TY - JOUR A1 - Sharma, Neha A1 - Ruelens, Philip A1 - Maggen, Thomas A1 - Dochy, Niklas A1 - Torfs, Sanne A1 - Kaufmann, Kerstin A1 - Rohde, Antje A1 - Geuten, Koen T1 - A Flowering Locus C Homolog Is a Vernalization-Regulated Repressor in Brachypodium and Is Cold Regulated in Wheat JF - Plant physiology : an international journal devoted to physiology, biochemistry, cellular and molecular biology, biophysics and environmental biology of plants N2 - Winter cereals require prolonged cold to transition from vegetative to reproductive development. This process, referred to as vernalization, has been extensively studied in Arabidopsis (Arabidopsis thaliana). In Arabidopsis, a key flowering repressor called FLOWERING LOCUS C (FLC) quantitatively controls the vernalization requirement. By contrast, in cereals, the vernalization response is mainly regulated by the VERNALIZATION genes, VRN1 and VRN2. Here, we characterize ODDSOC2, a recently identified FLC ortholog in monocots, knowing that it belongs to the FLC lineage. By studying its expression in a diverse set of Brachypodium accessions, we find that it is a good predictor of the vernalization requirement. Analyses of transgenics demonstrated that BdODDSOC2 functions as a vernalization-regulated flowering repressor. In most Brachypodium accessions BdODDSOC2 is down-regulated by cold, and in one of the winter accessions in which this down-regulation was evident, BdODDSOC2 responded to cold before BdVRN1. When stably down-regulated, the mechanism is associated with spreading H3K27me3 modifications at the BdODDSOC2 chromatin. Finally, homoeolog-specific gene expression analyses identify TaAGL33 and its splice variant TaAGL22 as the FLC orthologs in wheat (Triticum aestivum) behaving most similar to Brachypodium ODDSOC2. Overall, our study suggests that ODDSOC2 is not only phylogenetically related to FLC in eudicots but also functions as a flowering repressor in the vernalization pathway of Brachypodium and likely other temperate grasses. These insights could prove useful in breeding efforts to refine the vernalization requirement of temperate cereals and adapt varieties to changing climates. Y1 - 2016 U6 - https://doi.org/10.1104/pp.16.01161 SN - 0032-0889 SN - 1532-2548 VL - 173 IS - 2 SP - 1301 EP - 1315 PB - American Society of Plant Physiologists CY - Rockville ER - TY - JOUR A1 - Lah, Ljerka A1 - Löber, Ulrike A1 - Hsiang, Tom A1 - Hartmann, Stefanie T1 - A genomic comparison of putative pathogenicity-related gene families in five members of the Ophiostomatales with different lifestyles JF - Fungal biology N2 - Ophiostomatoid fungi are vectored by their bark-beetle associates and colonize different host tree species. To survive and proliferate in the host, they have evolved mechanisms for detoxification and elimination of host defence compounds, efficient nutrient sequestration, and, in pathogenic species, virulence towards plants. Here, we assembled a draft genome of the spruce pathogen Ophiostoma bicolor. For our comparative and phylogenetic analyses, we mined the genomes of closely related species (Ophiostoma piceae, Ophiostoma ulmi, Ophiostoma novo-ulmi, and Grosmannia clavigera). Our aim was to acquire a genomic and evolutionary perspective of gene families important in host colonization. Genome comparisons showed that both the nuclear and mitochondrial genomes in our assembly were largely complete. Our O. bicolor 25.3 Mbp draft genome had 10 018 predicted genes, 6041 proteins with gene ontology (GO) annotation, 269 carbohydrate-active enzymes (CAZymes), 559 peptidases and inhibitors, and 1373 genes likely involved in pathogen-host interactions. Phylogenetic analyses of selected protein families revealed core sets of cytochrome P450 genes, ABC transporters and backbone genes involved in secondary metabolite (SM) biosynthesis (polyketide synthases (PKS) and non-ribosomal synthases), and species-specific gene losses and duplications. Phylogenetic analyses of protein families of interest provided insight into evolutionary adaptations to host biochemistry in ophiostomatoid fungi. KW - Bark beetle KW - Bluestain fungi KW - Ips typographus Y1 - 2016 U6 - https://doi.org/10.1016/j.funbio.2016.12.002 SN - 1878-6146 SN - 1878-6162 VL - 121 SP - 234 EP - 252 PB - Elsevier CY - Oxford ER - TY - GEN A1 - Dortay, Hakan A1 - Müller-Röber, Bernd T1 - A highly efficient pipeline for protein expression in Leishmania tarentolae using infrared fluorescence protein as marker N2 - Background: Leishmania tarentolae, a unicellular eukaryotic protozoan, has been established as a novel host for recombinant protein production in recent years. Current protocols for protein expression in Leishmania are, however, time consuming and require extensive lab work in order to identify well-expressing cell lines. Here we established an alternative protein expression work-flow that employs recently engineered infrared fluorescence protein (IFP) as a suitable and easy-to-handle reporter protein for recombinant protein expression in Leishmania. As model proteins we tested three proteins from the plant Arabidopsis thaliana, including a NAC and a type-B ARR transcription factor. Results: IFP and IFP fusion proteins were expressed in Leishmania and rapidly detected in cells by deconvolution microscopy and in culture by infrared imaging of 96-well microtiter plates using small cell culture volumes (2 mu L - 100 mu L). Motility, shape and growth of Leishmania cells were not impaired by intracellular accumulation of IFP. In-cell detection of IFP and IFP fusion proteins was straightforward already at the beginning of the expression pipeline and thus allowed early pre-selection of well-expressing Leishmania clones. Furthermore, IFP fusion proteins retained infrared fluorescence after electrophoresis in denaturing SDS-polyacrylamide gels, allowing direct in-gel detection without the need to disassemble cast protein gels. Thus, parameters for scaling up protein production and streamlining purification routes can be easily optimized when employing IFP as reporter. Conclusions: Using IFP as biosensor we devised a protocol for rapid and convenient protein expression in Leishmania tarentolae. Our expression pipeline is superior to previously established methods in that it significantly reduces the hands-on-time and work load required for identifying well-expressing clones, refining protein production parameters and establishing purification protocols. The facile in-cell and in-gel detection tools built on IFP make Leishmania amenable for high-throughput expression of proteins from plant and animal sources. T3 - Zweitveröffentlichungen der Universität Potsdam : Mathematisch-Naturwissenschaftliche Reihe - 366 KW - System KW - Donovani Y1 - 2017 U6 - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:kobv:517-opus4-400876 ER - TY - GEN A1 - Kocyan, Alexander A1 - Wiland-Szymanska, Justyna T1 - A new name and a new combination for Friedmannia nom. illeg. (Hypoxidaceae) T2 - Phytotaxa : a rapid international journal for accelerating the publication of botanical taxonomy N2 - Recently, Kocyan & Wiland-Szymańska (2016) have published a thorough research article on one of the outstanding members of the family Hypoxidaceae on the Seychelles, which resulted in the raise of a new genus (Friedmannia Kocyan & Wiland-Szymańska 2016: 60) to accommodate the former Curculigo seychellensis Bojer ex Baker (1877: 368). However, it has turned out that the name Friedmannia Chantanachat & Bold (1962: 45) already exists in literature for a green alga, which renders the new hypoxid genus illegitimate (Melbourne Code; McNeill et al. 2012). Therefore, we assign a new generic epithet to Curculigo seychellensis. Y1 - 2017 U6 - https://doi.org/10.11646/phytotaxa.291.3.10 SN - 1179-3155 SN - 1179-3163 VL - 291 IS - 3 SP - 239 EP - 239 PB - Magnolia Press CY - Auckland ER - TY - JOUR A1 - Van den Wyngaert, Silke A1 - Seto, Kensuke A1 - Rojas-Jimenez, Keilor A1 - Kagami, Maiko A1 - Grossart, Hans-Peter T1 - A New Parasitic Chytrid, Staurastromyces oculus (Rhizophydiales, Staurastromy-cetaceae fam. nov.), Infecting the Freshwater Desmid Staurastrum sp. JF - Protist N2 - Chytrids are a diverse group of ubiquitous true zoosporic fungi. The recent molecular discovery of a large diversity of undescribed chytrids has raised awareness on their important, but so far understudied ecological role in aquatic ecosystems. In the pelagic zone, of both freshwater and marine ecosystems, many chytrid species have been morphologically described as parasites on almost all major groups of phytoplankton. However, the majority of these parasitic chytrids has rarely been isolated and lack DNA sequence data, resulting in a large proportion of "dark taxa" in databases. Here, we report on the isolation and in-depth morphological, molecular and host range characterization of a chytrid infecting the common freshwater desmid Staurastrum sp. We provide first insights on the metabolic activity of the different chytrid development stages by using the vital dye FUN (R)-1 (2-chloro-4-[2,3-dihydro-3-methyl-[benzo-1,3-thiazol-2-yl]-methylidene]-1-phenylquinolinium iodide). Cross infection experiments suggest that this chytrid is an obligate parasite and specific for the genus Staurastrum sp. Phylogenetic analysis, based on ITS1-5.8S-ITS2 and 28S rDNA sequences, placed it in the order Rhizophydiales. Based on the unique zoospore ultrastructure, combined with thallus morphology, and molecular phylogenetic placement, we describe this parasitic chytrid as a new genus and species Staurastromyces oculus, within a new family Staurastromycetaceae. (C) 2017 Elsevier GmbH. All rights reserved. KW - Chytrids KW - parasite KW - phytoplankton KW - Staurastromyces oculus KW - Staurastrum sp. Y1 - 2017 U6 - https://doi.org/10.1016/j.protis.2017.05.001 SN - 1434-4610 VL - 168 SP - 392 EP - 407 PB - Elsevier CY - Jena ER - TY - JOUR A1 - Lütkecosmann, Steffi A1 - Warsinke, Axel A1 - Tschöpe, Winfried A1 - Eichler, Rüdiger A1 - Hanack, Katja T1 - A novel monoclonal antibody suitable for the detection of leukotriene B4 JF - Biochemical and biophysical research communications N2 - Leukotriene B4 as an inflammatory mediator is an important biomarker for different respiratory diseases like asthma, chronic obstructive pulmonary disease or cystic lung fibrosis. Therefore the detection of LTB4 is helpful in the diagnosis of these pulmonary diseases. However, until now its determination in exhaled breath condensates suffers from problems of accuracy. Reasons for that could be improper sample collection and preparation methods of condensates and the lack of consistently assay specificity and reproducibility of the used immunoassay detection system. In this study we describe the development and the characterization of a specific monoclonal antibody (S27BC6) against LTB4, its use as molecular recognition element for the development of an enzyme-linked immunoassay to detect LTB4 and discuss possible future diagnostic applications. KW - Leukotriene B4 KW - Monoclonal antibody KW - Immunosensor KW - Chronic obstructive pulmonary disease (COPD) KW - Hapten Y1 - 2017 U6 - https://doi.org/10.1016/j.bbrc.2016.11.157 SN - 0006-291X SN - 1090-2104 VL - 482 IS - 4 SP - 1054 EP - 1059 PB - Elsevier CY - San Diego ER - TY - JOUR A1 - Hoehn, Richard S. A1 - Jernigan, Peter L. A1 - Japtok, Lukasz A1 - Chang, Alex L. A1 - Midura, Emily F. A1 - Caldwell, Charles C. A1 - Kleuser, Burkhard A1 - Lentsch, Alex B. A1 - Edwards, Michael J. A1 - Gulbins, Erich A1 - Pritts, Timothy A. T1 - Acid sphingomyelinase inhibition in stored erythrocytes reduces transfusion-associated lung inflammation JF - Annals of surgery : a monthly review of surgical science and practice N2 - Objective: We aimed to identify the role of the enzyme acid sphingomyelinase in the aging of stored units of packed red blood cells (pRBCs) and subsequent lung inflammation after transfusion. Summary Background Data: Large volume pRBC transfusions are associated with multiple adverse clinical sequelae, including lung inflammation. Microparticles are formed in stored pRBCs over time and have been shown to contribute to lung inflammation after transfusion. Methods: Human and murine pRBCs were stored with or without amitriptyline, a functional inhibitor of acid sphingomyelinase, or obtained from acid sphingomyelinase-deficient mice, and lung inflammation was studied in mice receiving transfusions of pRBCs and microparticles isolated from these units. Results: Acid sphingomyelinase activity in pRBCs was associated with the formation of ceramide and the release of microparticles. Treatment of pRBCs with amitriptyline inhibited acid sphingomyelinase activity, ceramide accumulation, and microparticle production during pRBC storage. Transfusion of aged pRBCs or microparticles isolated from aged blood into mice caused lung inflammation. This was attenuated after transfusion of pRBCs treated with amitriptyline or from acid sphingomyelinase-deficient mice. Conclusions: Acid sphingomyelinase inhibition in stored pRBCs offers a novel mechanism for improving the quality of stored blood. KW - acid sphingomyelinase KW - blood banking KW - ceramide KW - lung inflammation KW - microparticle Y1 - 2017 U6 - https://doi.org/10.1097/SLA.0000000000001648 SN - 0003-4932 SN - 1528-1140 VL - 265 IS - 1 SP - 218 EP - 226 PB - Lippincott Williams & Wilkins CY - Philadelphia ER - TY - JOUR A1 - McVey, Mark J. A1 - Kim, Michael A1 - Tabuchi, Arata A1 - Srbely, Victoria A1 - Japtok, Lukasz A1 - Arenz, Christoph A1 - Rotstein, Ori A1 - Kleuser, Burkhard A1 - Semple, John W. A1 - Kuebler, Wolfgang M. T1 - Acid sphingomyelinase mediates murine acute lung injury following transfusion of aged platelets JF - American journal of physiology : Lung cellular and molecular physiology N2 - Pulmonary complications from stored blood products are the leading cause of mortality related to transfusion. Transfusion-related acute lung injury is mediated by antibodies or bioactive mediators, yet underlying mechanisms are incompletely understood. Sphingolipids such as ceramide regulate lung injury, and their composition changes as a