ALBERT

Beschreibung: 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.

Beschreibung: 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 .

Beschreibung: Archaeomagnetic field models cover longer timescales than historical models and may therefore resolve the motion of geomagnetic features on the core–mantle boundary (CMB) in a more meaningful statistical sense. Here we perform a detailed appraisal of archaeomagnetic field models to infer some aspects of the physics of the outer core. We characterize and compare the identification and tracking of reversed flux patches (RFPs) in order to assess the RFPs robustness. We find similar behaviour within a family of models but differences among different families, suggesting that modelling strategy is more influential than data set. Similarities involve recurrent positions of RFPs, but no preferred direction of motion is found. The tracking of normal flux patches shows similar qualitative behaviour confirming that RFPs identification and tracking is not strongly biased by their relative weakness. We also compare the tracking of RFPs with that of the historical field model gufm1 and with seismic anomalies of the lowermost mantle to explore the possibility that RFPs have preferred locations prescribed by lower mantle lateral heterogeneity. The archaeomagnetic field model that most resembles the historical field is interpreted in terms of core dynamics and core–mantle thermal interactions. This model exhibits correlation between RFPs and low seismic shear velocity in co-latitude and a shift in longitude. These results shed light on core processes, in particular we infer toroidal field lines with azimuthal orientation below the CMB and large fluid upwelling structures with a width of about 80° (Africa) and 110° (Pacific) at the top of the core. Finally, similar preferred locations of RFPs in the past 9 and 3 kyr of the same archaeomagnetic field model suggest that a 3 kyr period is sufficiently long to reliably detect mantle control on core dynamics. This allows estimating an upper bound of 220–310 km for the magnetic boundary layer thickness below the CMB.

Beschreibung: 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.

Beschreibung: 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.

Beschreibung: Post-depositional reductive diagenesis usually results in partial or entire cleansing of the pristine palaeomagnetic signal, therefore, its intensity is important to be assessed for sediments that are in the purpose of retrieving palaeomagnetic information. Grain size, rock magnetic and geochemical studies on the entire core, along with scanning electron microscope observations and X-ray diffraction analyses for representative samples were carried out on a Holocene sediment core retrieved from the deep water part of Huguangyan maar lake (HGY), southeast China. The pristine magnetic mineral assemblage of the studied core is domianted by superparamagnetic (SP) and stable single domain titanomagnetite, and high coercivity minerals are not detectable. Based on down-core variations of the average grain size ( M Z ), total organic carbon (TOC), detrital elements (Al, Ti, Fe and Mn) and the concentration and mineralogy of magnetic minerals, the studied core could be divided into three subsections. The uppermost subsection is the least affected by diagenesis, with detrital titanomagnetite as the dominant magnetic mineral. This is owing to low TOC contents, but high detrital input generated by weak Asian summer monsoon intensity during the late Holocene. The intermediate subsection shows down-core progressively enhanced dissolution of detrital titanomagnetite, and concomitant formation of authigenic pyrite and siderite, which indicates down-core progressively enhanced diagenesis generated by down-core progressive increasing TOC content, but decreasing detrital input as the result of down-core progressively strengthened Asian summer monsoon intensity. The pristine magnetic mineral assemblage has been profoundly modified in the lowermost subsection. At certain positions of the lowermost subsection, detrital titanomagnetite has been even completely dissolved via diagenesis, giving place to authigenic pyrite and siderite. High TOC content, but low detrital input generated from strong Asian summer monsoon intensity during the early Holocene are accountable for intensive diagenesis in the lowermost subsection. Complete erasing of detrital magnetic input signal at certain positions of the lowermost subsection, and considerable formation of authigenic siderite indicate that palaeomagnetic records of the studied core have been significantly compromised. The studied core has relatively higher TOC content, lower detrital matter content, calmer sedimentary environments, and less DO available at its water–sediment interface than the cores retrieved at relatively shallower water depths, which all contribute to its relatively stronger diagenesis. Progressive thickening of the upper two subsections with increasing water depth is owing to progressive increase in sedimentation rate with increasing water depth, which is the key factor in determining the thickness of each diagenetic subsection of cores from HGY. It would be better that lake sediments for palaeomagnetic investigations collected at a water depth shallower than the depth of its thermocline.

Beschreibung: 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.

Beschreibung: 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.

Beschreibung: The rocks in the crust and the upper mantle of the Earth are believed to exhibit electrical anisotropy to some extent. It is beneficial to further understand and recognize the propagation of the electromagnetic waves in the Earth by investigating the magnetotelluric (which is one of the main geophysical techniques to probe the deep structures in the Earth) responses of the media with anisotropic conductivity structures. In this study, we examine the magnetotelluric fields over an idealized 2-D model consisting of two segments with axially anisotropic conductivity structures overlying a perfect conductor basement by a quasi-static analytic approach. The resulting analytic solution could not only contribute to the electromagnetic induction theory in the anisotropic Earth but also serve as at least an initial standard solution which could be used to validate the reliability and accuracy of the numerical algorithms developed for modelling the magnetotelluric responses of the 2-D media with much more general anisotropic conductivity.

Beschreibung: Several works have reported that haematite has non-linear initial susceptibility at room temperature, like pyrrhotite or titanomagnetite, but there is no explanation for the observed behaviours yet. This study sets out to determine which physical property (grain size, foreign cations content and domain walls displacements) controls the initial susceptibility. The performed measurements include microprobe analysis to determine magnetic phases different to haematite; initial susceptibility (300 K); hysteresis loops, SIRM and backfield curves at 77 and 300 K to calculate magnetic parameters and minor loops at 77 K, to analyse initial susceptibility and magnetization behaviours below Morin transition. The magnetic moment study at low temperature is completed with measurements of zero field cooled–field cooled and AC susceptibility in a range from 5 to 300 K. The minor loops show that the non-linearity of initial susceptibility is closely related to Barkhausen jumps. Because of initial magnetic susceptibility is controlled by domain structure it is difficult to establish a mathematical model to separate magnetic subfabrics in haematite-bearing rocks.

Beschreibung: 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 .

Beschreibung: 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 .

Beschreibung: 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.

Beschreibung: 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.

Beschreibung: A series of important geological events occurred in the Tibetan Plateau area during the Jurassic, such as the collision of the Lhasa and Qiangtang Terranes, the closure of the Meso-Tethyan Ocean, the opening of the Neo-Tethyan Ocean and the cessation of the mega-monsoon. The ~3000 m thick Jurassic sedimentary sequence in the Qiangtang Basin on the central Tibetan Plateau, which is called the Yanshiping (YSP) Group, recorded these geological events. However, the chronology of the sequence is surprisingly poorly constrained. Here, we perform a detailed palaeomagnetic analysis on the ~1060 m thick middle and upper portions of the YSP Group (the Xiali and Suowa Formations) in the YSP section of the eastern Qiangtang Basin. Three bivalve zones at stratigraphic intervals of ~40–140, 640–800 and 940–1040 m are identified, which yield a Bathonian–Callovian age for the Lower Xiali Fm., a Callovian–Oxfordian age for the Lower Suowa Fm. and an Oxfordian–Kimmeridgian age for the Upper Suowa Fm. A total of 544 oriented palaeomagnetic samples were collected from the section. By combining thermal and alternating field demagnetizations, clear characteristic remanent magnetization (ChRM) directions are isolated for most of the samples. The robust ChRM directions pass fold and reversals tests, which support the primary nature of the ChRMs and yield a palaeopole at 76.8°N/297.2°E (dp = 2.2°, dm = 3.7°). A total of 27 normal and 26 reversed polarity zones were successfully recorded in the section. Combined with fossil age constraints, results suggest that the section is plausibly composed of a Callovian-Early Kimmeridgian age sedimentary sequence.

Beschreibung: Data on the evolution of Earth's magnetic field intensity are important for understanding the geodynamo and planetary evolution. However, the paleomagnetic record in rocks may be adversely affected by many physical processes, which must be taken into account when analysing the palaeointensity database. This is especially important in the light of an ongoing debate regarding core thermal conductivity values, and how these relate to the Precambrian geodynamo. Here, we demonstrate that several data sets in the Precambrian palaeointensity database overestimate the true paleofield strength due to the presence of non-ideal carriers of palaeointensity signals and/or viscous re-magnetizations. When the palaeointensity overestimates are removed, the Precambrian database does not indicate a robust change in geomagnetic field intensity during the Mesoproterozoic. These findings call into question the recent claim that the solid inner core formed in the Mesoproterozoic, hence constraining the thermal conductivity in the core to ‘moderate’ values. Instead, our analyses indicate that the presently available palaeointensity data are insufficient in number and quality to constrain the timing of solid inner core formation, or the outstanding problem of core thermal conductivity. Very young or very old inner core ages (and attendant high or low core thermal conductivity values) are consistent with the presently known history of Earth's field strength. More promising available data sets that reflect long-term core structure are geomagnetic reversal rate and field morphology. The latter suggests changes that may reflect differences in Archean to Proterozoic core stratification, whereas the former suggest an interval of geodynamo hyperactivity at ca. 550 Ma.

