TY - JOUR A1 - Schmidt, Andreas A1 - Rabsch, Wolfgang A1 - Broeker, Nina K. A1 - Barbirz, Stefanie T1 - Bacteriophage tailspike protein based assay to monitor phase variable glucosylations in Salmonella O-antigens JF - BMC microbiology N2 - Background Non-typhoid Salmonella Typhimurium (S. Typhimurium) accounts for a high number of registered salmonellosis cases, and O-serotyping is one important tool for monitoring epidemiology and spread of the disease. Moreover, variations in glucosylated O-antigens are related to immunogenicity and spread in the host. However, classical autoagglutination tests combined with the analysis of specific genetic markers cannot always reliably register phase variable glucose modifications expressed on Salmonella O-antigens and additional tools to monitor O-antigen glucosylation phenotypes of S. Typhimurium would be desirable. Results We developed a test for the phase variable O-antigen glucosylation state of S. Typhimurium using the tailspike proteins (TSP) of Salmonella phages 9NA and P22. We used this ELISA like tailspike adsorption (ELITA) assay to analyze a library of 44 Salmonella strains. ELITA was successful in discriminating strains that carried glucose 1-6 linked to the galactose of O-polysaccharide backbone (serotype O1) from non-glucosylated strains. This was shown by O-antigen compositional analyses of the respective strains with mass spectrometry and capillary electrophoresis. The ELITA test worked rapidly in a microtiter plate format and was highly O-antigen specific. Moreover, TSP as probes could also detect glucosylated strains in flow cytometry and distinguish multiphasic cultures differing in their glucosylation state. Conclusions Tailspike proteins contain large binding sites with precisely defined specificities and are therefore promising tools to be included in serotyping procedures as rapid serotyping agents in addition to antibodies. In this study, 9NA and P22TSP as probes could specifically distinguish glucosylation phenotypes of Salmonella on microtiter plate assays and in flow cytometry. This opens the possibility for flow sorting of cell populations for subsequent genetic analyses or for monitoring phase variations during large scale O-antigen preparations necessary for vaccine production. KW - Salmonella Typhimurium KW - O-antigen KW - Tailspike protein KW - Bacteriophage KW - Phase variation KW - O-serotyping KW - Flow cytometry Y1 - 2016 U6 - https://doi.org/10.1186/s12866-016-0826-0 SN - 1471-2180 VL - 16 PB - BioMed Central CY - London ER - TY - JOUR A1 - Yan, Wenhao A1 - Chen, Dijun A1 - Kaufmann, Kerstin T1 - Efficient multiplex mutagenesis by RNA-guided Cas9 and its use in the characterization of regulatory elements in the AGAMOUS gene JF - Plant methods N2 - Background The efficiency of multiplex editing in plants by the RNA-guided Cas9 system is limited by efficient introduction of its components into the genome and by their activity. The possibility of introducing large fragment deletions by RNA-guided Cas9 tool provides the potential to study the function of any DNA region of interest in its ‘endogenous’ environment. Results Here, an RNA-guided Cas9 system was optimized to enable efficient multiplex editing in Arabidopsis thaliana. We demonstrate the flexibility of our system for knockout of multiple genes, and to generate heritable large-fragment deletions in the genome. As a proof of concept, the function of part of the second intron of the flower development gene AGAMOUS in Arabidopsis was studied by generating a Cas9-free mutant plant line in which part of this intron was removed from the genome. Further analysis revealed that deletion of this intron fragment results 40 % decrease of AGAMOUS gene expression without changing the splicing of the gene which indicates that this regulatory region functions as an activator of AGAMOUS gene expression. Conclusions Our modified RNA-guided Cas9 system offers a versatile tool for the functional dissection of coding and non-coding DNA sequences in plants. KW - RNA-guided Cas9 KW - Multiplex mutagenesis KW - Large fragment deletion KW - Germline transmission Y1 - 2016 U6 - https://doi.org/10.1186/s13007-016-0125-7 SN - 1746-4811 VL - 12 SP - 1 EP - 9 PB - BioMed Central CY - London ER - TY - JOUR A1 - Wacker, Alexander A1 - Piepho, Maike A1 - Harwood, John L. A1 - Guschina, Irina A. A1 - Arts, Michael