TY - THES A1 - Zwettler, Fabian Ulrich T1 - Expansionsmikroskopie kombiniert mit hochauflösender Fluoreszenzmikroskopie T1 - Expansion Microscopy combined with Super-Resolution Fluorescence Microscopy N2 - Fluorescence microscopy is a form of light microscopy that has developed during the 20th century and is nowadays a standard tool in Molecular and Cell biology for studying the structure and function of biological molecules. High-resolution fluorescence microscopy techniques, such as dSTORM (direct Stochastic Optical Reconstruction Microscopy) allow the visualization of cellular structures at the nanometre scale (10−9 m). This has already made it possible to decipher the composition and function of various biopolymers, such as proteins, lipids and nucleic acids, up to the three-dimensional (3D) structure of entire organelles. In practice, however, it has been shown that these imaging methods and their further developments still face great challenges in order to achieve an effective resolution below ∼ 10 nm. This is mainly due to the nature of labelling biomolecules. For the detection of molecular structures, immunostaining is often performed as a standard method. Antibodies to which fluorescent molecules are coupled, recognize and bind specifcally and with high affnity to the molecular section of the target structure, also called epitope or antigen. The fluorescent molecules serve as reporter molecules which are imaged with the use of a fluorescence microscope. However, the size of these labels with a length of about 10-15 nm in the case of immunoglobulin G (IgG) antibodies, cause a detection of the fluorescent molecules shifted to the real position of the studied antigen. In dense regions where epitopes are located close to each other, steric hindrance between antibodies can also occur and leads to an insuffcient label density. Together with the shifted detection of fluorescent molecules, these factors can limit the achievable resolution of a microscopy technique. Expansion microscopy (ExM) is a recently developed technique that achieves a resolution improvement by physical expansion of an investigated object. Therefore, biological samples such as cultured cells, tissue sections, whole organs or isolated organelles are chemically anchored into a swellable polymer. By absorbing water, this so-called superabsorber increases its own volume and pulls the covalently bound biomolecules isotropically apart. Routinely, this method achieves a magnifcation of the sample by about four times its volume. But protocol variants have already been developed that result in higher expansion factors of up to 50-fold. Since the ExM technique includes in the frst instance only the sample treatment for anchoring and magnifcation of the sample, it can be combined with various standard methods of fluorescence microscopy. In theory, the resolution of the used imaging technique improves linearly with the expansion factor of the ExM treated sample. However, an insuffcient label density and the size of the antibodies can here again impair the effective achievable resolution. The combination of ExM with high-resolution fluorescence microscopy methods represents a promising strategy to increase the resolution of light microscopy. In this thesis, I will present several ExM variants I developed which show the combination of ExM with confocal microscopy, SIM (Structured Illumination Microscopy), STED (STimulated Emission Depletion) and dSTORM. I optimized existing ExM protocols and developed different expansion strategies, which allow the combination with the respective imaging technique. Thereby, I gained new structural insights of isolated centrioles from the green algae Chlamydomonas reinhardtii by combining ExM with STED and confocal microscopy. In another project, I combined 3D-SIM imaging with ExM and investigated the molecular structure of the so-called synaptonemal complex. This structure is formed during meiosis in eukaryotic cells and contributes to the exchange of genetic material between homologous chromosomes. Especially in combination with dSTORM, the ExM method showed its high potential to overcome the limitations of modern fluorescence microscopy