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Here we present an exome-wide rare genetic variant association study for 30 blood biomarkers in 191,971 individuals in the UK Biobank. We compare gene- based association tests for separate functional variant categories to increase interpretability and identify 193 significant gene-biomarker associations. Genes associated with biomarkers were ~ 4.5-fold enriched for conferring Mendelian disorders. In addition to performing weighted gene-based variant collapsing tests, we design and apply variant-category-specific kernel-based tests that integrate quantitative functional variant effect predictions for mis- sense variants, splicing and the binding of RNA-binding proteins. For these tests, we present a computationally efficient combination of the likelihood- ratio and score tests that found 36% more associations than the score test alone while also controlling the type-1 error. Kernel-based tests identified 13% more associations than their gene-based collapsing counterparts and had advantages in the presence of gain of function missense variants. We introduce local collapsing by amino acid position for missense variants and use it to interpret associations and identify potential novel gain of function variants in PIEZO1. Our results show the benefits of investigating different functional mechanisms when performing rare-variant association tests, and demonstrate pervasive rare-variant contribution to biomarker variability.
X-ray observations of star-planet systems are important to grow our understanding of exoplanets; these observations allow for studies of photoevaporation of the exoplanetary atmosphere, and in some cases even estimations of the size of the outer planetary atmosphere. The German-Russian eROSITA instrument onboard the SRG (Spectrum Roentgen Gamma) mission is performing the first all-sky X-ray survey since the 1990s, and provides X-ray fluxes and spectra of exoplanet host stars over a much larger volume than was accessible before. Using new eROSITA data as well as archival data from XMM-Newton, Chandra, and ROSAT, we estimate mass-loss rates of exoplanets under an energy-limited escape scenario and identify several exoplanets with strong X-ray irradiation and expected mass loss that are amenable to follow-up observations at other wavelengths. We model sample spectra using a toy model of an exoplanetary atmosphere to predict what exoplanet transit observations with future X-ray missions such as Athena will look like and estimate the observable X-ray transmission spectrum for a typical hot Jupiter-type exoplanet.
We present a technique that identifies truly interacting subsystems of a complex system from multichannel data if the recordings are an unknown linear and instantaneous mixture of the true sources. The method is valid for arbitrary noise structure. For this, a blind source separation technique is proposed that diagonalizes antisymmetrized cross- correlation or cross-spectral matrices. The resulting decomposition finds truly interacting subsystems blindly and suppresses any spurious interaction stemming from the mixture. The usefulness of this interacting source analysis is demonstrated in simulations and for real electroencephalography data
Arctic lakes located in permafrost regions are susceptible to catastrophic drainage. In this study, we reconstructed historical lake drainage events on the western Arctic Coastal Plain of Alaska between 1955 and 2017 using USGS topographic maps, historical aerial photography (1955), and Landsat Imagery (ca. 1975, ca. 2000, and annually since 2000). We identified 98 lakes larger than 10 ha that partially (>25% of area) or completely drained during the 62-year period. Decadal-scale lake drainage rates progressively declined from 2.0 lakes/yr (1955-1975), to 1.6 lakes/yr (1975-2000), and to 1.2 lakes/yr (2000-2017) in the ~30,000-km(2) study area. Detailed Landsat trend analysis between 2000 and 2017 identified two years, 2004 and 2006, with a cluster (five or more) of lake drainages probably associated with bank overtopping or headward erosion. To identify future potential lake drainages, we combined the historical lake drainage observations with a geospatial dataset describing lake elevation, hydrologic connectivity, and adjacent lake margin topographic gradients developed with a 5-m-resolution digital surface model. We identified ~1900 lakes likely to be prone to drainage in the future. Of the 20 lakes that drained in the most recent study period, 85% were identified in this future lake drainage potential dataset. Our assessment of historical lake drainage magnitude, mechanisms and pathways, and identification of potential future lake drainages provides insights into how arctic lowland landscapes may change and evolve in the coming decades to centuries.
Flood damage estimation is a core task in flood risk assessments and requires reliable flood loss models. Identifying the driving factors of flood loss at residential buildings and gaining insight into their relations is important to improve our understanding of flood damage processes. For that purpose, we learn probabilistic graphical models, which capture and illustrate (in-)dependencies between the considered variables. The models are learned based on postevent surveys with flood-affected residents after six flood events, which occurred in Germany between 2002 and 2013. Besides the sustained building damage, the survey data contain information about flooding parameters, early warning and emergency measures, property-level mitigation measures and preparedness, socioeconomic characteristics of the household, and building characteristics. The analysis considers the entire data set with a total of 4,468 cases as well as subsets of the data set partitioned into single flood events and flood types: river floods, levee breaches, surface water flooding, and groundwater floods, to reveal differences in the damaging processes. The learned networks suggest that the flood loss ratio of residential buildings is directly influenced by hydrological and hydraulic aspects as well as by building characteristics and property-level mitigation measures. The study demonstrates also that for different flood events and process types the building damage is influenced by varying factors. This suggests that flood damage models need to be capable of reproducing these differences for spatial and temporal model transfers.
Objective
To improve consumer decision making, the results of risk assessments on food, feed, consumer products or chemicals need to be communicated not only to experts but also to non-expert audiences. The present study draws on evidence from literature reviews and focus groups with diverse stakeholders to identify content to integrate into an existing risk assessment communication (Risk Profile).
Methods
A combination of rapid literature reviews and focus groups with experts (risk assessors (n = 15), risk managers (n = 8)), and non-experts (general public (n = 18)) were used to identify content and strategies for including information about risk assessment results in the “Risk Profile” from the German Federal Institute for Risk Assessment. Feedback from initial focus groups was used to develop communication prototypes that informed subsequent feedback rounds in an iterative process. A final prototype was validated in usability tests with experts.
Results
Focus group feedback and suggestions from risk assessors were largely in line with findings from the literature. Risk managers and lay persons offered similar suggestions on how to improve the existing communication of risk assessment results (e.g., including more explanatory detail, reporting probabilities for individual health impairments, and specifying risks for subgroups in additional sections). Risk managers found information about quality of evidence important to communicate, whereas people from the general public found this information less relevant. Participants from lower educational backgrounds had difficulties understanding the purpose of risk assessments. User tests found that the final prototype was appropriate and feasible to implement by risk assessors.
Conclusion
An iterative and evidence-based process was used to develop content to improve the communication of risk assessments to the general public while being feasible to use by risk assessors. Remaining challenges include how to communicate dose-response relationships and standardise quality of evidence ratings across disciplines.
High Mountain Asia (HMA) – encompassing the Tibetan Plateau and surrounding mountain ranges – is the primary water source for much of Asia, serving more than a billion downstream users. Many catchments receive the majority of their yearly water budget in the form of snow, which is poorly monitored by sparse in situ weather networks. Both the timing and volume of snowmelt play critical roles in downstream water provision, as many applications – such as agriculture, drinking-water generation, and hydropower – rely on consistent and predictable snowmelt runoff. Here, we examine passive microwave data across HMA with five sensors (SSMI, SSMIS, AMSR-E, AMSR2, and GPM) from 1987 to 2016 to track the timing of the snowmelt season – defined here as the time between maximum passive microwave signal separation and snow clearance. We validated our method against climate model surface temperatures, optical remote-sensing snow-cover data, and a manual control dataset (n = 2100, 3 variables at 25 locations over 28 years); our algorithm is generally accurate within 3–5 days. Using the algorithm-generated snowmelt dates, we examine the spatiotemporal patterns of the snowmelt season across HMA. The climatically short (29-year) time series, along with complex interannual snowfall variations, makes determining trends in snowmelt dates at a single point difficult. We instead identify trends in snowmelt timing by using hierarchical clustering of the passive microwave data to determine trends in self-similar regions. We make the following four key observations. (1) The end of the snowmelt season is trending almost universally earlier in HMA (negative trends). Changes in the end of the snowmelt season are generally between 2 and 8 days decade−1 over the 29-year study period (5–25 days total). The length of the snowmelt season is thus shrinking in many, though not all, regions of HMA. Some areas exhibit later peak signal separation (positive trends), but with generally smaller magnitudes than trends in snowmelt end. (2) Areas with long snowmelt periods, such as the Tibetan Plateau, show the strongest compression of the snowmelt season (negative trends). These trends are apparent regardless of the time period over which the regression is performed. (3) While trends averaged over 3 decades indicate generally earlier snowmelt seasons, data from the last 14 years (2002–2016) exhibit positive trends in many regions, such as parts of the Pamir and Kunlun Shan. Due to the short nature of the time series, it is not clear whether this change is a reversal of a long-term trend or simply interannual variability. (4) Some regions with stable or growing glaciers – such as the Karakoram and Kunlun Shan – see slightly later snowmelt seasons and longer snowmelt periods. It is likely that changes in the snowmelt regime of HMA account for some of the observed heterogeneity in glacier response to climate change. While the decadal increases in regional temperature have in general led to earlier and shortened melt seasons, changes in HMA's cryosphere have been spatially and temporally heterogeneous.
Water-soluble heteroglycans (SHG) were isolated from leaves of wild-type Arabidopsis thaliana L. and from two starch-deficient mutants. Major constituents of the SHG are arabinose, galactose, rhamnose, and glucose. SHG was separated into low (< 10 kDa; SHG(S)) and high (> 10 kDa; SHG(L)) molecular weight compounds. SHG(S) was resolved into approximately 25 distinct oligoglycans by ion exchange chromatography. SHG(L) was further separated into two subfractions, designated as subfraction I and II, by field flow fractionation. For the intracellular localization of the various SHG compounds several approaches were chosen: first, leaf material was subjected to non-aqueous fractionation. The apolar gradient fractions were characterized by monitoring markers and were used as starting material for the SHG isolation. Subfraction I and SHG(S) exhibited a distribution similar to that of cytosolic markers whereas subfraction II cofractionated with crystalline cellulose. Secondly, intact organelles were isolated and used for SHG isolation. Preparations of intact organelles (mitochondria plus peroxisomes) contained no significant amount of any heteroglycan. In isolated intact microsomes a series of oligoglycans was recovered but neither subfraction I nor II. In in vitro assays using glucose 1-phosphate and recombinant cytosolic (Pho 2) phosphorylase both SHG(S) and subfraction I acted as glucosyl acceptor whereas subfraction II was essentially inactive. Rabbit muscle phosphorylase a did not utilize any of the plant glycans indicating a specific Pho 2-glycan interaction. As revealed by in vivo labeling experiments using (CO2)-C-14 carbon fluxes into subfraction I and II differed. Furthermore, in leaves the pool size of subfraction I varied during the light-dark regime
The Norway lobster, Nephrops norvegicus, is a burrowing decapod with a rhythmic burrow emergence (24 h) governed by the circadian system. It is an important resource for European fisheries and its behavior deeply affects its availability. The current knowledge of Nephrops circadian biology is phenomenological as it is currently the case for almost all crustaceans. In attempt to elucidate the putative molecular mechanisms underlying circadian gene regulation in Nephrops, we used a transcriptomics approach on cDNA extracted from the eyestalk, a structure playing a crucial role in controlling behavior of decapods. We studied 14 male lobsters under 12-12 light-darkness blue light cycle. We used the Hiseq 2000 Illumina platform to sequence two eyestalk libraries (under light and darkness conditions) obtaining about 90 millions 100-bp paired-end reads. Trinity was used for the de novo reconstruction of transcriptomes; the size at which half of all assembled bases reside in contigs (N50) was equal to 1796 (light) and 2055 (darkness). We found a list of candidate clock genes and focused our attention on canonical ones: timeless, period, clock and bmal1. The cloning of assembled fragments validated Trinity outputs. The putative Nephrops clock genes showed high levels of identity (blastx on NCBI) with known crustacean clock gene homologs such as Eurydice pulchra (period: 47%, timeless: 59%, bmal1: 79%) and Macrobrachium rosenbergii (clock: 100%). We also found a vertebrate-like cryptochrome 2. RT-qPCR showed that only timeless had a robust diel pattern of expression. Our data are in accordance with the current knowledge of the crustacean circadian clock, reinforcing the idea that the molecular clockwork of this group shows some differences with the established model in Drosophila melanogaster.