Beschreibung: 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.

Beschreibung: Red Clay underlying the loess-palaeosol sequences on the Chinese Loess Plateau is an eolian deposit. There is a controversy over whether magnetic susceptibility ( ) variations in Red Clay sequence can be used as an indicator of summer palaeomonsoon intensity. This study investigates the magnetic mineralogy, magnetic concentration and magnetic grain size distribution of Jiaxian Red Clay with multimagnetic methods. Our results indicate that the magnetic properties of Jiaxian Red Clay are similar to those of the Quaternary loess-palaeosol sequences, and ultrafine ferrimagnetic grains produced during pedogenesis are responsible for an increase in susceptibility, therefore the enhancement mechanism of Red Clay is similar to that of the overlying loess-palaeosol sequences. This paper explores variations in the Red Clay sequence through spatial and temporal analysis. The susceptibility variation of six sites along a NNE to SSW transect correlate to palaeoclimatic cycles, so can be used to trace the summer palaeomonsoon intensity from a spatial perspective. However, a simple loess-derived calibration function cannot be used to quantitative reconstruct the palaeomonsoon intensity variations thought time. An adjusted calibration function for palaeosols from Red Clay sequence needs to be developed, so that can be used to quantitative reconstruct palaeomonsoon intensity. Further study is necessary to develop such a transfer function.

Beschreibung: 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.

Beschreibung: Palaeointensity experiments were carried out to a sample collection from two sections of basalt lava flow sequences of Pliocene age in north central Iceland (Chron C2An) to further refine the knowledge of the behaviour of the palaeomagnetic field. Selection of samples was mainly based on their stability of remanence to thermal demagnetization as well as good reversibility in variations of magnetic susceptibility and saturation magnetization with temperature, which would indicate the presence of magnetite as a product of deuteric oxidation of titanomagnetite. Among 167 lava flows from two sections, 44 flows were selected for the Königsberger–Thellier–Thellier experiment in vacuum. In spite of careful pre-selection of samples, an Arai plot with two linear segments, or a concave-up appearance, was often encountered during the experiments. This non-ideal behaviour was probably caused by an irreversible change in the domain state of the magnetic grains of the pseudo-single-domain (PSD) range. This is assumed because an ideal linear plot was obtained in the second run of the palaeointensity experiment in which a laboratory thermoremanence acquired after the final step of the first run was used as a natural remanence. This experiment was conducted on six selected samples, and no clear difference between the magnetic grains of the experimented and pristine sister samples was found by scanning electron microscope and hysteresis measurements, that is, no occurrence of notable chemical/mineralogical alteration, suggesting that no change in the grain size distribution had occurred. Hence, the two-segment Arai plot was not caused by the reversible multidomain/PSD effect in which the curvature of the Arai plot is dependent on the grain size. Considering that the irreversible change in domain state must have affected data points at not only high temperatures but also low temperatures, f v ≥ 0.5 was adopted as one of the acceptance criteria where f v is a vectorially defined fraction of the linear segment. A measure of curvature k ' was also used to check the linearity of the selected linear segment. It was avoided, however, to reject the result out of hand by the large curvature k of the entire data points because it might still include a linear segment with a large fraction. Combining with the results of Shaw's experiments, 52 palaeointensities were obtained out of 192 specimens, or 11 flow means were obtained out of the 44 lava flows. Most of the palaeointensities were from the upper part of the lava section (Chron C2An.1n) and ranged between 30 and 66 μT. Including two results from the bottom part of the lava section, the mean virtual dipole moment for 2.5–3.5 Ma is 6.3 ± 1.4  x  10 22 Am 2 ( N  = 11), which is ~19 per cent smaller than the present-day dipole moment.

Beschreibung: 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.

Beschreibung: 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.

Beschreibung: 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 .

Beschreibung: 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.

Beschreibung: 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.

Beschreibung: Understanding the contribution of biogenic magnetic particles into sedimentary assemblages is a current challenge in palaeomagnetism. It has been demonstrated recently that magnetic particles produced through biologically controlled mineralization processes, such as magnetosomes from magnetotactic bacteria, contribute to the recording of natural remanent magnetization in marine and lacustrian sediments. Contributions from other, biologically induced, mineralization types, which are known from multiple laboratory experiments to include magnetic minerals, remain largely unknown. Here, we report magnetic properties of iron minerals formed by a community of iron- and manganese-reducing bacteria isolated from a natural groundwater deposit during a 2 yr long incubation experiment. The main iron phases of the biomineralized mass are lepidocrocite, goethite and magnetite, each of which has environmental significance. Unlike the majority of the previous studies that reported superparamagnetic grain size, and thus no remanence carrying capacity of biologically induced magnetite, hysteresis and first-order reversal curves measurements in our study have not detected significant superparamagnetic contribution. The biomineralized mass, instead, contains a mixture of single-domain to pseudo-single-domain and multidomain magnetite particles that are capable of carrying a stable chemical remanent magnetization. Isothermal remanent magnetization acquisition parameters and first-order reversal curves signatures of the biomineralized samples deviate from previously proposed criteria for the distinction of extracellular (biologically induced) magnetic particles in mixtures. Given its potential significance as a carrier of natural remanent magnetization, environmental requirements, distribution in nature and the efficiency in the geomagnetic field recording by biologically induced mineralization need comprehensive investigation.

Beschreibung: 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.

Beschreibung: 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 .

Beschreibung: 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.

Beschreibung: 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 .

Beschreibung: 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.

Beschreibung: Monthly means of the magnetic field measurements at ground observatories are a key data source for studying temporal changes of the core magnetic field. However, when they are calculated in the usual way, contributions of external (magnetospheric and ionospheric) origin may remain, which make them less favourable for studying the field generated by dynamo action in the core. We remove external field predictions, including a new way of characterizing the magnetospheric ring current, from the data and then calculate revised monthly means using robust methods. The geomagnetic secular variation (SV) is calculated as the first annual differences of these monthly means, which also removes the static crustal field. SV time-series based on revised monthly means are much less scattered than those calculated from ordinary monthly means, and their variances and correlations between components are smaller. On the annual to decadal timescale, the SV is generated primarily by advection in the fluid outer core. We demonstrate the utility of the revised monthly means by calculating models of the core surface advective flow between 1997 and 2013 directly from the SV data. One set of models assumes flow that is constant over three months; such models exhibit large and rapid temporal variations. For models of this type, less complex flows achieve the same fit to the SV derived from revised monthly means than those from ordinary monthly means. However, those obtained from ordinary monthly means are able to follow excursions in SV that are likely to be external field contamination rather than core signals. Having established that we can find models that fit the data adequately, we then assess how much temporal variability is required. Previous studies have suggested that the flow is consistent with torsional oscillations (TO), solid body-like oscillations of fluid on concentric cylinders with axes aligned along the Earth's rotation axis. TO have been proposed to explain decadal timescale changes in the length-of-day. We invert for flow models where the only temporal changes are consistent with TO, but such models have an unacceptably large data misfit. However, if we relax the TO constraint to allow a little more temporal variability, we can fit the data as well as with flows assumed constant over three months, demonstrating that rapid SV changes can be reproduced by rather small flow changes. Although the flow itself changes slowly, its time derivative can be locally (temporally and spatially) large, in particular when and where core surface secular acceleration peaks. Spherical harmonic expansion coefficients of the flows are not well resolved, and many of them are strongly correlated. Averaging functions, a measure of our ability to determine the flow at a given location from the data distribution available, are poor approximations to the ideal, even when centred on points of the core surface below areas of high observatory density. Both resolution and averaging functions are noticeably worse for the toroidal flow component, which dominates the flow, than the poloidal flow component, except around the magnetic equator where averaging functions for both components are poor.

Beschreibung: 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.

Beschreibung: 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.

Beschreibung: 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.

Beschreibung: 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.

Beschreibung: 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.

Beschreibung: 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.

Beschreibung: 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 .

Beschreibung: 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.

Beschreibung: 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.

Beschreibung: 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.

Beschreibung: We derive scaling relationships for planetary dynamos based on a balance between energy production and Joule dissipation, and between the curl of the buoyancy and Coriolis forces. These scaling relationships are deduced for the particular case of dynamos driven by helical waves, but are shown to have a much broader applicability. They are consistent with the evidence of the numerical dynamos, yielding predictions consistent with published empirical scaling laws and also with the observed transition from dipolar to multipolar dynamos. A direct comparison with the observational evidence for the planets is hampered by the fact that we do not know what sets the smallest scale of the motion in the planets. Nevertheless, we use our scaling relationships to show that the traditional assumption that the Elsasser number is of order unity is inconsistent with the observation that the gas-giant dynamos are dipolar dynamos, as is the more recent suggestion that the strength of the dipole is independent of rotation rate and controlled by the buoyancy flux alone. On the other hand, we show that the observational data is consistent with the hypothesis that a dipolar dynamo saturates at the lowest permissible magnetic energy compatible with a given buoyancy flux.