T. T1 - Light-Induced Changes in Fatty Acid Profiles of Specific Lipid Classes in Several Freshwater Phytoplankton Species JF - Frontiers in plant science : FPLS N2 - We tested the influence of two light intensities [40 and 300 μmol PAR / (m2s)] on the fatty acid composition of three distinct lipid classes in four freshwater phytoplankton species. We chose species of different taxonomic classes in order to detect potentially similar reaction characteristics that might also be present in natural phytoplankton communities. From samples of the bacillariophyte Asterionella formosa, the chrysophyte Chromulina sp., the cryptophyte Cryptomonas ovata and the zygnematophyte Cosmarium botrytis we first separated glycolipids (monogalactosyldiacylglycerol, digalactosyldiacylglycerol, and sulfoquinovosyldiacylglycerol), phospholipids (phosphatidylcholine, phosphatidylethanolamine, phosphatidylglycerol, phosphatidylinositol, and phosphatidylserine) as well as non-polar lipids (triacylglycerols), before analyzing the fatty acid composition of each lipid class. High variation in the fatty acid composition existed among different species. Individual fatty acid compositions differed in their reaction to changing light intensities in the four species. Although no generalizations could be made for species across taxonomic classes, individual species showed clear but small responses in their ecologically-relevant omega-3 and omega-6 polyunsaturated fatty acids (PUFA) in terms of proportions and of per tissue carbon quotas. Knowledge on how lipids like fatty acids change with environmental or culture conditions is of great interest in ecological food web studies, aquaculture, and biotechnology, since algal lipids are the most important sources of omega-3 long-chain PUFA for aquatic and terrestrial consumers, including humans. KW - freshwater algae KW - light adaptation KW - lipid classes KW - fatty acid changes Y1 - 2016 U6 - https://doi.org/10.3389/fpls.2016.00264 SN - 1664-462X VL - 7 SP - 1 EP - 13 PB - Frontiers Media CY - Lausanne ER - TY - JOUR A1 - Gaubert, Philippe A1 - Patel, Riddhi P. A1 - Veron, Geraldine A1 - Goodman, Steven M. A1 - Willsch, Maraike A1 - Vasconcelos, Raquel A1 - Lourenco, Andre A1 - Sigaud, Marie A1 - Justy, Fabienne A1 - Joshi, Bheem Dutt A1 - Fickel, Jörns A1 - Wilting, Andreas T1 - Phylogeography of the Small Indian Civet and Origin of Introductions to Western Indian Ocean Islands JF - The journal of heredity : official journal of the American Genetic Association N2 - The biogeographic dynamics affecting the Indian subcontinent, East and Southeast Asia during the Plio-Pleistocene has generated complex biodiversity patterns. We assessed the molecular biogeography of the small Indian civet (Viverricula indica) through mitogenome and cytochrome b + control region sequencing of 89 historical and modern samples to (1) establish a time-calibrated phylogeography across the species’ native range and (2) test introduction scenarios to western Indian Ocean islands. Bayesian phylogenetic analyses identified 3 geographic lineages (East Asia, sister-group to Southeast Asia and the Indian subcontinent + northern Indochina) diverging 3.2–2.3 million years ago (Mya), with no clear signature of past demographic expansion. Within Southeast Asia, Balinese populations separated from the rest 2.6–1.3 Mya. Western Indian Ocean populations were assigned to the Indian subcontinent + northern Indochina lineage and had the lowest mitochondrial diversity. Approximate Bayesian computation did not distinguish between single versus multiple introduction scenarios. The early diversification of the small Indian civet was likely shaped by humid periods in the Late Pliocene–Early Pleistocene that created evergreen rainforest barriers, generating areas of intra-specific endemism in the Indian subcontinent, East, and Southeast Asia. Later, Pleistocene dispersals through drier conditions in South and Southeast Asia were likely, giving rise to the species’ current natural distribution. Our molecular data supported the delineation of only 4 subspecies in V. indica, including an endemic Balinese lineage. Our study also highlighted the influence of prefirst millennium AD introductions to western Indian Ocean islands, with Indian and/or Arab traders probably introducing the species for its civet oil. KW - Asia KW - biogeography KW - civet oil KW - mtDNA KW - Plio-Pleistocene KW - Viverridae Y1 - 2016 U6 - https://doi.org/10.1093/jhered/esw085 SN - 0022-1503 SN - 1465-7333 VL - 108 SP - 270 EP - 279 PB - Oxford Univ. Press CY - Cary ER - TY - JOUR A1 - Thieken, Annegret Henriette A1 - Kienzler, Sarah A1 - Kreibich, Heidi A1 - Kuhlicke, Christian A1 - Kunz, Michael A1 - Mühr, Bernhard A1 - Müller, Meike A1 - Otto, Antje A1 - Petrow, Theresia A1 - Pisi, Sebastian A1 - Schröter, Kai T1 - Review of the flood risk management system in Germany after the major flood in 2013 JF - Ecology and society : E&S ; a journal of integrative science for resilience and sustainability N2 - Widespread flooding in June 2013 caused damage costs of €6 to 8 billion in Germany, and awoke many memories of the floods in August 2002, which resulted in total damage of €11.6 billion and hence was the most expensive natural hazard event in Germany up to now. The event of 2002 does, however, also mark a reorientation toward an integrated flood risk management system in Germany. Therefore, the flood of 2013 offered the opportunity to review how the measures that politics, administration, and civil society have implemented since 2002 helped to cope with the flood and what still needs to be done to achieve effective and more integrated flood risk management. The review highlights considerable improvements on many levels, in particular (1) an increased consideration of flood hazards in spatial planning and urban development, (2) comprehensive property-level mitigation and preparedness measures, (3) more effective flood warnings and improved coordination of disaster response, and (4) a more targeted maintenance of flood defense systems. In 2013, this led to more effective flood management and to a reduction of damage. Nevertheless, important aspects remain unclear and need to be clarified. This particularly holds for balanced and coordinated strategies for reducing and overcoming the impacts of flooding in large catchments, cross-border and interdisciplinary cooperation, the role of the general public in the different phases of flood risk management, as well as a transparent risk transfer system. Recurring flood events reveal that flood risk management is a continuous task. Hence, risk drivers, such as climate change, land-use changes, economic developments, or demographic change and the resultant risks must be investigated at regular intervals, and risk reduction strategies and processes must be reassessed as well as adapted and implemented in a dialogue with all stakeholders. KW - August 2002 flood KW - Central Europe KW - Floods Directive KW - governance KW - June 2013 flood KW - risk management cycle Y1 - 2016 U6 - https://doi.org/10.5751/ES-08547-210251 SN - 1708-3087 SN - 1195-5449 VL - 21 IS - 2 PB - Resilience Alliance CY - Wolfville, NS ER - TY - JOUR A1 - Heinze, Johannes A1 - Gensch, Sabine A1 - Weber, Ewald A1 - Joshi, Jasmin Radha T1 - Soil temperature modifies effects of soil biota on plant growth JF - Journal of plant ecology N2 - Aims Plants directly and indirectly interact with many abiotic and biotic soil components. Research so far mostly focused on direct, individual abiotic or biotic effects on plant growth, but only few studies tested the indirect effects of abiotic soil factors on plant growth. Therefore, we investigated how abiotic soil conditions affect plant performance, via changes induced by soil biota. Methods In a full-factorial experiment, we grew the widespread grass Dactylis glomerata either with or without soil biota and investigated the impact of soil temperature, fertility and moisture on the soil biota effects on plant growth. We measured biomass production, root traits and colonization by arbuscular mycorrhizal fungi as well as microbial respiration. Important Findings We found significant interaction effects between abiotic soil conditions and soil biota on plant growth for fertility, but especially for soil temperature, as an increase of 10 degrees C significantly changed the soil biota effects on plant growth from positive to neutral. However, if tested individually, an increase in soil temperature and fertility per se positively affected plant biomass production, whereas soil biota per se did not affect overall plant growth, but both influenced root architecture. By affecting soil microbial activity and root architecture, soil temperature might influence