The Norway lobster, Nephrops norvegicus, is a burrowing decapod with a rhythmic burrow emergence (24 h) governed by the circadian system. It is an important resource for European fisheries and its behavior deeply affects its availability. The current knowledge of Nephrops circadian biology is phenomenological as it is currently the case for almost all crustaceans. In attempt to elucidate the putative molecular mechanisms underlying circadian gene regulation in Nephrops, we used a transcriptomics approach on cDNA extracted from the eyestalk, a structure playing a crucial role in controlling behavior of decapods. We studied 14 male lobsters under 12–12 light-darkness blue light cycle. We used the Hiseq 2000 Illumina platform to sequence two eyestalk libraries (under light and darkness conditions) obtaining about 90 millions 100-bp paired-end reads. Trinity was used for the de novo reconstruction of transcriptomes; the size at which half of all assembled bases reside in contigs (N50) was equal to 1796 (light) and 2055 (darkness). We found a list of candidate clock genes and focused our attention on canonical ones: timeless, period, clock and bmal1. The cloning of assembled fragments validated Trinity outputs. The putative Nephrops clock genes showed high levels of identity (blastx on NCBI) with known crustacean clock gene homologs such as Eurydice pulchra (period: 47%, timeless: 59%, bmal1: 79%) and Macrobrachium rosenbergii (clock: 100%). We also found a vertebrate-like cryptochrome 2. RT-qPCR showed that only timeless had a robust diel pattern of expression. Our data are in accordance with the current knowledge of the crustacean circadian clock, reinforcing the idea that the molecular clockwork of this group shows some differences with the established model in Drosophila melanogaster.
Identification of YdhV as the First Molybdoenzyme Binding a Bis-Mo-MPT Cofactor in Escherichia coli
(2019)
The oxidoreductase YdhV in Escherichia coli has been predicted to belong to the family of molybdenum/tungsten cofactor (Moco/Wco)-containing enzymes. In this study, we characterized the YdhV protein in detail, which shares amino acid sequence homology with a tungsten-containing benzoyl-CoA reductase binding the bis-W-MPT (for metal-binding pterin) cofactor. The cofactor was identified to be of a bis-Mo-MPT type with no guanine nucleotides present, which represents a form of Moco that has not been found previously in any molybdoenzyme. Our studies showed that YdhV has a preference for bis-Mo-MPT over bis-W-MPT to be inserted into the enzyme. In-depth characterization of YdhV by X-ray absorption and electron paramagnetic resonance spectroscopies revealed that the bis-Mo-MPT cofactor in YdhV is redox active. The bis-Mo-MPT and bis-W-MPT cofactors include metal centers that bind the four sulfurs from the two dithiolene groups in addition to a cysteine and likely a sulfido ligand. The unexpected presence of a bis-Mo-MPT cofactor opens an additional route for cofactor biosynthesis in E. coli and expands the canon of the structurally highly versatile molybdenum and tungsten cofactors.
We selected the immunogenic cell wall beta-(1,3)-glucosyltransferase Bgl2p from Candida albicans as a target protein for the production of antibodies. We identified a unique peptide sequence in the protein and generated monoclonal anti- C. albicans Bgl2p antibodies, which bound in particular to whole C. albicans cells.
Primary carbohydrate metabolism in plants includes several sugar and sugar-derivative transport processes. Over recent years, evidences have shown that in starch-related transport processes, in addition to glucose 6-phosphate, maltose, glucose and triose-phosphates, glucose 1-phosphate also plays a role and thereby increases the possible fluxes of sugar metabolites in planta. In this study, we report the characterization of two highly similar transporters, At1g34020 and At4g09810, in Arabidopsis thaliana, which allow the import of glucose 1-phosphate through the plasma membrane. Both transporters were expressed in yeast and were biochemically analyzed to reveal an antiport of glucose 1-phosphate/phosphate. Furthermore, we showed that the apoplast of Arabidopsis leaves contained glucose 1-phosphate and that the corresponding mutant of these transporters had higher glucose 1-phosphate amounts in the apoplast and alterations in starch and starch-related metabolism.
We utilized femtosecond time-resolved resonant inelastic X-ray scattering and ab initio theory to study the transient electronic structure and the photoinduced molecular dynamics of a model metal carbonyl photocatalyst Fe(CO)(5) in ethanol solution. We propose mechanistic explanation for the parallel ultrafast intra-molecular spin crossover and ligation of the Fe(CO)(4) which are observed following a charge transfer photoexcitation of Fe(CO)(5) as reported in our previous study [ Wernet et al., Nature 520, 78 (2015)]. We find that branching of the reaction pathway likely happens in the (1)A(1) state of Fe(CO)(4). A sub-picosecond time constant of the spin crossover from B-1(2) to B-3(2) is rationalized by the proposed B-1(2) -> (1)A(1) -> B-3(2) mechanism. Ultrafast ligation of the B-1(2) Fe(CO)(4) state is significantly faster than the spin-forbidden and diffusion limited ligation process occurring from the B-3(2) Fe(CO)(4) ground state that has been observed in the previous studies. We propose that the ultrafast ligation occurs via B-1(2) -> (1)A(1) -> (1)A'Fe(CO)(4)EtOH pathway and the time scale of the (1)A(1) Fe(CO)(4) state ligation is governed by the solute-solvent collision frequency. Our study emphasizes the importance of understanding the interaction of molecular excited states with the surrounding environment to explain the relaxation pathways of photoexcited metal carbonyls in solution. (C) 2016 Author(s).
Sperm proteins of the marine sessile mussels of the Mytilus edulis species complex are models to investigate reproductive isolation and speciation. This study aimed at identifying sperm proteins and their corresponding genes. This was aided by the use of monoclonal antibodies that preferentially bind to yet unknown sperm molecules. By identifying their target molecules, this approach identified proteins with relevance to Mytilus sperm function. This procedure identified 16 proteins, for example, enkurin, laminin, porin and heat shock proteins. The potential use of these proteins as genetic markers to study reproductive isolation is exemplified by analysing the enkurin locus. Enkurin evolution is driven by purifying selection, the locus displays high levels of intraspecific variation and species-specific alleles group in distinct phylogenetic clusters. These findings characterize enkurin as informative candidate biomarker for analyses of clinal variation and differential introgression in hybrid zones, for example, to understand determinants of reproductive isolation in Baltic Mytilus populations.
Type 2 diabetes (T2D) is a complex metabolic disease regulated by an interaction of genetic predisposition and environmental factors. To understand the genetic contribution in the development of diabetes, mice varying in their disease susceptibility were crossed with the obese and diabetes-prone New Zealand obese (NZO) mouse. Subsequent whole-genome sequence scans revealed one major quantitative trait loci (QTL),Nidd/DBAon chromosome 4, linked to elevated blood glucose and reduced plasma insulin and low levels of pancreatic insulin. Phenotypical characterization of congenic mice carrying 13.6 Mbp of the critical fragment of DBA mice displayed severe hyperglycemia and impaired glucose clearance at week 10, decreased glucose response in week 13, and loss of beta-cells and pancreatic insulin in week 16. To identify the responsible gene variant(s), further congenic mice were generated and phenotyped, which resulted in a fragment of 3.3 Mbp that was sufficient to induce hyperglycemia. By combining transcriptome analysis and haplotype mapping, the number of putative responsible variant(s) was narrowed from initial 284 to 18 genes, including gene models and non-coding RNAs. Consideration of haplotype blocks reduced the number of candidate genes to four (Kti12,Osbpl9,Ttc39a, andCalr4) as potential T2D candidates as they display a differential expression in pancreatic islets and/or sequence variation. In conclusion, the integration of comparative analysis of multiple inbred populations such as haplotype mapping, transcriptomics, and sequence data substantially improved the mapping resolution of the diabetes QTLNidd/DBA. Future studies are necessary to understand the exact role of the different candidates in beta-cell function and their contribution in maintaining glycemic control.
Neisseria gonorrhoeae is one of the most prevalent sexually transmitted diseases worldwide with more than 100 million new infections per year. A lack of intense research over the last decades and increasing resistances to the recommended antibiotics call for a better understanding of gonococcal infection, fast diagnostics and therapeutic measures against N. gonorrhoeae. Therefore, the aim of this work was to identify novel immunogenic proteins as a first step to advance those unresolved problems. For the identification of immunogenic proteins, pHORF oligopeptide phage display libraries of the entire N. gonorrhoeae genome were constructed. Several immunogenic oligopeptides were identified using polyclonal rabbit antibodies against N. gonorrhoeae. Corresponding full-length proteins of the identified oligopeptides were expressed and their immunogenic character was verified by ELISA. The immunogenic character of six proteins was identified for the first time. Additional 13 proteins were verified as immunogenic proteins in N. gonorrhoeae.
Current attempts to prevent and manage type 2 diabetes have been moderately effective, and a better understanding of the molecular roots of this complex disease is important to develop more successful and precise treatment options.
Recently, we initiated the collective diabetes cross, where four mouse inbred strains differing in their diabetes susceptibility were crossed with the obese and diabetes-prone NZO strain and identified the quantitative trait loci (QTL) Nidd13/NZO, a genomic region on chromosome 13 that correlates with hyperglycemia in NZO allele carriers compared to B6 controls.
Subsequent analysis of the critical region, harboring 644 genes, included expression studies in pancreatic islets of congenic Nidd13/NZO mice, integration of single-cell data from parental NZO and B6 islets as well as haplotype analysis.
Finally, of the five genes (Acot12, S100z, Ankrd55, Rnf180, and Iqgap2) within the polymorphic haplotype block that are differently expressed in islets of B6 compared to NZO mice, we identified the calcium-binding protein S100z gene to affect islet cell proliferation as well as apoptosis when overexpressed in MINE cells. In summary, we define S100z as the most striking gene to be causal for the diabetes QTL Nidd13/NZO by affecting beta-cell proliferation and apoptosis. Thus, S100z is an entirely novel diabetes gene regulating islet cell function.
Industrialized food production is in urgent search for alternative packaging materials, which can serve the requirements of a globalized world in terms of longer product shelf lives, reduced freight weight to decrease transport costs, and better barrier functionality to preserve its freshness. Polymer materials containing organically modified nano clay particles as additives are one example for a new generation of packaging materials with specific barrier functionality to actually hit the market. Clay types used for these applications are aluminosilicates, which belong to the mineral group of phyllosilicates. These consist of nano-scaled thin platelets, which are organically modified with quaternary ammonium compounds acting as spacers between the different clay layers, thereby increasing the hydrophobicity of the mineral additive. A variety of different organically modified clays are already available, and the use as additive for food packaging materials is one important application. To ensure valid risk assessments of emerging nano composite polymers used in the food packaging industry, exact analytical characterization of the organically modified clay within the polymer matrix is of paramount importance. Time-of-flight SIMS in combination with multivariate statistical analysis was used to differentiate modified clay reference materials from another. Time-of-flight SIMS spectra of a reference polymer plate, which contained one specific nano clay composite, were acquired. For each modified clay additive, a set of characteristic diagnostic ions could be identified, which then was used to successfully assign unknown clay additives to the corresponding reference material. Thus, the described methodology could be used to define and characterize nano clay within polymer matrices. Copyright (c) 2014 John Wiley & Sons, Ltd.
The spatial magnetic properties, through-space NMR shieldings (TSNMRS), of amino-substituted heteraromatic six-membered ring systems such as pyrylium/thiopyrylium analogues have been calculated using the GIAO perturbation method employing the nucleus independent chemical shift (NICS) concept and visualized as iso-chemical-shielding surfaces (ICSS) of various size and direction. The TSNMRS values were employed to quantify and visualize the existing aromaticity of the studied compounds. Due to strong conjugation of six-membered ring pi-electrons and lone pairs of the exo-cyclic amino substituents (restricted rotation about partial C,N double bonds) the interplay of still aromatic and already dominating trimethine cyanine/merocyanine-like substructures can be estimated. (C) 2017 Elsevier Ltd. All rights reserved.
Two mapping populations of a cross between the Arabidopsis thaliana accessions Col-0 and C24 were cultivated and analyzed with respect to the levels of 181 metabolites to elucidate the biological phenomenon of heterosis at the metabolic level. The relative mid-parent heterosis in the F-1 hybrids was <20% for most metabolic traits. The first mapping population consisting of 369 recombinant inbred lines (RILs) and their test cross progeny with both parents allowed us to determine the position and effect of 147 quantitative trait loci (QTL) for metabolite absolute mid-parent heterosis (aMPH). Furthermore, we identified 153 and 83 QTL for augmented additive (Z(1)) and dominance effects (Z(2)), respectively. We identified putative candidate genes for these QTL using the ARACYC database (http://www.arabidopsis.org/ biocyc), and calculated the average degree of dominance, which was within the dominance and over-dominance range for most metabolites. Analyzing a second population of 41 introgression lines (ILs) and their test crosses with the recurrent parent, we identified 634 significant differences in metabolite levels. Nine per cent of these effects were classified as over-dominant, according to the mode of inheritance. A comparison of both approaches suggested epistasis as a major contributor to metabolite heterosis in Arabidopsis. A linear combination of metabolite levels was shown to significantly correlate with biomass heterosis (r = 0.62).
