TY - BOOK A1 - Raters, Marie-Luise T1 - Das moralische Dilemma im Ethik-Unterricht : moralphilosophische Überlegungen zur Dilemma-Methode nach Lawrence Kohlberg Y1 - 2011 SN - 978-3-942411-29-5 PB - Thelem CY - Dresden ER - TY - JOUR A1 - Valdivia Orozco, Pablo Emilio T1 - Configuraciones del convivir: algunos apuntes sobre el cruce teorico de la novela y el Caribe Y1 - 2011 SN - 978-3-938944-54-7 ER - TY - JOUR A1 - Röder, Katrin T1 - Mobiles Glück in den englischen Romanen des 18. Jahrhunderts JF - Mobilisierte Kulturen Y1 - 2011 U6 - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:kobv:517-opus-57356 SN - 2192-3019 SN - 2192-3027 IS - 1 SP - 91 EP - 133 ER - TY - JOUR A1 - Sammler, Svenja A1 - Bleidorn, Christoph A1 - Tiedemann, Ralph T1 - Full mitochondrial genome sequences of two endemic Philippine hornbill species (Aves: Bucerotidae) provide evidence for pervasive mitochondrial DNA recombination JF - BMC genomics N2 - Background: Although nowaday it is broadly accepted that mitochondrial DNA (mtDNA) may undergo recombination, the frequency of such recombination remains controversial. Its estimation is not straightforward, as recombination under homoplasmy (i.e., among identical mt genomes) is likely to be overlooked. In species with tandem duplications of large mtDNA fragments the detection of recombination can be facilitated, as it can lead to gene conversion among duplicates. Although the mechanisms for concerted evolution in mtDNA are not fully understood yet, recombination rates have been estimated from "one per speciation event" down to 850 years or even "during every replication cycle". Results: Here we present the first complete mt genome of the avian family Bucerotidae, i.e., that of two Philippine hornbills, Aceros waldeni and Penelopides panini. The mt genomes are characterized by a tandemly duplicated region encompassing part of cytochrome b, 3 tRNAs, NADH6, and the control region. The duplicated fragments are identical to each other except for a short section in domain I and for the length of repeat motifs in domain III of the control region. Due to the heteroplasmy with regard to the number of these repeat motifs, there is some size variation in both genomes; with around 21,657 bp (A. waldeni) and 22,737 bp (P. panini), they significantly exceed the hitherto longest known avian mt genomes, that of the albatrosses. We discovered concerted evolution between the duplicated fragments within individuals. The existence of differences between individuals in coding genes as well as in the control region, which are maintained between duplicates, indicates that recombination apparently occurs frequently, i. e., in every generation. Conclusions: The homogenised duplicates are interspersed by a short fragment which shows no sign of recombination. We hypothesize that this region corresponds to the so-called Replication Fork Barrier (RFB), which has been described from the chicken mitochondrial genome. As this RFB is supposed to halt replication, it offers a potential mechanistic explanation for frequent recombination in mitochondrial genomes. Y1 - 2011 U6 - https://doi.org/10.1186/1471-2164-12-35 SN - 1471-2164 VL - 12 IS - 2 PB - BioMed Central CY - London ER - TY - GEN A1 - Pokorny, Ina A1 - Sharma, Reeta A1 - Goyal, Surendra Prakash A1 - Mishra, Sudanshu A1 - Tiedemann, Ralph T1 - MHC class I and MHC class II DRB gene variability in wild and captive Bengal tigers (Panthera tigris tigris) (vol 10, pg 667, 2010) T2 - Immunogenetics Y1 - 2011 U6 - https://doi.org/10.1007/s00251-010-0496-2 SN - 0093-7711 VL - 63 IS - 2 SP - 121 EP - 121 PB - Springer CY - New York ER - TY - JOUR A1 - Wiesner, Kerstin R. A1 - Loxdale, Hugh D. A1 - Köhler, Günter A1 - Schneider, Anja R. R. A1 - Tiedemann, Ralph A1 - Weisser, Wolfgang W. T1 - Patterns of local and regional genetic structuring in the meadow grasshopper, Chorthippus parallelus (Orthoptera: Acrididae), in Central Germany revealed using microsatellite markers JF - Biological journal of the Linnean Society : a journal of evolution N2 - The meadow grasshopper, Chorthippus parallelus (Zetterstedt), is common and widespread in Central