techniques. In this project, I expanded microtubules in mammalian cells, a polymer of the cytoskeleton as well as isolated centrioles from C. reinhardtii. By labelling after expansion of the samples, I was able to signifcantly reduce the linkage error of the label and achieve an improved label density. In future, these advantages together with the single molecule sensitivity and high resolution obtained by the dSTORM method could pave the way for achieving molecular resolution in fluorescence microscopy N2 - Die Fluoreszenzmikroskopie ist eine Form der Lichtmikroskopie, die sich im Laufe des 20. Jahrhunderts entwickelt hat und heutzutage standardmäßig in der Molekular-und Zellbiologie zur Erforschung von Aufbau und Funktion biologischer Moleküle eingesetzt wird. Hochauflösende Verfahren der Fluoreszenzmikroskopie, wie die dSTORM (direct Stochastic Optical Reconstruction Microscopy) Technik, ermöglichen die Visualisierung zellulärer Strukturen im Nanometer-Größenbereich (10−9 m). Dadurch konnte bereits die Zusammensetzung und Funktion unterschiedlicher Biopolymere, wie die von Proteinen, Lipiden und Nukleinsäuren, bis hin zum dreidimensionalen (3D) Aufbau ganzer Organellen entschlüsselt werden. In der Praxis zeigt sich jedoch, dass diese Bildgebungsverfahren und ihre Weiterentwicklungen immer noch vor großen Herausforderungen stehen, bevor eine effektive Auflösung von unter ∼10 nm erreicht werden kann. Die größte Hürde stellt die Art und Weise der Markierung von Biomolekülen dar. Bei dieser wird zum Nachweis molekularer Strukturen häufig die sogenannte Immunfärbung als Standardmethode eingesetzt. Antikörper, welche mit Fluoreszenzmolekülen gekoppelt werden, erkennen und binden hierbei spezifisch und mit hoher Affinität den Molekülabschnitt der Zielstruktur, auch Epitop oder Antigen genannt. Die Fluoreszenzmoleküle dienen als Reportermoleküle, welche mit Hilfe eines Fluoreszenzmikroskops abgebildet werden. Die Größe der Antikörper, mit einer Länge von etwa 10-15 nm im Falle von Immunglobulin G (IgG) Antikörpern, bewirkt jedoch eine Detektion der fluoreszierenden Moleküle verschoben zur eigentlichen Lage des untersuchten Antigens. In Regionen mit räumlich dicht nebeneinander liegenden Epitopen kann es zusätzlich zur sterischen Hinderung zwischen den Antikörpern kommen. Dies führt zu einer unzureichenden Markierungsdichte und stellt - zusammen mit der verschobenen Detektion der Fluoreszenzmoleküle - eine Limitierung der zu erreichenden Auflösung dar. Die Expansionsmikroskopie (ExM) ist ein neu entwickeltes Verfahren, welches eine Auflösungsverbesserung durch die physikalische Expansion eines untersuchten Objekts erreicht. Hierbei werden biologische Proben, wie beispielsweise kultivierte Zellen, Gewebeschnitte, ganze Organe oder isolierte Organellen, chemisch in ein quellbares Polymer verankert. Durch Absorption von Wasser vergrößert dieser sogenannte Superabsorber sein eigenes Volumen und zieht während der räumlichen Expansion die kovalent gebundenen Biomoleküle isotrop auseinander. Standardmäßig wird durch dieses Verfahren eine Vergrößerung der Proben um etwa das vierfache Volumen erreicht, wobei bereits Protokollvarianten entwickelt wurden, die eine bis zu 50-fache Expansion erzielt haben. Da die ExM-Technik zunächst nur die Probenbehandlung zur Verankerung und Vergrößerung der Probe selbst beinhaltet, kann sie mit unterschiedlichen Standardmethoden der Fluoreszenzmikroskopie kombiniert werden. Dadurch verbessert sich die Auflösung des verwendeten Bildgebungsverfahrens theoretisch linear um den Faktor der Volumenvergrößerung der ExM behandelten Probe. Eine unzureichende Markierungsdichte und die Größe der verwendeten Antikörper können auch hier die effektiv erreichbare Auflösung beeinträchtigen. Die Kombination der ExM mit hochauflösenden Verfahren der Fluoreszenzmikroskopie stellt eine vielversprechende Strategie zur Erhöhung der bisher erreichbaren Auflösung in der Lichtmikroskopie dar. In dieser Arbeit werde ich mehrere von mir entwickelte ExM Varianten vorstellen, welche die Kombination von ExM mit konfokaler Mikroskopie, SIM (Structured Illumination Microscopy), STED (STimulated Emission Depletion) und dSTORM zeigen. Um die Verbindung