Groundwater transit time is an essential hydrologic metric for groundwater resources management. However, especially in tropical environments, studies on the transit time distribution (TTD) of groundwater infiltration and its corresponding mean transit time (mTT) have been extremely limited due to data sparsity. In this study, we primarily use stable isotopes to examine the TTDs and their mTTs of both vertical and horizontal infiltration at a riverbank infiltration area in the Vietnamese Mekong Delta (VMD), representative of the tropical climate in Asian monsoon regions. Precipitation, river water, groundwater, and local ponding surface water were sampled for 3 to 9 years and analysed for stable isotopes (delta O-18 and delta H-2), providing a unique data set of stable isotope records for a tropical region. We quantified the contribution that the two sources contributed to the local shallow groundwater by a novel concept of two-component lumped parameter models (LPMs) that are solved using delta O-18 records. The study illustrates that two-component LPMs, in conjunction with hydrological and isotopic measurements, are able to identify subsurface flow conditions and water mixing at riverbank infiltration systems. However, the predictive skill and the reliability of the models decrease for locations farther from the river, where recharge by precipitation dominates, and a low-permeable aquitard layer above the highly permeable aquifer is present. This specific setting impairs the identifiability of model parameters. For river infiltration, short mTTs (<40 weeks) were determined for sites closer to the river (<200 m), whereas for the precipitation infiltration, the mTTs were longer (>80 weeks) and independent of the distance to the river. The results not only enhance the understanding of the groundwater recharge dynamics in the VMD but also suggest that the highly complex mechanisms of surface-groundwater interaction can be conceptualized by exploiting two-component LPMs in general. The model concept could thus be a powerful tool for better understanding both the hydrological functioning of mixing processes and the movement of different water components in riverbank infiltration systems.
Carbon (C) inputs and nutrient availability are known to affect soil organic carbon (SOC) stocks. However, general rules regarding the operation of these factors across a range of soil nutrient availabilities and substrate qualities are unidentified. "Priming" (stimulated decomposition by labile C inputs) and 'preferential substrate utilization' (retarded decomposition due to shifts in community composition towards microbes that do not mineralize SOC) are two hypotheses to explain effects of labile C additions on SOC dynamics. For effects of nutrient additions (nitrogen and phosphorus) on SOC dynamics, the stoichiometric (faster decomposition of materials of low carbon-to-nutrient ratios) and 'microbial mining' (that is, reduced breakdown of recalcitrant C forms for nutrients under fertile conditions) hypotheses have been proposed. Using the natural gradient of soil nutrient availability and substrate quality of a chronosequence, combined with labile C and nutrient amendments, we explored the support for these contrasting hypotheses. Additions of labile C, nitrogen (N), phosphorus (P), and combinations of C and N and C and P were applied to three sites: 2-year fallow grassland, mature grassland and forest, and the effects of site and nutrient additions on litter decomposition and soil C dynamics were assessed. The response to C addition supported the preferential substrate hypothesis for easily degradable litter C and the priming hypothesis for SOC, but only in nitrogen-enriched soils of the forest site. Responses to N addition supported the microbial mining hypothesis irrespective of C substrate (litter or SOC), but only in the forest site. Further, P addition effects on SOC support the stoichiometric hypothesis; P availability appeared key to soil C release (priming) in the forest site if labile C and N is available. These results clearly link previously contrasting hypotheses of the factors controlling SOC with the natural gradient in litter quality and nutrient availability that exists in ecosystems at different successional stages. A holistic theory that incorporates this variability of responses, due to different mechanisms, depending on nutrient availability and substrate quality is essential for devising management strategies to safeguard soil C stocks.
Objective: Aging is accompanied by loss of brown adipocytes and a decline in their thermogenic potential, which may exacerbate the development of adiposity and other metabolic disorders. Presently, only limited evidence exists describing the molecular alterations leading to impaired brown adipogenesis with aging and the contribution of these processes to changes of systemic energy metabolism.
Methods: Samples of young and aged murine brown and white adipose tissue were used to compare age-related changes of brown adipogenic gene expression and thermogenesis-related lipid mobilization. To identify potential markers of brown adipose tissue aging, non-targeted proteomic and metabolomic as well as targeted lipid analyses were conducted on young and aged tissue samples. Subsequently, the effects of several candidate lipid classes on brown adipocyte function were examined.
Results: Corroborating previous reports of reduced expression of uncoupling protein-1, we observe impaired signaling required for lipid mobilization in aged brown fat after adrenergic stimulation. Omics analyses additionally confirm the age-related impairment of lipid homeostasis and reveal the accumulation of specific lipid classes, including certain sphingolipids, ceramides, and dolichols in aged brown fat. While ceramides as well as enzymes of dolichol metabolism inhibit brown adipogenesis, inhibition of sphingosine 1-phosphate receptor 2 induces brown adipocyte differentiation.
Conclusions: Our functional analyses show that changes in specific lipid species, as observed during aging, may contribute to reduced thermogenic potential. They thus uncover potential biomarkers of aging as well as molecular mechanisms that could contribute to the degradation of brown adipocytes, thereby providing potential treatment strategies of age-related metabolic conditions.
Trade-offs are inherent to biochemical networks governing diverse cellular functions, from gene expression to metabolism. Yet, trade-offs between fluxes of biochemical reactions in a metabolic network have not been formally studied. Here, we introduce the concept of absolute flux trade-offs and devise a constraint-based approach, termed FluTO, to identify and enumerate flux trade-offs in a given genome-scale metabolic network. By employing the metabolic networks of Escherichia coli and Saccharomyces cerevisiae, we demonstrate that the flux trade-offs are specific to carbon sources provided but that reactions involved in the cofactor and prosthetic group biosynthesis are present in trade-offs across all carbon sources supporting growth. We also show that absolute flux trade-offs depend on the biomass reaction used to model the growth of Arabidopsis thaliana under different carbon and nitrogen conditions. The identified flux trade-offs reflect the tight coupling between nitrogen, carbon, and sulphur metabolisms in leaves of C-3 plants. Altogether, FluTO provides the means to explore the space of alternative metabolic routes reflecting the constraints imposed by inherent flux trade-offs in large-scale metabolic networks.
We combine sensitivity to atomic number, chemical shifts, probing depth, and magnetic order in a field- dependent magnetic circular X-ray dichroism study at the Mn L-edge of the diluted ferromagnetic semiconductor Ga1-xMnxAs and observe different Mn constituents: ferromagnetic Mn with an n(d) > 5 lineshape and paramagnetic Mn with distinct n(d) = 5 lineshape. The paramagnetic Mn is assigned to interstitials with surface segregation tendency. (c) 2005 Elsevier B.V. All rights reserved
Identification of Endodormancy Release for Cherries (Prunus Avium L.) by
Abscisic Acid and Sugars
(2017)
In order to develop reliable and physiologically sound models for the plant development in spring, the date of endodormancy release is always a crucial and mostly unknown model parameter. Until present, classical approaches - such as climate chamber experiments - are used to derive this unknown parameter. In these experiments, progressive plant development or significant changes in bud’s fresh weight or water content are measurable markers for dormancy release. This study presents an alternative approach, which is based on four well-known metabolites. For 5 seasons (2011/12-2015/16), the content of abscisic acid (ABA) and sugars such as fructose, sucrose and glucose in sweet cherry flower buds (cultivar ‘Summit’) were weekly analysed between beginning of October and April. These data allow comparing the annual course of these metabolites with the date of endodormancy release, derived from a classical climate chamber experiment, published in a previous study. Results showed that ABA and sucrose are two important metabolites which can help to identify the date of endodormancy release of sweet cherries. On average, ABA content reached a plateau of 5.65 μg g-1 DW-1 during endodormancy, which was maintained for 3-6 weeks. The significant reduction of the ABA content after this period to 4.41 μg g-1 DW-1 on average during ecodormancy was nearly in agreement with the date of endodormancy release of ‘Summit’ on 28 November (332 DOY).
The annual cycle of sucrose, which has a cryoprotective effect during winter, is well comprehensible and showed a close relationship to
the annual course of minimum air temperature after leaf fall(r=-0.90). The nearly constant level of sucrose during ecodormancy (21.0 mg g-1 DW-1, 5 yr. mean) did not only allow deriving the date of endodormancy release but can also be helpful to define the beginning of ontogenetic development.
The analysis of palaeoclimate time series is usually affected by severe methodological problems, resulting primarily from non-equidistant sampling and uncertain age models. As an alternative to existing methods of time series analysis, in this paper we argue that the statistical properties of recurrence networks - a recently developed approach - are promising candidates for characterising the system's nonlinear dynamics and quantifying structural changes in its reconstructed phase space as time evolves. In a first order approximation, the results of recurrence network analysis are invariant to changes in the age model and are not directly affected by non-equidistant sampling of the data. Specifically, we investigate the behaviour of recurrence network measures for both paradigmatic model systems with non-stationary parameters and four marine records of long-term palaeoclimate variations. We show that the obtained results are qualitatively robust under changes of the relevant parameters of our method, including detrending, size of the running window used for analysis, and embedding delay. We demonstrate that recurrence network analysis is able to detect relevant regime shifts in synthetic data as well as in problematic geoscientific time series. This suggests its application as a general exploratory tool of time series analysis complementing existing methods.
With the jABC it is possible to realize workflows for numerous questions in different fields. The goal of this project was to create a workflow for the identification of differentially expressed genes. This is of special interest in biology, for it gives the opportunity to get a better insight in cellular changes due to exogenous stress, diseases and so on. With the knowledge that can be derived from the differentially expressed genes in diseased tissues, it becomes possible to find new targets for treatment.
Resolving Earth's surface at the meter scale is essential for an improved understanding of the dynamics of mass-movement processes. In this study, we explore the applicability and potential of digital elevation models (DEMs) derived from stereophotogrammetry to detect debris-flow channels in the Quebrada del Toro in the northwestern Argentine Andes. Our analysis relies on a high-resolution (3 m) DEM created from SPOT-7 tri-stereo satellite data. We carefully validated DEM quality with ∼6,000 differential GPS points and identified optimal parameters for DEM generation in high-relief terrain. After multiple processing steps, we achieved an accuracy of 0.051 ± 1.915 m (1σ) using n = 3,139 control points with cm precision. Previous studies have used the drainage area and slope framework to identify topographic signatures of debris flows within a catchment. We built upon this and investigated individual river-channel segments using connected-component (CC) analysis on meter-scale topographic data. We define CC as segments of similar slope along the channel profile. Based on seven manually identified debris-flow catchments, we developed a debris-flow similarity index using component length and mean channel-segment slope and identified channel segments that have likely been shaped by debris flows. The presented approach has the potential to resolve intra-catchment variability of transport processes, allows to constrain the extent of debris-flow channels more precisely than slope-area analysis, and highlights the versatility of combined space- and field-based observations for natural-hazard assessments.
In order to elucidate factors that determine substrate specificity and activity of mammalian molybdo-flavoproteins we performed site directed mutagenesis of mouse aldehyde oxidase 3 (mAOX3). The sequence alignment of different aldehyde oxidase (AOX) isoforms identified variations in the active site of mAOX3 in comparison to other AOX proteins and xanthine oxidoreductases (XOR). Based on the structural alignment of mAOX3 and bovine XOR, differences in amino acid residues involved in substrate binding in XORs in comparison to AOXs were identified. We exchanged several residues in the active site to the ones found in other AOX homologues in mouse or to residues present in bovine XOR in order to examine their influence on substrate selectivity and catalytic activity. Additionally we analyzed the influence of the [2Fe-2S] domains of mAOX3 on its kinetic properties and cofactor saturation. We applied UV-VIS and EPR monitored redox-titrations to determine the redox potentials of wild type mAOX3 and mAOX3 variants containing the iron-sulfur centers of mAOX1. In addition, a combination of molecular docking and molecular dynamic simulations (MD) was used to investigate factors that modulate the substrate specificity and activity of wild type and AOX variants. The successful conversion of an AOX enzyme to an XOR enzyme was achieved exchanging eight residues in the active site of mAOX3. It was observed that the absence of the K889H exchange substantially decreased the activity of the enzyme towards all substrates analyzed, revealing that this residue has an important role in catalysis.
