum at W c = 20 per cent and higher sensitivity to W c , rather than T , suggesting that pronounced inelasticity of the clay has to be taken into account in geophysical modelling and analysis. The repository-model traces confirm that active seismic monitoring is, in principle, capable of depicting physical changes in the bentonite barrier. However, the locations of sources and receivers relative to the tunnel need to be optimized to achieve best sensitivity to the changes. It was also observed that first arrivals do not necessarily represent the most informative part of the signal; thus the time windows of the analysis need to be selected, accounting for the geometry of the acquisition system. This contribution should be considered as a next step towards the development of a numerical approach to aid in the design and optimization of a non-intrusive monitoring system in HLRW repositories.

Description: Presence of excess unaltered, wild-type (WT) DNA providing no information of biological or clinical value often masks rare alterations containing diagnostic or therapeutic clues in cancer, prenatal diagnosis, infectious diseases or organ transplantation. With the surge of high-throughput technologies there is a growing demand for removing unaltered DNA over large pools-of-sequences. Here we present nuclease-assisted minor-allele enrichment with probe-overlap (NaME-PrO), a single-step approach with broad genome coverage that can remove WT-DNA from numerous sequences simultaneously, prior to genomic analysis. NaME-PrO employs a double-strand-DNA-specific nuclease and overlapping oligonucleotide-probes interrogating WT-DNA targets and guiding nuclease digestion to these sites. Mutation-containing DNA creates probe-DNA mismatches that inhibit digestion, thus subsequent DNA-amplification magnifies DNA-alterations at all selected targets. We demonstrate several-hundred-fold mutation enrichment in diverse human samples on multiple clinically relevant targets including tumor samples and circulating DNA in 50-plex reactions. Enrichment enables routine mutation detection at 0.01% abundance while by adjusting conditions it is possible to sequence mutations down to 0.00003% abundance, or to scan tumor-suppressor genes for rare mutations. NaME-PrO introduces a simple and highly parallel process to remove un-informative DNA sequences and unmask clinically and biologically useful alterations.