Europe, with a low dispersal range per generation. A population study in Central Germany (Frankenwald and Thuringer Schiefergebirge) showed strong interpopulation differences in abundance and individual fitness. We examined genetic variability using microsatellite markers within and between 22 populations in a short-to long-distance sampling (19 populations, Frankenwald, Schiefergebirge, as well as a southern transect), and in the Erzgebirge region (three populations), with the latter aiming to check for effects as a result of historical forest cover. Of the 671 C. parallelus captured, none was macropterous (functionally winged). All populations showed a high level of expected and observed heterozygosity (mean 0.80-0.90 and 0.60-0.75, respectively), whereas there was evidence of inbreeding (F(IS) values all positive). Allelic richness for all locus-population combinations was high (mean 9.3-11.2), whereas alleles per locus ranged from 15-62. At a local level, genic and genotypic differences were significant. Pairwise F(ST) values were in the range 0.00-0.04, indicating little interpopulation genetic differentiation. Similarly, the calculated gene flow was very high, based on the respective F(ST) (19.5) and using private alleles (7.7). A Neighbour-joining tree using Nei's D(A) and principal coordinate analysis separated two populations that were collected in the Erzgebirge region. Populations from this region may have escaped the effects of the historical forest cover. The visualization of the spatial arrangement of genotypes revealed one geographical barrier to gene flow in the short-distance sampling. KW - adaptation KW - gene flow KW - diversity KW - landscape structure KW - wing polyphenism Y1 - 2011 U6 - https://doi.org/10.1111/j.1095-8312.2011.01698.x SN - 0024-4066 VL - 103 IS - 4 SP - 875 EP - 890 PB - Wiley-Blackwell CY - Malden ER - TY - JOUR A1 - Stoof-Leichsenring, Kathleen Rosemarie A1 - Junginger, Annett A1 - Olaka, Lydia A. A1 - Tiedemann, Ralph A1 - Trauth, Martin H. T1 - Environmental variability in Lake Naivasha, Kenya, over the last two centuries JF - Journal of paleolimnolog N2 - Lake Naivasha, Kenya, is one of a number of freshwater lakes in the East African Rift System. Since the beginning of the twentieth century, it has experienced greater anthropogenic influence as a result of increasingly intensive farming of coffee, tea, flowers, and other horticultural crops within its catchment. The water-level history of Lake Naivasha over the past 200 years was derived from a combination of instrumental records and sediment data. In this study, we analysed diatoms in a lake sediment core to infer past lacustrine conductivity and total phosphorus concentrations. We also measured total nitrogen and carbon concentrations in the sediments. Core chronology was established by (210)Pb dating and covered a similar to 186-year history of natural (climatic) and human-induced environmental changes. Three stratigraphic zones in the core were identified using diatom assemblages. There was a change from littoral/epiphytic diatoms such as Gomphonema gracile and Cymbella muelleri, which occurred during a prolonged dry period from ca. 1820 to 1896 AD, through a transition period, to the present planktonic Aulacoseira sp. that favors nutrient-rich waters. This marked change in the diatom assemblage was caused by climate change, and later a strong anthropogenic overprint on the lake system. Increases in sediment accumulation rates since 1928, from 0.01 to 0.08 g cm(-2) year(-1) correlate with an increase in diatom-inferred total phosphorus concentrations since the beginning of the twentieth century. The increase in phosphorus accumulation suggests increasing eutrophication of freshwater Lake Naivasha. This study identified two major periods in the lake's history: (1) the period from 1820 to 1950 AD, during which the lake was affected mainly by natural climate variations, and (2) the period since 1950, during which the effects of anthropogenic activity overprinted those of natural climate variation. KW - Lake sediments KW - Diatoms KW - Conductivity KW - Lake Naivasha KW - Human impact KW - Eutrophication Y1 - 2011 U6 - https://doi.org/10.1007/s10933-011-9502-4 SN - 0921-2728 VL - 45 IS - 3 SP - 353 EP - 367 PB - Springer CY - Dordrecht ER - TY - JOUR A1 - Schwarte, Sandra A1 - Tiedemann, Ralph T1 - A Gene Duplication/Loss Event in the Ribulose-1,5-Bisphosphate-Carboxylase/Oxygenase (Rubisco) Small Subunit Gene Family among Accessions of Arabidopsis thaliana JF - Molecular biology and evolution N2 - Rubisco (ribulose-1,5-bisphosphate carboxylase/oxygenase; EC 4.1.1.39), the most abundant protein in nature, catalyzes the assimilation of CO(2) (worldwide about 10(11) t each year) by carboxylation of ribulose-1,5-bisphosphate. It is a hexadecamer consisting of eight large and eight small subunits. Although the Rubisco large subunit (rbcL) is encoded by a single gene on the multicopy chloroplast genome, the Rubisco small subunits (rbcS) are encoded by a family of nuclear genes. In Arabidopsis thaliana, the rbcS gene family comprises four members, that is, rbcS-1a, rbcS-1b, rbcS-2b, and rbcS-3b. We sequenced all Rubisco genes in 26 worldwide distributed A. thaliana accessions. In three of these accessions, we detected a gene duplication/loss event, where rbcS-1b was lost and substituted by a duplicate of rbcS-2b (called rbcS-2b*). By screening 74 additional accessions using a specific polymerase chain reaction assay, we detected five additional accessions with this duplication/loss event. In summary, we found the gene duplication/loss in 8 of 100 A. thaliana accessions, namely, Bch, Bu, Bur, Cvi, Fei, Lm, Sha, and Sorbo. We sequenced an about 1-kb promoter region for all Rubisco genes as well. This analysis revealed that the gene duplication/loss event was associated with promoter alterations (two insertions of 450 and 850 bp, one deletion of 730 bp) in rbcS-2b and a promoter deletion (2.3 kb) in rbcS-2b* in all eight affected accessions. The substitution of rbcS-1b by a duplicate of rbcS-2b (i.e., rbcS-2b*) might be caused by gene conversion. All four Rubisco genes evolve under purifying selection, as expected for central genes of the highly conserved photosystem of green plants. We inferred a single positive selected site, a tyrosine to aspartic acid substitution at position 72 in rbcS-1b. Exactly the same substitution compromises carboxylase activity in the cyanobacterium Anacystis nidulans. In A. thaliana, this substitution is associated with an inferred recombination. Functional implications of the substitution remain to be evaluated. KW - Arabidopsis thaliana KW - Arabidopsis lyrata KW - Rubisco KW - gene duplication KW - positive selection Y1 - 2011 U6 - https://doi.org/10.1093/molbev/msr008 SN - 0737-4038 VL - 28 IS - 6 SP - 1861 EP - 1876 PB - Oxford Univ. Press CY - Oxford ER - TY - JOUR A1 - Tiedemann, Ralph A1 - Paulus, Kirsten B. A1 - Havenstein, Katja A1 - Thorstensen, Sverrir A1 - Petersen, Aevar A1 - Lyngs, Peter A1 - Milinkovitch, Michel C. T1 - Alien eggs in duck nests brood parasitism or a help from Grandma? JF - Molecular ecology N2 - Intraspecific brood parasitism (IBP) is a remarkable phenomenon by which parasitic females can increase their reproductive output by laying eggs in conspecific females' nests in addition to incubating eggs in their own nest. Kin selection could explain the tolerance, or even the selective advantage, of IBP, but different models of IBP based on game theory yield contradicting predictions. Our analyses of seven polymorphic autosomal microsatellites in two eider duck colonies indicate that relatedness between host and parasitizing females is significantly higher than the background relatedness within the colony. This result is unlikely to be a by-product of relatives nesting in close vicinity, as nest distance and genetic identity are not correlated. For eider females that had been ring-marked during the decades prior to our study, our analyses indicate that (i) the average age of parasitized females is higher than the age of nonparasitized females, (ii) the percentage of nests with alien eggs increases with the age of nesting females, (iii) the level of IBP increases with the host females' age, and (iv) the number of own eggs