mit dem jeweiligen Bildgebungsverfahren zu ermöglichen, optimierte ich bestehende ExM-Protokolle und entwickelte unterschiedliche Expansionsstrategien. Dadurch konnte ich neue strukturelle Erkenntnisse von isolierten Zentriolen aus der Grünalge Chlamydomonas reinhardtii durch die Verbindung von ExM mit STED und konfokaler Mikroskopie gewinnen. In einem weiteren Projekt kombinierte ich 3D-SIM mit ExM und untersuchte den molekularen Aufbau des sogenannten synaptonemalen Komplexes. Diese Struktur bildet sich in eukaryotischen Zellen während der Reifeteilung (Meiose) aus und trägt zum Austausch des genetischen Materials zwischen homologen Chromosomen bei. Vor allem in Verbindung mit dSTORM zeigte sich das hohe Potential der ExM-Methode, die bisherigen Limitierungen moderner Techniken der Fluoreszenzmikroskopie zu überwinden. In diesem Projekt expandierte ich Mikrotubuli in Säugetierzellen, ein Polymer des Zytoskeletts, sowie isolierte Zentriolen aus C. reinhardtii. Dadurch, dass die Markierung erst nach dem Expandieren der Proben erfolgte, gelang es, den Abstandsfehler der Markierung deutlich zu verringern und eine verbesserte Markierungsdichte zu erreichen. Diese Vorteile könnten in Verbindung mit der Einzelmolekülsensititvität und hohen Auflösung der dSTORM Methode Wegbereiter zur Erreichung einer molekularen Auflösung sein KW - Fluoreszenzmikroskopie KW - Expansionsmikroskopie KW - Einzelmolekül-Lokalisationsmikroskopie KW - Zentriolen KW - Synaptonemaler Komplex Y1 - 2021 U6 - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-212362 ER - TY - THES A1 - Zachary, Marie T1 - Functional characterization of small non-coding RNAs of \(Neisseria\) \(gonorrhoeae\) T1 - Funktionelle Charakterisierung kleiner nicht-kodierender RNAs in \(Neisseria\) \(gonorrhoeae\) N2 - During infection, bacteria need to adapt to a changing environment and have to endure various stress conditions. Small non-coding RNAs are considered as important regulators of bacterial gene expression and so allow quick adaptations by altering expression of specific target genes. Regulation of gene expression in the human-restricted pathogen Neisseria gonorrhoeae, the causative agent of the sexually transmitted disease gonorrhoea, is only poorly understood. The present study aims a better understanding of gene regulation in N. gonorrhoeae by studying small non-coding RNAs. The discovery of antisense RNAs for all opa genes led to the hypothesis of asRNA-mediated degradation of out-of-frame opa transcripts. Analysis of asRNA expression revealed a very low abundance of the transcripts and inclusion of another phase-variable gene in the study indicates that the asRNAs are not involved in degradation of out-of-frame transcripts. This doctoral thesis focuses on the analysis of trans-acting sRNAs. The sibling sRNAs NgncR_162 and NgncR_163 were discovered as post-transcriptional regulators altering expression of genes involved in metabolic processes, amino acid uptake and transcriptional regulation. A more detailed analysis by in silico and transcriptomic approaches showed that the sRNAs regulate a broad variety of genes coding for proteins of central metabolism, amino acid biosynthesis and degradation and several transport processes. Expression levels of the sibling sRNAs depend on the growth phase of the bacteria and on the growth medium. This indicates that NgncR_162 and NgncR_163 are involved in the adaptation of the gonococcal metabolism to specific growth conditions. This work further initiates characterisation of the sRNA NgncR_237. An in silico analysis showed details on sequence conservation and a possible secondary structure. A combination of in silico target prediction and differential RNA sequencing resulted in the identification of several target genes involved in type IV pilus biogenesis and DNA recombination. However, it was not successful to find induction conditions for sRNA expression. Interestingly, a possible sibling sRNA could be identified that shares the target interaction sequence with NgncR_237 and could therefore target the same mRNAs. In conclusion, this thesis provides further insights in gene regulation by non-coding RNAs in N. gonorrhoeae by analysing