Beschreibung: 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.

Beschreibung: We present a detailed palaeomagnetic study from 35 sites on Holocene lava flows of the Tongariro Volcanic Centre, central North Island, New Zealand. Prior to the study the eruption ages of these flows were constrained to within a few thousand years by recently published high-precision 40 Ar/ 39 Ar geochronological data and tephrostratigraphic controls. Correlation of flow mean palaeomagnetic directions with a recently published continuous sediment record from Lake Mavora, Fiordland, allows us to reduce the age uncertainty to 300–500 yr in some cases. Our refined ages significantly improve the chronology of Holocene effusive eruptions of the volcanoes of the Tongariro Volcanic Centre. For instance, differences in the palaeomagnetic directions recorded by lavas from the voluminous Iwikau and Rangataua members suggest that individual effusive periods lasted up to thousands of years and that these bursts have been irregularly spaced over time. While over the last few millennia the effusive eruptive activity from Mt Ruapehu has been relatively quiet, the very young age (200–500 BP) of a Red Crater sourced flow suggests that effusive activity around Mt Tongariro lasted into the past few centuries. This adds an important hazard context to the historical record, which has otherwise comprised frequent relatively small, tephra producing, explosive eruptions without the production of lava flows.

Beschreibung: The inversion of magnetotelluric data into subsurface electrical conductivity poses an ill-posed problem. Smoothing constraints are widely employed to estimate a regularized solution. Here, we present an alternative inversion scheme that estimates a sparse representation of the model in a wavelet basis. The objective of the inversion is to determine the few non-zero wavelet coefficients which are required to fit the data. This approach falls into the class of sparsity constrained inversion schemes and minimizes the combination of the data misfit in a least-squares 2 sense and of a model coefficient norm in an 1 sense ( 2 - 1 minimization). The 1 coefficient norm renders the solution sparse in a suitable representation such as the multiresolution wavelet basis, but does not impose explicit structural penalties on the model as it is the case for 2 regularization. The presented numerical algorithm solves the mixed 2 - 1 norm minimization problem for the nonlinear magnetotelluric inverse problem. We demonstrate the feasibility of our algorithm on synthetic 2-D MT data as well as on a real data example. We found that sparse models can be estimated by inversion and that the spatial distribution of non-vanishing coefficients indicates regions in the model which are resolved.

Beschreibung: Earth's liquid core hosts a diverse set of waves with periods ranging from days to thousands of years. One class of waves with periods of several decades is known to arise from an interplay between magnetic, Archimedes and Coriolis forces. These so-called MAC waves are thought to be relevant for interpreting historical fluctuations in the geomagnetic field. In this study, we show that MAC waves provide a good description of time-dependent zonal flow at the top of the core. The same collection of waves also offers a simple explanation for observed fluctuations in the dipole field. Both of these predictions require a stratified layer at the top of the core with a thickness of 130–140 km and a buoyancy frequency comparable to Earth's rotation rate. We extend these predictions to include changes in the length of day (LOD) and find that MAC waves can account for about half of the observed fluctuation at decadal periods. Larger fluctuations are possible when electromagnetic stresses couple MAC waves to flow in the interior of the core. In fact, an idealized model for the coupled motion overestimates the LOD fluctuations, probably reflecting limitations in this idealized model. Our results offer support for stable stratification at the top of the core and suggest a common origin for decadal fluctuations in the geomagnetic field and the LOD.

Beschreibung: 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.

Beschreibung: 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.

Beschreibung: 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.

Beschreibung: Surface nuclear magnetic resonance technique, also called magnetic resonance sounding (MRS), is an emerging geophysical method that can detect the presence and spatial variations of the subsurface water content directly. In this paper, we introduce the MRS central loop geometry, in which the receiver loop is smaller than the transmitter loop and placed in its centre. In addition, using a shielded receiver coil we show how this configuration greatly increases signal-to-noise ratio and improves the resolution of the subsurface layers compared to the typically used coincident loop configuration. We compare sensitivity kernels for different loop configurations and describe advantages of the MRS central loop geometry in terms of superior behaviour of the sensitivity function, increased sensitivity values, reduced noise level of the shielded receiver coil, improved resolution matrix and reduced instrument dead time. With no extra time and effort in the field, central-loop MRS makes it possible to reduce measurement time and to measure data in areas with high anthropogenic noise. The results of our field example agree well with the complementary data, namely airborne electromagnetics, borehole data, and the hydrologic model of the area.

Beschreibung: Frictional heating during earthquake rupture reveals important information on earthquake mechanisms and energy dissipation. The amount of annealing varies widely and is, as yet, poorly constrained. Here we use magnetic susceptibility versus temperature measurements during cycling to increasingly elevated temperatures to constrain the maximum temperature a slip zone has experienced. The case study comprises sheared clay cored from the Japan Trench subduction plate-boundary fault zone (décollement), which accommodated the large slip of the 2011 M w 9.0 Tohoku-oki earthquake. The décollement was cored during the Integrated Ocean Drilling Program (IODP) Expedition 343, the Japan Trench Fast Drilling Project (JFAST). Heating signatures with estimated maximum temperatures ranging from ~300 to over 500 °C are determined close to the multiple slip surfaces within the décollement. Since it is impossible to tie a specific slip surface to a certain earthquake, thermal evidence for the cumulative effect of several earthquakes is unveiled. This as yet preliminary rock magnetic ‘geothermometer’ would be a useful tool to detect seismic heating along faults that experienced medium temperature rise, a range which is difficult to assess with other approaches.

Beschreibung: Feldspars are the most abundant rock-forming minerals in the Earth's crust, but their magnetic properties have not been rigorously studied. This work focuses on the intrinsic magnetic anisotropy of 31 feldspar samples with various chemical compositions. Because feldspar is often twinned or shows exsolution textures, measurements were performed on twinned and exsolved samples as well as single crystals. The anisotropy is controlled by the diamagnetic susceptibility and displays a consistent orientation of principal susceptibility axes; the most negative or minimum susceptibility is parallel to [010], and the maximum (least negative) is close to the crystallographic [001] axis. However, the magnetic anisotropy is weak when compared to other rock-forming minerals, 1.53 x 10 –9 m 3 kg –1 at maximum. Therefore, lower abundance minerals, such as augite, hornblende or biotite, often dominate the bulk paramagnetic anisotropy of a rock. Ferromagnetic anisotropy is not significant in most samples. In the few samples that do show ferromagnetic anisotropy, the principal susceptibility directions of the ferromagnetic subfabric do not display a systematic orientation with respect to the feldspar lattice. These results suggest that palaeointensity estimates of the geomagnetic field made on single crystals of feldspar will not be affected by a systematic orientation of the ferromagnetic inclusions within the feldspar lattice.

Beschreibung: Feldspars are the most abundant rock-forming minerals in the Earth's crust, but their magnetic properties have not been rigorously studied. This work focuses on the intrinsic magnetic anisotropy of 31 feldspar samples with various chemical compositions. Because feldspar is often twinned or shows exsolution textures, measurements were performed on twinned and exsolved samples as well as single crystals. The anisotropy is controlled by the diamagnetic susceptibility and displays a consistent orientation of principal susceptibility axes; the most negative or minimum susceptibility is parallel to [010], and the maximum (least negative) is close to the crystallographic [001] axis. However, the magnetic anisotropy is weak when compared to other rock-forming minerals, 1.53 x 10 –9 m 3 kg –1 at maximum. Therefore, lower abundance minerals, such as augite, hornblende or biotite, often dominate the bulk paramagnetic anisotropy of a rock. Ferromagnetic anisotropy is not significant in most samples. In the few samples that do show ferromagnetic anisotropy, the principal susceptibility directions of the ferromagnetic subfabric do not display a systematic orientation with respect to the feldspar lattice. These results suggest that palaeointensity estimates of the geomagnetic field made on single crystals of feldspar will not be affected by a systematic orientation of the ferromagnetic inclusions within the feldspar lattice.

Beschreibung: 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/ .

Beschreibung: 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 .