function of time in stored blood. Here, we tested the hypothesis that aged platelets may induce lung injury via a sphingolipid-mediated mechanism. To assess this hypothesis, a two-hit mouse model was devised. Recipient mice were treated with 2 mg/kg intraperitoneal lipopolysaccharide (priming) 2 h before transfusion of 10 ml/kg stored (1-5 days) platelets treated with or without addition of acid sphingomyelinase inhibitor ARC39 or platelets from acid sphingomyelinase-deficient mice, which both reduce ceramide formation. Transfused mice were examined for signs of pulmonary neutrophil accumulation, endothelial barrier dysfunction, and histological evidence of lung injury. Sphingolipid profiles in stored platelets were analyzed by mass spectrophotometry. Transfusion of aged platelets into primed mice induced characteristic features of lung injury, which increased in severity as a function of storage time. Ceramide accumulated in platelets during storage, but this was attenuated by ARC39 or in acid sphingomyelinase-deficient platelets. Compared with wild-type platelets, transfusion of ARC39-treated or acid sphingomyelinase-deficient aged platelets alleviated lung injury. Aged platelets elicit lung injury in primed recipient mice, which can be alleviated by pharmacological inhibition or genetic deletion of acid sphingomyelinase. Interventions targeting sphingolipid formation represent a promising strategy to increase the safety and longevity of stored blood products. KW - transfusion-related acute lung injury KW - ceramide KW - acid sphingomyelinase KW - platelets KW - storage Y1 - 2017 U6 - https://doi.org/10.1152/ajplung.00317.2016 SN - 1040-0605 SN - 1522-1504 VL - 312 IS - 5 SP - 625 EP - 637 PB - American Physiological Society CY - Bethesda ER - TY - JOUR A1 - Zou, Jie A1 - Wang, Weiwei A1 - Neffe, Axel T. A1 - Xu, Xun A1 - Li, Zhengdong A1 - Deng, Zijun A1 - Sun, Xianlei A1 - Ma, Nan A1 - Lendlein, Andreas T1 - Adipogenic differentiation of human adipose derived mesenchymal stem cells in 3D architectured gelatin based hydrogels (ArcGel) JF - Clinical hemorheology and microcirculation : blood flow and vessels N2 - Polymeric matrices mimicking multiple functions of the ECM are expected to enable a material induced regeneration of tissues. Here, we investigated the adipogenic differentiation of human adipose derived mesenchymal stem cells (hADSCs) in a 3D architectured gelatin based hydrogel (ArcGel) prepared from gelatin and L-lysine diisocyanate ethyl ester (LDI) in an one-step process, in which the formation of an open porous morphology and the chemical network formation were integrated. The ArcGel was designed to support adipose tissue regeneration with its 3D porous structure, high cell biocompatibility, and mechanical properties compatible with human subcutaneous adipose tissue. The ArcGel could support initial cell adhesion and survival of hADSCs. Under static culture condition, the cells could migrate into the inner part of the scaffold with a depth of 840 +/- 120 mu m after 4 days, and distributed in the whole scaffold (2mm in thickness) within 14 days. The cells proliferated in the scaffold and the fold increase of cell number after 7 days of culture was 2.55 +/- 0.08. The apoptotic rate of hADSCs in the scaffold was similar to that of cells maintained on tissue culture plates. When cultured in adipogenic induction medium, the hADSCs in the scaffold differentiated into adipocytes with a high efficiency (93 +/- 1%). Conclusively, this gelatin based 3D scaffold presented high cell compatibility for hADSC cultivation and differentiation, which could serve as a potential implant material in clinical applications for adipose tissue reparation and regeneration. KW - Mesenchymal stem cells KW - gelatin based scaffold KW - adipose tissue regeneration KW - adipogenic differentiation Y1 - 2017 U6 - https://doi.org/10.3233/CH-179210 SN - 1386-0291 SN - 1875-8622 VL - 67 IS - 3-4 SP - 297 EP - 307 PB - IOS Press CY - Amsterdam ER - TY - JOUR A1 - Heunisch, Fabian A1 - Chaykovska, Lyubov A1 - von Einem, Gina A1 - Alter, Markus A1 - Dschietzig, Thomas A1 - Kretschmer, Axel A1 - Kellner, Karl-Heinz A1 - Hocher, Berthold T1 - ADMA predicts major adverse renal events in patients with mild renal impairment and/or diabetes mellitus undergoing coronary angiography JF - Medicine N2 - Asymmetric dimethylarginine (ADMA) is a competitive inhibitor of the nitric oxide (NO)-synthase and a biomarker of endothelial dysfunction (ED). ED plays an important role in the pathogenesis of contrast-induced nephropathy (CIN). The aim of our study was to evaluate serum ADMA concentration as a biomarker of an acute renal damage during the follow-up of 90 days after contrast medium (CM) application. Blood samples were obtained from 330 consecutive patients with diabetes mellitus or mild renal impairment immediately before, 24 and 48 hours after the CM application for coronary angiography. The patients were followed for 90 days. The composite endpoints were major adverse renal events (MARE) defined as occurrence of death, initiation of dialysis, or a doubling of serum creatinine concentration. Overall, ADMA concentration in plasma increased after CM application, although, there was no differences between ADMA levels in patients with and without CIN. ADMA concentration 24 hours after the CM application was predictive for dialysis with a specificity of 0.889 and sensitivity of 0.653 at values higher than 0.71 mu mol/L (area under the curve: 0.854, 95% confidential interval: 0.767-0.941, P<0.001). This association remained significant in multivariate Cox regression models adjusted for relevant factors of long-term renal outcome. 24 hours after the CM application, ADMA concentration in plasma was predictive for MARE with a specificity of 0.833 and sensitivity of 0.636 at a value of more than 0.70 mu mol/L (area under the curve: 0.750, 95% confidence interval: 0.602-0.897, P=0.004). Multivariate logistic regression analysis confirmed that ADMA and anemia were significant predictors of MARE. Further analysis revealed that increased ADMA concentration in plasma was highly significant predictor of MARE in patients with CIN. Moreover, patients with CIN and MARE had the highest plasma ADMA levels 24 hours after CM exposure in our study cohort. The impact of ADMA on MARE was independent of such known CIN risk factors as anemia, pre-existing renal failure, pre-existing heart failure, and diabetes. ADMA concentration in plasma is a promising novel biomarker of major contrast-induced nephropathy-associated events 90 days after contrast media exposure. KW - asymmetric dimethylarginine (ADMA) KW - biomarkers of renal failure KW - contrast-induced nephropathy Y1 - 2017 U6 - https://doi.org/10.1097/MD.0000000000006065 SN - 0025-7974 SN - 1536-5964 VL - 96 IS - 6 PB - Lippincott Williams & Wilkins CY - Philadelphia ER - TY - GEN A1 - Koziel, Slawomir A1 - Hermanussen, Michael A1 - Gomula, Alexandra A1 - Swanson, James A1 - Kaczmarek, Maria A1 - El-Shabrawi, Mortada A1 - Elhusseini, Mona A1 - Satake, Takashi A1 - Martinovic Klaric, Irena A1 - Scheffler, Christiane A1 - Morkuniene, Ruta A1 - Godina, Elena A1 - Sasa, Missoni A1 - Tutkuviene, Janina A1 - Siniarska, Anna A1 - Nieczuja-Dwojacka, Joanna A1 - Nunez, Javier A1 - Groth, Detlef A1 - Barbieri, Davide T1 - Adolescence - a Transition to Adulthood Proceedings of the 24th Aschauer Soiree, held at Jurata, Poland, November 5th 2016 T2 - Pediatric Endocrinology Reviews N2 - Eighteen scientists met at Jurata, Poland, to discuss various aspects of the transition from adolescence to adulthood. This transition is a delicate period facing complex interactions between the adolescents and the social group they belong to. Social identity, group identification and identity signalling, but also stress affecting basal salivary cortisol rhythms, hypertension, inappropriate nutrition causing latent and manifest obesity, moreover, in developing and under-developed countries, parasitosis causing anaemia thereby impairing growth and development, are issues to be dealt with during this period of the human development. In addition, some new aspects of the association between weight, height and head circumference in the newborns were discussed, as well as intrauterine head growth and head circumference as health risk indicators. KW - Strategic growth adjustment KW - BMI KW - Growth faltering KW - Secular trend KW - Obesity KW - Growth modelling Y1 - 2017 SN - 1565-4753 VL - 14 IS - 3 SP - 326 EP - 334 PB - Medical Media CY - Netanya ER - TY - JOUR A1 - Fischbach, Jens A1 - Loh, Qiuting A1 - Bier, Frank Fabian A1 - Lim, Theam Soon A1 - Frohme, Marcus A1 - Glökler, Jörn T1 - Alizarin Red S for Online Pyrophosphate Detection Identified by a Rapid Screening Method JF - Scientific reports N2 - We identified Alizarin Red S and other well known fluorescent dyes useful for the online detection of pyrophosphate in enzymatic assays, including the loop mediated isothermal amplification (LAMP) and polymerase chain reaction (PCR) assays. An iterative screening was used for a selected set of compounds to first secure enzyme compatibility, evaluate inorganic pyrophosphate sensitivity in the presence of manganese as quencher and optimize conditions for an online detection. Of the selected dyes, the inexpensive alizarin red S was found to selectively detect pyrophosphate under LAMP and PCR conditions and is superior with respect to its defined red-shifted spectrum, long shelf life and low toxicity. In addition, the newly identified properties may also be useful in other enzymatic assays which do not generate nucleic acids but are based on inorganic pyrophosphate. Finally, we propose that our screening method may provide a blueprint for rapid screening of compounds for detecting inorganic pyrophosphate. Y1 - 2017 U6 - https://doi.org/10.1038/srep45085 SN - 2045-2322 VL - 7 PB - Nature Publ. Group CY - London ER - TY - JOUR A1 - Salleh, Faezah Mohd A1 - Ramos-Madrigal, Jazmin A1 - Penaloza, Fernando A1 - Liu, Shanlin A1 - Sinding, Mikkel-Holger S. A1 - Patel, Riddhi P. A1 - Martins, Renata A1 - Lenz, Dorina A1 - Fickel, Jörns A1 - Roos, Christian A1 - Shamsir, Mohd Shahir A1 - Azman, Mohammad Shahfiz A1 - Lim, Burton K. A1 - Rossiter, Stephen J. A1 - Wilting, Andreas A1 - Gilbert, M. Thomas P. T1 - An expanded mammal mitogenome dataset from Southeast Asia JF - Gigascience N2 - Background: Findings: Approximately 55 gigabases of raw sequence were generated. From this data we assembled 72 complete mitogenome sequences, with an average depth of coverage of 102.9x and 55.2x for modern samples and historical samples, respectively. This dataset represents 52 species, of which 30 species had no previous mitogenome data available. The mitogenomes were geotagged to their sampling location, where known, to display a detailed geographical distribution of the species. Conclusion: KW - invertebrate-derived (iDNA) KW - metabarcoding KW - GenBank KW - Taxonomic assignment Y1 - 2017 SN - 2047-217X VL - 6 IS - 8 SP - 1 EP - 19 PB - Oxford Univ. Press CY - Oxford ER - TY - JOUR A1 - Thomas, Jessica E. A1 - Carvalho, Gary R. A1 - Haile, James A1 - Martin, Michael D. A1 - Castruita, Jose A. Samaniego A1 - Niemann, Jonas A1 - Sinding, Mikkel-Holger S. A1 - Sandoval-Velasco, Marcela A1 - Rawlence, Nicolas J. A1 - Fuller, Errol A1 - Fjeldsa, Jon A1 - Hofreiter, Michael A1 - Stewart, John R. A1 - Gilbert, M. Thomas P. A1 - Knapp, Michael T1 - An ‛Aukward’ tale BT - a genetic approach to discover the whereabouts of the Last Great Auks JF - Genes N2 - One hundred and seventy-three years ago, the last two Great Auks, Pinguinus impennis, ever reliably seen were killed. Their internal organs can be found in the collections of the Natural History Museum of Denmark, but the location of their skins has remained a mystery. In 1999, Great Auk expert Errol Fuller proposed a list of five potential candidate skins in museums around the world. Here we take a palaeogenomic approach to test which—if any—of Fuller’s candidate skins likely belong to either of the two birds. Using mitochondrial genomes from the five candidate birds (housed in museums in Bremen, Brussels, Kiel, Los Angeles, and Oldenburg) and the organs of the last two known individuals, we partially solve the mystery that has been on Great Auk scholars’ minds for generations and make new suggestions as to the whereabouts of the still-missing skin from these two birds. KW - ancient DNA KW - extinct birds KW - mitochondrial genome KW - museum specimens KW - palaeogenomics Y1 - 2017 U6 - https://doi.org/10.3390/genes8060164 SN - 2073-4425 VL - 8 IS - 6 SP - 164 PB - MDPI CY - Basel ER - TY - THES A1 - Radon, Christin T1 - Analyse der Funktion der dualen Lokalisation der 3-Mercaptopyruvat Sulfurtransferase im Menschen Y1 - 2017 ER - TY - JOUR A1 - Czernitzki, Anna-Franziska A1 - Pospisil, Christina A1 - Musalek, Martin A1 - Mumm, Rebekka A1 - Scheffler, Christiane T1 - Analysis of longitudinal data of height z-scores in kindergarten children BT - a pilot study JF - Journal of biological and clinical anthropology : Anthropologischer Anzeiger ; Mitteilungsorgan der Gesellschaft für Anthropologie N2 - Changes in body height throughout extended historic periods are very complex and dynamic processes. Thispilot study aimed to investigate the pattern of longitudinal height z-scores changes in children before and after entering kindergarten. In summer 2016, we measured height and weight of 32 children from 4 groups of two kindergartens aged 3–6 years. All ages were centered according to the age of entry into the kindergarten. For each child we determined mean z-scores for height before and after entering the kindergarten, and assessed the variances for each kindergarten group. Twenty-two children targeted in height z-scores towards average height of their respective kindergarten group, 10 children did not. Due to the small numbers, the convergence in height variance however, remained insignificant (chi-squared independence test, p = 0.127). Additional studies with larger sample sizes are needed to confirm this pilot study. KW - Height z-score KW - kindergarten