The comprehensive systems-biology database (CSB.DB) was used to reveal brassinosteroid (BR)-related genes from expression profiles based on co-response analyses. Genes exhibiting simultaneous changes in transcript levels are candidates of common transcriptional regulation. Combining numerous different experiments in data matrices allows ruling out outliers and conditional changes of transcript levels. CSB.DB was queried for transcriptional co-responses with the BR-signalling components BRI1 and BAK1: 301 out of 9694 genes represented in the nasc0271 database showed co-responses with both genes. As expected, these genes comprised pathway-involved genes (e.g. 72 BR-induced genes), because the BRI1 and BAK1 proteins are required for BR-responses. But transcript co-response takes the analysis a step further compared with direct approaches because BR-related non BR-responsive genes were identified. Insights into networks and the functional context of genes are provided, because factors determining expression patterns are reflected in correlations. Our findings demonstrate that transcript co-response analysis presents a valuable resource to uncover common regulatory patterns of genes. Different data matrices in CSB.DB allow examination of specific biological questions. All matrices are publicly available through CSB.DB. This work presents one possible roadmap to use the CSB.DB resources
Identification of benzenoid and quinonoid structures by through-space NMR shieldings (TSNMRS)
(2010)
The continuous expansion of nosocomial infections around the globe has become a precarious situation. Key challenges include mounting dissemination of multiple resistances to antibiotics, the easy transmission and the growing mortality rates of hospital-acquired bacterial diseases. Thus, new ways to rapidly detect these infections are vital. Consequently, researchers around the globe pursue innovative approaches for point-of-care devices. In many cases the specific interaction of an antigen and a corresponding antibody is pivotal. However, the knowledge about suitable antigens is lacking. The aim of this study was to identify novel antigens as specific diagnostic markers. Additionally, these proteins might be aptly used for the generation of vaccines to improve current treatment options. Hence, a cDNA-based expression library was constructed and screened via microarrays to detect novel antigens of Klebsiella pneumoniae, a prominent agent of nosocomial infections well-known for its extensive antibiotics resistance, especially by extended-spectrum beta-lactamases (ESBL). After screening 1536 clones, 14 previously unknown immunogenic proteins were identified. Subsequently, each protein was expressed in full-length and its immunodominant character examined by ELISA and microarray analyses. Consequently, six proteins were selected for epitope mapping and three thereof possessed linear epitopes. After specificity analysis, homology survey and 3d structural modelling, one epitope sequence GAVVALSTTFA of KPN_00363, an ion channel protein, was identified harboring specificity for K. pneumoniae. The remaining epitopes showed ambiguous results regarding the specificity for K. pneumoniae. The approach adopted herein has been successfully utilized to discover novel antigens of Campylobacter jejuni and Salmonella enterica antigens before. Now, we have transferred this knowledge to the key nosocomial agent, K. pneumoniae. By identifying several novel antigens and their linear epitope sites, we have paved the way for crucial future research and applications including the design of point-of-care devices, vaccine development and serological screenings for a highly relevant nosocomial pathogen.
Identification of a super-functional Tfh-like subpopulation in murine lupus by pattern perception
(2020)
Dysregulated cytokine expression by T cells plays a pivotal role in the pathogenesis of autoimmune diseases. However, the identification of the corresponding pathogenic subpopulations is a challenge, since a distinction between physiological variation and a new quality in the expression of protein markers requires combinatorial evaluation. Here, we were able to identify a super-functional follicular helper T cell (Tfh)-like subpopulation in lupus-prone NZBxW mice with our binning approach "pattern recognition of immune cells (PRI)". PRI uncovered a subpopulation of IL-21(+) IFN-gamma(high) PD-1(low) CD40L(high) CXCR5(-) Bcl-6(-) T cells specifically expanded in diseased mice. In addition, these cells express high levels of TNF-alpha and IL-2, and provide B cell help for IgG production in an IL-21 and CD40L dependent manner. This super-functional T cell subset might be a superior driver of autoimmune processes due to a polyfunctional and high cytokine expression combined with Tfh-like properties.
hEP4-R (human prostaglandin E2 receptor, subtype EP4) is a G(s)-linked heterotrimeric GPCR (G-protein-coupled receptor). It undergoes agonist-induced desensitization and internalization that depend on the presence of its C- terminal domain. Desensitization and internalization of GPCRs are often linked to agonist-induced beta-arrestin complex formation, which is stabilized by phosphorylation. Subsequently beta-arrestin uncouples the receptor from its G-protein and links it to the endocytotic machinery. The C-terminal domain of hEP4-R contains 38 Ser/Thr residues that represent potential phosphorylation sites. The present study aimed to analyse the relevance of these Ser/Thr residues for agonist- induced phosphorylation, interaction with beta-arrestin and internalization. In response to agonist treatment, hEP4-R was phosphorylated. By analysis of proteolytic phosphopeptides of the wild-type receptor and mutants in which groups of Ser/Thr residues had been replaced by Ala, the principal phosphorylation site was mapped to a Ser/Thr-containing region comprising residues 370-382, the presence of which was necessary and sufficient to obtain full agonist-induced phosphorylation. A cluster of Ser/Thr residues (Ser-389-Ser-390-Thr-391-Ser-392) distal to this site, but not the principal phosphorylation site, was essential to allow agonist-induced recruitment of beta-arrestin1. However, phosphorylation greatly enhanced the stability of the beta-arrestin1-receptor complexes. For maximal agonist-induced internalization, phosphorylation of the principal phosphorylation site was not required, but both beta-arrestin1 recruitment and the presence of Ser/Thr residues in the distal half of the C-terminal domain were necessary.
Identification of a novel heteroglycan-interacting protein, HIP 1.3, from Arabidopsis thaliana
(2011)
Plastidial degradation of transitory starch yields mainly maltose and glucose. Following the export into the cytosol, maltose acts as donor for a glucosyl transfer to cytosolic heteroglycans as mediated by a cytosolic transglucosidase (DPE2; EC 2.4.1.25) and the second glucosyl residue is liberated as glucose. The cytosolic phosphorylase (Pho2/PHS2; EC 2.4.1.1) also interacts with heteroglycans using the same intramolecular sites as DPE2. Thus, the two glucosyl transferases interconnect the cytosolic pools of glucose and glucose 1-phosphate. Due to the complex monosaccharide pattern, other heteroglycan-interacting proteins (Hips) are expected to exist.
Identification of those proteins was approached by using two types of affinity chromatography. Heteroglycans from leaves of Arabidopsis thaliana (Col-0) covalently bound to Sepharose served as ligands that were reacted with a complex mixture of buffer-soluble proteins from Arabidopsis leaves. Binding proteins were eluted by sodium chloride. For identification, SDS-PAGE, tryptic digestion and MALDI-TOF analyses were applied. A strongly interacting polypeptide (approximately 40 kDa; designated as HIP1.3) was observed as product of locus At1g09340. Arabidopsis mutants deficient in HIP1.3 were reduced in growth and contained heteroglycans displaying an altered monosaccharide pattern. Wild type plants express HIP1.3 most strongly in leaves. As revealed by immuno fluorescence, HIP1.3 is located in the cytosol of mesophyll cells but mostly associated with the cytosolic surface of the chloroplast envelope membranes. In an HIP1.3-deficient mutant the immunosignal was undetectable. Metabolic profiles from leaves of this mutant and wild type plants as well were determined by GC-MS. As compared to the wild type control, more than ten metabolites, such as ascorbic acid, fructose, fructose bisphosphate, glucose, glycine, were elevated in darkness but decreased in the light. Although the biochemical function of HIP1.3 has not yet been elucidated, it is likely to possess an important function in the central carbon metabolism of higher plants.
The phosphorylation of amylopectin by the glucan, water dikinase (GWD; EC 2.7.9.4) is an essential step within starch metabolism. This is indicated by the starch excess phenotype of GWD-deficient plants, such as the sex1-3 mutant of Arabidopsis (Arabidopsis thaliana). To identify starch-related enzymes that rely on glucan-bound phosphate, we studied the binding of proteins extracted from Arabidopsis wild-type leaves to either phosphorylated or nonphosphorylated starch granules. Granules prepared from the sex1-3 mutant were prephosphorylated in vitro using recombinant potato (Solanum tuberosum) GWD. As a control, the unmodified, phosphate free granules were used. An as-yet uncharacterized protein was identified that preferentially binds to the phosphorylated starch. The C-terminal part of this protein exhibits similarity to that of GWD. The novel protein phosphorylates starch granules, but only following prephosphorylation with GWD. The enzyme transfers the beta-P of ATP to the phosphoglucan, whereas the gamma-P is released as orthophosphate. Therefore, the novel protein is designated as phosphoglucan, water dikinase (PWD). Unlike GWD that phosphorylates preferentially the C6 position of the glucose units, PWD phosphorylates predominantly (or exclusively) the C3 position. Western-blot analysis of protoplast and chloroplast fractions from Arabidopsis leaves reveals a plastidic location of PWD. Binding of PWD to starch granules strongly increases during net starch breakdown. Transgenic Arabidopsis plants in which the expression of PWD was reduced by either RNAi or a T-DNA insertion exhibit a starch excess phenotype. Thus, in Arabidopsis leaves starch turnover requires a close collaboration of PWD and GWD
The molybdenum cofactor is an important cofactor, and its biosynthesis is essential for many organisms, including humans. Its basic form comprises a single molybdopterin (MPT) unit, which binds a molybdenum ion bearing three oxygen ligands via a dithiolene function, thus forming Mo-MPT. In bacteria, this form is modified to form the bis-MPT guanine dinucleotide cofactor with two MPT units coordinated at one molybdenum atom, which additionally contains GMPs bound to the terminal phosphate group of the MPTs (bis-MGD). The MobA protein catalyzes the nucleotide addition to MPT, but the mechanism of the biosynthesis of the bis-MGD cofactor has remained enigmatic. We have established an in vitro system for studying bis-MGD assembly using purified compounds. Quantification of the MPT/molybdenum and molybdenum/phosphorus ratios, time-dependent assays for MPT and MGD detection, and determination of the numbers and lengths of Mo-S and Mo-O bonds by X-ray absorption spectroscopy enabled identification of a novel bis-Mo-MPT intermediate on MobA prior to nucleotide attachment. The addition of Mg-GTP to MobA loaded with bis-Mo-MPT resulted in formation and release of the final bis-MGD product. This cofactor was fully functional and reconstituted the catalytic activity of apo-TMAO reductase (TorA). We propose a reaction sequence for bis-MGD formation, which involves 1) the formation of bis-Mo-MPT, 2) the addition of two GMP units to form bis-MGD on MobA, and 3) the release and transfer of the mature cofactor to the target protein TorA, in a reaction that is supported by the specific chaperone TorD, resulting in an active molybdoenzyme.
14-Hydroxy-retro-retinol was previously described as an in vivo and in vitro metabolite of retinol. Furthermore, the retinoid 4-hydroxy-retinol was identified as an endogenous occurring retinoid in the amphibian organism and an in vitro metabolite of retinol. We describe in the present study that 14-hydroxy-retro-retinol and 4-hydroxy- retinol are present in normal neonatal rat serum as endogenous occurring retinoids in normal non-vitamin A supplemented mammals (rats). Both retinoids were detected in serum and liver of neonatal rats at days 3 and 11 after birth. The respective concentrations at day 11 after birth were 41.8 +/- 2.8 ng/ml (serum)/ 104 +/- 6 ng/g (liver) for 4-hydroxy- retinol and 23 +/- 4.6 ng/ml (serum)/ 285 +/- 5 ng/g (liver) for 14-hydroxy-retro-retinol. Both retinoids could not be detected in adult rat serum and liver. From our experiments important physiological functions of these retinoids during postnatal development could be postulated. (C) 2004 Elsevier Inc. All rights reserved
Prostanoid receptors belong to the class of heptahelical plasma membrane receptors. For the five prostanoids, eight receptor subtypes have been identified. They display an overall sequence similarity of roughly 30%. Based on sequence comparison, single amino acids in different subtypes of different species have previously been identified by site-directed mutagenesis or in hybrid receptors that appear to be essential for ligand binding or G-protein coupling. Based on this information, a series of mutants of the human FP receptor was generated and characterized in ligand- binding and second-messenger-formation studies. It was found that mutation of His-81 to Ala in transmembrane domain 2 and of Arg-291 to Leu in transmembrane domain 7, which are putative interaction partners for the prostanoid's carboxyl group, abolished ligand binding. Mutants in which Ser-263 in transmembrane domain 6 or Asp-300 in transmembrane domain 7 had been replaced by Ala or Gln, respectively, no longer discriminated between prostaglandins PGF(2alpha) and PGD(2). Thus distortion of the topology of transmembrane domains 6 and 7 appears to interfere with the cyclopentane ring selectivity of the receptor. PGF(2alpha)-induced inositol formation was strongly reduced in the mutant Asp-300Gln, inferring a role for this residue in agonist-induced G-protein activation.