Beschreibung: Palaeomagnetic data obtained from archaeological materials are used for reconstructions of the Earth's magnetic field of the past millennia. While many studies tested the reliability of this recorder for palaeointensity only a few studies did this for direction. The study presents an archaeomagnetic and rock magnetic investigation applied to an experimental pottery kiln, which was operated in 2003 to produce stone ware. This kind of high-quality pottery needs a temperature of at least 1160 °C. Shortly before heating of the kiln direct absolute measurements of the absolute geomagnetic field vector have been carried out close to it. After cooling of the kiln 24 oriented palaeomagnetic samples have been taken. Although Curie temperatures are about 580 °C, that is the typical temperature for magnetite, thermal as well as alternating field demagnetisations reveal also a considerable amount of hematite as magnetic carrier. This mixture of magnetite and hematite is dominated by pseudo-single domain grains. Demagnetisation removed in some cases weak secondary components, but in most cases the specimens carried a single component thermoremanent magnetisation. The mean characteristic remanent magnetisation direction agrees on 95 per cent confidence level with the directly measured field direction. Archaeointensity was obtained from five specimens with the Thellier–Coe method and with the multiple-specimen palaeointensity domain-state corrected method. Six of these specimens also provided a result of the Dekkers–Böhnel method, which overestimated the archaeointensity by about 9 per cent compared to the direct value, while after correction for fraction the value agrees very well. For the multiple-specimen palaeointensity domain-state corrected method only fractions between 25 and 75 per cent have been used and specimens showing alteration have been excluded. Above 450 °C many specimens showed alteration of the magnetic grains. Because median destructive temperatures were often above this value in most cases the fraction was less than 50 per cent. Nevertheless the obtained intensity (48.48 ± 0.24 μ) is on 95 per cent confidence level in agreement with the direct observation. Behaviour of the specimens during the Thellier-experiments was not ideal because of narrow unblocking temperature spectra and alteration. Nevertheless, the obtained mean archaeointensity is also in agreement with the direct field observation. Here the relative palaeointensity error is about 6 per cent and very high compared the multiple-specimen palaeointensity domain-state corrected method. The investigation demonstrates that a pottery kiln can provide a very precise estimate of the ancient geomagnetic field vector.

Beschreibung: In this work, we carry out numerical simulations of the seismo-electromagnetic fields associated with a fault in a porous medium by considering the electrokinetic effect. In addition to porous materials, the adopted layered models comprise solid materials in which the electrokinetic effect is inoperative. First, sensitivity study is performed for the evanescent and direct radiation electromagnetic (EM) waves generated by a double couple point source embedded in a porous half-space below a solid half-space. Results suggest that both the evanescent and direct radiation EM waves are sensitive to some medium properties, for example porosity, salinity, fluid viscosity, and conductivity of solid layer. Then, adopting an eight-layer half-space model, we simulate the seismic and EM wavefields generated by the rupture process of a finite fault. It is shown that the electrokinetic effect is able to generate observable corupture EM signals, but the observability depends on some factors such as the epicentral distance, properties of the medium where the fault is located, and local activity levels of EM noise. Time synchronization coseismic EM signals are recorded when the receiver is close to the ground water level but located in a solid medium. In addition to the post-seismic electric field, our results also show the post-seismic magnetic field which has not been identified in previous simulation studies on the electrokinetic effect. The generation of the post-seismic magnetic field probably requires a sufficiently strong medium heterogeneity or fluid-pressure gradient.

Beschreibung: Fault ruptures in the Earth's crust generate both elastic and electromagnetic (EM) waves. If the corresponding EM signals can be observed, then earthquakes could be detected before the first seismic waves arrive. In this study, I consider the piezomagnetic effect as a mechanism that converts elastic waves to EM energy, and I derive analytical formulas for the conversion process. The situation considered in this study is a whole-space model, in which elastic and EM properties are uniform and isotropic. In this situation, the governing equations of the elastic and EM fields, combined with the piezomagnetic constitutive law, can be solved analytically in the time domain by ignoring the displacement current term. Using the derived formulas, numerical examples are investigated, and the corresponding characteristics of the expected magnetic signals are resolved. I show that temporal variations in the magnetic field depend strongly on the electrical conductivity of the medium, meaning that precise detection of signals generated by the piezomagnetic effect is generally difficult. Expected amplitudes of piezomagnetic signals are estimated to be no larger than 0.3 nT for earthquakes with a moment magnitude of ≥7.0 at a source distance of 25 km; however, this conclusion may not extend to the detection of real earthquakes, because piezomagnetic stress sensitivity is currently poorly constrained.

Beschreibung: 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.

Beschreibung: 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.

Beschreibung: 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.

Beschreibung: This paper presents a methodology to sample equivalence domain (ED) in nonlinear partial differential equation (PDE)-constrained inverse problems. For this purpose, we first applied state-of-the-art stochastic optimization algorithm called Covariance Matrix Adaptation Evolution Strategy (CMAES) to identify low-misfit regions of the model space. These regions were then randomly sampled to create an ensemble of equivalent models and quantify uncertainty. CMAES is aimed at exploring model space globally and is robust on very ill-conditioned problems. We show that the number of iterations required to converge grows at a moderate rate with respect to number of unknowns and the algorithm is embarrassingly parallel. We formulated the problem by using the generalized Gaussian distribution. This enabled us to seamlessly use arbitrary norms for residual and regularization terms. We show that various regularization norms facilitate studying different classes of equivalent solutions. We further show how performance of the standard Metropolis–Hastings Markov chain Monte Carlo algorithm can be substantially improved by using information CMAES provides. This methodology was tested by using individual and joint inversions of magneotelluric, controlled-source electromagnetic (EM) and global EM induction data.

Beschreibung: 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.

Beschreibung: 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.

Beschreibung: 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).

Beschreibung: 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.

Beschreibung: 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.

Beschreibung: 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.

Beschreibung: Recent high-resolution aeromagnetic surveys in South Norway have revealed numerous remanent anomalies over Mesoproterozoic metamorphic rocks. Studies on the nature of the minerals that are the remanent carriers has led to discoveries of titanohematite samples with unusual magnetic properties caused by nanoscale exsolution lamellae with their related lamellar magnetism. Here we focus on a rock unit dominated by quartz-plagioclase-biotite granulite containing titanohematite grains with a strong lattice-preferred orientation parallel to regional foliation. When samples with their natural remanent magnetization (NRM), acquired nearly 1 billion years ago, are cooled to 10 K and hysteresis loops measured, these loops show bi-modal exchange bias caused by the magnetism induced within the ilmenite by antiferromagnetic coupling with the adjacent lamellar NRM. By contrast when the samples are cooled in a strong magnetic field (1.5 Tesla), this results in unimodal lamellar magnetism, and, below the T N of ilmenite it adopts a consistent negative orientation, giving rise to unimodal negative exchange bias of 〉500 mT. The results presented here cover the chemical and magnetic properties, Mossbauer results and transmission electron microscopy of the titanohematite and ilmenite lamellae. Initial magnetic experiments indicated the shifts found in the exchange-bias experiments were directly related to the orientation of the sample to the applied field and the initial state of the NRM. In most samples with these unusual magnetic properties, ilmenite lamellae could not be seen in an optical or a scanning electron microscope. However magnetic experiments gave proof of the presence of ilmenite, later confirmed by Mössbauer spectroscopy. Several attempts were made to identify ilmenite in TEM studies, finally successful in showing ilmenite lamellae parallel to (001) of hematite with thicknesses ~1.2 to 1.7 nm and aspect ratios 7–13. Here we compare new TEM images and the magnetic behaviour of these samples to the MOD2 samples that previously showed extraordinary exchange bias properties, and investigate further the nature of these magnetic minerals.

Beschreibung: 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 .

Beschreibung: 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 .

Beschreibung: Rock-magnetic and geochemical characteristics of three Vertisol profiles with different degree of textural differentiation have been studied. Thermomagnetic analyses, thermal demagnetization of laboratory remanences and acquisition of isothermal remanence curves are applied for identification of iron oxide mineralogy. The main magnetic minerals in Vertisols are ferrihydrite, single-domain magnetite, maghemite and hematite. Variations in magnetic susceptibility, anhysteretic remanent magnetization, isothermal remanent magnetization, as well as different ratios (Xarm/X, ARM/SIRM, S-ratio) along depth are studied. Concentration of magnetic minerals in Vertisols is low, influenced by the intense reductomorphic processes. The lowest magnetic susceptibility is found in the most texturally differentiated soil. However, rock-magnetic data suggest the presence of small, but well defined fraction of single domain-like magnetite with relatively wide grain-size distribution found in those parts of the profiles, which are subjected to most intense and frequent seasonal changes in oxidation-reduction conditions. It is suggested that this fraction is formed as a result of transformations of ferrihydrite under repeated cycles of anaerobic/aerobic conditions. Based on geochemical data, CALMAG weathering index was calculated for the three Vertisols. Using the established relation between CALMAG and mean annual precipitation (MAP), palaeo-MAP was evaluated for the studied profiles. The obtained MAP estimations fall in the range 1000–1200 mm and are much higher compared to contemporary precipitation in the area (MAP in the interval 540–770 mm). This finding confirms the relict character of Vertisols on Bulgarian territory and gives more information about the palaeoclimate during the initial stages of Vertisol formation.