in the nest of parasitized females significantly decreases with age. IBP may allow those older females unable to produce as many eggs as they can incubate to gain indirect fitness without impairing their direct fitness: genetically related females specialize in their energy allocation, with young females producing more eggs than they can incubate and entrusting these to their older relatives. Intraspecific brood parasitism in ducks may constitute cooperation among generations of closely related females. KW - eider duck KW - indirect fitness KW - intraspecific brood parasitism KW - microsatellites KW - relatedness KW - Somateria mollissima Y1 - 2011 U6 - https://doi.org/10.1111/j.1365-294X.2011.05158.x SN - 0962-1083 VL - 20 IS - 15 SP - 3237 EP - 3250 PB - Wiley-Blackwell CY - Hoboken ER - TY - JOUR A1 - Epp, Laura Saskia A1 - Stoof-Leichsenring, Kathleen Rosemarie A1 - Trauth, Martin H. A1 - Tiedemann, Ralph T1 - Molecular profiling of diatom assemblages in tropical lake sediments using taxon-specific PCR and Denaturing High-Performance Liquid Chromatography (PCR-DHPLC) JF - Molecular ecology resources N2 - Here we present a protocol to genetically detect diatoms in sediments of the Kenyan tropical Lake Naivasha, based on taxon-specific PCR amplification of short fragments (approximately 100 bp) of the small subunit ribosomal (SSU) gene and subsequent separation of species-specific PCR products by PCR-based denaturing high-performance liquid chromatography (DHPLC). An evaluation of amplicons differing in primer specificity to diatoms and length of the fragments amplified demonstrated that the number of different diatom sequence types detected after cloning of the PCR products critically depended on the specificity of the primers to diatoms and the length of the amplified fragments whereby shorter fragments yielded more species of diatoms. The DHPLC was able to discriminate between very short amplicons based on the sequence difference, even if the fragments were of identical length and if the amplicons differed only in a small number of nucleotides. Generally, the method identified the dominant sequence types from mixed amplifications. A comparison with microscopic analysis of the sediment samples revealed that the sequence types identified in the molecular assessment corresponded well with the most dominant species. In summary, the PCR-based DHPLC protocol offers a fast, reliable and cost-efficient possibility to study DNA from sediments and other environmental samples with unknown organismic content, even for very short DNA fragments. KW - diatoms KW - environmental DNA KW - lake sediments KW - PCR-DHPLC Y1 - 2011 U6 - https://doi.org/10.1111/j.1755-0998.2011.03022.x SN - 1755-098X VL - 11 IS - 5 SP - 842 EP - 853 PB - Wiley-Blackwell CY - Hoboken ER - TY - JOUR A1 - Pavesi, Laura A1 - De Matthaeis, Elvira A1 - Tiedemann, Ralph A1 - Ketmaier, Valerio T1 - Temporal population genetics and COI phylogeography of the sandhopper macarorchestia remyi (Amphipoda: Talitridae) JF - Zoological studies N2 - Laura Pavesi, Elvira De Matthaeis, Ralph Tiedemann, and Valerio Ketmaier (2011) Temporal population genetics and COI phylogeography of the sandhopper Macarorchestia remyi (Amphipoda: Talitridae). Zoological Studies 50(2): 220-229. In this study we assessed levels of genetic divergence and variability in 208 individuals of the supralittoral sandhopper Macarorchestia remyi, a species strictly associated with rotted wood stranded on sand beaches, by analyzing sequence polymorphisms in a fragment of the mitochondrial DNA (mtDNA) gene coding cytochrome oxidase subunit I (COI). The geographical distribution and ecology of the species are poorly known. The study includes 1 Tyrrhenian and 2 Adriatic populations sampled along the Italian peninsula plus a single individual found on Corfu Is. (Greece). The Tyrrhenian population was sampled monthly for 1 yr. Genetic data revealed a deep phylogeographic break between the Tyrrhenian and Adriatic populations with no shared haplotypes. The single individual collected on Corfu Is. carried the most common haplotype found in the Tyrrhenian population. A mismatch