Marketing of adulterated beeswax foundation has recently become a major economic problem for beekeepers. Paraffin contamination leads to collapse of combs, and stearic acid has a negative influence on the development of bee brood. The quality of beeswax for beekeeping has not been standardized in EU regulations. Recently, it was shown that attenuated total reflectance Fourier-transform infrared spectroscopy (FTIR-ATR) can be used to determine beeswax adulteration. Differences in the IR spectra of authentic beeswax can be identified and calculated through comparison with authentic beeswax. In this study, the method is further validated by employing a high number of samples of authentic beeswax from different origins. Low quantification and detection limits are achieved for paraffin, stearic acid, tallow, carnauba wax, and candelilla wax. Furthermore, the FTIR-ATR analytical conditions are verified by analyzing 358 samples of commercial and beekeeper-produced beeswax foundations. Multi-adulterated samples with as many as five different additives in beeswax mixtures are identified with the same accuracy as single substances. Additionally, the spectra of a further 14 different natural and synthetic waxes and hardened fats are analyzed and are compared with beeswax. Finally, a spectral library is established that can be used for further studies. Practical Applications: FTIR-ATR is a fast and cost-efficient tool in beeswax analysis for accurately monitoring a high sample volume. Analysis of 358 beeswax foundations showed an adulteration of 21.8% of the samples with paraffin, stearic acid, tallow, and combinations. Based on the results of this study, it is possible to detect beeswax adulteration of less than 3% of these adulterants and their combinations by FTIR-ATR spectroscopy. This method can be used for monitoring beeswax foundations to identify adulterated materials, exclude these materials from the recycling process, and produce high-quality beeswax, which is essential for bee health.
A large-scale metabolic quantitative trait loci (mQTL) analysis was performed on the well-characterized Solanum pennellii introgression lines to investigate the genomic regions associated with secondary metabolism in tomato fruit pericarp. In total, 679 mQTLs were detected across the 76 introgression lines. Heritability analyses revealed that mQTLs of secondary metabolism were less affected by environment than mQTLs of primary metabolism. Network analysis allowed us to assess the interconnectivity of primary and secondary metabolism as well as to compare and contrast their respective associations with morphological traits. Additionally, we applied a recently established real-time quantitative PCR platform to gain insight into transcriptional control mechanisms of a subset of the mQTLs, including those for hydroxycinnamates, acyl-sugar, naringenin chalcone, and a range of glycoalkaloids. Intriguingly, many of these compounds displayed a dominant-negative mode of inheritance, which is contrary to the conventional wisdom that secondary metabolite contents decreased on domestication. We additionally performed an exemplary evaluation of two candidate genes for glycolalkaloid mQTLs via the use of virus-induced gene silencing. The combined data of this study were compared with previous results on primary metabolism obtained from the same material and to other studies of natural variance of secondary metabolism.
In order to broaden our understanding of the eukaryotic CO2- concentrating mechanism the occurrence and localization of a thylakoid-asssocaate carbonic anhydrase (EC 4.2.1.1) were studied in the green algae Tetraedron on minimum and Chlamydomonas noctigama. Both algae induce a CO2-concentrating mechanism when grown under limiting CO2 conditions. Using mass- spectrometric measurements of O-18 exchange from doubly labelled CO2, the presence of a thylakoid-associated carbonic anhydrase was confirmed for both species. From purified thylakoid membranes, photosystem I (PSI), photosystem II (PSII) and the light-harvesting complex of the photosynthetic apparatus were isolated by mild detergent gel. The protein fractions were identified by 77 K fluorescence spectroscopy and immunological studies. A polypeptide was found to immunoreact with an antibody raised against thylakoid carbonic anhydrase (CAH3) from Chlamydomonas reinhardtii. It was found that this polypeptide was mainly associated with PSII, although a certain proportion was also connected to light harvesting complex II. This was confirmed by activity measurements of carbonic anhydrase in isolated bands extracted from the mild detergent gel. The thylakoid carbonic anhydrase isolated from T. minimum had an isoelectric point between 5.4 and 4.8. Together the results are consistent with the hypothesis that thylakoid carbonic anhydrase resides within the lumen where it is associated with the PSII complex.
Identification and LC-MS/MS-based analyses of technical enzymes in wheat flour and baked products
(2016)
The use of technical enzymes in bakery industry is necessary for a consistent and good quality of baked products. Since the cultivation of cereals leads to low amounts of endogenous enzymes being present, a need of their commercial alternatives is becoming a routine process in order to meet the consumer quality demands. Targeted quantification proteomics-based methods are necessary for their detection to meet the regulatory criteria. Here, we initially report on the identification of Lipase FE-01, a lipase from fungus Thermomyces lanuginosus, as analyzed by SDS-PAGE, in-Gel digestion, and MALDI-TOF-MS. In further experiments, the focus of the study was directed toward an extensive use and optimization of in-solution enzymatic digestion in combination with LC-MS/MS techniques in identification of specific peptide markers and finally in utilization of the latter in delivering reproducible quantification data for several different technical enzymes (alpha-amylases, xylanase, and lipases from microbial origin) in complex matrices such as baked bread and wheat flour. Two digestion protocols (a fast option using thermocycler program and the well-established overnight method) were tested, and both of these can be successfully applied. The application of isotopically labeled analogs of the MRM targeted peptides as internal standards and the addition of an internal protein standard during the extraction/digestion experiment were compared to determine the optimal quantification algorithm of the recovered enzyme concentrations. Thus, a standardized sensitive LC-MS/MS method could be developed to determine technical enzymes as forthcoming ingredients in the prefabricated food formulations in concentrations lower than 10 ppm.
Identification and Characterization of Three Epithiospecifier Protein Isoforms in Brassica oleracea
(2019)
Glucosinolates present in Brassicaceae play a major role in herbivory defense. Upon tissue disruption, glucosinolates come into contact with myrosinase, which initiates their breakdown to biologically active compounds. Among these, the formation of epithionitriles is triggered by the presence of epithiospecifier protein (ESP) and a terminal double bond in the glucosinolate side chain. One ESP gene is characterized in the model plant Arabidopsis thaliana (AtESP; At1g54040.2). However, Brassica species underwent genome triplication since their divergence from the Arabidopsis lineage. This indicates the presence of multiple ESP isoforms in Brassica crops that are currently poorly characterized. We identified three B. oleracea ESPs, specifically BoESP1 (LOC106296341), BoESP2 (LOC106306810), and BoESP3 (LOC106325105) based on in silico genome analysis. Transcript and protein abundance were assessed in shoots and roots of four B. oleracea vegetables, namely broccoli, kohlrabi, white, and red cabbage, because these genotypes showed a differential pattern for the formation of glucosinolate hydrolysis products as well for their ESP activity. BoESP1 and BoESP2 were expressed mainly in shoots, while BoESP3 was abundant in roots. Biochemical characterization of heterologous expressed BoESP isoforms revealed different substrate specificities towards seven glucosinolates: all isoforms showed epithiospecifier activity on alkenyl glucosinolates, but not on non-alkenyl glucosinolates. The pH-value differently affected BoESP activity: while BoESP1 and BoESP2 activities were optimal at pH 6-7, BoESP3 activity remained relatively stable from pH 4 to 7. In order test their potential for the in vivo modification of glucosinolate breakdown, the three isoforms were expressed in A. thaliana Hi-0, which lacks AtESP expression, and analyzed for the effect on their respective hydrolysis products. The BoESPs altered the hydrolysis of allyl glucosinolate in the A. thaliana transformants to release 1-cyano-2,3-epithiopropane and reduced formation of the corresponding 3-butenenitrile and allyl isothiocyanate. Plants expressing BoESP2 showed the highest percentage of released epithionitriles. Given these results, we propose a model for isoform-specific roles of B. oleracea ESPs in glucosinolate breakdown.
IntroductionTo date, most studies of natural variation and metabolite quantitative trait loci (mQTL) in tomato have focused on fruit metabolism, leaving aside the identification of genomic regions involved in the regulation of leaf metabolism.ObjectiveThis study was conducted to identify leaf mQTL in tomato and to assess the association of leaf metabolites and physiological traits with the metabolite levels from other tissues.MethodsThe analysis of components of leaf metabolism was performed by phenotypying 76 tomato ILs with chromosome segments of the wild species Solanum pennellii in the genetic background of a cultivated tomato (S. lycopersicum) variety M82. The plants were cultivated in two different environments in independent years and samples were harvested from mature leaves of non-flowering plants at the middle of the light period. The non-targeted metabolite profiling was obtained by gas chromatography time-of-flight mass spectrometry (GC-TOF-MS). With the data set obtained in this study and already published metabolomics data from seed and fruit, we performed QTL mapping, heritability and correlation analyses.ResultsChanges in metabolite contents were evident in the ILs that are potentially important with respect to stress responses and plant physiology. By analyzing the obtained data, we identified 42 positive and 76 negative mQTL involved in carbon and nitrogen metabolism.ConclusionsOverall, these findings allowed the identification of S. lycopersicum genome regions involved in the regulation of leaf primary carbon and nitrogen metabolism, as well as the association of leaf metabolites with metabolites from seeds and fruits.
Identification and characterization of growing large-scale en-echelon fractures in a salt mine
(2014)
The spatiotemporal seismicity of acoustic emission (AE) events recorded in the Morsleben salt mine is investigated. Almost a year after backfilling of the cavities from 2003, microevents are distributed with distinctive stripe shapes above cavities at different depth levels. The physical forces driving the creation of these stripes are still unknown. This study aims to find the active stripes and track fracture developments over time by combining two different temporal and spatial clustering techniques into a single methodological approach. Anomalous seismicity parameters values like sharp b-value changes for two active stripes are good indicators to explain possible stress accumulation at the stripe tips. We identify the formation of two new seismicity stripes and show that the AE activities in active clusters are migrated mostly unidirectional to eastward and upward. This indicates that the growth of underlying macrofractures is controlled by the gradient of extensional stress. Studying size distribution characteristic in terms of frequency-magnitude distribution and b-value in active phase and phase with constant seismicity rate show that deviations from the Gutenberg-Richter power law can be explained by the inclusion of different activity phases: (1) the inactive period before the formation of macrofractures, which is characterized by a deficit of larger events (higher b-values) and (2) the period of fracture growth characterized by the occurrence of larger events (smaller b-values).
Hemidiaptomus diaptomid copepods are known to be excellent biological indicators for the highly biodiverse crustacean communities inhabiting Mediterranean temporary ponds (MTPs), an endangered inland water habitat whose conservation is considered a priority according to the "Habitat Directive" of the European Union. This study reports on the characterization of five polymorphic microsatellite loci in Hemidiaptomus gurneyi, to be used as markers for fine-scale studies on the population genetic structure and metapopulation dynamics of a typical and obligate MTP dweller. The five selected loci proved to be polymorphic in the species, with three to five polymorphic loci per studied population. Overall, mean heterozygosity scored for all loci and populations was lower than that reported for the few other diaptomid species for which microsatellite loci have been to date described; this is possibly due to the intrinsically fragmented and isolated peculiar habitat inhabited by the species. Furthermore, the presence of indels within the flanking regions of selected loci was scored. This study, albeit confirming the technical difficulties in finding proper microsatellite markers in copepods, provides for the first time a set of useful polymorphic microsatellite loci for a Hemidiaptomus species, thus allowing the realization of fine-scale phylogeographic and population genetics studies of this flagship crustacean taxon for MTPs.
The centrosome is the main microtubule-organizing center and constitutes the largest protein complex in a eukaryotic cell. The Dictyostelium centrosome is an established model for acentriolar centrosomes and it consists of a layered core structure Surrounded by a so-called corona, which harbors microtubule nucleation complexes. We have identified 34 new centrosomal candidate proteins through mass spectrometrical analysis of the proteome of isolated Dictyostelium centrosomes. Here we present a characterization of 12 centrosomal candidate proteins all featuring coiled coil regions and low expression levels, which are the most common attributes of centrosomal proteins. We used GFP fusion proteins to localize the candidate proteins in whole cells and on microtubule-free, isolated centrosomes. Thus we were able to identify nine new genuine centrosomal proteins including a putative orthologue of Cep192, an interaction partner of polo-like kinase 4 in human centriole biogenesis. In this respect, centrosomal localization of the only polo-like kinase in Dictyostelium, Pik, is also shown in this work. Using confocal deconvolution microscopy, four components, CP39, CP55, CP75, and CP91 could be clearly assigned to the so far almost uncharacterized centrosomal core structure, while CP148 and Cep192 localized to a zone between that of corona marker and core proteins. Finally, CP103 and CP248 were constituents of the corona. In contrast, NE81 was localized at the nuclear envelope and three others, an orthologue of the spindle checkpoint component Mad1, the novel Cenp68, and the centrosomal CP248 were observed at the centromeres, which are clustered and linked to the centrosome throughout the entire cell cycle. Cell Motil. Cytoskeleton 66: 915-928, 2009.