two pairs of sibling sRNAs modulating bacterial metabolism or possibly type IV pilus biogenesis. N2 - Bakterien müssen sich während des Infektionsprozesses an eine sich veränderte Umgebung anpassen und sind dabei zahlreichen Stressfaktoren ausgesetzt. Kleine, nicht-kodierende RNAs gelten als wichtige Regulatoren der bakteriellen Genexpression und ermöglichen daher eine schnelle Anpassung durch eine Veränderung der Expression spezifischer Ziel-Gene. Die Regulation der Genexpression des Humanpathogens Neisseria gonorrhoeae, Auslöser der Geschlechtskrankheit Gonorrhö, ist bis jetzt kaum verstanden. Die vorliegende Studie soll durch die Analyse kleiner, nicht-kodierender RNAs zum besseren Verständnis der Genregulation in Gonokokken beitragen. Durch die Entdeckung von antisense-RNAs für alle opa Gene wurde die Hypothese entwickelt, dass diese für den Abbau von opa Transkripten außerhalb des Leserahmens verantwortlich sind. Eine Analyse der asRNA Expression zeigte jedoch, dass diese sehr wenig exprimiert werden und auch die Untersuchung eines anderen phasenvariablen Gens weist darauf hin, dass die asRNAs keine Bedeutung für den Abbau von Transkripten außerhalb des Leserahmens haben. Der Schwerpunkt der Doktorarbeit liegt auf der Untersuchung trans-codierter sRNAs. Die Zwillings-sRNAs NgncR_162 und NgncR_163 agieren als post-transkriptionelle Regulatoren, die die Expression von Genen verändern, die bei Stoffwechselprozessen, Aminosäureaufnahme und transkriptioneller Regulation eine Rolle spielen. Eine detailliertere Analyse durch in silico- und Transkriptom-Studien zeigte, dass die sRNAs ein großes Spektrum an Genen regulieren, die für Proteine des Zentralstoffwechsels, der Aminosäurebiosynthese und des –abbaus, sowie zahlreicher Transportprozesse kodieren. Die Expressionslevel der Zwillings-sRNAs hängen von der Wachstumsphase der Bakterien und dem Wachstumsmedium ab. Das weist darauf hin, dass NgncR_162 und NgncR_163 eine Rolle bei der Adaptation des Stoffwechsels von Gonokokken zu bestimmten Wachstumsbedingungen spielen. In dieser Arbeit wird zudem die Charakterisierung der sRNA NgncR_237 initiiert. Im Rahmen von in silico Analysen wurde die Sequenzkonservierung und mögliche Sekundärstruktur untersucht. Eine Kombination aus in silico Zielgen-Vorhersage und differentieller RNA Sequenzierung führte zur Identifizierung zahlreicher Zielgene, die in der Biogenese von Typ IV Pili und DNA Rekombination eine Rolle spielen. Allerdings konnten keine Induktionsbedingungen für die sRNA Expression gefunden werden. Interessanterweise konnte eine mögliche Zwillings-sRNA identifiziert werden, die dieselbe Targetinteraktionsdomäne wie NgncR_237 hat und somit dieselben Zielgene regulieren könnte. Zusammenfassend ermöglicht diese Arbeit neue Einblicke in die Genregulation durch nicht-kodierende RNAs in Gonokokken, indem zwei Paare Zwillings-sRNAs analysiert wurden, die den bakteriellen Stoffwechsel anpassen oder möglicherweise eine Rolle in der Typ IV Pilus Biogenese spielen. KW - Neisseria gonorrhoeae KW - Non-coding RNA KW - Genregulation KW - regulation of gene expression Y1 - 2021 U6 - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-245826 ER - TY - JOUR A1 - Yu, Yidong A1 - Wolf, Ann-Katrin A1 - Thusek, Sina A1 - Heinekamp, Thorsten A1 - Bromley, Michael A1 - Krappmann, Sven A1 - Terpitz, Ulrich A1 - Voigt, Kerstin A1 - Brakhage, Axel A. A1 - Beilhack, Andreas T1 - Direct Visualization of Fungal Burden in Filamentous Fungus-Infected Silkworms JF - Journal of Fungi N2 - Invasive fungal infections (IFIs) are difficult to diagnose and to treat and, despite several available antifungal drugs, cause high mortality rates. In the past decades, the incidence of IFIs has continuously increased. More recently, SARS-CoV-2-associated lethal IFIs have been reported worldwide in critically ill patients. Combating IFIs requires a more profound understanding of fungal pathogenicity to facilitate the development of novel antifungal strategies. Animal models are indispensable for studying fungal infections and to develop new antifungals. However, using mammalian animal models faces various hurdles including ethical issues and high costs, which makes large-scale