analysis could not reject the hypothesis of a sudden demographic expansion in almost all but 2 monthly samples. When compared to previous genetic data centered on a variety of Mediterranean talitrids, our results place M. remyi among those species with profound intraspecific divergence (sandhoppers) and dissimilar from beachfleas, which generally display little population genetic structuring. KW - Macarorchestia remyi KW - Talitridae KW - Cytochrome oxidase I KW - Population genetics Y1 - 2011 SN - 1021-5506 VL - 50 IS - 2 SP - 220 EP - 229 PB - Institute of Zoology, Academia Sinica CY - Taipei ER - TY - INPR A1 - Walz, Norbert A1 - Adrian, Rita A1 - Gilbert, John J. A1 - Monaghan, Michael T. A1 - Weithoff, Guntram A1 - Zimmermann-Timm, Heike T1 - Preface T2 - Hydrobiologia : acta hydrobiologica, hydrographica, limnologica et protistologica Y1 - 2011 U6 - https://doi.org/10.1007/s10750-010-0514-2 SN - 0018-8158 VL - 662 IS - 1 SP - 1 EP - 4 PB - Springer CY - Dordrecht ER - TY - JOUR A1 - Sharma, Reeta A1 - Stuckas, Heiko A1 - Bhaskar, Ranjana A1 - Khan, Imran A1 - Goyal, Surendra Prakash A1 - Tiedemann, Ralph T1 - Genetically distinct population of Bengal tiger (Panthera tigris tigris) in Terai Arc Landscape (TAL) of India JF - Mammalian biology = Zeitschrift für Säugetierkunde N2 - We analyzed mtDNA polymorphisms (a total of 741 bp from a part of conserved control region, ND5, ND2, Cyt b and 12S) in 91 scats and 12 tissue samples of Bengal tiger (Panthera tigris tigris) populations across Terai Arc Landscape (TAL) located at the foothills of Himalayas in North Western India, Buxa Tiger Reserve (BTR), and North East India. In TAL and BTR, we found a specific haplotype at high frequency, which was absent elsewhere, indicating a genetically distinct population in these regions. Within the TAL region, there is some evidence for genetic isolation of the tiger populations west of river Ganges, i.e., in the western part of Rajaji National Park (RNP). Although the river itself might not constitute a significant barrier for tigers, recent human-induced changes in habitat and degradation of the Motichur-Chilla Corridor connecting the two sides of the tiger habitat of RNP might effectively prevent genetic exchange. A cohesive population is observed for the rest of the TAL. Even the more eastern BTR belongs genetically to this unit, despite the present lack of a migration corridor between BTR and TAL. In spite of a close geographic proximity, Chitwan (Nepal) constitutes a tiger population genetically different from TAL. Moreover, it is observed that the North East India tiger populations are genetically different from TAL and BTR, as well as from the other Bengal tiger populations in India. KW - Bengal tiger KW - Population genetics KW - mtDNA haplotype KW - Terai Arc Y1 - 2011 U6 - https://doi.org/10.1016/j.mambio.2010.10.005 SN - 1616-5047 VL - 76 IS - 4 SP - 484 EP - 490 PB - Elsevier CY - Jena ER - TY - JOUR A1 - Nahavandi, Nahid A1 - Plath, Martin A1 - Tiedemann, Ralph A1 - Mirzajani, Ali R. T1 - Sexual and natural selection on morphological traits in a marine amphipod, Pontogammarus maeoticus (Sowinsky, 1894) JF - Marine biology research N2 - Sexual selection often leads to sexual dimorphism, where secondary sexual traits are more expressed in the male sex. This may be due, for example, to increased fighting or mate-guarding abilities of males expressing those traits. We investigated sexually dimorphic traits in four populations of a marine amphipod, Pontogammarus maeoticus (Gammaridea: Pontogammaridae), the most abundant amphipod species in the sublittoral zone along the southern shoreline of the Caspian Sea. Male amphipods are typically larger in body size than females, and have relatively larger posterior gnathopods and antennae. However, it remains to be studied for most other body appendages whether or not, and to what extent, they are sexually dimorphic. Using Analysis of Covariance (ANCOVA), we compared the relationships between body size and trait expression for 35 metric characters between males and females, and among the four populations examined by performing three different