Beschreibung: 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.

Beschreibung: 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.

Beschreibung: 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.

Beschreibung: This work presents MARE2DEM, a freely available code for 2-D anisotropic inversion of magnetotelluric (MT) data and frequency-domain controlled-source electromagnetic (CSEM) data from onshore and offshore surveys. MARE2DEM parametrizes the inverse model using a grid of arbitrarily shaped polygons, where unstructured triangular or quadrilateral grids are typically used due to their ease of construction. Unstructured grids provide significantly more geometric flexibility and parameter efficiency than the structured rectangular grids commonly used by most other inversion codes. Transmitter and receiver components located on topographic slopes can be tilted parallel to the boundary so that the simulated electromagnetic fields accurately reproduce the real survey geometry. The forward solution is implemented with a goal-oriented adaptive finite-element method that automatically generates and refines unstructured triangular element grids that conform to the inversion parameter grid, ensuring accurate responses as the model conductivity changes. This dual-grid approach is significantly more efficient than the conventional use of a single grid for both the forward and inverse meshes since the more detailed finite-element meshes required for accurate responses do not increase the memory requirements of the inverse problem. Forward solutions are computed in parallel with a highly efficient scaling by partitioning the data into smaller independent modeling tasks consisting of subsets of the input frequencies, transmitters and receivers. Non-linear inversion is carried out with a new Occam inversion approach that requires fewer forward calls. Dense matrix operations are optimized for memory and parallel scalability using the ScaLAPACK parallel library. Free parameters can be bounded using a new non-linear transformation that leaves the transformed parameters nearly the same as the original parameters within the bounds, thereby reducing non-linear smoothing effects. Data balancing normalization weights for the joint inversion of two or more data sets encourages the inversion to fit each data type equally well. A synthetic joint inversion of marine CSEM and MT data illustrates the algorithm's performance and parallel scaling on up to 480 processing cores. CSEM inversion of data from the Middle America Trench offshore Nicaragua demonstrates a real world application. The source code and MATLAB interface tools are freely available at http://mare2dem.ucsd.edu .

Beschreibung: The spectral complex conductivity of a water-bearing sand during interaction with carbon dioxide (CO 2 ) is influenced by multiple, simultaneous processes. These processes include partial saturation due to the replacement of conductive pore water with CO 2 and chemical interaction of the reactive CO 2 with the bulk fluid and the grain-water interface. We present a laboratory study on the spectral induced polarization of water-bearing sands during exposure to and flow-through by CO 2 . Conductivity spectra were measured successfully at pressures up to 30 MPa and 80 °C during active flow and at steady-state conditions concentrating on the frequency range between 0.0014 and 100 Hz. The frequency range between 0.1 and 100 Hz turned out to be most indicative for potential monitoring applications. The presented data show that the impact of CO 2 on the electrolytic conductivity may be covered by a model for pore-water conductivity, which depends on salinity, pressure and temperature and has been derived from earlier investigations of the pore-water phase. The new data covering the three-phase system CO 2 -brine-sand further show that chemical interaction causes a reduction of surface conductivity by almost 20 per cent, which could be related to the low pH-value in the acidic environment due to CO 2 dissolution and the dissociation of carbonic acid. The quantification of the total CO 2 effect may be used as a correction during monitoring of a sequestration in terms of saturation. We show that this leads to a correct reconstruction of fluid saturation from electrical measurements. In addition, an indicator for changes of the inner surface area, which is related to mineral dissolution or precipitation processes, can be computed from the imaginary part of conductivity. The low frequency range between 0.0014 and 0.1 Hz shows additional characteristics, which deviate from the behaviour at higher frequencies. A Debye decomposition approach is applied to isolate the feature dominating the data at low frequencies. We conclude from our study that electrical conductivity is not only a highly sensitive indicator for CO 2 saturation in pore space. When it is measured in its full spectral and complex form it contains additional information on the chemical state of the system, which holds the potential of getting access to both saturation and interface properties with one monitoring method.

Beschreibung: Our study attempts to identify a characteristic magnetic signature of overbank sediments exhibiting anthropogenically induced magnetic enhancement and thereby to distinguish them from unenhanced sediments with weak magnetic background values, using a novel approach based on data mining methods, thus providing a mean of rapid pollution determination. Data were obtained from 539 bulk samples from vertical profiles through overbank sediment, collected on seven rivers in the eastern Czech Republic and three rivers in northwest England. k -Means clustering and hierarchical clustering methods, paired group (UPGMA) and Ward's method, were used to divide the samples to natural groups according to their attributes. Interparametric ratios: SIRM/; SIRM/ARM; and S –0.1T were chosen as attributes for analyses making the resultant model more widely applicable as magnetic concentration values can differ by two orders. Division into three clusters appeared to be optimal and corresponded to inherent clusters in the data scatter. Clustering managed to separate samples with relatively weak anthropogenically induced enhancement, relatively strong anthropogenically induced enhancement and samples lacking enhancement. To describe the clusters explicitly and thus obtain a discrete magnetic signature, classification rules (JRip method) and decision trees (J4.8 and Simple Cart methods) were used. Samples lacking anthropogenic enhancement typically exhibited an S –0.1T 〈 c. 0.5, SIRM/ARM 〈 c. 150 and SIRM/ 〈 c. 6000 A m –1 . Samples with magnetic enhancement all exhibited an S –0.1T 〉 0.5. Samples with relatively stronger anthropogenic enhancement were unequivocally distinguished from the samples with weaker enhancement by an SIRM/ARM 〉 c. 150. Samples with SIRM/ARM in a range c. 126–150 were classified as relatively strongly enhanced when their SIRM/ 〉 18 000 A m –1 and relatively less enhanced when their SIRM/ 〈 18 000 A m –1 . An additional rule was arbitrary added to exclude samples with fd% 〉 6 per cent from anthropogenically enhanced clusters as samples with natural magnetic enhancement. The characteristics of the clusters resulted mainly from the relationship between SIRM/ARM and the S –0.1T , and SIRM/ and the S –0.1T . Both SIRM/ARM and SIRM/ increase with increasing S –0.1T values reflecting a greater level of anthropogenic magnetic particles. Overall, data mining methods demonstrated good potential for utilization in environmental magnetism.

Beschreibung: The construction of geomagnetic, archaeomagnetic or palaeomagnetic field models requires some prior knowledge about the actual field, which can be gathered from the statistical properties of the field over a variety of length-scales and timescales. However, available geomagnetic data on centennial to millennial periods are too sparse to infer directly these statistical properties. We thus use high-resolution numerical simulations of the geodynamo to test a method for estimating the temporal power spectra (or equivalently the autocovariance functions) of the individual Gauss coefficients that describe the geomagnetic field outside the Earth's fluid outer core. Based on the spectral analysis of our simulations, we argue that a prior for the observational geomagnetic field over decennial to millennial periods can be constructed from the statistics of the field during the short satellite era. The method rests on the assumption that time-series of spherical harmonic coefficients can be considered as realizations of stationary and differentiable stochastic processes, namely order 2 autoregressive (AR2) processes. In the framework of these processes, the statistics of Gauss coefficients are well constrained by their variance and one or two timescales. We find that the time spectra in the dynamo simulations of all Gauss coefficients but the axial dipole are well approximated by the spectra of AR2 processes characterized by only one timescale. The process parameters can simply be deduced from instantaneous estimates of the spatial power spectra of the magnetic field and of its first time derivative. Some deviations of the Gauss coefficients statistics from this minimal model are also discussed. Characterizing the axial dipole clearly requires a more sophisticated AR2 process, with a second distinct timescale.

Beschreibung: 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.

Beschreibung: The hydrocarbonoclastic bacterium Acinetobacter venetianus RAG-1 has attracted substantial attention due to its powerful oil-degrading capabilities and its potential to play an important ecological role in the cleanup of alkanes. In this study, we compare the transcriptome of the strain RAG-1 grown in dodecane, the corresponding alkanol (dodecanol), and sodium acetate for the characterization of genes involved in dodecane uptake and utilization. Comparison of the transcriptional responses of RAG-1 grown on dodecane led to the identification of 1074 genes that were differentially expressed relative to sodium acetate. Of these, 622 genes were upregulated when grown in dodecane. The highly upregulated genes were involved in alkane catabolism, along with stress response. Our data suggest AlkMb to be primarily involved in dodecane oxidation. Transcriptional response of RAG-1 grown on dodecane relative to dodecanol also led to the identification of permease, outer membrane protein and thin fimbriae coding genes potentially involved in dodecane uptake. This study provides the first model for key genes involved in alkane uptake and metabolism in A. venetianus RAG-1.