Firms increasingly use ideas from online innovation communities to solve problems or to better address customer needs. However, in many cases the number of submitted ideas has exploded, it leads to an information overload that firms hardly can handle considering their limited cognitive resources. Therefore, we use the Elaboration Likelihood Model to distinguish between the quick and lean idea preselection process as a peripheral route of information processing and the subsequent idea review process as a central route of information processing. In our empirical study with a sample of more than 163,000 ideas collected from the Xiaomi MIUI community, we analyse influencing factors that increase the likelihood of ideas being preselected or reviewed. Results show that user status, user initiative contribution, and community recognition have a significantly positive influence on idea preselction, whereas user response contribution has no influence. Idea presentation characteristics have an inverted U-curve relationship with idea adoption. Community absorptive capacity has a moderate effect on the curvilinear relationship between idea description length and idea adoption.
Immigrant integration has become a primary political concern for leaders in Germany and the United States. The information systems (IS) community has begun to research how information and communications technologies can assist immigrants and refugees, such as by examining how countries can facilitate social-inclusion processes. Migrants face the challenge of joining closed communities that cannot integrate or fear doing so. We conducted a panel discussion at the 2019 Americas Conference on Information Systems (AMCIS) in Cancun, Mexico, to introduce multiple viewpoints on immigration. In particular, the panel discussed how technology can both support and prevent immigrants from succeeding in their quest. We conducted the panel to stimulate a thoughtful and dynamic discussion on best practices and recommendations to enhance the discipline's impact on alleviating the challenges that occur for immigrants in their host countries. In this panel report, we introduce the topic of using ICT to help immigrants integrate and identify differences between North/Central America and Europe. We also discuss how immigrants (particularly refugees) use ICT to connect with others, feel that they belong, and maintain their identity. We also uncover the dark and bright sides of how governments use ICT to deter illegal immigration. Finally, we present recommendations for researchers and practitioners on how to best use ICT to assist with immigration.
This paper discusses results from a small-scale research
study, together with some recently published research into student
perceptions of ICT for learning in schools, to consider relevant skills
that do not appear to currently being taught. The paper concludes by
raising three issues relating to learning with and through ICT that need
to be addressed in school curricula and classroom teaching.
Ten ice-sheet models are used to study sensitivity of the Greenland and Antarctic ice sheets to prescribed changes of surface mass balance, sub-ice-shelf melting and basal sliding. Results exhibit a large range in projected contributions to sea-level change. In most cases, the ice volume above flotation lost is linearly dependent on the strength of the forcing. Combinations of forcings can be closely approximated by linearly summing the contributions from single forcing experiments, suggesting that nonlinear feedbacks are modest. Our models indicate that Greenland is more sensitive than Antarctica to likely atmospheric changes in temperature and precipitation, while Antarctica is more sensitive to increased ice-shelf basal melting. An experiment approximating the Intergovernmental Panel on Climate Change's RCP8.5 scenario produces additional first-century contributions to sea level of 22.3 and 8.1 cm from Greenland and Antarctica, respectively, with a range among models of 62 and 14 cm, respectively. By 200 years, projections increase to 53.2 and 26.7 cm, respectively, with ranges of 79 and 43 cm. Linear interpolation of the sensitivity results closely approximates these projections, revealing the relative contributions of the individual forcings on the combined volume change and suggesting that total ice-sheet response to complicated forcings over 200 years can be linearized.
Neodymium isotopic composition (epsilon Nd) has enjoyed widespread use as a palaeotracer, principally because it behaves quasi-conservatively in the modern ocean. However, recent bottom water epsilon Nd reconstructions from the eastern North Atlantic are difficult to interpret under assumptions of conservative behaviour. The observation that this apparent departure from conservative behaviour increases with enhanced ice-rafted debris (IRD) fluxes has resulted in the suggestion that IRD leads to the overprinting of bottom water epsilon Nd through reversible scavenging. In this study, a simple water column model successfully reproduces epsilon Nd reconstructions from the eastern North Atlantic at the Last Glacial Maximum and Heinrich Stadial 1, and demonstrates that the changes in scavenging intensity required for good model-data fit is in good agreement with changes in the observed IRD flux. Although uncertainties in model parameters preclude a more definitive conclusion, the results indicate that the suggestion of IRD as a source of non-conservative behaviour in the epsilon Nd tracer is reasonable and that further research into the fundamental chemistry underlying the marine neodymium cycle is necessary to increase confidence in assumptions of conservative epsilon Nd behaviour in the past.
Forest and steppe communities in the Altai region of Central Asia are threatened by changing climate and anthropogenic pressure. Specifically, increasing drought and grazing pressure may cause collapses of moisture-demanding plant communities, particularly forests. Knowledge about past vegetation and fire responses to climate and land use changes may help anticipating future ecosystem risks, given that it has the potential to disclose mechanisms and processes that govern ecosystem vulnerability. We present a unique paleoecological record from the high-alpine Tsambagarav glacier in the Mongolian Altai that provides novel large-scale information on vegetation, fire and pollution with an exceptional temporal resolution and precision. Our palynological record identifies several late-Holocene boreal forest expansions, contractions and subsequent recoveries. Maximum forest expansions occurred at 3000-2800 BC, 2400-2100 BC, and 1900-1800 BC. After 1800 BC mixed boreal forest communities irrecoverably declined. Fires reached a maximum at 1600 BC, 200 years after the final forest collapse. Our multiproxy data suggest that burning peaked in response to dead biomass accumulation resulting from forest diebacks. Vegetation and fire regimes partly decoupled from climate after 1700 AD, when atmospheric industrial pollution began, and land use intensified. We conclude that moisture availability was more important than temperature for past vegetation dynamics, in particular for forest loss and steppe expansion. The past Mongolian Altai evidence implies that in the future forests of the Russian Altai may collapse in response to reduced moisture availability.
Changes in ice discharge from Antarctica constitute the largest uncertainty in future sea-level projections, mainly because of the unknown response of its marine basins(1). Most of West Antarctica's marine ice sheet lies on an inland-sloping bed(2) and is thereby prone to a marine ice sheet instability(3-5). A similar topographic configuration is found in large parts of East Antarctica, which holds marine ice equivalent to 19 m of global sea-level rise(6), that is, more than five times that of West Antarctica. Within East Antarctica, the Wilkes Basin holds the largest volume of marine ice that is fully connected by subglacial troughs. This ice body was significantly reduced during the Pliocene epoch(7). Strong melting underneath adjacent ice shelves with similar bathymetry(8) indicates the ice sheet's sensitivity to climatic perturbations. The stability of the Wilkes marine ice sheet has not been the subject of any comprehensive assessment of future sea level. Using recently improved topographic data(6) in combination with ice-dynamic simulations, we show here that the removal of a specific coastal ice volume equivalent to less than 80 mm of global sea-level rise at the margin of the Wilkes Basin destabilizes the regional ice flow and leads to a self-sustained discharge of the entire basin and a global sea-level rise of 3-4 m. Our results are robust with respect to variation in ice parameters, forcing details and model resolution as well as increased surface mass balance, indicating that East Antarctica may become a large contributor to future sea-level rise on timescales beyond a century.
Some of the largest and most erosive floods on Earth result from the failure of glacial dams. While potentially cataclysmic ice dams are recognized to have repeatedly formed along ice-sheet margins, much less is known about the frequency and longevity of ice dams caused by mountain glaciers, and their impact on landscape evolution. Here we present field observations and results from cosmogenic nuclide dating that allow reconstructing a > 100-k.y.-long history of glacial damming in the Shyok Valley, eastern Karakoram (South Asia). Our field observations provide evidence that Asia's second-longest glacier, the Siachen, once extended for over 180 km and blocked the Shyok River during the penultimate glacial period, leading to upstream deposition of a more than 400-m-thick fluvio-lacustrine valley fill. Be-10-depth profile modeling indicates that glacial damming ended with the onset of the Eemian interglacial and that the Shyok River subsequently incised the valley fill at an average rate of similar to 4-7 m k.y.(-1). Comparison with contemporary ice-dammed lakes in the Karakoram and elsewhere suggests recurring outburst floods during the aggradation period, while over 25 cycles of fining-upward lake deposits within the valley fill indicate impounding of floods from farther upstream. Despite prolonged damming, the net effect of this and probably earlier damming episodes by the Siachen Glacier is dominated by glacial erosion in excess of fluvial incision, as evidenced by a pronounced overdeepening that follows the glaciated valley reach. Strikingly similar overdeepened valleys at all major confluences of the Shyok and Indus Rivers with Karakoram tributaries indicate that glacial dams and subsequent outburst floods have been widespread and frequent in this region during the Quaternary. Our study suggests that the interaction of Karakoram glaciers with the Shyok and Indus Rivers promoted valley incision and headward erosion into the western margin of the Tibetan Plateau.
Late Quaternary landscapes of unglaciated Beringia were largely shaped by ice-wedge polygon tundra. Ice Complex (IC) strata preserve such ancient polygon formations. Here we report on the Yukagir IC from Bol'shoy Lyakhovsky Island in northeastern Siberia and suggest that new radioisotope disequilibria (230Th/U) dates of the Yukagir IC peat confirm its formation during the Marine Oxygen Isotope Stage (MIS) 7a–c interglacial period. The preservation of the ice-rich Yukagir IC proves its resilience to last interglacial and late glacial–Holocene warming. This study compares the Yukagir IC to IC strata of MIS 5, MIS 3, and MIS 2 ages exposed on Bol'shoy Lyakhovsky Island. Besides high intrasedimental ice content and syngenetic ice wedges intersecting silts, sandy silts, the Yukagir IC is characterized by high organic matter (OM) accumulation and low OM decomposition of a distinctive Drepanocladus moss-peat. The Yukagir IC pollen data reveal grass-shrub-moss tundra indicating rather wet summer conditions similar to modern ones. The stable isotope composition of Yukagir IC wedge ice is similar to those of the MIS 5 and MIS 3 ICs pointing to similar atmospheric moisture generation and transport patterns in winter. IC data from glacial and interglacial periods provide insights into permafrost and climate dynamics since about 200 ka.
The new in situ geodynamic laboratory established in the framework of the ICDP Eger project aims to develop the most modern, comprehensive, multiparameter laboratory at depth for studying earthquake swarms, crustal fluid flow, mantle-derived CO2 and helium degassing, and processes of the deep biosphere. In order to reach a new level of high-frequency, near-source and multiparameter observation of earthquake swarms and related phenomena, such a laboratory comprises a set of shallow boreholes with high-frequency 3-D seismic arrays as well as modern continuous real-time fluid monitoring at depth and the study of the deep biosphere.
This laboratory is located in the western part of the Eger Rift at the border of the Czech Republic and Germany (in the West Bohemia–Vogtland geodynamic region) and comprises a set of five boreholes around the seismoactive zone. To date, all monitoring boreholes have been drilled. This includes the seismic monitoring boreholes S1, S2 and S3 in the crystalline units north and east of the major Nový Kostel seismogenic zone, borehole F3 in the Hartoušov mofette field and borehole S4 in the newly discovered Bažina maar near Libá. Supplementary borehole P1 is being prepared in the Neualbenreuth maar for paleoclimate and biological research. At each of these sites, a borehole broadband seismometer will be installed, and sites S1, S2 and S3 will also host a 3-D seismic array composed of a vertical geophone chain and surface seismic array. Seismic instrumenting has been completed in the S1 borehole and is in preparation in the remaining four monitoring boreholes. The continuous fluid monitoring site of Hartoušov includes three boreholes, F1, F2 and F3, and a pilot monitoring phase is underway. The laboratory also enables one to analyze microbial activity at CO2 mofettes and maar structures in the context of changes in habitats. The drillings into the maar volcanoes contribute to a better understanding of the Quaternary paleoclimate and volcanic activity.
The new in situ geodynamic laboratory established in the framework of the ICDP Eger project aims to develop the most modern, comprehensive, multiparameter laboratory at depth for studying earthquake swarms, crustal fluid flow, mantle-derived CO2 and helium degassing, and processes of the deep biosphere. In order to reach a new level of high-frequency, near-source and multiparameter observation of earthquake swarms and related phenomena, such a laboratory comprises a set of shallow boreholes with high-frequency 3-D seismic arrays as well as modern continuous real-time fluid monitoring at depth and the study of the deep biosphere.