Discriminant Function Analyses (DFA). We detected several thus far undescribed sexual dimorphic traits such as the seventh peraeopods or the epimeral plates. We also found that the size of the propodus of the first and second gnathopods increases with increasing body size, and this allometric increase was stronger in males than in females. Finally, we found that the degree of sexual dimorphism in the expression of the width of the third epimeral plate varies across sites, suggesting that differences in ecology might affect the strength of sexual selection in different populations. KW - Caspian Sea KW - crustacean KW - dimorphism KW - scramble competition Y1 - 2011 U6 - https://doi.org/10.1080/17451001003713589 SN - 1745-1000 VL - 7 IS - 2 SP - 135 EP - 146 PB - Taylor & Francis Group CY - Oslo ER - TY - JOUR A1 - Virchow, Fabian T1 - Die „Demonstrationspolitik“ der extremen Rechten im Bundesgebiet und im Land Brandenburg JF - Die Grenzen der Toleranz : rechtsextremes Milieu und demokratische Gesellschaft in Brandenburg ; Bilanz und Perspektiven Y1 - 2011 U6 - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:kobv:517-opus-52208 SN - 978-3-86956-038-0 SP - 109 EP - 128 ER - TY - JOUR A1 - Schnittcher, Gerd T1 - Bekämpfung des Rechtsextremismus (PMK) aus Sicht einer brandenburgischen Staatsanwaltschaft JF - Die Grenzen der Toleranz : rechtsextremes Milieu und demokratische Gesellschaft in Brandenburg ; Bilanz und Perspektiven Y1 - 2011 U6 - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:kobv:517-opus-52223 SN - 978-3-86956-038-0 SP - 151 EP - 165 ER - TY - THES A1 - Schatz, Daniela T1 - LNA-clamp-PCR zum sensitiven Nachweis von Punktmutationen im Rahmen der Entwicklung eines Darmkrebsfrüherkennungstests T1 - LNA-clamp-PCR as a method for sensitive detection of point mutations as part of the development of an assay for the early diagnosis of colon cancer N2 - Darmkrebs ist die zweithäufigste malignombedingte Todesursache in den westlichen Industrieländern. Durch eine frühzeitige Diagnose besteht jedoch eine hohe Chance auf Heilung. Der Goldstandard zur Darmkrebsfrüherkennung ist gegenwärtig die Koloskopie. Eine Darmspiegelung ist jedoch invasiv und mit Unannehmlichkeiten für den Patienten verbunden. Die Akzeptanz in der Bevölkerung ist daher gering. Ziel des BMBF- Projektes „Entwicklung eines nichtinvasiven Nachweissystems zur Früherkennung von humanem Darmkrebs“, in dessen Rahmen diese Arbeit entstand, ist die Bereitstellung eines nichtinvasiven Nachweisverfahrens zur Darmkrebsfrüherkennung. Der Nachweis soll über die Detektion von aus neoplastischen Zellen stammender DNA in Stuhl erfolgen. Die Entartung dieser Zellen beruht auf Veränderungen im Erbgut, welches unter anderem Mutationen sind. Im ersten Teil des BMBF-Projektes wurde ein Set von Mutationen zusammengestellt, welches eine hohe Sensitivität für Vorstufen von Darmkrebs aufweist. Ziel dieser Arbeit war es, eine Nachweismethode für die zuvor identifizierten Punktmutationen zu entwickeln. Das Nachweisverfahren musste dabei unempfindlich gegen einen hohen Hintergrund nichtmutierter DNA sein, da im Stuhl geringe Mengen DNA aus neoplastischen Zellen bei einem hohen Hintergrund von DNA aus gesunden Zellen vorliegen. Hierzu wurden Plasmidmodellsysteme für die aus dem Marker-Set stammenden Genfragmente BRAF und dessen Mutante V600E, CTNNB1 und T41I, T41A, S45P und K-ras G12C hergestellt. Mit Hilfe dieser Plasmidmodellsysteme wurde dann das Nachweissystem entwickelt. Der entscheidende Schritt für die Detektion von Punktmutationen bei hohem Wildtypüberschuss ist eine vorhergehende Anreicherung. In der vorliegenden Arbeit wurde dazu die Methode der LNA-clamp-PCR (locked nucleic acid) etabliert. Die Bewertung der erzielten Anreicherung erfolgte über das relative Detektionslimit. Zur Bestimmung des Detektionslimits wurde die Schmelzkurvenanalyse von Hybridisierungssonden eingesetzt; diese wurde im Rahmen dieser Arbeit für die drei oben genannten Genfragmente und ihre Mutanten entwickelt. Die LNA-clamp-PCR wird in Anwesenheit eines LNA-Blockers durchgeführt. Das Nukleotidanalogon LNA weist im Vergleich zu DNA eine erhöhte Affinität