both mutualistic and pathogenic interactions between plants and soil biota. Such soil temperature effects should be considered in soil feedback studies to ensure greater transferability of results from artificial and experimental conditions to natural environmental conditions. KW - plant-soil interaction KW - soil biota KW - abiotic soil factors KW - root traits KW - plant growth Y1 - 2016 U6 - https://doi.org/10.1093/jpe/rtw097 SN - 1752-9921 SN - 1752-993X VL - 10 SP - 808 EP - 821 PB - Oxford Univ. Press CY - Oxford ER - TY - JOUR A1 - Edlich, Alexander A1 - Gerecke, Christian A1 - Giulbudagian, Michael A1 - Neumann, Falko A1 - Hedtrich, Sarah A1 - Schaefer-Korting, Monika A1 - Ma, Nan A1 - Calderon, Marcelo A1 - Kleuser, Burkhard T1 - Specific uptake mechanisms of well-tolerated thermoresponsive polyglycerol-based nanogels in antigen-presenting cells of the skin JF - European Journal of Pharmaceutics and Biopharmaceutics N2 - Engineered nanogels are of high value for a targeted and controlled transport of compounds due to the ability to change their chemical properties by external stimuli. As it has been indicated that nanogels possess a high ability to penetrate the stratum corneum, it cannot be excluded that nanogels interact with dermal dendritic cells, especially in diseased skin. In this study the potential crosstalk of the thermore-sponsive nanogels (tNGs) with the dendritic cells of the skin was investigated with the aim to determine the immunotoxicological properties of the nanogels. The investigated tNGs were made of dendritic polyglycerol (dPG) and poly(glycidyl methyl ether-co-ethyl glycidyl ether) (p(GME-co-EGE)), as polymer conferring thermoresponsive properties. Although the tNGs were taken up, they displayed neither cytotoxic and genotoxic effects nor any induction of reactive oxygen species in the tested cells. Interestingly, specific uptake mechanisms of the tNGs by the dendritic cells were depending on the nanogels cloud point temperature (Tcp), which determines the phase transition of the nanoparticle. The study points to caveolae-mediated endocytosis as being the major tNGs uptake mechanism at 37 degrees C, which is above the Tcp of the tNGs. Remarkably, an additional uptake mechanism, beside caveolae-mediated endocytosis, was observed at 29 degrees C, which is the Tcp of the tNGs. At this temperature, which is characterized by two different states of the tNGs, macropinocytosis was involved as well. In summary, our study highlights the impact of thermoresponsivity on the cellular uptake mechanisms which has to be taken into account if the tNGs are used as a drug delivery system. KW - Dendritic cells KW - Drug delivery systems KW - Nanogel KW - Nanoparticle KW - Nanoparticle uptake KW - Nanotoxicology Y1 - 2017 U6 - https://doi.org/10.1016/j.ejpb.2016.12.016 SN - 0939-6411 SN - 1873-3441 VL - 116 SP - 155 EP - 163 PB - Elsevier CY - Amsterdam ER - TY - JOUR A1 - Batsios, Petros A1 - Ren, Xiang A1 - Baumann, Otto A1 - Larochelle, Denis A. A1 - Gräf, Ralph T1 - Src1 is a Protein of the Inner Nuclear Membrane Interacting with the Dictyostelium Lamin NE81 JF - Cells N2 - The nuclear envelope (NE) consists of the outer and inner nuclear membrane (INM), whereby the latter is bound to the nuclear lamina. Src1 is a Dictyostelium homologue of the helix-extension-helix family of proteins, which also includes the human lamin-binding protein MAN1. Both endogenous Src1 and GFP-Src1 are localized to the NE during the entire cell cycle. Immuno-electron microscopy and light microscopy after differential detergent treatment indicated that Src1 resides in the INM. FRAP experiments with GFP-Src1 cells suggested that at least a fraction of the protein could be stably engaged in forming the nuclear lamina together with the Dictyostelium lamin NE81. Both a BioID proximity assay and mis-localization of soluble, truncated mRFP-Src1 at cytosolic clusters consisting of an intentionally mis-localized mutant of GFP-NE81 confirmed an interaction of Src1 and NE81. Expression GFP-Src11–646, a fragment C-terminally truncated after the first transmembrane domain, disrupted interaction of nuclear membranes with the nuclear lamina, as cells formed protrusions of the NE that were dependent on cytoskeletal pulling forces. Protrusions were dependent on intact microtubules but not actin filaments. Our results indicate that Src1 