children KW - secular trend KW - strategic growth adjustment KW - social signal Y1 - 2017 U6 - https://doi.org/10.1127/anthranz/2017/0708 SN - 0003-5548 VL - 74 IS - 2 SP - 109 EP - 112 PB - Schweizerbart science publishers CY - Stuttgart ER - TY - THES A1 - Diez Cocero, Mercedes T1 - Analysis of Rubisco - carbonic anhydrase fusions in tobacco as an approach to reduce photorespiration Y1 - 2017 ER - TY - THES A1 - Diez Cocero, Mercedes T1 - Analysis of Rubisco – carbonic anhydrase fusions in tobacco as an approach to reduce photorespiration N2 - Rubisco catalyses the first step of CO2 assimilation into plant biomass. Despite its crucial role, it is notorious for its low catalytic rate and its tendency to fix O2 instead of CO2, giving rise to a toxic product that needs to be recycled in a process known as photorespiration. Since almost all our food supply relies on Rubisco, even small improvements in its specificity for CO2 could lead to an improvement of photosynthesis and ultimately, crop yield. In this work, we attempted to improve photosynthesis by decreasing photorespiration with an artificial CCM based on a fusion between Rubisco and a carbonic anhydrase (CA). A preliminary set of plants contained fusions between one of two CAs, bCA1 and CAH3, and the N- or C-terminus of RbcL connected by a small flexible linker of 5 amino acids. Subsequently, further fusion proteins were created between RbcL C-terminus and bCA1/CAH3 with linkers of 14, 23, 32, and 41 amino acids. The transplastomic tobacco plants carrying fusions with bCA1 were able to grow autotrophically even with the shortest linkers, albeit at a low rate, and accumulated very low levels of the fusion protein. On the other hand, plants carrying fusions with CAH3 were autotrophic only with the longer linkers. The longest linker permitted nearly wild-type like growth of the plants carrying fusions with CAH3 and increased the levels of fusion protein, but also of smaller degradation products. The fusion of catalytically inactive CAs to RbcL did not cause a different phenotype from the fusions with catalytically active CAs, suggesting that the selected CAs were not active in the fusion with RbcL or their activity did not have an effect on CO2 assimilation. However, fusions to RbcL did not abolish RbcL catalytic activity, as shown by the autotrophic growth, gas exchange and in vitro activity measurements. Furthermore, Rubisco carboxylation rate and specificity for CO2 was not altered in some of the fusion proteins, suggesting that despite the defect in RbcL folding or assembly caused by the fusions, the addition of 60-150 amino acids to RbcL does not affect its catalytic properties. On the contrary, most growth defects of the plants carrying RbcL-CA fusions are related to their reduced Rubisco content, likely caused by impaired RbcL folding or assembly. Finally, we found that fusions with RbcL C-terminus were better tolerated than with the N-terminus, and increasing the length of the linker relieved the growth impairment imposed by the fusion to RbcL. Together, the results of this work constitute considerable relevant findings for future Rubisco engineering. N2 - Rubisco katalysiert den ersten Schritt der CO2-Assimilierung. Trotz seiner bedeutenden Rolle, zeichnet sich Rubisco durch eine niedrige katalytische Geschwindigkeit aus. Außerdem, entsteht bei der Bindung von O2 anstatt CO2 ein toxisches Zwischenprodukt, welches in einem Prozess, genannt Photorespiration, aufbereitet wird. Da fast die gesamte Nahrungsmittelversorgung auf der Aktivität von Rubisco basiert, könnten schon kleine Verbesserungen in der Spezifität für CO2 zu einem großen Effekt in der Photosysntheserate und letztendlich größeren Ernteerträgen führen. In dieser Arbeit wurde versucht die Effizienz der Photosynthese zu verbessern, indem ein künstlicher CO2 konzentrierender Mechanismus aus einer Fusion von RbcL und einer Carboanhydrase (CA) gebildet wird. Als Vorversuch wurden je bCA1 und CAH3 an Rubiscos C- beziehungsweise N-Terminus mittels eines kleinen, flexiblen Linkers aus 5 Aminosäuren fusioniert. Anschließend wurden weitere Fusionsproteine zwischen dem C-Terminus von RbcL und bCA1/CAH3 mittels Linkern von 14, 23, 32 und 41 Aminosäuren Länge in Chloroplasten von Tabak eingebracht. Die entstandenen transplastomischen Pflanzen mit bCA1-Fusionen waren trotz ihres sehr langsamen Wachstums dazu fähig schon bei kurzen Linkern autotroph zu wachsen und geringe Mengen an Fusionsproteinen zu akkumulieren. Pflanzen mit CAH3 Fusionsproteinen hingegen waren nur mit längeren Linkern autotroph, zeigten aber dafür ähnliche Wachstumsraten zum Wildtyp bei Nutzung des längsten Linkers. Außerdem enthielten diese Pflanzen größere Mengen an Fusionsproteinen aber auch eine erhöhte Anreicherung von kleineren Abbauprodukten. Bei den in dieser Arbeit gewählten CA als Fusionsprotein mit RbcL konnte im Vergleich mit katalytisch inaktiven Varianten kein Effekt auf die CO2-Assimilierung gefunden werden. Wie das autotroph Wachstum sowie die Gaswechsel- und in-vitro-Aktivitätsmessungen zeigen, haben die Fusionen allerdings nicht die katalytische Aktivität von Rubisco blockiert. Ebenso verhielt sich die Carboxylierungsrate von Rubisco und deren Spezifität für CO2 unverändert. Dies weist darauf hin, dass trotz Rubiscos Faltungs- oder Assemblierungsdefekten das Anfügen von 60-150 Aminosäure an den C-Terminus von RbcL nicht die katalytische Leistung des Enzyms beeinträchtigt. Im Gegenteil, die Wachstumsdefekte waren durch die geringe Menge an Rubisco begründet, vermutlich verursacht durch Defekte in der Faltung oder Assemblierung von RbcL. Schlussendlich konnten wichtige Erkenntnisse für zukünftige gentechnische Veränderungen von Rubisco gemacht werden: Fusionen mit dem C-Terminus von RbcL wurden besser toleriert als mit dem N-Terminus und längere Linker verringerten die von der Fusion ausgelösten Wachstumsdefekte. KW - Rubisco KW - fusion Y1 - 2017 ER - TY - THES A1 - Bajdzienko, Krzysztof T1 - Analysis of Target of Rapamycin (Tor) induced changes of the Arabidopsis thaliana proteome using sub-cellular resolution Y1 - 2017 ER - TY - THES A1 - Neuber, Corinna T1 - Analytik zur Biotransformation des Sphingosin 1-phosphat-abbauproduktes (2E)-Hexadecenal Y1 - 2017 ER - TY - JOUR A1 - Kang, Mi-Sun A1 - Lim, Hae-Soon A1 - Oh, Jong-Suk A1 - Lim, You-jin A1 - Wuertz-Kozak, Karin A1 - Harro, Janette M. A1 - Shirtliff, Mark E. A1 - Achermann, Yvonne T1 - Antimicrobial activity of Lactobacillus salivarius and Lactobacillus fermentum against Staphylococcus aureus JF - Pathogens and disease / Federation of European Microbiology Societies N2 - The increasing prevalence of methicillin-resistant Staphylococcus aureus has become a major public health threat. While lactobacilli were recently found useful in combating various pathogens, limited data exist on their therapeutic potential for S. aureus infections. The aim of this study was to determine whether Lactobacillus salivarius was able to produce bactericidal activities against S. aureus and to determine whether the inhibition was due to a generalized reduction in pH or due to secreted Lactobacillus product(s). We found an 8.6-log10 reduction of planktonic and a 6.3-log10 reduction of biofilm S. aureus. In contrast, the previously described anti-staphylococcal effects of L. fermentum only caused a 4.0-log10 reduction in planktonic S. aureus cells, with no effect on biofilm S. aureus cells. Killing of S. aureus was partially pH dependent, but independent of nutrient depletion. Cell-free supernatant that was pH neutralized and heat inactivated or proteinase K treated had significantly reduced killing of L. salivarius than with pH-neutralized supernatant alone. Proteomic analysis of the L. salivarius secretome identified a total of five secreted proteins including a LysM-containing peptidoglycan binding protein and a protein peptidase M23B. These proteins may represent potential novel anti-staphylococcal agents that could be effective against S. aureus biofilms. KW - antibacterial activity KW - biofilm KW - Lactobacillus fermentum KW - Lactobacillus salivarius KW - LysM KW - Staphylococcus aureus Y1 - 2017 U6 - https://doi.org/10.1093/femspd/ftx009 SN - 2049-632X VL - 75 IS - 2 PB - Oxford Univ. Press CY - Oxford ER - TY - JOUR A1 - Shahnejat-Bushehri, Sara A1 - Allu, Annapurna Devi A1 - Mehterov, Nikolay A1 - Thirumalaikumar, Venkatesh P. A1 - Alseekh, Saleh A1 - Fernie, Alisdair R. A1 - Mueller-Roeber, Bernd A1 - Balazadeh, Salma T1 - Arabidopsis NAC Transcription Factor JUNGBRUNNEN1 Exerts Conserved Control Over Gibberellin and Brassinosteroid Metabolism and Signaling Genes in Tomato JF - Frontiers in plant science N2 - The Arabidopsis thaliana NAC transcription factor JUNGBRUNNEN1 (AtJUB1) regulates growth by directly repressing GA3ox1 and DWF4, two key genes involved in gibberellin (GA) and brassinosteroid (BR) biosynthesis, respectively, leading to GA and BR deficiency phenotypes. AtJUB1 also reduces the expression of PIF4, a bHLH transcription factor that positively controls cell elongation, while it stimulates the expression of DELLA genes, which are important repressors of growth. Here, we extend our previous findings by demonstrating that AtJUB1 induces similar GA and BR deficiency phenotypes and changes in gene expression when overexpressed in tomato (Solanum lycopersicum). Importantly, and in accordance with the growth phenotypes observed, AtJUB1 inhibits the expression of growth-supporting genes, namely the tomato orthologs of GA3ox1, DWF4 and PIF4, but activates the expression of DELLA orthologs, by directly binding to their promoters. Overexpression of AtJUB1 in tomato delays fruit ripening, which is accompanied by reduced expression of several ripeningrelated genes, and leads to an increase in the levels of various amino acids (mostly proline, beta-alanine, and phenylalanine), gamma-aminobutyric acid (GABA), and major organic acids including glutamic acid and aspartic acid. The fact that AtJUB1 exerts an inhibitory effect on the GA/BR biosynthesis and PIF4 genes but acts as a direct activator of DELLA genes in both, Arabidopsis and tomato, strongly supports the model that the molecular constituents of the JUNGBRUNNEN1 growth control module are considerably conserved across species. KW - Arabidopsis KW - tomato KW - fruit KW - growth KW - transcription factor KW - gibberellic acid KW - brassinosteroid KW - DELLA proteins Y1 - 2017 U6 - https://doi.org/10.3389/fpls.2017.00214 SN - 1664-462X VL - 8 PB - Frontiers Research Foundation CY - Lausanne ER - TY - JOUR A1 - Nakamura, Moritaka A1 - Claes, Andrea R. A1 - Grebe, Tobias A1 - Hermkes, Rebecca A1 - Viotti, Corrado A1 - Ikeda, Yoshihisa A1 - Grebe, Markus T1 - Auxin and ROP GTPase Signaling of Polar Nuclear Migration in Root Epidermal Hair Cells JF - Plant physiology : an international journal devoted to physiology, biochemistry, cellular and molecular biology, biophysics and environmental biology of plants N2 - Polar nuclear migration is crucial during the development of diverse eukaryotes. In plants, root hair growth requires polar nuclear migration into the outgrowing hair. However, knowledge about the dynamics and the regulatory mechanisms underlying nuclear movements in root epidermal cells remains limited. Here, we show that both auxin and Rho-of-Plant (ROP) signaling modulate polar nuclear position at the inner epidermal plasma membrane domain oriented to the cortical cells during cell elongation as well as subsequent polar nuclear movement to the outer domain into the emerging hair bulge in Arabidopsis (Arabidopsis thaliana). Auxin signaling via the nuclear AUXIN RESPONSE FACTOR7 (ARF7)/ARF19 and INDOLE ACETIC ACID7 pathway ensures correct nuclear placement toward the inner membrane domain. Moreover, precise inner nuclear placement relies on SPIKE1 Rho-GEF, SUPERCENTIPEDE1 Rho-GDI, and ACTIN7 (ACT7) function and to a lesser extent on VTI11 vacuolar SNARE activity. Strikingly, the directionality and/or velocity of outer polar nuclear migration into the hair outgrowth along actin strands also are ACT7 dependent, auxin sensitive, and regulated by ROP signaling. Thus, our findings provide a founding framework revealing auxin and ROP signaling of inner polar nuclear position with some contribution by vacuolar morphology and of actin-dependent outer polar nuclear migration in root epidermal hair cells. Y1 - 2017 U6 - https://doi.org/10.1104/pp.17.00713 SN - 0032-0889 SN - 1532-2548 VL - 176 IS - 1 SP - 378 EP - 391 PB - American Society of Plant Physiologists CY - Rockville ER - TY - JOUR A1 - Leimkühler, Silke A1 - Lemaire, Olivier N. A1 - Iobbi-Nivol, Chantal T1 - Bacterial Molybdoenzymes BT - Chaperones, Assembly and Insertion JF - Molybdenum and tungsten enzymes : biochemistry N2 - The biogenesis of molybdoenzymes is a cytoplasmic event requiring both the folded apoenzymes and the matured molybdenum cofactor. The structure and the complexity of the molybdenum cofactor varies in each molybdoenzyme family and consequently different accessory proteins are required for the maturation of the respective enzymes. Thus, for enzymes of both the DMSO reductase and xanthine oxidase families, specific chaperones exist which are dedicated to increase the stability and the folding of specific members of each family. In this review, we describe the role of these chaperones for molybdoenzyme maturation. We present a model which describes step by step the mechanism of the maturation of representative molybdoenzymes from each family. Y1 - 2016 SN - 978-1-78262-391-5 SN - 978-1-78262-089-1 U6 - https://doi.org/10.1039/9781782623915-00117 VL - 5 SP - 117 EP - 142 PB - Royal Society of Chemistry CY - Cambridge ER - TY - GEN 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 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. T3 - Zweitveröffentlichungen der Universität Potsdam : Mathematisch-Naturwissenschaftliche Reihe - 313 KW - Bacteriophage KW - Flow cytometry KW - O-antigen KW - O-serotyping KW - Phase variation KW - Salmonella Typhimurium KW - Tailspike protein Y1 - 2017 U6 - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:kobv:517-opus4-103769 