zu komplementären DNA-Strängen auf. Gleichzeitig kommt es bei Anwesenheit einer Basenfehlpaarung zu einer größeren Destabilisierung der Bindung. Als Blocker werden kurze LNA-DNA-Hybridoligonukleotide eingesetzt, die den mutierten Sequenzbereich überspannen und selbst der Wildtypsequenz entsprechen. Durch Bindung an die Wildtypsequenz wird deren Amplifikation während der PCR verhindert (clamp = arretieren, festklemmen). Der Blocker selbst wird dabei nicht verlängert. Der Blocker bindet unter optimalen Bedingungen jedoch nicht an die mutierte Sequenz. Die Mutante wird daher ungehindert amplifiziert und somit gegenüber dem Wildtyp-Fragment angereichert. Die Position des Blockers kann im Bindungsbereich eines der Primer sein und hier dessen Hybridisierung an dem Wildtyp-Fragment verhindern oder zwischen den beiden Primern liegen und so die Synthese durch die Polymerase inhibieren. Die Anwendbarkeit beider Systeme wurde in dieser Arbeit gezeigt. Die LNA-clamp-PCR mit Primerblocker wurde für BRAF etabliert. Es wurde ein Detektionslimit von mindestens 1:100 erzielt. Die LNA-clamp-PCR mit Amplifikationsblocker wurde erfolgreich für BRAF, K-ras und CTNNB1: T41I, T41A mit einem Detektionslimit von 1:1000 bis 1:10 000 entwickelt. In Stuhlproben liegt DNA aus neoplastischen Zellen nach Literaturangaben zu einem Anteil von 1% bis 0,1% vor. Die LNA-clamp-PCR weist also mit Amplifikationsblockern ein ausreichend hohes Detektionslimit für die Analyse von Stuhlproben auf. Durch die erfolgreiche Etablierung der Methode auf drei verschiedenen Genfragmenten und vier unterschiedlichen Punktmutationen konnte deren universelle Einsetzbarkeit gezeigt werden. Für die Ausweitung der LNA-clamp-PCR auf die übrigen Mutationen des Marker-Sets wurden Richtlinien ausgearbeitet und die Blockereffizienz als Kennzahl eingeführt. Die LNA-clamp-PCR ist ein schnelles, kostengünstiges Verfahren, welches einen geringen Arbeitsaufwand erfordert und wenig fehleranfällig ist. Sie ist somit ein geeignetes Anreicherungsverfahren für Punktmutationen in einem diagnostischen System zur Darmkrebsfrüherkennung. Darüber hinaus kann die LNA-clamp-PCR auch in anderen Bereichen, in denen die Detektion von Punktmutationen in einem hohen Wildtyphintergrund erforderlich ist, eingesetzt werden. N2 - Colon cancer is the second leading cause of cancer related deaths in the western world. However if diagnosed early there is a great chance curing the disease. Coloscopy is the gold standard for early detection of colorectal cancer today. Its greatest disadvantage is the fact that it is an invasive technique and provides some discomfort for the patients. Therefore, the compliance to undergo such a procedure is extremely low. This work was generated in the context of the BMBF-project „Development of a non-invasive assay for the early detection of preneoplastic and neoplastic lesions in the human colon“. The aim of the work described here is the development of a non-invasive assay for the early detection of colon cancer. The assay should detect DNA from neoplastic cells in feces samples. The transformation of these cells is based on alterations in the genome predominantly mutations. In the first part of the BMBF-project a mutation panel with high sensitivity for preneoplastic lesions of colon cancer was determined. The aim of this work was to develop a detection method for the point mutations of the determined mutation panel. The rare mutant DNA needs to be detected in the presence of a great amount of wild-type DNA shed from healthy tissue. The assay system needs to be insensitive to this high background of healthy DNA. Therefore a model system of plasmid DNA containing gene fragments of BRAF and its mutation V600E, CTNNB1 and T41I, T41A, S45P and K-ras G12C obtained from the marker panel was established. Using these plasmid system the detection method was developed. The most critical parameter for the detection of rare point mutations is an enrichment of these rare DNA molecules. In this work LNA-clamp-PCR (locked nucleic acid) technology was used to enrich the mutant DNA.. For the estimation of the achieved enrichment the relative detection