infection experiments extremely challenging. To overcome these limitations, we optimized an invertebrate model and introduced a simple calcofluor white (CW) staining protocol to macroscopically and microscopically monitor disease progression in silkworms (Bombyx mori) infected with the human pathogenic filamentous fungi Aspergillus fumigatus and Lichtheimia corymbifera. This advanced silkworm A. fumigatus infection model could validate knockout mutants with either attenuated, strongly attenuated or unchanged virulence. Finally, CW staining allowed us to efficiently visualize antifungal treatment outcomes in infected silkworms. Conclusively, we here present a powerful animal model combined with a straightforward staining protocol to expedite large-scale in vivo research of fungal pathogenicity and to investigate novel antifungal candidates. KW - fungal infection model KW - calcofluor white staining KW - Aspergillus KW - Lichtheimia KW - silkworm Y1 - 2021 U6 - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-228855 SN - 2309-608X VL - 7 IS - 2 ER - TY - JOUR A1 - Ye, Mingyu A1 - Wilhelm, Martina A1 - Gentschev, Ivaylo A1 - Szalay, Aladár T1 - A modified limiting dilution method for monoclonal stable cell line selection using a real-time fluorescence imaging system: A practical workflow and advanced applications JF - Methods and Protocols N2 - Stable cell lines are widely used in laboratory research and pharmaceutical industry. They are mainly applied in recombinant protein and antibody productions, gene function studies, drug screens, toxicity assessments, and for cancer therapy investigation. There are two types of cell lines, polyclonal and monoclonal origin, that differ regarding their homogeneity and heterogeneity. Generating a high-quality stable cell line, which can grow continuously and carry a stable genetic modification without alteration is very important for most studies, because polyclonal cell lines of multicellular origin can be highly variable and unstable and lead to inconclusive experimental results. The most commonly used technologies of single cell originate monoclonal stable cell isolation in laboratory are fluorescence-activated cell sorting (FACS) sorting and limiting dilution cloning. Here, we describe a modified limiting dilution method of monoclonal stable cell line selection using the real-time fluorescence imaging system IncuCyte\(^®\)S3. KW - monoclonal stable cell KW - limiting dilution cloning KW - ncuCyte\(^®\)S3 Y1 - 2021 U6 - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-228896 VL - 4 IS - 1 ER - TY - JOUR A1 - Ye, Mingyu A1 - Keicher, Markus A1 - Gentschev, Ivaylo A1 - Szalay, Aladar A. T1 - Efficient selection of recombinant fluorescent vaccinia virus strains and rapid virus titer determination by using a multi-well plate imaging system JF - Biomedicines N2 - Engineered vaccinia virus (VACV) strains are used extensively as vectors for the development of novel cancer vaccines and cancer therapeutics. In this study, we describe for the first time a high-throughput approach for both fluorescent rVACV generation and rapid viral titer measurement with the multi-well plate imaging system, IncuCyte\(^®\)S3. The isolation of a single, well-defined plaque is critical for the generation of novel recombinant vaccinia virus (rVACV) strains. Unfortunately, current methods of rVACV engineering via plaque isolation are time-consuming and laborious. Here, we present a modified fluorescent viral plaque screening and selection strategy that allows one to generally obtain novel fluorescent rVACV strains in six days, with a minimum of just four days. The standard plaque assay requires chemicals for fixing and staining cells. Manual plaque counting based on visual inspection of the cell culture plates is time-consuming. Here, we developed a fluorescence-based plaque assay for quantifying the vaccinia virus that does not require a cell staining step. This approach is less toxic to researchers and is reproducible; it is thus an improvement over the traditional assay. Lastly, plaque counting by virtue of a fluorescence-based image is very convenient, as it can be performed directly on the computer. KW - fluorescent