ER - TY - JOUR A1 - Bendjeddou, Mohammed Lamine A1 - Loumassine, Hibat Allah A1 - Scheffler, Ingo A1 - Bouslama, Zihad A1 - Amr, Zuhair T1 - Bat ectoparasites (Nycteribiidae, Streblidae, Siphonaptera, Heteroptera, Mesostigmata, Argasidae, and Ixodidae) from Algeria JF - Journal of Vector Ecology N2 - Twenty two species of ectoparasites (Family Nycteribiidae: Nycteribia (Listropoda) schmidlii schmidlii, Nycteribia (Nycteribia) latreillii, Nycteribia (Nycteribia) pedicularia, Penicillidia (Penicillidia) dufourii, and Phthiridium biarticulatum; Family Streblidae: Brachytarsina (Brachytarsina) flavipennis and Raymondia huberi; Order Siphonaptera: Rhinolophopsylla unipectinata arabs, Nycteridopsylla longiceps, Araeopsylla gestroi, Ischnopsyllus intermedius, and Ischnopsyllus octactenus; Order Heteroptera: Cimex pipistrelli, Cimex lectularius, and Cacodmus vicinus; Class Arachnida: Order Mesostigmata: Spinturnix myoti and Eyndhovenia euryalis; Order Ixodida: Family Argasidae: Argas transgariepinus and Argas vespertilionis; Family Ixodidae: Hyalomma dromedarii, Ixodes ricinus, and Ixodes vespertilionis) were recovered from 19 bat species in Algeria. New host records for bats are recorded for the first time: N. schmidlii from Rh. clivosus and R. cystops; N. latreillii from Rh. blasii and P. gaisleri; R. huberi from Rh. clivosus; C. pipistrelli from E. isabellinus and H. savii; C. vicinus from E. isabellinus; S. myoti from P. gaisleri; E. euryalis from P. gaisleri and Rh. blasii; A. vespertilionis from P. gaisleri; I. ricinus from T. teniotis and Rh. hipposideros and H. dromedarii from P. kuhlii. Raymondia huberi is recorded for the first time from Algeria. KW - Bats KW - Nycteribiidae KW - Streblidae KW - Siphonaptera KW - Heteroptera KW - Mesostigmata KW - Argasidae KW - Ixodidae KW - Algeria Y1 - 2017 U6 - https://doi.org/10.1111/jvec.12235 SN - 1948-7134 VL - 42 SP - 13 EP - 23 PB - Wiley Interscience CY - Hoboken, NJ ER - TY - JOUR A1 - Meyer, Sebastian Tobias A1 - Ptacnik, Robert A1 - Hillebrand, Helmut A1 - Bessler, Holger A1 - Buchmann, Nina A1 - Ebeling, Anne A1 - Eisenhauer, Nico A1 - Engels, Christof A1 - Fischer, Markus A1 - Halle, Stefan A1 - Klein, Alexandra-Maria A1 - Oelmann, Yvonne A1 - Roscher, Christiane A1 - Rottstock, Tanja A1 - Scherber, Christoph A1 - Scheu, Stefan A1 - Schmid, Bernhard A1 - Schulze, Ernst-Detlef A1 - Temperton, Vicky M. A1 - Tscharntke, Teja A1 - Voigt, Winfried A1 - Weigelt, Alexandra A1 - Wilcke, Wolfgang A1 - Weisser, Wolfgang W. T1 - Biodiversity-multifunctionality relationships depend on identity and number of measured functions JF - Nature Ecology & Evolution N2 - Biodiversity ensures ecosystem functioning and provisioning of ecosystem services, but it remains unclear how biodiversity-ecosystem multifunctionality relationships depend on the identity and number of functions considered. Here, we demonstrate that ecosystem multifunctionality, based on 82 indicator variables of ecosystem functions in a grassland biodiversity experiment, increases strongly with increasing biodiversity. Analysing subsets of functions showed that the effects of biodiversity on multifunctionality were stronger when more functions were included and that the strength of the biodiversity effects depended on the identity of the functions included. Limits to multifunctionality arose from negative correlations among functions and functions that were not correlated with biodiversity. Our findings underline that the management of ecosystems for the protection of biodiversity cannot be replaced by managing for particular ecosystem functions or services and emphasize the need for specific management to protect biodiversity. More plant species from the experimental pool of 60 species contributed to functioning when more functions were considered. An individual contribution to multifunctionality could be demonstrated for only a fraction of the species. Y1 - 2017 U6 - https://doi.org/10.1038/s41559-017-0391-4 SN - 2397-334X VL - 2 IS - 1 SP - 44 EP - 49 PB - Nature Publ. Group CY - London ER - TY - JOUR A1 - Neumann, Bettina A1 - Kielb, Patrycja A1 - Rustam, Lina A1 - Fischer, Anna A1 - Weidinger, Inez M. A1 - Wollenberger, Ulla T1 - Bioelectrocatalytic Reduction of Hydrogen Peroxide by Microperoxidase-11 Immobilized on Mesoporous Antimony-Doped Tin Oxide JF - ChemElectrChem N2 - The heme-undecapeptide microperoxidase-11 (MP-11) was immobilized on mesoporous antimony-doped tin oxide (ATO) thin-film electrodes modified with the positively charged binding promotor polydiallyldimethylammonium chloride. Surface concentrations of MP-11 of 1.5 nmol cm(-2) were sufficiently high to enable spectroelectrochemical analyses. UV/Vis spectroscopy and resonance Raman spectroscopy revealed that immobilized MP-11 adopts a six-coordinated low-spin conformation, as in solution in the presence of a polycation. Cathodic reduction of hydrogen peroxide at potentials close to +500mV versus Ag/AgCl indicates that the reaction proceeds via a Compound I-type like intermediate, analogous to natural peroxidases, and confirms mesoporous ATO as a suitable host material for adsorbing the heme-peptide in its native state. A hydrogen peroxide sensor is proposed by using the bioelectrocatalytic properties of the MP-11-modified ATO. KW - electrochemistry KW - enzyme catalysis KW - mesoporous materials KW - microperoxidase KW - spectroelectrochemistry Y1 - 2017 U6 - https://doi.org/10.1002/celc.201600776 SN - 2196-0216 VL - 4 IS - 4 SP - 913 EP - 919 PB - Wiley-VCH CY - Weinheim ER - TY - GEN A1 - Dammhahn, Melanie A1 - Randriamoria, Toky M. A1 - Goodman, Steven M. T1 - Broad and flexible stable isotope niches in invasive non-native Rattus spp. in anthropogenic and natural habitats of central eastern Madagascar T2 - Postprints der Universität Potsdam Mathematisch-Naturwissenschaftliche Reihe N2 - Background: Rodents of the genus Rattus are among the most pervasive and successful invasive species, causing major vicissitudes in native ecological communities. A broad and flexible generalist diet has been suggested as key to the invasion success of Rattus spp. Here, we use an indirect approach to better understand foraging niche width, plasticity, and overlap within and between introduced Rattus spp. in anthropogenic habitats and natural humid forests of Madagascar. Results: Based on stable carbon and nitrogen isotope values measured in hair samples of 589 individual rodents, we found that Rattus rattus had an extremely wide foraging niche, encompassing the isotopic space covered by a complete endemic forest-dwelling Malagasy small mammal community. Comparisons of Bayesian standard ellipses, as well as (multivariate) mixed-modeling analyses, revealed that the stable isotope niche of R. rattus tended to change seasonally and differed between natural forests and anthropogenic habitats, indicating plasticity in feeding niches. In co-occurrence, R. rattus and Rattus norvegicus partitioned feeding niches. Isotopic mismatch of signatures of individual R. rattus and the habitat in which they were captured, indicate frequent dispersal movements for this species between natural forest and anthropogenic habitats. Conclusions: Since R. rattus are known to transmit a number of zoonoses, potentially affecting communities of endemic small mammals, as well as humans, these movements presumably increase transmission potential. Our results suggest that due to their generalist diet and potential movement between natural forest and anthropogenic habitats, Rattus spp. might affect native forest-dependent Malagasy rodents as competitors, predators, and disease vectors. The combination of these effects helps explain the invasion success of Rattus spp. and the detrimental effects of this genus on the endemic Malagasy rodent fauna. T3 - Zweitveröffentlichungen der Universität Potsdam : Mathematisch-Naturwissenschaftliche Reihe - 722 KW - Bayesian standard ellipse KW - coexistence KW - habitat use KW - humid forest KW - invasion ecology KW - invasive species KW - Rattus rattus KW - Rattus norvegicus KW - rodents KW - fur KW - stable carbon isotope KW - stable nitrogen isotope Y1 - 2019 U6 - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:kobv:517-opus4-429419 SN - 1866-8372 IS - 722 ER - TY - JOUR A1 - Zibulski, Romy A1 - Wesener, Felix A1 - Wilkes, Heinz A1 - Plessen, Birgit A1 - Pestryakova, Luidmila Agafyevna A1 - Herzschuh, Ulrike T1 - C / N ratio, stable isotope (delta C-13, delta N-15), and n-alkane patterns of brown mosses along hydrological gradients of low-centred polygons of the Siberian Arctic JF - Biogeosciences N2 - Mosses are a major component of the arctic vegetation, particularly in wetlands. We present C / N atomic ratio, delta C-13 and delta N-15 data of 400 brown-moss samples belonging to 10 species that were collected along hydrological gradients within polygonal mires located on the southern Taymyr Peninsula and the Lena River delta in northern Siberia. Additionally, n-alkane patterns of six of these species (16 samples) were investigated. The aim of the study is to see whether the inter-and intraspecific differences in C / N, isotopic compositions and n-alkanes are indicative of habitat, particularly with respect to water level. Overall, we find high variability in all investigated parameters for two different moisture-related groups of moss species. The C / N ratios range between 11 and 53 (median: 32) and show large variations at the intraspecific level. However, species preferring a dry habitat (xero-mesophilic mosses) show higher C / N ratios than those preferring a wet habitat (meso-hygrophilic mosses). The delta C-13 values range between 37.0 and 22.5% (median D 27.8 %). The delta N-15 values range between 6.6 and C 1.7%(median D 2.2 %). We find differences in delta C-13 and delta N-15 compositions between both habitat types. For some species of the meso-hygrophilic group, we suggest that a relationship between the individ-ual habitat water level and isotopic composition can be inferred as a function of microbial symbiosis. The n-alkane distribution also shows differences primarily between xeromesophilic and meso-hygrophilic mosses, i. e. having a dominance of n-alkanes with long (n-C29, n-C31 /and intermediate (n-C25 /chain lengths, respectively. Overall, our results reveal that C / N ratios, isotopic signals and n-alkanes of studied brown-moss taxa from polygonal wetlands are characteristic of their habitat. Y1 - 2017 U6 - https://doi.org/10.5194/bg-14-1617-2017 SN - 1726-4170 SN - 1726-4189 VL - 14 SP - 1617 EP - 1630 PB - Copernicus CY - Göttingen ER - TY - GEN A1 - Zibulski, Romy A1 - Wesener, Felix A1 - Wilkes, Heinz A1 - Plessen, Birgit A1 - Pestryakova, Luidmila Agafyevna A1 - Herzschuh, Ulrike T1 - C / N ratio, stable isotope (δ 13 C, δ 15 N), and n-alkane patterns of brown mosses along hydrological gradients of low-centred polygons of the Siberian Arctic T2 - Postprints der Universität Potsdam : Mathematisch-Naturwissenschaftliche Reihe N2 - Mosses are a major component of the arctic vegetation, particularly in wetlands. We present C / N atomic ratio, delta C-13 and delta N-15 data of 400 brown-moss samples belonging to 10 species that were collected along hydrological gradients within polygonal mires located on the southern Taymyr Peninsula and the Lena River delta in northern Siberia. Additionally, n-alkane patterns of six of these species (16 samples) were investigated. The aim of the study is to see whether the inter-and intraspecific differences in C / N, isotopic compositions and n-alkanes are indicative of habitat, particularly with respect to water level. Overall, we find high variability in all investigated parameters for two different moisture-related groups of moss species. The C / N ratios range between 11 and 53 (median: 32) and show large variations at the intraspecific level. However, species preferring a dry habitat (xero-mesophilic mosses) show higher C / N ratios than those preferring a wet habitat (meso-hygrophilic mosses). The delta C-13 values range between 37.0 and 22.5% (median D 27.8 %). The delta N-15 values range between 6.6 and C 1.7%(median D 2.2 %). We find differences in delta C-13 and delta N-15 compositions between both habitat types. For some species of the meso-hygrophilic group, we suggest that a relationship between the individ-ual habitat water level and isotopic composition can be inferred as a function of microbial symbiosis. The n-alkane distribution also shows differences primarily between xeromesophilic and meso-hygrophilic mosses, i. e. having a dominance of n-alkanes with long (n-C29, n-C31 /and intermediate (n-C25 /chain lengths, respectively. Overall, our results reveal that C / N ratios, isotopic signals and n-alkanes of studied brown-moss taxa from polygonal wetlands are characteristic of their habitat. T3 - Zweitveröffentlichungen der Universität Potsdam : Mathematisch-Naturwissenschaftliche Reihe - 672 KW - atmospheric nitrogen deposition KW - Lena River delta KW - free amino-acids KW - ombrotrophic peat KW - carbon isotopes KW - aquatic macrophytes KW - methane oxidation KW - organic matter KW - soil-nitrogen KW - plants Y1 - 2019 U6 - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:kobv:517-opus4-417104 SN - 1866-8372 IS - 672 ER - TY - JOUR A1 - Bäurle, Isabel T1 - Can't remember to forget you BT - Chromatin-based priming of somatic stress responses JF - Seminars in cell & developmental biology N2 - In nature plants are exposed to frequent changes in their abiotic and biotic environment. While some environmental cues are used to gauge the environment and align growth and development, others are beyond the regularly encountered spectrum of a species and trigger stress responses. Such stressful conditions provide a potential threat to survival and integrity. Plants adapt to extreme environmental conditions through physiological adaptations that are usually transient and are maintained until stressful environments subside. It is increasingly appreciated that in some cases environmental cues activate a stress memory that persists for some time after the extreme condition has subsided. Recent research