This laboratory is located in the western part of the Eger Rift at the border of the Czech Republic and Germany (in the West Bohemia-Vogtland geodynamic region) and comprises a set of five boreholes around the seismoactive zone. To date, all monitoring boreholes have been drilled. This includes the seismic monitoring boreholes S1, S2 and S3 in the crystalline units north and east of the major Novy Kostel seismogenic zone, borehole F3 in the Hartousov mofette field and borehole S4 in the newly discovered Bazina maar near Liba. Supplementary borehole P1 is being prepared in the Neualbenreuth maar for paleoclimate and biological research. At each of these sites, a borehole broadband seismometer will be installed, and sites S1, S2 and S3 will also host a 3-D seismic array composed of a vertical geophone chain and surface seismic array. Seismic instrumenting has been completed in the S1 borehole and is in preparation in the remaining four monitoring boreholes. The continuous fluid monitoring site of Hartousov includes three boreholes, F1, F2 and F3, and a pilot monitoring phase is underway. The laboratory also enables one to analyze microbial activity at CO2 mofettes and maar structures in the context of changes in habitats. The drillings into the maar volcanoes contribute to a better understanding of the Quaternary paleoclimate and volcanic activity.
Accelerograms are the primary source for characterizing strong ground motion. It is therefore of paramount interest to have high-quality recordings free from any nonphysical contamination. Frequently, accelerograms are affected by baseline jumps and drifts, either related to the instrument and/or a major earthquake. In this work, I propose a correction method for these undesired baseline drifts based on segmented linear least squares. The algorithm operates on the integrated waveforms and combines all three instrument components to estimate a model that modifies the baseline to be at zero continuously. The procedure consists of two steps: first a suite of models with variable numbers of discontinuities is derived for all three instrument components. During this process, the number of discontinuities is reduced in a parsimonious way, for example, two very close discontinuities are merged into a single one. In the second step, the optimal model is selected on the basis of the Bayesian information criterion. I exemplify the application on synthetic waveforms with known discontinuities and on observed waveforms from a unified strong-motion database of the Japan Meteorological Agency (JMA) and the National Research Institute for Earth Science and Disaster Prevention (NIED, Japan) networks for the major events of the 2016 Kumamoto earthquakes. After the baseline jump correction, the waveforms are furthermore corrected for displacement according to Wang et al.(2011). The resulting displacements are comparable to the Interferometric Synthetic Aperture Radar-derived displacement estimates for the Kumamoto earthquake sequence.
In five studies, we evaluated the psychometric properties of a revised German version of the State Self-Esteem Scale (SSES; Heatherton & Polivy, 1991). In Study 1, the results of a confirmatory factor analysis on the original scale revealed poor model fit and poor construct validity in a student sample that resembled those in the literature; thus, a revised 15-item version was developed (i.e., the SSES-R) and thoroughly validated. Study 2 showed a valid three-factor structure (Performance, Social, and Appearance) and good internal consistency of the SSES-R. Correlations between subscales of trait and state SE empirically supported the scale's construct validity. Temporal stability and intrapersonal sensitivity of the scale to naturally occurring events were investigated in Study 3. Intrapersonat sensitivity of the scale to experimentally induced changes in state SE was uncovered in Study 4 via social feedback (acceptance vs. rejection) and performance feedback (positive vs. negative). In Study 5, the scale's interpersonal sensitivity was confirmed by comparing depressed and healthy individuals. Finally, the usefulness of the SSES-R was demonstrated by assessing SE instability as calculated from repeated measures of state SE.
I Can See It in Your Face.
(2019)
The purpose of this study was to illustrate that people’s affective valuation of exercise can be identified in their faces. The study was conducted with a software for automatic facial expression analysis and it involved testing the hypothesis that positive or negative affective valuation occurs spontaneously when people are reminded of exercise. We created a task similar to an emotional Stroop task, in which participants responded to exercise-related and control stimuli with a positive or negative facial expression (smile or frown) depending on whether the photo was presented upright or tilted. We further asked participants how much time they would normally spend for physical exercise, because we assumed that the affective valuation of those who exercise more would be more positive. Based on the data of 86 participants, regression analysis revealed that those who reported less exercise and a more negative reflective evaluation of exercise initiated negative facial expressions on exercise-related stimuli significantly faster than those who reported exercising more often. No significant effect was observed for smile responses. We suspect that responding with a smile to exercise-related stimuli was the congruent response for the majority of our participants, so that for them no Stroop interference occurred in the exercise-related condition. This study suggests that immediate negative affective reactions to exercise-related stimuli result from a postconscious automatic process and can be detected in the study participants’ faces. It furthermore illustrates how methodological paradigms from social–cognition research (here: the emotional Stroop paradigm) can be adapted to collect and analyze biometric data for the investigation of exercisers’ and non-exercisers’ automatic valuations of exercise.
I can see it in your eyes
(2021)
Over the past years, extensive research has been dedicated to developing robust platforms and data-driven dialog models to support long-term human-robot interactions. However, little is known about how people's perception of robots and engagement with them develop over time and how these can be accurately assessed through implicit and continuous measurement techniques. In this paper, we explore this by involving participants in three interaction sessions with multiple days of zero exposure in between. Each session consists of a joint task with a robot as well as two short social chats with it before and after the task. We measure participants' gaze patterns with a wearable eye-tracker and gauge their perception of the robot and engagement with it and the joint task using questionnaires. Results disclose that aversion of gaze in a social chat is an indicator of a robot's uncanniness and that the more people gaze at the robot in a joint task, the worse they perform. In contrast with most HRI literature, our results show that gaze toward an object of shared attention, rather than gaze toward a robotic partner, is the most meaningful predictor of engagement in a joint task. Furthermore, the analyses of gaze patterns in repeated interactions disclose that people's mutual gaze in a social chat develops congruently with their perceptions of the robot over time. These are key findings for the HRI community as they entail that gaze behavior can be used as an implicit measure of people's perception of robots in a social chat and of their engagement and task performance in a joint task.
I can see clearly now
(2017)
Past research has revealed that narcissists lack forgiveness. However, little is known about factors that might either buffer or, more critically, enhance the link between narcissism and lack of forgiveness. To address this gap in the literature, the present studies focused on the moderating role of clarity of transgression-related motivations. In an original and a replication study (Ns = 509 and 532, respectively), participants rated their levels of narcissistic admiration and rivalry and recalled a personal episode in which someone had hurt them. Subsequently, participants reported on their lack of forgiveness toward their transgressor. Response speed to these ratings served as an indirect clarity measure, with faster responses indicating greater clarity. In both studies, narcissistic rivalry (but not admiration) was positively related to lack of forgiveness and this relationship was stronger among individuals who were clear about their transgression-related motivations. Results inform our understanding of socio-emotional factors that contribute to narcissists' lack of forgiveness following interpersonal hurt.
In her writings on ancient myth, the British author Natalie Haynes moves women to the centre of attention. Her two latest books, A Thousand Ships and Pandora’s Jar – a fiction novel and a non-fiction one – approach this topic from two different perspectives. This interview takes stock of Haynes’ motives and methodology as well as of the challenges she faces in the process of writing.
Hysteresis in the pinning-depinning transitions of spiral waves rotating around a hole in a circular shaped two- dimensional excitable medium is studied both by use of the continuation software AUTO and by direct numerical integration of the reaction-diffusion equations for the FitzHugh-Nagumo model. In order to clarify the role of different factors in this phenomenon, a kinematical description is applied. It is found that the hysteresis phenomenon computed for the reaction-diffusion model can be reproduced qualitatively only when a nonlinear eikonal equation (i.e. velocity- curvature relationship) is assumed. However, to obtain quantitative agreement, the dispersion relation has to be taken into account.
Bifurcations of dynamos in rotating and buoyancy-driven spherical Rayleigh-Benard convection in an electrically conducting fluid are investigated numerically. Both nonmagnetic and magnetic solution branches comprised of rotating waves are traced by path-following techniques, and their bifurcations and interconnections for different Ekman numbers are determined. In particular, the question of whether the dynamo branches bifurcate super- or sub-critically and whether a direct link to the primary pure convective states exists is answered.
Inflammatory cytokines play an important role in intervertebral disc degeneration. Although largely produced by immune cells, nucleus pulposus (NP) cells can also secrete them under various conditions, for example, under free swelling. Thus, tissue hypotonicity may be an inflammatory trigger for NP cells. The aim of this study was to investigate whether decreased tonicity under restricted swelling conditions (as occurring in early disc degeneration) could initiate an inflammatory cascade that mediates further degeneration. Healthy bovine NP tissue was balanced against different PEG concentrations (0-30%) to obtain various tissue tonicities. Samples were then placed in an artificial annulus (fixed volume) and were cultured for 3, 7, or 21 days, with free swelling NP as control. Tissue content (water, glycosaminoglycan, collagen) was analyzed, and both the tissue and medium were screened for tumor necrosis factor alpha (TNF-alpha), interleukin-1 beta (IL-1 beta), interleukin-6 (IL-6), interleukin-8 (IL-8), prostaglandin-E-2 (PGE(2)), and nitric oxide (NO). A range of tonicities (isotonic to hypotonic) was present at day 3 in the PEG-treated samples. However, during culture, the tonicity range narrowed as GAGs leached from the tissue. TNF-alpha and IL-1 beta were below detection limits in all conditions, while mid- and downstream inflammatory cytokines were detected. This may suggest that the extracellular environment directly affects NP cells instead of inducing a classical inflammatory cascade. Furthermore, IL-8 increased in swelling restricted samples, while IL-6 and PGE(2) were elevated in free swelling controls. These findings may suggest the involvement of different mechanisms in disc degeneration with intact AF compared to herniation, and encourage further investigation. (c) 2019 The Authors. Journal of Orthopaedic Research (R) Published by Wiley Periodicals, Inc. on behalf of Orthopaedic Research Society. J Orthop Res
The method of twin surrogates has been introduced to test for phase synchronization of complex systems in the case of passive experiments. In this paper we derive new analytical expressions for the number of twins depending on the size of the neighborhood, as well as on the length of the trajectory. This allows us to determine the optimal parameters for the generation of twin surrogates. Furthermore, we determine the quality of the twin surrogates with respect to several linear and nonlinear statistics depending on the parameters of the method. In the second part of the paper we perform a hypothesis test for phase synchronization in the case of experimental data from fixational eye movements. These miniature eye movements have been shown to play a central role in neural information processing underlying the perception of static visual scenes. The high number of data sets (21 subjects and 30 trials per person) allows us to compare the generated twin surrogates with the "natural" surrogates that correspond to the different trials. We show that the generated twin surrogates reproduce very well all linear and nonlinear characteristics of the underlying experimental system. The synchronization analysis of fixational eye movements by means of twin surrogates reveals that the synchronization between the left and right eye is significant, indicating that either the centers in the brain stem generating fixational eye movements are closely linked, or, alternatively that there is only one center controlling both eyes.
Adult-child interactions can support children's development and are established as predictors of program quality in early childhood settings. However, the linguistic components that constitute positive interactions have not yet been studied in detail. This study investigates the effects of hypotheses proposed by adults on children's responses in a dyadic picture-book viewing situation. In 2 experiments, adults' use of hypotheses (e.g., "Maybe this is a dwarf's door") was tested against the use of instructive statements ("This is a dwarf's door") and in combination with open questions ("What do you think, why is the door so small?"). In Experiment 1, hypotheses differed from instructions only by the modal marker "maybe". Children's responses to hypotheses were longer and contained more self-generated explanations as compared to responses to instructions. The use of hypotheses also seemed to encourage children to attach more importance to their own explanations. In Experiment 2, combining hypotheses with open-ended why questions elicited longer responses but no more self-generated explanations in children than openended questions alone. Results indicate that subtle differences in adults' utterances can directly influence children's reasoning and children's contributions to dialogues.