Beschreibung: Dissimilatory sulfate reduction is the central microbial metabolism in global sulfur cycling. Understanding the importance of sulfate reduction to Earth's biogeochemical S cycle requires aggregating single-cell processes with geochemical signals. For sulfate reduction, these signals include the ratio of stable sulfur isotopes preserved in minerals, as well as the hydrogen isotope ratios and structures of microbial membrane lipids preserved in organic matter. In this study, we cultivated the model sulfate reducer, Desulfovibrio vulgaris DSM 644 T , to investigate how these parameters were perturbed by changes in expression of the protein DsrC. DsrC is critical to the final metabolic step in sulfate reduction to sulfide. S and H isotopic fractionation imposed by the wild type was compared to three mutants. Discrimination against 34 S in sulfate, as calculated from the residual reactant, did not discernibly differ among all strains. However, a closed-system sulfur isotope distillation model, based on accumulated sulfide, produced inconsistent results in one mutant strain IPFG09. Lipids produced by IPFG09 were also slightly enriched in 2 H. These results suggest that DsrC alone does not have a major impact on sulfate-S, though may influence sulfide-S and lipid-H isotopic compositions. While intriguing, a mechanistic explanation requires further study under continuous culture conditions.

Beschreibung: Lacticin Q is an unmodified leaderless bacteriocin produced by Lactococcus lactis QU 5. It has been revealed that the production and self-immunity of lacticin Q are facilitated by a gene cluster lnqQBCDEF . The gene for a putative TetR-family transcriptional regulator, termed lnqR , was found nearby the lnqQBCDEF cluster, but its involvement in lacticin Q biosynthesis remained unknown. In this study, we created an LnqR-overexpressing QU 5 recombinant by using lactococcal constitutive promoter P 32 . The recombinant QU 5 showed enhanced production of and self-immunity to lacticin Q. RT-PCR analysis has revealed that an overexpression of LnqR increases the amounts of lnqQBCDEF transcripts, and these six genes are transcribed as an operon in a single transcriptional unit. Interestingly, LnqR expression and thus lacticin Q production by L. lactis QU 5 was found temperature dependent, while LnzR, an LnqR-homologue, in L. lactis QU 14 was expressed in a similar but not identical manner to LnqR, resulting in dissimilar bacteriocin productivities by these strains. This report demonstrates LnqR as the first TetR-family transcriptional regulator involved in LAB bacteriocin biosynthesis and that, as an exceptional case of TetR-family regulators, LnqR positively regulates the transcription of these biosynthetic genes.

Beschreibung: The Luonan Basin, located in the transitional zone between temperate and subtropical China, is an important locality for human evolution during the early to middle Pleistocene. The loess deposits in the Luonan Basin contain numerous in situ lithic artefacts; the deposits also constitute suitable material for dating the artefacts and are potentially useful for reconstructing the climatic fluctuations which is important for studying the adaptation and occupation of the area by early humans. We carried out a combined rock magnetic and geochemical investigation of a loess sequence from the Liuwan Palaeolithic site in the Luonan Basin. The results indicate a mixture of magnetic minerals, including magnetite/maghemite and hematite/goethite. Magnetic susceptibility was used as a palaeoclimate proxy on the Chinese Loess Plateau; however, its application to the Luonan Basin may be problematic because the provenance of the loess parent material, as well as the depositional environment, differs from that of the Chinese Loess Plateau. We found that rock magnetic parameters related to the grain size of magnetic minerals, such as SIRM/ and ARM /SIRM, are better palaeoclimatic indicators than magnetic susceptibility. Overall, the magnetic results, together with the results of bulk grain-size and chemical index of alteration, indicate that the interglacial environment of early humans in Luonan Basin was warmer and more humid than the coeval environment of the Chinese Loess Plateau.

Beschreibung: Cytosolic, globular droplets with an average diameter of 50 nm were observed in vitrified Thermoplasma acidophilum cells by means of cryo-electron tomography. These droplets were isolated by column chromatography and immunoprecipitation protein purification methods. Subsequent chemical and biochemical analyses identified lipid and protein components, respectively. Two major lipid components, comigrating menaquinones at the solvent front and the slower migrating Thermoplasma polar lipid U4, were detected by TLC experiments. The major protein component was identified as the 153 amino acid long Ta0547 vitellogenin-N domain protein. This domain has been found so far exclusively in large lipid transport proteins of vertebrates and non-vertebrates. Blast protein database homology searches with Ta0547 did not return any eukaryal hits; homologous sequences were found only in thermo-acidophilic archaeons. However, a profile-sequence domain search performed with the vitellogenin-N domain (PF01347) hmm-profile against the T. acidophilum proteome returned Ta0547 as hit. Electron microscopy appearance of isolated droplets resembled to lipoprotein particles. However, no (tetraether) lipid layer could be detected on the droplets surface, rather hydrophobic compounds of the electron dense lumen were surrounded by a denser discontinuous protein boundary. Based on described features, these particles qualify for a novel lipoprotein particle category, what we nominated Thermoplasma Quinone Droplet.

Beschreibung: Glyoxal (GO) belongs to the reactive electrophilic species generated in vivo in all organisms. In order to identify targets of GO and their response mechanisms, we attempted to screen for GO-sensitive mutants by random insertions of Tn phoA -132. The genes responsible for GO susceptibility were functionally classified as the following: (i) tRNA modification; trmE , gidA and truA , (ii) DNA repair; recA and recC , (iii) toxin–antitoxin; mqsA and (iv) redox metabolism; yqhD and caiC . In addition, an insertion in the crp gene, encoding the cAMP responsive transcription factor, exhibits a GO-resistant phenotype, which is consistent with the phenotype of adenylate cyclase ( cya ) mutant showing GO resistance. This suggests that global regulation involving cAMP is operated in a stress response to GO. To further characterize the CRP-regulated genes directly associated with GO resistance, we created double mutants deficient in both crp and one of the candidate genes including yqhD , gloA and sodB . The results indicate that these genes are negatively regulated by CRP as confirmed by real-time RT-PCR. We propose that tRNA as well as DNA are the targets of GO and that toxin/antitoxin, antioxidant and cAMP are involved in cellular response to GO.

Beschreibung: We develop the theory for the magnetic to magnetic and kinetic to magnetic energy transfer between different spherical harmonic degrees due to the interaction of fluid flow and radial magnetic field at the top of the Earth's core. We show that non-zero secular variation of the total magnetic energy could be significant and may provide evidence for the existence of stretching secular variation, which suggests the existence of radial motions at the top of the Earth's core—whole core convection or MAC waves. However, the uncertainties of the small scales of the geomagnetic field prevent a definite conclusion. Combining core field and flow models we calculate the detailed magnetic to magnetic and kinetic to magnetic energy transfer matrices. The magnetic to magnetic energy transfer shows a complex behaviour with local and non-local transfers. The spectra of magnetic to magnetic energy transfers show clear maxima and minima, suggesting an energy cascade. The kinetic to magnetic energy transfers, which are much weaker due to the weak poloidal flow, are either local or non-local between degree one and higher degrees. The patterns observed in the matrices resemble energy transfer patterns that are typically found in 3-D MHD numerical simulations.

Beschreibung: The magnetic susceptibility measured in alternating field can in general be resolved into a component that is in-phase with the applied field and a component that is out-of-phase. While in non-conductive diamagnetic, paramagnetic and many ferromagnetic materials the phase is effectively zero, in some ferromagnetic minerals, such as pyrrhotite, hematite, titanomagnetite or small magnetically viscous grains of magnetite, it is clearly non-zero. The anisotropy of out-of-phase susceptibility (opAMS) can then be used as a tool for the direct determination of the magnetic subfabrics of the minerals with non-zero phase. The error in determination of out-of-phase susceptibility non-linearly increases with decreasing phase angle. This may result in imprecise determination of the opAMS in specimens with very low phase angle. The degree of opAMS is higher than that of ipAMS, which may in contrast result in slightly increasing precision n the opAMS determination. It is highly recommended to inspect the results of the statistical tests of each specimen and to exclude the specimens whose opAMS is determined with insufficient precision from further processing. In rocks, whose magnetism is dominated by the mineral with non-zero out-of-phase susceptibility, the principal directions of the opAMS and ipAMS are virtually coaxial, while the degree of opAMS is higher than that of ipAMS. In some specific cases, the opAMS provides us with similar data to those provided by anisotropies of low-field dependent susceptibility and frequency-dependent susceptibility. The advantage of the opAMS compared to the other two anisotropies is its simultaneous measurement with the ipAMS during one measuring process, while the other two anisotropies require the AMS measurements in several fields or at least at two operating frequencies.