recombinant vaccinia virus KW - plaque isolation KW - IncuCyte\(^®\)S3 KW - plaque assay Y1 - 2021 U6 - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-245104 SN - 2227-9059 VL - 9 IS - 8 ER - TY - JOUR A1 - Wolf, Matthias T1 - How to teach about what is a species JF - Biology N2 - To ask students what a species is always has something rhetorical about it. Too quickly comes the rote answer, often learned by heart without ever thinking about it: “A species is a reproductive community of populations (reproductively isolated from others), which occupies a specific niche in nature” (Mayr 1982). However, do two people look alike because they are twins or are they twins because they look alike? “Two organisms do not belong to the same species because they mate and reproduce, but they only are able to do so because they belong to the same species” (Mahner and Bunge 1997). Unfortunately, most biology (pre-university) teachers have no opinion on whether species are real or conceptual, simply because they have never been taught the question themselves, but rather one answer they still pass on to their students today, learned by heart without ever thinking about it. Species are either real or conceptual and, in my opinion, it is this “or” that we should teach about. Only then can we discuss those fundamental questions such as who or what is selected, who or what evolves and, finally, what is biodiversity and phylogenetics all about? Individuals related to each other by the tree of life. KW - biospecies KW - species as individuals KW - species as natural kinds KW - species concept KW - species problem Y1 - 2021 U6 - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-241052 SN - 2079-7737 VL - 10 IS - 6 ER - TY - JOUR A1 - Wohlwend, Michael R. A1 - Craven, Dylan A1 - Weigelt, Patrick A1 - Seebens, Hanno A1 - Winter, Marten A1 - Kreft, Holger A1 - Zurell, Damaris A1 - Sarmento Cabral, Juliano A1 - Essl, Franz A1 - van Kleunen, Mark A1 - Pergl, Jan A1 - Pyšek, Petr A1 - Knight, Tiffany M. T1 - Anthropogenic and environmental drivers shape diversity of naturalized plants across the Pacific JF - Diversity and Distributions N2 - Aim The Pacific exhibits an exceptional number of naturalized plant species, but the drivers of this high diversity and the associated compositional patterns remain largely unknown. Here, we aim to (a) improve our understanding of introduction and establishment processes and (b) evaluate whether this information is sufficient to create scientific conservation tools, such as watchlists. Location Islands in the Pacific Ocean, excluding larger islands such as New Zealand, Japan, the Philippines and Indonesia. Methods We combined information from the most up‐to‐date data sources to quantify naturalized plant species richness and turnover across island groups and investigate the effects of anthropogenic, biogeographic and climate drivers on these patterns. In total, we found 2,672 naturalized plant species across 481 islands and 50 island groups, with a total of 11,074 records. Results Most naturalized species were restricted to few island groups, and most island groups have a low number of naturalized species. Island groups with few naturalized species were characterized by a set of widespread naturalized species. Several plant families that contributed many naturalized species globally also did so in the Pacific, particularly Fabaceae and Poaceae. However, many families were significantly over‐ or under‐represented in the Pacific naturalized flora compared to other regions of the world. Naturalized species richness increased primarily with increased human activity and island altitude/area, whereas similarity between island groups in temperature along with richness differences was most important for beta diversity. Main conclusions The distribution and richness of naturalized species can be explained by a small set of drivers. The Pacific region contains many naturalized plant species also naturalized in other regions in the world, but our results highlight key differences such as a stronger role of anthropogenic drivers in shaping diversity patterns. Our results establish a basis for predicting and preventing future