has shown that this stress-induced environmental memory is mediated by epigenetic and chromatin-based mechanisms and both histone methylation and nucleosome occupancy are associated with it. KW - Priming KW - Transcriptional memory KW - Chromatin KW - H3K4 methylation KW - Nucleosome occupancy Y1 - 2017 U6 - https://doi.org/10.1016/j.semcdb.2017.09.032 SN - 1084-9521 VL - 83 SP - 133 EP - 139 PB - Elsevier CY - London ER - TY - THES A1 - Hethey, Christoph Philipp T1 - Cell physiology based pharmacodynamic modeling of antimicrobial drug combinations T1 - Zellphysiologie-basierte pharmakodynamische Modellierung von antimikrobiellen Wirkstoffkombinationen N2 - Mathematical models of bacterial growth have been successfully applied to study the relationship between antibiotic drug exposure and the antibacterial effect. Since these models typically lack a representation of cellular processes and cell physiology, the mechanistic integration of drug action is not possible on the cellular level. The cellular mechanisms of drug action, however, are particularly relevant for the prediction, analysis and understanding of interactions between antibiotics. Interactions are also studied experimentally, however, a lacking consent on the experimental protocol hinders direct comparison of results. As a consequence, contradictory classifications as additive, synergistic or antagonistic are reported in literature. In the present thesis we developed a novel mathematical model for bacterial growth that integrates cell-level processes into the population growth level. The scope of the model is to predict bacterial growth under antimicrobial perturbation by multiple antibiotics in vitro. To this end, we combined cell-level data from literature with population growth data for Bacillus subtilis, Escherichia coli and Staphylococcus aureus. The cell-level data described growth-determining characteristics of a reference cell, including the ribosomal concentration and efficiency. The population growth data comprised extensive time-kill curves for clinically relevant antibiotics (tetracycline, chloramphenicol, vancomycin, meropenem, linezolid, including dual combinations). The new cell-level approach allowed for the first time to simultaneously describe single and combined effects of the aforementioned antibiotics for different experimental protocols, in particular different growth phases (lag and exponential phase). Consideration of ribosomal dynamics and persisting sub-populations explained the decreased potency of linezolid on cultures in the lag phase compared to exponential phase cultures. The model captured growth rate dependent killing and auto-inhibition of meropenem and - also for vancomycin exposure - regrowth of the bacterial cultures due to adaptive resistance development. Stochastic interaction surface analysis demonstrated the pronounced antagonism between meropenem and linezolid to be robust against variation in the growth phase and pharmacodynamic endpoint definition, but sensitive to a change in the experimental duration. Furthermore, the developed approach included a detailed representation of the bacterial cell-cycle. We used this representation to describe septation dynamics during the transition of a bacterial culture from the exponential to stationary growth phase. Resulting from a new mechanistic understanding of transition processes, we explained the lag time between the increase in cell number and bacterial biomass during the transition from the lag to exponential growth phase. Furthermore, our model reproduces the increased intracellular RNA mass fraction during long term exposure of bacteria to chloramphenicol. In summary, we contribute a new approach to disentangle the impact of drug effects, assay readout and experimental protocol on antibiotic interactions. In the absence of a consensus on the corresponding experimental protocols, this disentanglement is key to translate information between heterogeneous experiments and also ultimately to the clinical setting. N2 - Der Zusammenhang zwischen antibiotischer Exposition und antibakterieller Wirkung wird derzeitlich erfolgreich mithilfe von mathematischen Bakterienwachstumsmodellen studiert. Üblicherweise ignorieren diese Modelle jedoch die bakterielle Physiologie und Prozesse auf Zellebene. Es folgt, dass das mechanistische Einbinden von Wirkstoffeffekten auf Zellebene nicht möglich ist. Jedoch ist der zelluläre Wirkmechanismus besonders relevant für die Vorhersage, die Analyse und das Verständnis von Antibiotikainteraktionen. Leider gibt es keinen Konsens bezüglich des experimentellen Protokolls, um diese Interaktionen zu untersuchen. Das ist einer der Gründe, warum wir in der Literatur widersprüchliche Klassifizierungen von Antibiotikainteraktionen als additiv, synergistisch oder antagonistisch finden. In der vorliegenden Arbeit entwickelten wir ein neuartiges mathematisches Bakterienwachstumsmodel, welches Prozesse auf Zellebene in das Populationswachstum einbindet. Der Anwendungszweck dieses Models ist die Vorhersage bakteriellen Wachstums unter antimikrobieller Mehrfachexposition in vitro. Um das zu erreichen, kombinierten wir die Zellebene beschreibende Daten aus der Literatur mit Wachstumsdaten für Bacillus subtilis, Escherichia coli und Staphylococcus aureus. Die die Zellebene beschreibenden Daten bezogen sich auf Wachstums-bestimmende Charakteristika einer Referenzzelle, unter anderem auf die ribosomale Konzentration und Effizienz. Die Wachstumsdaten beinhalteten umfangreiche Zeit-Absterbe-Kurven für klinisch relevante Antibiotika (Tetracyclin, Chloramphenicol, Vancomycin, Meropenem, Linezolid) und Zweifachkombinationen aus diesen. Der neue Zellebenen-Ansatz erlaubt es erstmalig, einzelne und kombinierte Effekte der erwähnten Antibiotika für unterschiedliche experimentelle Protokolle gleichzeitig zu beschreiben. Insbesondere beziehen sich diese Unterschiede auf die Wachstumsphasen (Lag oder exponentiellen Phase). Die Berücksichtigung der ribosomalen Konzentration und persistenter Subpopulationen erklärte die verminderte Potenz von Linezolid gegen Kulturen in der Lag Phase im Vergleich zu Kulturen, die sich in der exponentiellen Phase befanden. Das Model erfasst Wachstumsraten-abhängiges Zelltöten und die Selbstinhibierung von Meropenem und - ebenso für Vancomycin - ein Wiederanwachsen der bakteriellen Kulturen aufgrund von adaptiver Resistenzentwicklung. Stochastische Analysen der Interaktionsoberflächen zeigen, dass der ausgeprägte Antagonismus zwischen Meropenem und Linezolid zwar robust gegenüber Variation der Wachstumsphase und der Definition des pharmakodynamischen Endpunktes reagiert, jedoch empfindlich von der Zeitspanne des Experiments beeinflusst wird. Desweiteren enthält der entwickelte Ansatz eine detaillierte Repräsentation des bakteriellen Zellzyklus. Wir nutzten diese Repräsentation, um Septierungsdynamiken während des Übergangs einer bakteriellen Kultur aus der exponentiellen Phase in die stationäre Phase zu beschreiben. Basierend auf einem neugewonnenen mechanistischen Verständnis für diese Übergänge, konnten wir außerdem die zeitliche Verzögerung erklären, die zwischen dem Anstieg der Zellanzahl und der Biomasse während des Übergangs von Lag zu exponentieller Phase auftritt. Außerdem reproduziert unser Modell den erhöhten intrazellulären RNA Massenanteil, der auftritt, wenn Bakterien Chloramphenikol ausgesetzt werden. Zusammenfassend steuern wir einen neuen Ansatz bei, der es erlaubt, die Einflüsse von Wirkstoffeffekten, Endpunktdefinitionen und des experimentellen Protokolls zu entflechten. Da kein Konsens hinsichtlich eines entsprechenden experimentellen Protokolls existiert, ist eine solche Entflechtung der Schlüssel, um Informationen zwischen unterschiedlichen Experimenten - und letztendlich auch in die Klinik - zu transferieren. KW - antibiotic combinations KW - bacterial population growth KW - pharmacodynamics KW - drug drug interactions KW - time-kill curves KW - ribosomal dynamics KW - Antibiotika KW - Wirkstoffinteraktionen KW - Pharmakodynamik KW - mathematische Modellierung KW - mathematical modelling Y1 - 2017 U6 - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:kobv:517-opus4-401056 ER - TY - JOUR A1 - Ietswaart, Robert A1 - Rosa, Stefanie A1 - Wu, Zhe A1 - Dean, Caroline A1 - Howard, Martin T1 - Cell-Size-Dependent Transcription of FLC and Its Antisense Long Non-coding RNA COOLAIR Explain Cell-to-Cell Expression Variation JF - Cell systems N2 - Single-cell quantification of transcription kinetics and variability promotes a mechanistic understanding of gene regulation. Here, using single-molecule RNA fluorescence in situ hybridization and mathematical modeling, we dissect cellular RNA dynamics for Arabidopsis FLOWERING LOCUS C (FLC). FLC expression quantitatively determines flowering time and is regulated by antisense (COOLAIR) transcription. In cells without observable COOLAIR expression, we quantify FLC transcription initiation, elongation, intron processing, and lariat degradation, as well as mRNA release from the locus and degradation. In these heterogeneously sized cells, FLC mRNA number increases linearly with cell size, resulting in a large cell-to-cell variability in transcript level. This variation is accounted for by cell-sizedependent, Poissonian FLC mRNA production, but not by large transcriptional bursts. In COOLAIRexpressing cells, however, antisense transcription increases with cell size and contributes to FLC transcription decreasing with cell size. Our analysis therefore reveals an unexpected role for antisense transcription in modulating the scaling of transcription with cell size. Y1 - 2017 U6 - https://doi.org/10.1016/j.cels.2017.05.010 SN - 2405-4712 SN - 2405-4720 VL - 4 SP - 622 EP - 635 PB - Cell Press CY - Cambridge ER - TY - JOUR A1 - Braig, Friederike A1 - Kriegs, Malte A1 - Voigtlaender, Minna A1 - Habel, Beate A1 - Grob, Tobias A1 - Biskup, Karina A1 - Blanchard, Veronique A1 - Sack, Markus A1 - Thalhammer, Anja A1 - Ben Batalla, Isabel A1 - Braren, Ingke A1 - Laban, Simon A1 - Danielczyk, Antje A1 - Goletz, Steffen A1 - Jakubowicz, Elzbieta A1 - Maerkl, Bruno A1 - Trepel, Martin A1 - Knecht, Rainald A1 - Riecken, Kristoffer A1 - Fehse, Boris A1 - Loges, Sonja A1 - Bokemeyer, Carsten A1 - Binder, Mascha T1 - Cetuximab Resistance in Head and Neck Cancer Is Mediated by EGFR-K-521 Polymorphism JF - Cancer research N2 - Head and neck squamous cell carcinomas (HNSCC) exhibiting resistance to the EGFR-targeting drug cetuximab poses a challenge to their effective clinical management. Here, we report a specific mechanism of resistance in this setting based upon the presence of a single nucleotide polymorphism encoding EGFR-K-521 (K-allele), which is expressed in > 40% of HNSCC cases. Patients expressing the K-allele showed significantly shorter progressionfree survival upon palliative treatment with cetuximab plus chemotherapy or radiation. In several EGFR-mediated cancer models, cetuximab failed to inhibit downstream signaling or to kill cells harboring a high K-allele frequency. Cetuximab affinity for EGFR-K-521 was reduced slightly, but ligand-mediated EGFR acti-vation was intact. We found a lack of glycan sialyation on EGFR-K-521 that associated with reduced protein stability, suggesting a structural basis for reduced cetuximab efficacy. CetuGEX, an antibody with optimized Fc glycosylation targeting the same epitope as cetuximab, restored HNSCC sensitivity in a manner associated with antibody-dependent cellular cytotoxicity rather than EGFR pathway inhibition. Overall, our results highlight EGFR-K-521 expression as a key mechanism of cetuximab resistance to evaluate prospectively as a predictive biomarker in HNSCC patients. Further, they offer a preclinical rationale for the use of ADCC-optimized antibodies to treat tumors harboring this EGFR isoform. Y1 - 2017 U6 - https://doi.org/10.1158/0008-5472.CAN-16-0754 SN - 0008-5472 SN - 1538-7445 VL - 77 IS - 5 SP - 1188 EP - 1199 PB - American Association for Cancer Research CY - Philadelphia ER - TY - THES A1 - Suchoszek, Monika T1 - Characterization of inducible galactolipid biosynthesis mutants in tobacco N2 - Chloroplast membranes have a unique composition characterized by very high contents of the galactolipids, MGDG and DGDG. Many studies on constitutive, galactolipid-deficient mutants revealed conflicting results about potential functions of galactolipids in photosynthetic membranes. Likely, this was caused by pleiotropic effects such as starvation artefacts because of impaired photosynthesis from early developmental stages of the plants onward. Therefore, an ethanol inducible RNAi-approach has been taken to suppress two key enzymes of galactolipid biosynthesis in the chloroplast, MGD1 and DGD1. Plants were allowed to develop fully functional source leaves prior to induction, which then could support plant growth. Then, after the ethanol induction, both young and mature leaves were investigated over time. Our studies revealed similar changes in both MGDG- and DGDG-deficient lines, however young and mature leaves of transgenic lines showed a different response to galactolipid deficiency. While no changes of photosynthetic parameters and minor changes in lipid content were observed in mature leaves of transgenic lines, strong reductions in total chlorophyll content and in the accumulation of all photosynthetic complexes and significant changes in contents of various lipid groups occurred in young leaves. Microscopy studies revealed an appearance of lipid droplets in the cytosol of young leaves in all transgenic lines which correlates with significantly higher levels of TAGs. Since in young leaves the production of membrane lipids is lowered, the excess of fatty acids is used for storage lipids production, resulting in the accumulation of TAGs. Our data indicate that both investigated galactolipids serve as structural lipids since changes in photosynthetic parameters were mainly the result of reduced amounts of all photosynthetic constituents. In response to restricted galactolipid synthesis, thylakoid biogenesis is precisely readjusted to keep the proper stoichiometry