Beschreibung: The temperature dependence of magnetic susceptibility ( –T curves) and magnetization ( M–T curves) has been used as a routine rock magnetic tool to characterize the magnetic mineralogy and magnetic granulometry of Chinese loess/palaeosols. However, palaeoclimatic interpretation of these thermomagnetic analyses remains controversial. In the present study, total organic carbon (TOC), thermomagnetic and low-temperature magnetic experiments on Mid-Late Pleistocene loess/palaeosols in Central Asia and the Chinese Loess Plateau (CLP) have been conducted. We found that the M ( T ) cooling curves at room temperature were mostly lower than the corresponding heating curves, whereas for the ( T ) analyses the cooling curves at room temperature were always much higher than the heating curves. Low-temperature magnetic measurements demonstrated that a large amount of superparamagnetic ferrimagnetic particles were produced during the thermal treatment and resulted in the aforementioned differences. This finding further indicated that the use of the M–T curves to estimate the relative content of maghemite in the loess/palaeosols from the CLP was problematic. In addition, a positive correlation exists between the TOC and the frequency-dependent susceptibility ( FD ) in the CLP, suggesting that stronger pedogenesis would result in the simultaneous increase in the content of both maghemite and organic matter. Consequently, the parameters 1 (representing the relative content of pedogenic maghemite), 2 ([ ph – ] + 1 ) and ph (related to the organic matter concentration), which can be calculated from the –T analyses, can potentially be used as new indicators of pedogenesis and palaeoclimate in Central Asia and the CLP.

Beschreibung: This is the second of three papers investigating properties of titanohematite-bearing quartzo-feldspathic rocks that create a significant remanent magnetic anomaly in the Modum District, South Norway. The first paper provided initial magnetic results, mineralogical characterization and evidence for the presence of lamellar magnetism. In this paper, knowledge of lamellar magnetic properties is explored through experiments where ilmenite lamellae were magnetized below 57 K, and interact magnetically along interfaces with the titanohematite host. Samples with known NRM directions were placed in specific orientations in an MPMS then cooled in zero field to 5 K, where hysteresis loops were measured in fields up to 5 Tesla. This assured that results were ultimately related to the natural lamellar magnetism produced during cooling ~1 billion years ago. In a second set of experiments the same oriented samples, were subjected to a +5 Tesla (T) field then field cooled to 5 K before hysteresis experiments. The first experiments consistently produced asymmetric shifted hysteresis loops with two loop separations, one in a positive field and one in a negative field. Without exception, when the NRM was oriented toward the negative field end of the MPMS, the bimodal loop showed a dominant loop separation in a positive field. By contrast, when the NRM was oriented toward the positive field end of the MPMS, the bimodal loop showed a dominant loop separation in a negative field. Both observations are consistent with antiferromagnetic coupling between the hard magnetization of ilmenite and the more easily shifted lamellar magnetism of the hematite. The bimodal nature of the loops indicates that the NRMs are vector sums of natural lamellar moments, which are oriented both positively and negatively, and that these opposite moments control the orientations of ilmenite magnetizations when cooling through 57 K. Here, extreme exchange biases up to 1.68 T were measured. The second set of experiments produced asymmetric shifted hysteresis loops with one opening always in the negative field. These observations indicate that the +5 T field applied at room temperature rotated the hematite lamellar magnetism in a positive direction, so that upon cooling all the ilmenite lamellae acquired negative magnetic moments, thus causing unimodal negatively shifted loops. Here, the largest exchange bias among the unimodal loops was only 0.7 T. These results will be used in paper III to build a better understanding of lamellar magnetism at the atomic layer scale.

Beschreibung: The culturability of Escherichia coli , Ralstonia eutropha and Bacillus subtilis after incubation in phosphate-buffered saline at either 5°C or 30°C was determined. The culturability of B. subtilis showed little dependence on temperature. The culturability of E. coli rapidly decreased at 30°C but remained almost constant at 5°C. In contrast, the culturability of R. eutropha decreased by three orders of magnitude at 5°C within 24 h but only moderately decreased (one order of magnitude) at 30°C. Remarkably, prolonged incubation of R. eutropha at 30°C resulted in a full recovery of colony forming units in contrast to only a partial recovery at 5°C. Ralstonia eutropha cells at 30°C remained culturable for 3 weeks while culturability at 5°C constantly decreased. The effect of temperature was significantly stronger in a polyhydroxybutyrate-negative mutant. Our data show that accumulated polyhydroxybutyrate has a cold-protective function and can prevent R. eutropha entering the viable but not culturable state.

Beschreibung: Non-heating palaeointensity methods are a vital tool to explore magnetic field strength variations recorded by thermally sensitive materials of both terrestrial and extraterrestrial origin. One such method is the calibrated pseudo-Thellier method in which a specimen's natural remanent magnetization is alternating field demagnetized and replaced with a laboratory induced anhysteretic remanent magnetization (as an analogue of a thermoremanent magnetization, TRM). Using a set of 56 volcanic specimens given laboratory TRMs in fields of 10–130 μT, we refine the calibration of the pseudo-Thellier method and better define the uncertainty associated with its palaeointensity estimates. Our new calibration, obtained from 32 selected specimens, resolves the issue of non-zero intercept, which is theoretically predicted, but not satisfied by any previous calibration. The range of individual specimen calibration factors, however, is relatively large, but consistent with the variability expected for SD magnetite. We explore a number of rock magnetic parameters in an attempt to identify selection thresholds for reducing the calibration scatter, but fail to find a suitable choice. We infer that our careful selection process, which incorporates more statistics then previous studies, may be largely screening out any strong rock magnetic dependence. Some subtle grain size or mineralogical dependencies, however, remain after selection, but cannot be discerned from the scatter expected for grain size variability of SD magnetite. As a consequence of the variability in the calibration factor, the uncertainty associated with pseudo-Thellier results is much larger than previously indicated. The scatter of the calibration is ~25 per cent of the mean value, which implies that, when combined with the scatter of results typically obtained from a single site, the uncertainty of averaged pseudo-Thellier results will always be 〉25 per cent. As such, pseudo-Thellier results should be complementary to, and cross-validated with results from other methods. Nevertheless, the pseudo-Thellier method remains a valuable tool for obtaining palaeointensity estimates from thermally sensitive terrestrial and extraterrestrial materials and with careful data selection and analysis can yield results that are accurate to within a factor of 4 or better.

Beschreibung: As a consequence of measuring time variations of the electric and the magnetic field, which are related to current flow and charge distribution, magnetotelluric (MT) data in 2-D and 3-D environments are not only sensitive to the geoelectrical structures below the measuring points but also to any lateral anomalies surrounding the acquisition site. This behaviour complicates the characterization of the electrical resistivity distribution of the subsurface, particularly in complex areas. In this manuscript we assess the main advantages of complementing the standard MT impedance tensor ( Z ) data with interstation horizontal magnetic tensor ( H ) and geomagnetic transfer function ( T ) data in constraining the subsurface in a 3-D environment beneath a MT profile. Our analysis was performed using synthetic responses with added normally distributed and scattered random noise. The sensitivity of each type of data to different resistivity anomalies was evaluated, showing that the degree to which each site and each period is affected by the same anomaly depends on the type of data. A dimensionality analysis, using Z , H and T data, identified the presence of the 3-D anomalies close to the profile, suggesting a 3-D approach for recovering the electrical resistivity values of the subsurface. Finally, the capacity for recovering the geoelectrical structures of the subsurface was evaluated by performing joint inversion using different data combinations, quantifying the differences between the true synthetic model and the models from inversion process. Four main improvements were observed when performing joint inversion of Z , H and T data: (1) superior precision and accuracy at characterizing the electrical resistivity values of the anomalies below and outside the profile; (2) the potential to recover high electrical resistivity anomalies that are poorly recovered using Z data alone; (3) improvement in the characterization of the bottom and lateral boundaries of the anomalies with low electrical resistivity; and (4) superior imaging of the horizontal continuity of structures with low electrical resistivity. These advantages offer new opportunities for the MT method by making the results from a MT profile in a 3-D environment more convincing, supporting the possibility of high-resolution studies in 3-D areas without expending a large amount of economical and computational resources, and also offering better resolution of targets with high electrical resistivity.

Beschreibung: Corynebacterium glutamicum is able to metabolize different nitrogen and carbon sources. In standard minimal media, ammonium and urea typically serve as nitrogen source and glucose or sucrose as carbon and energy source; however, amino acids might also play a role as nitrogen, carbon and energy source. In this study, the function of the putative glutaminase GlsK was investigated. A glsK deletion strain showed impaired growth with L-glutamine as carbon and energy source, while growth was improved upon glsK overexpression. GlsK possesses a carboxy-terminal domain that seems to be restricted to Corynebacterium species . A truncated GlsK lacking this extension led to faster growth, indicating a regulatory function of this domain. In fact, GlsK activity is regulated in a pH-dependent manner depending on the carboxy-terminal extension, and is positively influenced by cAMP. Furthermore, the C-terminal extension seems to be important for oligomerization of GlsK.