Stable carbon isotopes of sediment organic matter (delta C-13(OM)) are widely applied in paleoenvironmental studies. Interpretations of delta C-13(OM), however, remain challenging and factors that influence delta C-13(OM) may not apply across all lakes. Common explanations for stratigraphic shifts in delta C-13(OM) include changes in lake productivity or changes in inputs of allochthonous OM. We investigated the influence of different oxygen conditions (oxic versus anoxic) on the delta C-13(OM) values in the sediments of Lake Tiefer See. We analysed (1) a long sediment core from the deepest part of the lake, (2) two short, sediment-water interface cores from shallower water depths, and (3) OM in the water column, i.e. from sediment traps. Fresh OM throughout the entire water column showed a relatively constant delta C-13(OM) value of approximately -30.5 parts per thousand. Similar values, about -31 parts per thousand, were obtained for well-varved sediments in both the long and short, sediment-water interface cores. In contrast, delta C-13(OM) values from non-varved sediments in all cores were significantly less negative (-29 parts per thousand). The delta C-13(OM) values in the sediment-water interface cores from different water depths differ for sediments of the same age, if oxygen conditions at the time of deposition were different at these sites, as suggested by the state of varve preservation. Sediments deposited from AD 1924 to 1980 at 62m water depth are varved and exhibit delta C-13(OM) values around -31 parts per thousand, whereas sediments of the same age in the core from 35m water depth are not varved and show less negative delta C-13(OM) values of about -29 parts per thousand. The relation between varve occurrence and delta C-13(OM) values suggests that delta C-13(OM) is associated with oxygen conditions because varve preservation depends on hypolimnetic anoxia. A mechanism that likely influences delta C-13(OM) is selective degradation of OM under oxic conditions, such that organic components with more negative delta C-13(OM) are preferably decomposed, leading to less negative delta C-13(OM) values in the remaining, undegraded OM pool. Greater decomposition of OM in non-varved sediments is supported by lower TOC concentrations in these deposits (similar to 5%) compared to well-varved sediments (similar to 15%). Even in lakes that display small variations in productivity and terrestrial OM input through time, large spatial and temporal differences in hypolimnetic oxygen concentrations may be an important factor controlling sediment delta C-13(OM).
Hyperspectral REE (Rare Earth Element) Mapping of Outcrops-Applications for Neodymium Detection
(2015)
In this study, an in situ application for identifying neodymium (Nd) enriched surface materials that uses multitemporal hyperspectral images is presented (HySpex sensor). Because of the narrow shape and shallow absorption depth of the neodymium absorption feature, a method was developed for enhancing and extracting the necessary information for neodymium from image spectra, even under illumination conditions that are not optimal. For this purpose, the two following approaches were developed: (1) reducing noise and analyzing changing illumination conditions by averaging multitemporal image scenes and (2) enhancing the depth of the desired absorption band by deconvolving every image spectrum with a Gaussian curve while the rest of the spectrum remains unchanged (Richardson-Lucy deconvolution). To evaluate these findings, nine field samples from the Fen complex in Norway were analyzed using handheld X-ray fluorescence devices and by conducting detailed laboratory-based geochemical rare earth element determinations. The result is a qualitative outcrop map that highlights zones that are enriched in neodymium. To reduce the influences of non-optimal illumination, particularly at the studied site, a minimum of seven single acquisitions is required. Sharpening the neodymium absorption band allows for robust mapping, even at the outer zones of enrichment. From the geochemical investigations, we found that iron oxides decrease the applicability of the method. However, iron-related absorption bands can be used as secondary indicators for sulfidic ore zones that are mainly enriched with rare earth elements. In summary, we found that hyperspectral spectroscopy is a noninvasive, fast and cost-saving method for determining neodymium at outcrop surfaces.
Military areas are valuable habitats and refuges for rare and endangered plants and animals. We developed a new approach applying innovative methods of hyperspectral remote sensing to bridge the existing gap between remote sensing technology and the demands of the nature conservation community. Remote sensing has already proven to be a valuable monitoring instrument. However, the approaches lack the consideration of the demands of applied nature conservation which includes the legal demands of the EU Habitat Directive. Following the idea of the Vital Signs Monitoring in the USA, we identified a subset of the highest priority monitoring indicators for our study area. We analyzed continuous spectral response curves and tested the measurability of N=19 indicators on the basis of complexity levels aggregated from extensive vegetation assemblages. The spectral differentiability for the floristic as well as faunistic indicators revealed values up to 100% accuracy. We point out difficulties when it comes to distinguishing faunistic habitat requirements of several species adapted to dry open landscapes, which in this case results in OVERALL ACCURACY of 67, 87-95, and 35% in the error matrix. In summary, we provide an applicable and feasible method to facilitating monitoring military areas by hyperspectral remote sensing in the following. (C) 2014 Elsevier Ltd. All rights reserved.
The fusion of hyperspectral imaging (HSI) sensor and airborne lidar scanner (ALS) data provides promising potential for applications in environmental sciences. Standard fusion approaches use reflectance information from the HSI and distance measurements from the ALS to increase data dimen-sionality and geometric accuracy. However, the potential for data fusion based on the respective intensity information of the complementary active and passive sensor systems is high and not yet fully exploited. Here, an approach for the rigorous illumination correction of HSI data, based on the radiometric cross-calibrated return intensity information of ALS data, is presented. The cross calibration utilizes a ray tracing-based fusion of both sensor measurements by intersecting their particular beam shapes. The developed method is capable of compensating for the drawbacks of passive HSI systems, such as cast and cloud shadowing effects, illumination changes over time, across track illumination, and partly anisotropy effects. During processing, spatial and temporal differences in illumination patterns are detected and corrected over the entire HSI wavelength domain. The improvement in the classification accuracy of urban and vegetation surfaces demonstrates the benefit and potential of the proposed HSI illumination correction. The presented approach is the first step toward the rigorous in-flight fusion of passive and active system characteristics, enabling new capabilities for a variety of applications.
Plasma secretion of acid sphingomyelinase is a hallmark of cellular stress response resulting in the formation of membrane embedded ceramide-enriched lipid rafts and the reorganization of receptor complexes. Consistently, decompartmentalization of ceramide formation from inert sphingomyelin has been associated with signaling events and regulation of the cellular phenotype. Herein, we addressed the question of whether the secretion of acid sphingomyelinase is involved in host response during sepsis. We found an exaggerated clinical course in mice genetically deficient in acid sphingomyelinase characterized by an increased bacterial burden, an increased phagocytotic activity, and a more pronounced cytokine storm. Moreover, on a functional level, leukocyte-endothelial interaction was found diminished in sphingomyelinase-deficient animals corresponding to a distinct leukocytes' phenotype with respect to rolling and sticking as well as expression of cellular surface proteins.(jlr) We conclude that hydrolysis of membrane-embedded sphingomyelin, triggered by circulating sphingomyelinase, plays a pivotal role in the first line of defense against invading microorganisms. This function might be essential during the early phase of infection leading to an adaptive response of remote cells and tissues.-Jbeily, N., I. Suckert, F. A. Gonnert, B. Acht, C. L. Bockmeyer, S. D. Grossmann, M. F. Blaess, A. Lueth, H.-P. Deigner, M. Bauer, and R. A. Claus. Hyperresponsiveness of mice deficient in plasma-secreted sphingomyelinase reveals its pivotal role in early phase of host response. J. Lipid Res. 2013. 54: 410-424.
Hyperpolarizability of donor-acceptor azines subject to push-pull character and steric hindrance
(2009)
The push-pull character of two series of donor-acceptor azines has been quantified by C-13, N-15 chemical shift differences of the partial C(1)=N(1) and N(2)=C(2) double bonds in the central linking C(1)=N(1)-N(2)=C(2) unit and by the quotient of the occupations of the bonding pi and anti-bonding pi* orbitals of these bonds. Excellent correlation of the latter push-pull parameter with the corresponding bond lengths d(C=N) strongly recommend both the occupation quotients pi*/pi and the corresponding bond lengths as reasonable sensors for quantifying the push, pull character along the C=N-N=C linking unit, for the donor-acceptor quality of the two series of azines and for the molecular hyperpolarizability beta(0) of these compounds. Within this context, reasonable conclusions concerning the interplay of steric hindrance in the chromophore, push-pull character and hyperpolarizability of the azines and their application as NLO materials will be drawn.
Laforin or malin deficiency causes Lafora disease, characterized by altered glycogen metabolism and teenage-onset neurodegeneration with intractable and invariably fatal epilepsy. Plant starches possess small amounts of metabolically essential monophosphate esters. Glycogen contains similar phosphate amounts, which are thought to originate from a glycogen synthase error side reaction and therefore lack any specific function. Glycogen is also believed to lack monophosphates at glucosyl carbon C6, an essential phosphorylation site in plant starch metabolism. We now show that glycogen phosphorylation is not due to a glycogen synthase side reaction, that C6 is a major glycogen phosphorylation site, and that C6 monophosphates predominate near centers of glycogen molecules and positively correlate with glycogen chain lengths. Laforin or malin deficiency causes C6 hyperphosphorylation, which results in malformed long-chained glycogen that accumulates in many tissues, causing neurodegeneration in brain. Our work advances the understanding of Lafora disease pathogenesis and suggests that glycogen phosphorylation has important metabolic function.
Epigenetic silencing of tumor suppressor genes is involved in early transforming events and has a high impact on colorectal carcinogenesis. Likewise, colon cancers that derive from chronically inflamed bowel diseases frequently exhibit epigenetic changes. But there is little data about epigenetic aberrations causing colorectal cancer in chronically inflamed tissue. The aim of the present study was to evaluate the aberrant gain of methylation in the gene promoters of VIM, TFPI2 and ITGA4 as putative early markers in the development from inflamed tissue via precancerous lesions toward colorectal cancer.
Initial screening of different cancer cell lines by using methylation-specific PCR revealed a putative colon cancer-specific methylation pattern. Additionally, a demethylation assay was performed to investigate the methylation-dependent gene silencing of ITGA4. The candidate markers were analyzed in colonic tissue specimens from patients with colorectal cancer (n = 15), adenomas (n = 76), serrated lesions (n = 13), chronic inflammation (n = 10) and normal mucosal samples (n = 9).
A high methylation frequency of VIM (55.6 %) was observed in normal colon tissue, whereas ITGA4 and TFPI2 were completely unmethylated in controls. A significant gain of methylation frequency with progression of disease as well as an age-dependent effect was detectable for TFPI2. ITGA4 methylation frequency was high in precancerous and cancerous tissues as well as in inflammatory bowel diseases (IBD).
The already established methylation marker VIM does not permit a specific and sensitive discrimination of healthy and neoplastic tissue. The methylation markers ITGA4 and TFPI2 seem to be suitable risk markers for inflammation-associated colon cancer.
The trans diesters of 1,4-cyclohexanediol with a number of acetic acid analogues, CX3COOH, of varying steric hindrance and polarity (CX3 = Me, Et, iso-Pr, tert-Bu, CF3, CH2Cl, CHCl2, CCl3, CH2Br, CHBr2, CBr3) were synthesized, and the axial, axial/equatorial, equatorial conformational equilibria were studied by low-temperature H-1 NMR spectroscopy in CD2Cl2. The structures and relative energies of the axial, axial and equatorial, equatorial conformers were calculated at both the MP2/6-311G* and the MP2/6-311+G* levels of theory, and it was only by including diffuse functions that a good correlation of Delta G degrees(calcd) vs Delta G(exptl) could be obtained. Both the structures and the energy differences of the axial, axial and equatorial, equatorial conformers are discussed with respect to the established models of conformational analysis, viz., steric 1,3-diaxial and hyperconjugative interactions. Interestingly, the hyperconjugative interactions sigma(C-C)/sigma(C-H)->sigma*(C-O), together with a steric effect which also destabilizes the equatorial, equatorial conformers on increasing bulk of the substituents, proved to dominate the position of the conformational equilibria. In addition, the preference of the axial, axial conformers with respect to their equatorial, equatorial analogues was greater than expected from the conformational energies of the corresponding substituents in the monosubstituted cyclohexyl esters. The reason for this very interesting and unexpected result is also discussed
We consider time evolution of Turing patterns in an extended system governed by an equation of the Swift-Hohenberg type, where due to an external periodic parameter modulation longwave and shortwave patterns with length scales related as 1:3 emerge in succession. We show theoretically and demonstrate numerically that the spatial phases of the patterns, being observed stroboscopically, are governed by an expanding circle map, so that the corresponding chaos of Turing patterns is hyperbolic, associated with a strange attractor of the Smale-Williams solenoid type. This chaos is shown to be robust with respect to variations of parameters and boundary conditions.
Hyperbolic chaos of standing wave patterns generated parametrically by a modulated pump source
(2013)
We outline a possibility of hyperbolic chaotic dynamics associated with the expanding circle map for spatial phases of parametrically excited standing wave patterns. The model system is governed by a one-dimensional wave equation with nonlinear dissipation. The phenomenon arises due to the pump modulation providing the alternating excitation of modes with the ratio of characteristic scales 1 : 3. DOI: 10.1103/PhysRevE.87.040901
We show that a hyperbolic chaos can be observed in resonantly coupled oscillators near a Hopf bifurcation, described by normal-form-type equations for complex amplitudes. The simplest example consists of four oscillators, comprising two alternatively activated, due to an external periodic modulation, pairs. In terms of the stroboscopic Poincare map, the phase differences change according to an expanding Bernoulli map that depends on the coupling type. Several examples of hyperbolic chaos for different types of coupling are illustrated numerically.