limit was used. The detection limit was determined by melting curve analysis of hybridization probes. These assays were established in the present work for the three above mentioned gene fragments. LNA-clamp-PCR is performed in the presence of an LNA blocker. LNA is a synthetic DNA analog. LNA nucleotide analog bind to complementary DNA strands with higher affinity. In addition a single mismatch in the LNA-DNA duplex causes a much greater destabilization compared to a DNA-DNA duplex. Short LNA-DNA-hybrids were used as clamp, which cover the mutated region and represent the wild-type sequence. Within an appropriate temperature range, LNA can specifically bind to wild type template and can inhibit its amplification. The clamp itself will not be elongated. Under optimal conditions the LNA clamp will not interfere with the amplification of the mismatched template. Therefore the mutated gene fragment will be enriched in comparison to the wild-type. The position of the LNA clamp can either be at the primer binding site inhibiting primer hybridization on the wild-type fragment or the LNA clamp is positioned between the two primer binding sites inhibiting chain elongation of the perfectly matched template. In the present work both systems were applied. For the gene fragment BRAF the LNA was used at the primer binding site. The achieved detection limit was at least 1:100. The LNA-clamp-PCR with LNA inhibiting the chain elongation were developed successfully for BRAF, K-ras and CTNNB1: T41I, T41A achieving a detection limit of 1:1000 to 1:10 000. According to the literature 1% to 0.1% of the DNA in feces derives from neoplastic cells. Therefore the detection limit achieved by LNA-clamp-PCR with LNA inhibiting chain elongation would be sufficient for analyzing feces samples. LNA-clamp-PCR protocols were established for three different gene fragments and four diverse point mutations indicating that the technology can generally be used for high sensitive detection of DNA mutations. For the development of LNA-clamp-PCR protocols for the other mutations of the marker panel development guidelines were established. Clamp efficiency was identified as a quantitative parameter for protocol optimization. The LNA-clamp-PCR is a robust, fast and cost-saving technique which needs low labor input. Therefore the method is adequate for enriching point mutated gene fragments in a diagnostic assay for the detection of early colon cancer stages. In addition LNA-clamp-PCR can be applied in other fields where rare sequence variations need to be detected in the presence of high wild-type DNA background. KW - LNA-clamp-PCR KW - Darmkrebsdiagnostik KW - Punktmutation KW - BRAF KW - K-ras KW - LNA- clamp-PCR KW - colon cancer diagnosis KW - point mutation KW - BRAF KW - K-ras Y1 - 2011 U6 - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:kobv:517-opus-52308 ER - TY - JOUR A1 - Butterwegge, Christoph T1 - Finanzmarktkrise, Armut und rechtsextreme Politik JF - Die Grenzen der Toleranz : rechtsextremes Milieu und demokratische Gesellschaft in Brandenburg ; Bilanz und Perspektiven Y1 - 2011 U6 - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:kobv:517-opus-52179 SN - 978-3-86956-038-0 SP - 41 EP - 55 ER - TY - JOUR A1 - Sturzbecher, Dietmar A1 - Burkert, Mathias A1 - Hoffmann, Lars T1 - Zwischen Akzeptanz und Absage BT - das Verhältnis brandenburgischer Jugendlicher zu Gewalt und Rechtsextremismus JF - Die Grenzen der Toleranz : rechtsextremes Milieu und demokratische Gesellschaft in Brandenburg ; Bilanz und Perspektiven Y1 - 2011 U6 - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:kobv:517-opus-52193 SN - 978-3-86956-038-0 SP - 83 EP - 107 ER - TY - JOUR A1 - Schubarth, Wilfried A1 - Speck, Karsten A1 - Seidel, Andreas A1 - Kamm, Caroline A1 - Kleinfeld, Merle A1 - Sarrar, Lea T1 - Evidenzbasierte Professionalisierung der Praxisphasen in außeruniversitären Lernorten BT - Erste Ergebnisse des Forschungsprojektes ProPrax JF - Potsdamer Beiträge zur Hochschulforschung Y1 - 2011 U6 - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:kobv:517-opus-52687 SN - 2192-1075 SN - 2192-1083 IS - 1 SP - 79 EP - 212 ER -