Beschreibung: Sulfur is an essential element for growth and many physiological functions. As sulfur sources for Escherichia coli and related bacteria, specific transporters import various sulfur-containing compounds from the environment. In this study, we identified and characterized an alternative function of the cystine transporter YdjN in E. coli as a transporter of S -sulfocysteine, a sulfur-containing intermediate in the assimilatory cysteine biosynthesis that is used as a sulfur source for the growth of E. coli . We also demonstrated that the transport of S -sulfocysteine via YdjN depends on the transcriptional regulator CysB, a master regulator that controls most of the genes involved in sulfur assimilation and cysteine metabolism. We found that the use of S -sulfocysteine as a sulfur source depends on glutathione because mutations in glutathione biosynthetic genes abolish growth when S -sulfocysteine is used as a sole sulfur source, thereby supporting the previous findings that the conversion of S -sulfocysteine to cysteine is catalyzed by glutaredoxins. To the best of our knowledge, this is the first report of a functional S -sulfocysteine transporter across organisms, which strongly supports the hypothesis that S -sulfocysteine is not only a metabolic intermediate but also a physiologically significant substance in specific natural environments.

Beschreibung: Inference of globally averaged eustatic sea level (ESL) rise since the Last Glacial Maximum (LGM) highly depends on the interpretation of relative sea level (RSL) observations at Barbados and Bonaparte Gulf, Australia, which are sensitive to the viscosity structure of Earth's mantle. Here we examine the RSL changes at the LGM for Barbados and Bonaparte Gulf ( ${\rm{RSL}}_{\rm{L}}^{{\rm{Bar}}}$ and ${\rm{RSL}}_{\rm{L}}^{{\rm{Bon}}}$ ), differential RSL for both sites ( $\Delta {\rm{RSL}}_{\rm{L}}^{{\rm{Bar}},{\rm{Bon}}})$ and rate of change of degree-two harmonics of Earth's geopotential due to glacial isostatic adjustment (GIA) process (GIA-induced 2 ) to infer the ESL component and viscosity structure of Earth's mantle. Differential RSL, $\Delta {\rm{RSL}}_{\rm{L}}^{{\rm{Bar}},{\rm{Bon}}}$ and GIA-induced 2 are dominantly sensitive to the lower-mantle viscosity, and nearly insensitive to the upper-mantle rheological structure and GIA ice models with an ESL component of about (120–130) m. The comparison between the predicted and observationally derived $\Delta {\rm{RSL}}_{\rm{L}}^{{\rm{Bar}},{\rm{Bon}}}$ indicates the lower-mantle viscosity higher than ~2  x  10 22 Pa s, and the observationally derived GIA-induced 2 of –(6.0–6.5)  x  10 –11 yr –1 indicates two permissible solutions for the lower mantle, ~10 22 and (5–10)  x  10 22 Pa s. That is, the effective lower-mantle viscosity inferred from these two observational constraints is (5–10)  x  10 22 Pa s. The LGM RSL changes at both sites, ${\rm{RSL}}_{\rm{L}}^{{\rm{Bar}}}$ and ${\rm{RSL}}_{\rm{L}}^{{\rm{Bon}}}$ , are also sensitive to the ESL component and upper-mantle viscosity as well as the lower-mantle viscosity. The permissible upper-mantle viscosity increases with decreasing ESL component due to the sensitivity of the LGM sea level at Bonaparte Gulf ( ${\rm{RSL}}_{\rm{L}}^{{\rm{Bon}}}$ ) to the upper-mantle viscosity, and inferred upper-mantle viscosity for adopted lithospheric thicknesses of 65 and 100 km is (1–3)  x  10 20 Pa s for ESL~130 m and (4–10)  x  10 20 Pa s for ESL~125 m. The former solution of (1–3)  x  10 20 Pa s is consistent with the inferences from the postglacial differential RSL changes in the Australian region and also inversion study of far-field sea-level data. The inference of the viscosity structure based on these four observational constraints is, however, relatively insensitive to the viscosity structure of D'' layer.

Beschreibung: We present the theory, algorithms and implementation of a parallel finite-volume algorithm for the solution of the incompressible magnetohydrodynamic (MHD) equations using unstructured grids that are applicable for a wide variety of geometries. Our method implements a mixed Adams-Bashforth/Crank-Nicolson scheme for the nonlinear terms in the MHD equations and we prove that it is stable independent of the time step. To ensure that the solenoidal condition is met for the magnetic field, we use a method whereby a pseudo-pressure is introduced into the induction equation; since we are concerned with incompressible flows, the resulting Poisson equation for the pseudo-pressure is solved alongside the equivalent Poisson problem for the velocity field. We validate our code in a variety of geometries including periodic boxes, spheres, spherical shells, spheroids and ellipsoids; for the finite geometries we implement the so-called ferromagnetic or pseudo-vacuum boundary conditions appropriate for a surrounding medium with infinite magnetic permeability. This implies that the magnetic field must be purely perpendicular to the boundary. We present a number of comparisons against previous results and against analytical solutions, which verify the code's accuracy. This documents the code's reliability as a prelude to its use in more difficult problems. We finally present a new simple drifting solution for thermal convection in a spherical shell that successfully sustains a magnetic field of simple geometry. By dint of its rapid stabilization from the given initial conditions, we deem it suitable as a benchmark against which other self-consistent dynamo codes can be tested.

Beschreibung: We consider an inviscid and perfectly conducting fluid sphere in rapid rotation and permeated by a background magnetic field. Such a system admits normal modes in the form of torsional oscillations, namely azimuthal motions of cylinders coaxial with the rotation axis. We analyse this system for a particular background magnetic field that provides a new closed form normal mode solution. We derive Wentzel–Kramers–Brillouin–Jeffreys (WKBJ) approximations to the normal modes, and focus particularly on the reflections that take place on the rotation axis and at the equator. We propose a procedure to calculate the reflection coefficients and we discuss the analogy of our findings with well-known seismological results. Our analytical results are tested against numerical calculations and show good agreement.

Beschreibung: The 69 Ma Prospector Mountain stock is located in southwestern Yukon in the northern Canadian Cordillera. This massive monzonite-syenogranite stock is thought to be the intrusive volcanic centre for the surrounding coeval Carmacks Group volcanics. Anomalous palaeomagnetic data from these volcanics have provided the only evidence for the commonly posited hypothesis that the Yukon-Tanana terrane (YTT) was part of the far-travelled (1950 ± 600 km northward) ‘Baja BC’ terrane from 70 to 50 Ma. All other geologic evidence and averaged palaeomagnetic data support a northward displacement of ~415 ± 15 km. This study provides a direct test of the Carmacks volcanics’ estimate and examines the possible causes of its anomalous results. Both the stock and volcanics are unmetamorphosed and rest unconformably on metamorphosed basement rocks of the YTT. Palaeomagnetic and mineral magnetic results from 17 of 19 tested sites (218 specimens) in the Prospector stock and its peripheral skarn isolated a stable thermoremanent magnetization (TRM) in magnetite or low-Ti titanomagnetite that was mostly determined on demagnetization between temperatures of 500 and 580 °C. The TRM has a direction of Decl. = 8.3°, Incl. = 82.4° ( N = 17, k = 71.9, α 95 = 4.2°), providing a non-significant northwards translation estimate of 70 ± 880 km for the YTT. The normal-polarity TRM direction at Prospector Mountain provides a highly significant palaeomagnetic reversals test with the reversed-polarity TRM of the 70 Ma Swede Dome stock, another volcanic centre of the Carmacks Group about 190 km to the north. The test affirms to a high probability that both stocks carry primary TRMs and have not been tectonically tilted significantly since emplacement. Combining the palaeopoles for these two stocks with that for the 75 Ma Mount Lorne volcanic centre stock about 210 km south of Prospector Mountain yields a combined northward translation estimate of 330 ± 400 km for the YTT since ~71 Ma. This estimate agrees closely with the 415 ± 15 km estimate from geological constraints on strike-slip motion along the inboard Tintina fault zone since ~70 Ma, and with the palaeomagnetic estimate of 650 ± 450 km northward since ~108 Ma from four mid-Cretaceous batholiths. In contrast, the palaeomagnetic directions from four areas of the ~70 Ma Carmacks volcanics are poorly clustered. Nonetheless their estimates of ~1900 km northward have been used for a quarter of a century to support a far-travelled ‘Baja BC’ tectonic model for the YTT. This paper discusses and concludes that the anomalous far-travelled estimates from the Carmacks volcanics are attributable to the unfortunate additive effects of inadequate averaging of secular variation, dipole offset error, unrecognized primary dip and other possible causes.