and functionality of the photosynthetic apparatus. Ultimately, the data revealed that downregulation of one galactolipid triggers changes not only in chloroplasts but also in the nucleus as shown by downregulation of nuclear encoded subunits of the photosynthetic complexes. KW - galactolipids KW - photosynthesis KW - tobacco Y1 - 2017 ER - TY - THES A1 - Foti, Alessandro T1 - Characterization of the human aldehyde oxidase T1 - Charakterisierung der menschlichen Aldehydoxidase BT - Studies on the FAD active site and ROS generation N2 - In this work the human AOX1 was characterized and detailed aspects regarding the expression, the enzyme kinetics and the production of reactive oxygen species (ROS) were investigated. The hAOX1 is a cytosolic enzyme belonging to the molybdenum hydroxylase family. Its catalytically active form is a homodimer with a molecular weight of 300 kDa. Each monomer (150 kDa) consists of three domains: a N-terminal domain (20 kDa) containing two [2Fe-2S] clusters, a 40 kDa intermediate domain containing a flavin adenine dinucleotide (FAD), and a C-terminal domain (85 kDa) containing the substrate binding pocket and the molybdenum cofactor (Moco). The hAOX1 has an emerging role in the metabolism and pharmacokinetics of many drugs, especially aldehydes and N- heterocyclic compounds. In this study, the hAOX1 was hetereogously expressed in E. coli TP1000 cells, using a new codon optimized gene sequence which improved the expressed protein yield of around 10-fold compared to the previous expression systems for this enzyme. To increase the catalytic activity of hAOX1, an in vitro chemical sulfuration was performed to favor the insertion of the equatorial sulfido ligand at the Moco with consequent increased enzymatic activity of around 10-fold. Steady-state kinetics and inhibition studies were performed using several substrates, electron acceptors and inhibitors. The recombinant hAOX1 showed higher catalytic activity when molecular oxygen was used as electron acceptor. The highest turn over values were obtained with phenanthridine as substrate. Inhibition studies using thioridazine (phenothiazine family), in combination with structural studies performed in the group of Prof. M.J. Romão, Nova Universidade de Lisboa, showed a new inhibition site located in proximity of the dimerization site of hAOX1. The inhibition mode of thioridazine resulted in a noncompetitive inhibition type. Further inhibition studies with loxapine, a thioridazine-related molecule, showed the same type of inhibition. Additional inhibition studies using DCPIP and raloxifene were carried out. Extensive studies on the FAD active site of the hAOX1 were performed. Twenty new hAOX1 variants were produced and characterized. The hAOX1 variants generated in this work were divided in three groups: I) hAOX1 single nucleotide polymorphisms (SNP) variants; II) XOR- FAD loop hAOX1 variants; III) additional single point hAOX1 variants. The hAOX1 SNP variants G46E, G50D, G346R, R433P, A439E, K1231N showed clear alterations in their catalytic activity, indicating a crucial role of these residues into the FAD active site and in relation to the overall reactivity of hAOX1. Furthermore, residues of the bovine XOR FAD flexible loop (Q423ASRREDDIAK433) were introduced in the hAOX1. FAD loop hAOX1 variants were produced and characterized for their stability and catalytic activity. Especially the variants hAOX1 N436D/A437D/L438I, N436D/A437D/L438I/I440K and Q434R/N436D/A437D/L438I/I440K showed decreased catalytic activity and stability. hAOX1 wild type and variants were tested for reactivity toward NADH but no reaction was observed. Additionally, the hAOX1 wild type and variants were tested for the generation of reactive oxygen species (ROS). Interestingly, one of the SNP variants, hAOX1 L438V, showed a high ratio of superoxide prodction. This result showed a critical role for the residue Leu438 in the mechanism of oxygen radicals formation by hAOX1. Subsequently, further hAOX1 variants having the mutated Leu438 residue were produced. The variants hAOX1 L438A, L438F and L438K showed superoxide overproduction of around 85%, 65% and 35% of the total reducing equivalent obtained from the substrate oxidation. The results of this work show for the first time a characterization of the FAD active site of the hAOX1, revealing the importance of specific residues involved in the generation of ROS and effecting the overall enzymatic activity of hAOX1. The hAOX1 SNP variants presented here indicate that those allelic variations in humans might cause alterations ROS balancing and clearance of drugs in humans. N2 - Aldehydoxidasen (AOX) sind Molybdo-enzyme, die durch breite Substratspezifität gekennzeichnet sind, aromatische/aliphatische Aldehyde in die entsprechenden Carbonsäuren oxidieren und verschiedene heteroaromatische Ringe hydroxylieren. Die Enzyme verwenden Sauerstoff als terminalen Elektronenakzeptor und produzieren reduzierte Sauerstoffspezies während des Umsatzes. Die physiologische Funktion von Säugetier-AOX-Isoenzymen ist noch unklar, aber menschliches AOX (hAOX1) ist ein Enzym von Phase-I-Wirkstoff-Metabolismus. Weiterhin, wurden zahlreiche Einzelnukleotidpolymorphismen (SNP) und weitere hAOX1-Mutanten im hAOX1-Gen identifiziert. SNPs sind eine Hauptquelle für die interindividuelle Variabilität in der menschlichen Population, und SNP-basierte Aminosäureaustausche in hAOX1 modulieren die katalytische Funktion des Enzyms entweder positiv oder negativ. In diesem Bericht haben wir zehn neue SNPs ausgewählt, die zu Aminosäureaustauschen in der Nähe der FAD-Cofaktor von hAOX1 führen und die gereinigten Enzyme nach heterologen Expression in Escherichia coli charakterisieren. Darüber hinaus haben wir zehn weitere FAD-Varianten produziert. Die hAOX1-Varianten wurden sorgfältig durch quantitative Unterschiede in ihrer Fähigkeit zur Herstellung von Superoxidradikal charakterisiert. ROS repräsentieren markante Schlüsselmoleküle in physiologischen und pathologischen Zuständen in der Zelle. Unsere Daten zeigen signifikante Veränderungen der Superoxid-Anionenproduktion unter den Varianten. Insbesondere führte der Rest L438 in der Nähe des Isoalloxanzinringes des FAD-Cofaktors zu einer erhöhten Superoxid-Radikalproduktion von 75-85%. In Anbetracht der hohen Toxizität des Superoxid-Anions in der Zelle ist die hAOX1-L438V SNP-Variante ein eventueller Kandidat für kritische oder pathologische Rollen dieser natürlichen Variante innerhalb der menschlichen Population. KW - aldehyde KW - oxidase KW - ROS KW - reactive oxygen species Y1 - 2017 U6 - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:kobv:517-opus4-410107 ER - TY - THES A1 - Nitezki, Tina T1 - Charakterisierung von Stereotypien bei der FVB/NJ-Maus hinsichtlich metabolischer und immunologischer Aspekte auf die Stoffwechselleistung T1 - Characterization of stereotypies in FVB/NJ mice and their impact on metabolism and immune system N2 - Im Sinne des Refinements von Tierversuchen sollen alle Bedingungen während der Zucht, der Haltung und des Transports von zu Versuchszwecken gehaltenen Tieren und alle Methoden während des Versuchs so verbessert werden, dass die verwendeten Tiere ein minimales Maß an potentiellem Distress, Schmerzen oder Leiden erfahren. Zudem soll ihr Wohlbefinden durch die Möglichkeit des Auslebens speziesspezifischer Verhaltensweisen und die Anwendung tierschonender Verfahren maximal gefördert werden. Zur Etablierung von Grundsätzen des Refinements sind grundlegende Kenntnisse über die physiologischen Bedürfnisse und Verhaltensansprüche der jeweiligen Spezies unabdingbar. Die Experimentatoren sollten das Normalverhalten der Tiere kennen, um potentielle Verhaltensabweichungen, wie Stereotypien, zu verstehen und interpretieren zu können. Standardisierte Haltungsbedingungen von zu Versuchszwecken gehaltenen Mäusen weichen in diversen Aspekten von der natürlichen Umgebung ab und erfordern eine gewisse Adaptation. Ist ein Tier über einen längeren Zeitraum unfähig, sich an die gegebenen Umstände anzupassen, können abnormale Verhaltensweisen, wie Stereotypien auftreten. Stereotypien werden definiert als Abweichungen vom Normalverhalten, die repetitiv und ohne Abweichungen im Ablauf ausgeführt werden, scheinbar keiner Funktion dienen und der konkreten Umweltsituation nicht immer entsprechen. Bisher war unklar, in welchem Ausmaß stereotypes Verhalten den metabolischen Phänotyp eines Individuums beeinflusst. Ziel dieser Arbeit war es daher, das stereotype Verhalten der FVB/NJ-Maus erstmals detailliert zu charakterisieren, systematisch zusammenzutragen, welche metabolischen Konsequenzen dieses Verhalten bedingt und wie sich diese auf das Wohlbefinden der Tiere und die Verwendung stereotyper Tiere in Studien mit tierexperimentellem Schwerpunkt auswirken. Der Versuch begann mit der Charakterisierung der mütterlichen Fürsorge in der Parentalgeneration. Insgesamt wurden 35 Jungtiere der F1-Generation vom Absatz an, über einen Zeitraum von 11 Wochen einzeln gehalten, kontinuierlich beobachtet, bis zum Versuchsende wöchentlich Kotproben gesammelt und das Körpergewicht bestimmt. Zusätzlich erfolgten begleitende Untersuchungen wie Verhaltenstests und die Erfassung der physischen Aktivität und metabolischer Parameter. Anschließend wurden u.a. die zerebralen Serotonin- und Dopamingehalte, fäkale Glucocorticoidlevels, hepatisches Glykogen und muskuläre Glykogen- und Triglyceridlevels bestimmt. Nahezu unabhängig von der mütterlichen Herkunft entwickelte sich bei mehr als der Hälfte der 35 Jungtiere in der F1-Generation stereotypes Verhalten. Diese Daten deuten darauf hin, dass es keine Anzeichen für das Erlernen oder eine direkte genetische Transmission stereotypen Verhaltens bei der FVB/NJ-Maus gibt. Über den gesamten Beobachtungszeitraum zeichneten sich die stereotypen FVB/NJ-Mäuse durch ein eingeschränktes Verhaltensrepertoire aus. Zu Gunsten der erhöhten Aktivität und des Ausübens stereotypen Verhaltens lebten sie insgesamt weniger andere Verhaltensweisen (Klettern, Graben, Nagen) aus. Darüber hinaus waren Stereotypien sowohl im 24-Stunden Open Field Test als auch in der Messeinrichtung der indirekten Tierkalorimetrie mit einer erhöhten Aktivität und Motilität assoziiert, während die circadiane Rhythmik nicht divergierte. Diese erhöhte körperliche Betätigung spiegelte sich in den niedrigeren Körpergewichtsentwicklungen der stereotypen Tiere wieder. Außerdem unterschieden sich die Körperfett- und Körpermuskelanteile. Zusammenfassend lässt sich sagen, dass das Ausüben stereotypen Verhaltens zu Differenzen im metabolischen Phänotyp nicht-stereotyper und stereotyper FVB/NJ-Mäuse führt. Im Sinne der „Guten Wissenschaftlichen Praxis“ sollte das zentrale Ziel jedes Wissenschaftlers sein, aussagekräftige und reproduzierbare Daten hervorzubringen. Jedoch können keine validen Resultate von Tieren erzeugt werden, die in Aspekten variieren, die für den vorgesehenen Zweck der Studie nicht berücksichtigt wurden. Deshalb sollten nicht-stereotype und stereotype Individuen nicht innerhalb einer Versuchsgruppe randomisiert werden. Stereotype Tiere demzufolge von geplanten Studien auszuschließen, würde allerdings dem Gebot des zweiten R’s – der Reduction – widersprechen. Um Refinement zu garantieren, sollte der Fokus auf der maximal erreichbaren Prävention stereotypen Verhaltens liegen. Diverse Studien haben bereits gezeigt, dass die Anreicherung der Haltungsumwelt (environmental enrichment) zu einer Senkung der Prävalenz von Stereotypien bei Mäusen führt, dennoch kommen sie weiterhin vor. Daher sollte environmental enrichment zukünftig weniger ein „Kann“, sondern ein „Muss“ sein – oder vielmehr: der Goldstandard. Zudem würde eine profunde phänotypische Charakterisierung dazu beitragen, Mausstämme zu erkennen, die zu Stereotypien neigen und den für den spezifischen Zweck am besten geeigneten Mausstamm zu identifizieren, bevor ein Experiment geplant wird. N2 - In the sense of refinement animal experimentation, all conditions during breeding, husbandry and transport of animals used for experimental purposes and all methods during the experiment should be improved to reduce the degree of potential distress, pain or suffering. In addition, their well-being should be guaranteed by the possibility of expressing natural and species-specific behavioural patterns and by the application of considerate procedures. In order to establish principles for refinement, basic knowledge about the physiological needs and behavioural requirements of the respective species is indispensable. The experimenters should know the normal behaviour of animals in order to understand and interpret potential behavioural deviations, such as stereotypies. Standardized housing conditions of laboratory mice deviate from the natural environment in various aspects and might require a certain adaptation. Behavioural adaptation allows animals to adjust to environmental changes and leads to species’ characteristic behaviour. If an animal is unable to adapt to environmental conditions, abnormal behaviours like stereotypies might occur. Stereotypies are defined as deviations from normal behaviour, which are executed repetitively and without deviations in the performance, seem to serve no function and do not always correspond to the concrete environmental situation. Since it remains unclear to what extend stereotypic behaviour influences the individual’s metabolic phenotype, this study investigated behaviour of FVB/NJ mice in detail, exemplarily for stereotypy-prone mouse strains, and compiled the impact of behavioural deviations on physical activity, animal metabolism, animal welfare and on results obtained from studies with an animal specific focus. To detect early indicators for the later development of stereotypic behaviour in the F1 generation, this study started with investigating maternal care in the parental generation. Overall, 35 animals of the F1 generation were kept individually from weaning age. For 11 weeks they were observed, faecal samples were obtained and body weight was determined. Additionally, behavioural tests, metabolic parameters and physical activity were