is required for integrity of the NE and highlight Dictyostelium as a promising model for the evolution of nuclear architecture. KW - Dictyostelium KW - lamin KW - nuclear lamina KW - nucleus KW - nucleolus KW - HeH-protein KW - LEM-domain protein Y1 - 2016 U6 - https://doi.org/10.3390/cells5010013 SN - 2073-4409 VL - 5 IS - 1 PB - MDPI CY - Basel ER - TY - JOUR A1 - Zabel, André A1 - Winter, Alette A1 - Kelling, Alexandra A1 - Schilde, Uwe A1 - Strauch, Peter T1 - Tetrabromidocuprates(II)-Synthesis, Structure and EPR JF - International journal of molecular sciences N2 - Metal-containing ionic liquids (ILs) are of interest for a variety of technical applications, e.g., particle synthesis and materials with magnetic or thermochromic properties. In this paper we report the synthesis of, and two structures for, some new tetrabromidocuprates(II) with several “onium” cations in comparison to the results of electron paramagnetic resonance (EPR) spectroscopic analyses. The sterically demanding cations were used to separate the paramagnetic Cu(II) ions for EPR measurements. The EPR hyperfine structure in the spectra of these new compounds is not resolved, due to the line broadening resulting from magnetic exchange between the still-incomplete separated paramagnetic Cu(II) centres. For the majority of compounds, the principal g values (g|| and gK) of the tensors could be determined and information on the structural changes in the [CuBr4]2- anions can be obtained. The complexes have high potential, e.g., as ionic liquids, as precursors for the synthesis of copper bromide particles, as catalytically active or paramagnetic ionic liquids. KW - tetrabromidocuprate(II) KW - X-ray structure KW - electron paramagnetic resonance KW - copper(II) Y1 - 2016 U6 - https://doi.org/10.3390/ijms17040596 VL - 17 IS - 4 PB - MDPI CY - Basel ER - TY - JOUR A1 - Klauschies, Toni A1 - Vasseur, David A. A1 - Gaedke, Ursula T1 - Trait adaptation promotes species coexistence in diverse predator and prey communities JF - Ecology and evolution N2 - Species can adjust their traits in response to selection which may strongly influence species coexistence. Nevertheless, current theory mainly assumes distinct and time-invariant trait values. We examined the combined effects of the range and the speed of trait adaptation on species coexistence using an innovative multispecies predator–prey model. It allows for temporal trait changes of all predator and prey species and thus simultaneous coadaptation within and among trophic levels. We show that very small or slow trait adaptation did not facilitate coexistence because the stabilizing niche differences were not sufficient to offset the fitness differences. In contrast, sufficiently large and fast trait adaptation jointly promoted stable or neutrally stable species coexistence. Continuous trait adjustments in response to selection enabled a temporally variable convergence and divergence of species traits; that is, species became temporally more similar (neutral theory) or dissimilar (niche theory) depending on the selection pressure, resulting over time in a balance between niche differences stabilizing coexistence and fitness differences promoting competitive exclusion. Furthermore, coadaptation allowed prey and predator species to cluster into different functional groups. This equalized the fitness of similar species while maintaining sufficient niche differences among functionally different species delaying or preventing competitive exclusion. In contrast to pre- vious studies, the emergent feedback between biomass and trait dynamics enabled supersaturated coexistence for a broad range of potential trait adaptation and parameters. We conclude that accounting for trait adaptation may explain stable and supersaturated species coexistence for a broad range of environmental conditions in natural systems when the absence of such adaptive changes would preclude it. Small trait changes, coincident with those that may occur within many natural populations, greatly enlarged the number of coexisting species. KW - Coadaptation KW - equalizing and stabilizing mechanisms KW - maintenance of functional diversity KW - niche and fitness differences KW - supersaturated species coexistence KW - trait convergence and divergence Y1 - 2016 U6 - https://doi.org/10.1002/ece3.2172 SN - 2045-7758 PB - John Wiley & Sons, Inc. ER -