investigated. Furthermore, among others, cerebral serotonin and dopamine contents, faecal glucocorticoid levels and hepatic glycogen, muscular triglyceride and glycogen levels were assessed. Almost independently of the mother's origin, more than half of the 35 pups developed stereotypic behavior in the F1 generation. Data suggest that there is obviously no evidence of learning or a direct genetic transmission of stereotypic behavior in the FVB/NJ-mouse. The predominant portion of stereotypic animals performed the stereotypy of back-flipping (backwards jumping), some animals demonstrated stereotypic circuit running (running in circles on the cage bottom) and wire gnawing (persistent gnawing on the cage grid while hanging with the forelimbs on it). Because of the increased activity and the performance of stereotypic behaviour, stereotypic mice displayed a restricted behavioural repertoire (reduced climbing, digging, gnawing). Moreover, stereotypies were associated with increased activity and motility, both in the 24-hours open field test and in the ITK system, while the circadian rhythm did not diverge. This elevated physical activity was reflected in the expected gender-dependent lower body weight development of stereotypic animals. In addition, stereotypic FVB/NJ-mice contained more relative muscle mass and less fat mass compared to non-stereotypic FVB/NJ-mice in experimental weeks 7 and 12. Besides, significant differences in relative organ weights were found. In conclusion, the performance of stereotypic behaviour leads to differences in the metabolic phenotype between non-stereotypic and stereotypic FVB/NJ mice. In the sense of "Good Scientific Practice", the central aim of any scientist should be to generate meaningful and reproducible data. However, no valid results can be generated with data derived from animals which differ in aspects that were not considered for the designated purpose of the study. Therefore, stereotypic and non-stereotypic individuals should not be randomized within one trial group. To generally exclude stereotypic animals from further studies, though, would interfere with the commandment of the second "R" - the reduction. To guarantee a maximum refinement, the focus should be the highest achievable prevention of stereotypies. Multiple studies indicate that environmental enrichment decreases the prevalence of stereotypic behaviour in mice, nevertheless they still occur. Thus, environmental enrichment of animal housing should not be a "can" but a "must", or rather the “golden standard”. Moreover, a profound phenotypic characterization would help to identify a stereotypy-prone mouse strain and to determine the mouse strain most suitable for the specific purpose before planning an experiment. KW - Stereotypien KW - Verhalten KW - FVB/NJ Maus KW - Versuchstierkunde KW - stereotypy KW - behaviour KW - FVB/NJ mouse KW - laboratory animal sciences Y1 - 2017 U6 - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:kobv:517-opus4-402265 ER - TY - JOUR A1 - Weyhenmeyer, Gesa A. A1 - Mackay, Murray A1 - Stockwell, Jason D. A1 - Thiery, Wim A1 - Grossart, Hans-Peter A1 - Augusto-Silva, Petala B. A1 - Baulch, Helen M. A1 - de Eyto, Elvira A1 - Hejzlar, Josef A1 - Kangur, Kuelli A1 - Kirillin, Georgiy A1 - Pierson, Don C. A1 - Rusak, James A. A1 - Sadro, Steven A1 - Woolway, R. Iestyn T1 - Citizen science shows systematic changes in the temperature difference between air and inland waters with global warming JF - Scientific reports N2 - Citizen science projects have a long history in ecological studies. The research usefulness of such projects is dependent on applying simple and standardized methods. Here, we conducted a citizen science project that involved more than 3500 Swedish high school students to examine the temperature difference between surface water and the overlying air (T-w-T-a) as a proxy for sensible heat flux (Q(H)). If Q(H) is directed upward, corresponding to positive T-w-T-a, it can enhance CO2 and CH4 emissions from inland waters, thereby contributing to increased greenhouse gas concentrations in the atmosphere. The students found mostly negative T-w-T-a across small ponds, lakes, streams/rivers and the sea shore (i.e. downward Q(H)), with T-w-T-a becoming increasingly negative with increasing T-a. Further examination of T-w-T-a using high-frequency temperature data from inland waters across the globe confirmed that T-w-T-a is linearly related to T-a. Using the longest available high-frequency temperature time series from Lake Erken, Sweden, we found a rapid increase in the occasions of negative T-w-T-a with increasing annual mean T-a since 1989. From these results, we can expect that ongoing and projected global warming will result in increasingly negative T-w-T-a, thereby reducing CO2 and CH4 transfer velocities from inland waters into the atmosphere. Y1 - 2017 U6 - https://doi.org/10.1038/srep43890 SN - 2045-2322 VL - 7 PB - Nature Publ. Group CY - London ER - TY - JOUR A1 - Saini, Jeetendra A1 - Guenther, Franziska A1 - Aichner, Bernhard A1 - Mischke, Steffen A1 - Herzschuh, Ulrike A1 - Zhang, Chengjun A1 - Maeusbacher, Roland A1 - Gleixner, Gerd T1 - Climate variability in the past similar to 19,000 yr in NE Tibetan Plateau inferred from biomarker and stable isotope records of Lake Donggi Cona JF - Quaternary science reviews : the international multidisciplinary research and review journal N2 - We investigated 4.84-m-long sediment record spanning over the Late Glacial and Holocene from Lake Donggi Cona to be able to reconstruct circulation pattern on the Tibetan Plateau (TP). Presently, Lake Donggi Cona is located at the boundaries of Westerlies and Asian monsoon circulations in the northeastern TP. However, the exact timing and stimulating mechanisms for climatic changes and monsoon shifts in this region are still debated. We used a 19-ka-long stable isotope record of sedimentary n-alkanes to address this discrepancy by providing insights into paleohydrological conditions. The SD of nC(23) is influenced by lake water evaporation; the BD. values of sedimentary nC(29) are mainly controlled by moisture source and temperature changes. Long-chain n-alkanes dominate over the core whereas three mean clusters (i.e. microbial, aquatic and terrestrial) can be inferred. Multi-proxies suggest five major episodes in the history of Lake Donggi Cona. The Lake Donggi Cona record indicates that the Late Glacial(18.4-14.8 cal ka BP) was dominated by low productivity of mainly microbial and aquatic organisms. Relatively low delta D values suggest low temperatures and moist conditions eventually caused by stronger Westerlies, winter monsoon and melt-water influence. Likely, the shift (similar to 17.9 cal ka BP) from microbial to enhanced aquatic input suggests either a change from deep to shallow water lake or a break in local stratification. Between 14.8 and 13.0 cal ka BP, variable climatic conditions prevailed. Although the Westerlies weekend, the increase in temperature enhanced the permafrost and snow melting (displayed by a high sedimentary accumulation rate). Higher delta D values indicate increasingly arid conditions with higher temperatures which eventually lead to high evaporative conditions and lowest lake levels. Low vegetation cover and high erosion rates led to high sediment accumulation resulting in stratification followed by anoxia in the terminal lake. From 13.0 to 9.2 cal ka BP, lowered values of 813 along with high contents of terrestrial organic matter marked the early-Holocene warming indicating a further strengthening of summer precipitation and higher lake levels. A cooling trend was observed in the mid-Holocene between 9.2 and 3.0 cal ka BP accompanied by higher moisture availability (displayed by lowered SD values) caused by reduced evaporative conditions due to a drop in temperature and recovering Westerlies. After 3.0 cal ka BP, a decrease in lake productivity and cold and semi-arid conditions prevailed suggesting lower lake levels and reduced moisture from recycled air masses and Westerlies. We propose that the summer monsoon was the predominant moisture source during the Belling-Allered warm complex and early -Holocene followed by Westerlies in mid-to-late Holocene period. Stable carbon isotope values-32%o indicate the absence of C-4 -type vegetation in the region contradicting with their presence in the Lake Qinghai record. The 81) record from lake Donggi Cona highlights the importance of the interplay between Westerlies and summer monsoon circulation at this location, which is highly dynamic in northeastern plateau compared to the North Atlantic circulation and insolation changes. Consequently lake Donggi Cona might be an important anchor point for environmental reconstructions on the Tibetan Plateau. (C) 2017 The Authors. Published by Elsevier Ltd. KW - n-alkanes KW - Hydrogen isotopes (delta D) KW - Carbon isotopes (delta C-13) KW - Carbon preference index (CPI) KW - Westerlies KW - Continental air masses KW - Precipitation KW - Late Glacial and Holocene Y1 - 2017 U6 - https://doi.org/10.1016/j.quascirev.2016.12.023 SN - 0277-3791 VL - 157 SP - 129 EP - 140 PB - Elsevier CY - Oxford ER - TY - GEN A1 - Laux, Eva-Maria A1 - Docoslis, A. A1 - Wenger, C. A1 - Bier, Frank Fabian A1 - Hölzel, Ralph T1 - Combination of dielectrophoresis and SERS for bacteria detection and characterization T2 - European biophysics journal : with biophysics letters ; an international journal of biophysics Y1 - 2017 SN - 0175-7571 SN - 1432-1017 VL - 46 SP - S331 EP - S331 PB - Springer CY - New York ER - TY - GEN A1 - Mohandesan, Elmira A1 - Speller, Camilla F. A1 - Peters, Joris A1 - Uerpmann, Hans-Peter A1 - Uerpmann, Margarethe A1 - De Cupere, Bea A1 - Hofreiter, Michael A1 - Burger, Pamela A. T1 - Combined hybridization capture and shotgun sequencing for ancient DNA analysis of extinct wild and domestic dromedary camel T2 - Postprints der Universität Potsdam Mathematisch-Naturwissenschaftliche Reihe N2 - The performance of hybridization capture combined with next-generation sequencing (NGS) has seen limited investigation with samples from hot and arid regions until now. We applied hybridization capture and shotgun sequencing to recover DNA sequences from bone specimens of ancient-domestic dromedary (Camelus dromedarius) and its extinct ancestor, the wild dromedary from Jordan, Syria, Turkey and the Arabian Peninsula, respectively. Our results show that hybridization capture increased the percentage of mitochondrial DNA (mtDNA) recovery by an average 187-fold and in some cases yielded virtually complete mitochondrial (mt) genomes at multifold coverage in a single capture experiment. Furthermore, we tested the effect of hybridization temperature and time by using a touchdown approach on a limited number of samples. We observed no significant difference in the number of unique dromedary mtDNA reads retrieved with the standard capture compared to the touchdown method. In total, we obtained 14 partial mitochondrial genomes from ancient-domestic dromedaries with 17-95% length coverage and 1.27-47.1-fold read depths for the covered regions. Using whole-genome shotgun sequencing, we successfully recovered endogenous dromedary nuclear DNA (nuDNA) from domestic and wild dromedary specimens with 1-1.06-fold read depths for covered regions. Our results highlight that despite recent methodological advances, obtaining ancient DNA (aDNA) from specimens recovered from hot, arid environments is still problematic. Hybridization protocols require specific optimization, and samples at the limit of DNA preservation need multiple replications of DNA extraction and hybridization capture as has been shown previously for Middle Pleistocene specimens. T3 - Zweitveröffentlichungen der Universität Potsdam : Mathematisch-Naturwissenschaftliche Reihe - 789 KW - ancient DNA KW - Camelus dromedarius KW - capture enrichment KW - degraded DNA KW - mitochondrial genome (mtDNA) KW - next-generation sequencing Y1 - 2019 U6 - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:kobv:517-opus4-439955 SN - 1866-8372 IS - 789 SP - 300 EP - 313 ER - TY - JOUR A1 - Mohandesan, Elmira A1 - Speller, Camilla F. A1 - Peters, Joris A1 - Uerpmann, Hans-Peter A1 - Uerpmann, Margarethe A1 - De Cupere, Bea A1 - Hofreiter, Michael A1 - Burger, Pamela A. T1 - Combined hybridization capture and shotgun sequencing for ancient DNA analysis of extinct wild and domestic dromedary camel JF - Molecular ecology resources N2 - The performance of hybridization capture combined with next-generation sequencing (NGS) has seen limited investigation with samples from hot and arid regions until now. We applied hybridization capture and shotgun sequencing to recover DNA sequences from bone specimens of ancient-domestic dromedary (Camelus dromedarius) and its extinct ancestor, the wild dromedary from Jordan, Syria, Turkey and the Arabian Peninsula, respectively. Our results show that hybridization capture increased the percentage of mitochondrial DNA (mtDNA) recovery by an average 187-fold and in some cases yielded virtually complete mitochondrial (mt) genomes at multifold coverage in a single capture experiment. Furthermore, we tested the effect of hybridization temperature and time by using a touchdown approach on a limited number of samples. We observed no significant difference in the number of unique dromedary mtDNA reads retrieved with the standard capture compared to the touchdown method. In total, we obtained 14 partial mitochondrial genomes from ancient-domestic dromedaries with 17-95% length coverage and 1.27-47.1-fold read depths for the covered regions. Using whole-genome shotgun sequencing, we successfully recovered endogenous dromedary nuclear DNA (nuDNA) from domestic and wild dromedary specimens with 1-1.06-fold read depths for covered regions. Our results highlight that despite recent methodological advances, obtaining ancient DNA (aDNA) from specimens recovered from hot, arid environments is still problematic. Hybridization protocols require specific optimization, and samples at the limit of DNA preservation need multiple replications of DNA extraction and hybridization capture as has been shown previously for Middle Pleistocene specimens. KW - ancient DNA KW - Camelus dromedarius KW - capture enrichment KW - degraded DNA KW - mitochondrial genome (mtDNA) KW - next-generation sequencing Y1 - 2017 U6 - https://doi.org/10.1111/1755-0998.12551 SN - 1755-098X SN - 1755-0998 VL - 17 IS - 2 SP - 300 EP - 313 PB - Wiley CY - Hoboken ER - TY - THES A1 - Schedina, Ina-Maria T1 - Comparative genetic and transcriptomic analyses of the amazon molly, poecilia formosa and its parental species, poecilia mexicana and poecilia latipinna Y1 - 2017 ER - TY - THES A1 - Pandey-Pant, Pooja T1 - Comparative transcriptomics and functional genomics during phosphorus limitation in plants Y1 - 2017 ER - TY - JOUR A1 - Dolotovskaya, Sofya A1 - Bordallo, Juan Torroba A1 - Haus, Tanja A1 - Noll, Angela A1 - Hofreiter, Michael A1 - Zinner, Dietmar A1 - Roos, Christian T1 - Comparing mitogenomic timetrees for two African savannah primate genera (Chlorocebus and Papio) JF - Zoological Journal of the Linnean Society N2 - Complete mitochondrial (mtDNA) genomes have proved to be useful in reconstructing primate phylogenies with higher resolution and confidence compared to reconstructions based on partial mtDNA sequences. Here, we analyse complete mtDNA genomes of African green monkeys (genus Chlorocebus), a widely distributed primate genus in Africa representing an interesting phylogeographical model for the evolution of savannah species. Previous studies on partial mtDNA sequences revealed nine major clades, suggesting several cases of para- and polyphyly among Chlorocebus species. However, in these studies, phylogenetic relationships among several clades were not resolved, and divergence times were not estimated. We analysed complete mtDNA genomes for ten Chlorocebus samples representing major mtDNA clades to find stronger statistical support in the phylogenetic reconstruction than in the previous studies and to estimate divergence times. Our results confirmed para- and polyphyletic relationships of most Chlorocebus species, while the support for the phylogenetic relationships between the mtDNA clades increased compared to the previous studies. Our results indicate an initial west-east division in the northern part of the Chlorocebus range with subsequent divergence into north-eastern and southern clades. This phylogeographic scenario contrasts with that for another widespread African savannah primate genus, the baboons (Papio), for which a dispersal from southern Africa into East and West Africa was suggested. KW - African green monkeys KW - baboons KW - mitochondrial genomes KW - phylogeny KW - phylogeography Y1 - 2017 U6 - https://doi.org/10.1093/zoolinnean/zlx001 SN - 0024-4082 SN - 1096-3642 VL - 181 IS - 2 SP - 471 EP - 483 PB - Oxford Univ. Press CY - Oxford ER - TY - JOUR A1 - Westbury, Michael V. A1 - Dalerumb, Fredrik A1 - Noren, Karin A1 - Hofreiter, Michael T1 - Complete mitochondrial genome of a bat-eared fox (Otocyon megalotis), along with phylogenetic considerations JF - Mitochondrial DNA. Part B N2 - The bat-eared fox, Otocyon megalotis, is the only member of its genus and is thought to occupy a basal position within the dog family. These factors can lead to challenges in complete mitochondrial reconstructions and accurate phylogenetic positioning. Here, we present the first complete mitochondrial genome of the bat-eared fox recovered using shotgun sequencing and iterative mapping to three distantly related species. Phylogenetic analyses placed the bat-eared fox basal in the Canidae family within the clade including true foxes (Vulpes) and the raccoon dog (Nyctereutes) with high support values. This position is in good agreement with previously published results based on short fragments of mitochondrial and nuclear genes, therefore adding more support to the basal positioning of the bat-eared fox within Canidae. KW - Phylogenetics KW - mitochondria KW - iterative mapping KW - Canidae Y1 - 2017 U6 - https://doi.org/10.1080/23802359.2017.1331325 SN - 2380-2359 VL - 2 IS - 1 SP - 298 EP - 299 PB - Routledge, Taylor & Francis Group CY - London ER - TY - JOUR A1 - Riedel, M. A1 - Sabir, N. A1 - Scheller, Frieder W. A1 - Parak, Wolfgang J. A1 - Lisdat, Fred T1 - Connecting quantum dots with enzymes BT - mediator-based approaches for the light-directed read-out of glucose and fructose oxidation JF - Nanoscale N2 - The combination of the biocatalytic features of enzymes with the unique physical properties of nanoparticles in a biohybrid system provides a promising approach for the development of advanced bioelectrocatalytic devices. This study describes the construction of photoelectrochemical signal chains based on CdSe/ZnS quantum dot (QD) modified gold electrodes as light switchable elements, and low molecular weight redox molecules for the combination with different biocatalysts. Photoelectrochemical and photoluminescence experiments verify that electron transfer can be achieved between the redox molecules hexacyanoferrate and ferrocene, and the QDs under illumination. Since for both redox mediators a concentration dependent photocurrent change has been found, light switchable enzymatic signal chains are built up with fructose dehydrogenase (FDH) and pyrroloquinoline quinone-dependent glucose dehydrogenase ((PQQ) GDH) for the detection of sugars. After immobilization of the enzymes at the QD electrode the biocatalytic oxidation of the substrates can be followed by conversion of the redox mediator in solution and subsequent detection at the QD electrode. Furthermore, (PQQ) GDH has been assembled together with ferrocenecarboxylic acid on top of the QD electrode for the construction of a funtional biohybrid architecture, showing that electron transfer can be realized from the enzyme over the redox mediator to the QDs and subsequently to the electrode in a completely immobilized fashion. The results obtained here do not only provide the basis for light-switchable biosensing and bioelectrocatalytic applications, but may also open the way for self-driven point-of-care systems by combination with solar cell approaches (power generation at the QD electrode by enzymatic substrate consumption). Y1 - 2017 U6 - https://doi.org/10.1039/c7nr00091j SN - 2040-3364 SN - 2040-3372 VL - 9 SP - 2814 EP - 2823 PB - Royal Society of Chemistry CY - Cambridge ER - TY - GEN A1 - Herde, Antje A1 - Eccard, Jana T1 - Consistency in boldness, activity and exploration at different stages of life N2 - Background: Animals show consistent individual behavioural patterns over time and over situations. This phenomenon has been referred to as animal personality or behavioural syndromes. Little is known about consistency of animal personalities over entire life times. We investigated the repeatability of behaviour in common voles (Microtus arvalis) at different life stages, with different time intervals, and in different situations. Animals were tested using four behavioural tests in three experimental groups: 1. before and after maturation over three months, 2. twice as adults during one week, and 3. twice as adult animals over three months, which resembles a substantial part of their entire adult life span of several months. Results: Different behaviours were correlated within and between tests and a cluster analysis showed three possible behavioural syndrome-axes, which we name boldness, exploration and activity. Activity and exploration behaviour in all tests was highly repeatable in adult animals tested over one week. In animals tested over maturation, exploration behaviour was consistent whereas activity was not. Voles that were tested as adults with a three-month interval showed the opposite pattern with stable activity but unstable exploration behaviour. Conclusions: The consistency in behaviour over time suggests that common voles do express stable personality over short time. Over longer periods however, behaviour is more flexible and depending on life stage (i.e. tested before/after maturation or as adults) of the tested individual. Level of boldness or activity does not differ between tested groups and maintenance of variation in behavioural traits can therefore not be explained by expected future assets as reported in other studies. T3 - Zweitveröffentlichungen der Universität Potsdam : Mathematisch-Naturwissenschaftliche Reihe - 376 KW - animal personality KW - behavioural type KW - Microtus arvalis KW - common vole KW - plasticity KW - consistency KW - repeatability Y1 - 2017 U6 - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:kobv:517-opus4-401395 ER - TY - THES A1 - Robaina Estevez, Semidan T1 - Context-specific metabolic predictions T1 - Kontextspezifische metabolische Vorhersagen BT - computational methods and applications BT - Berechnungsmethoden und Anwendungen N2 - All life-sustaining processes are ultimately driven by thousands of biochemical reactions occurring in the cells: the metabolism. These reactions form an intricate network which produces all required chemical compounds, i.e., metabolites, from a set of input molecules. Cells regulate the activity through metabolic reactions in a context-specific way; only reactions that are required in a cellular context, e.g., cell type, developmental stage or environmental condition, are usually active, while the rest remain inactive. The context-specificity of metabolism can be captured by several kinds of experimental data, such as by gene and protein expression or metabolite profiles. In addition, these context-specific data can be assimilated into computational models of metabolism, which then provide context-specific metabolic predictions. This thesis is composed of three individual studies focussing on context-specific experimental data integration into computational models of metabolism. The first study presents an optimization-based method to obtain context-specific metabolic predictions, and offers the advantage of being fully automated, i.e., free of user defined parameters. The second study explores the effects of alternative optimal solutions arising during the generation of context-specific metabolic predictions. These alternative optimal solutions are metabolic model predictions that represent equally well the integrated data, but that can markedly differ. This study proposes algorithms to analyze the space of alternative solutions, as well as some ways to cope with their impact in the predictions. Finally, the third study investigates the metabolic specialization of the guard cells of the plant Arabidopsis thaliana, and compares it with that of a different cell type, the mesophyll cells. To this end, the computational methods developed in this thesis are applied to obtain metabolic predictions specific to guard cell and mesophyll cells. These cell-specific predictions are then compared to explore the differences in metabolic activity between the two cell types. In addition, the effects of alternative optima are taken into consideration when comparing the two cell types. The computational results indicate a major reorganization of the primary metabolism in guard cells. These results are supported by an independent 13C labelling experiment. N2 - Alle lebenserhaltenden Prozesse werden durch tausende biochemische Reaktionen in der Zelle bestimmt, welche den Metabolismus charakterisieren. Diese Reaktionen bilden ein komplexes Netzwerk, welches alle notwendigen chemischen Verbindungen, die sogenannten Metabolite, aus einer bestimmten Menge an Ausgangsmolekülen produziert Zellen regulieren ihren Stoffwechsel kontextspezifisch, dies bedeutet, dass nur Reaktionen die in einem zellulären Kontext, zum Beispiel Zelltyp, Entwicklungsstadium oder verschiedenen Umwelteinflüssen, benötigt werden auch tatsächlich aktiv sind. Die übrigen Reaktionen werden als inaktiv betrachtet. Die Kontextspezifität des Metabolismus kann durch verschiedene experimentelle Daten, wie Gen- und Proteinexpressionen oder Metabolitprofile erfasst werden. Zusätzlich können diese Daten in Computersimulationen des Metabolismus integriert werden, um kontextspezifische (metabolische) Vorhersagen zu treffen. Diese Doktorarbeit besteht aus drei unabhängigen Studien, welche die Integration von kontextspezifischen experimentellen Daten in Computersimulationen des Metabolismus thematisieren. Die erste Studie beschreibt ein Konzept, basierend auf einem mathematischen Optimierungsproblem, welches es erlaubt kontextspezifische, metabolische Vorhersagen zu treffen. Dabei bietet diese vollautomatische Methode den Vorteil vom Nutzer unabhängige Parameter, zu verwenden. Die zweite Studie untersucht den Einfluss von alternativen optimalen Lösungen, welche bei kontextspezifischen metabolischen Vorhersagen generiert werden. Diese alternativen Lösungen stellen metabolische Modellvorhersagen da, welche die integrierten Daten gleichgut wiederspiegeln, sich aber grundlegend voneinander unterscheiden können. Diese Studie zeigt verschiedene Ansätze alternativen Lösungen zu analysieren und ihren Einfluss auf die Vorhersagen zu berücksichtigen. Schlussendlich, untersucht die dritte Studie die metabolische Spezialisierung der Schließzellen in Arabidopsis thaliana und vergleicht diese mit einer weiteren Zellart, den Mesophyllzellen. Zu diesem Zweck wurden die in dieser Doktorarbeit vorgestellten Methoden angewandt um metabolische Vorhersagen speziell für Schließzellen und Mesophyllzellen zu erhalten. Anschließend wurden die zellspezifischen Vorhersagen auf Unterschiede in der metabolischen Aktivität der Zelltypen, unter Berücksichtigung des Effekt von alternativen Optima, untersucht. Die Ergebnisse der Simulationen legen eine grundlegende Neuorganisation des Primärmetabolismus in Schließzellen verglichen mit Mesophyllzellen nahe. Diese Ergebnisse werden durch unabhängige 13C markierungs Experimente bestätigt. KW - systems biology KW - bioinformatics KW - metabolic networks KW - constraint-based modeling KW - data integration KW - Systemsbiologie KW - Bioinformatik KW - Stoffwechselnetze KW - Constraint-basierte Modellierung KW - Datenintegration Y1 - 2017 U6 - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:kobv:517-opus4-401365 ER -