naturalizations in a threatened biodiversity hotspot. KW - anthropogenic drivers KW - beta diversity KW - island biogeography KW - naturalized species KW - Pacific Ocean KW - plant invasion Y1 - 2021 U6 - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-239925 VL - 27 IS - 6 SP - 1120 EP - 1133 ER - TY - JOUR A1 - Welter, Nils A1 - Wagner, Angelo A1 - Furtwängler, Rhoikos A1 - Melchior, Patrick A1 - Kager, Leo A1 - Vokuhl, Christian A1 - Schenk, Jens-Peter A1 - Meier, Clemens Magnus A1 - Siemer, Stefan A1 - Gessler, Manfred A1 - Graf, Norbert T1 - Correction: Welter et al. Characteristics of nephroblastoma/nephroblastomatosis in children with a clinically reported underlying malformation or cancer predisposition syndrome. Cancers 2021, 13, 5016 JF - Cancers N2 - In the original article [1] there was a mistake in Table 2 as published. Table 2 contains wrong percentages in lines Bilateral disease and Patients with CPS or GU. For this reason the table should be replaced with the correct one as shown below. KW - nephroblastomatosis Y1 - 2021 U6 - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-250135 SN - 2072-6694 VL - 13 IS - 22 ER - TY - JOUR A1 - Welter, Nils A1 - Wagner, Angelo A1 - Furtwängler, Rhoikos A1 - Melchior, Patrick A1 - Kager, Leo A1 - Vokuhl, Christian A1 - Schenk, Jens-Peter A1 - Meier, Clemens Magnus A1 - Siemer, Stefan A1 - Gessler, Manfred A1 - Graf, Norbert T1 - Characteristics of nephroblastoma/nephroblastomatosis in children with a clinically reported underlying malformation or cancer predisposition syndrome JF - Cancers N2 - (1) Background: about 10% of Wilms Tumor (WT) patients have a malformation or cancer predisposition syndrome (CPS) with causative germline genetic or epigenetic variants. Knowledge on CPS is essential for genetic counselling. (2) Methods: this retrospective analysis focused on 2927 consecutive patients with WTs registered between 1989 and 2017 in the SIOP/GPOH studies. (3) Results: Genitourinary malformations (GU, N = 66, 2.3%), Beckwith-Wiedemann spectrum (BWS, N = 32, 1.1%), isolated hemihypertrophy (IHH, N = 29, 1.0%), Denys-Drash syndrome (DDS, N = 24, 0.8%) and WAGR syndrome (N = 20, 0.7%) were reported most frequently. Compared to others, these patients were younger at WT diagnosis (median age 24.5 months vs. 39.0 months), had smaller tumors (349.4 mL vs. 487.5 mL), less often metastasis (8.2% vs. 18%), but more often nephroblastomatosis (12.9% vs. 1.9%). WT with IHH was associated with blastemal WT and DDS with stromal subtype. Bilateral WTs were common in WAGR (30%), DDS (29%) and BWS (31%). Chemotherapy induced reduction in tumor volume was poor in DDS (0.4% increase) and favorable in BWS (86.9% reduction). The event-free survival (EFS) of patients with BWS was significantly (p = 0.002) worse than in others. (4) Conclusions: CPS should be considered in WTs with specific clinical features resulting in referral to a geneticist. Their outcome was not always favorable. KW - nephroblastoma KW - clinical malformations KW - cancer predisposition syndromes KW - tumor surveillance KW - outcome Y1 - 2021 U6 - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-248434 SN - 2072-6694 VL - 13 IS - 19 ER - TY - JOUR A1 - Walter, Thomas A1 - Degen, Jacqueline A1 - Pfeiffer, Keram A1 - Stöckl, Anna A1 - Montenegro, Sergio A1 - Degen, Tobias T1 - A new innovative real-time tracking method for flying insects applicable under natural conditions JF - BMC Zoology N2 - Background Sixty percent of all species are insects, yet despite global efforts to monitor animal movement patterns, insects are continuously underrepresented. This striking difference between species richness and the number of species monitored is not due to a lack of interest but rather to the lack of technical solutions. Often the accuracy and speed of established tracking methods is not high enough to record behavior and react to it experimentally in real-time, which applies in particular to small flying animals. Results Our new method of real-time tracking relates to frequencies of solar radiation which are almost completely absorbed by traveling through the atmosphere. For tracking, photoluminescent tags with a peak emission (1400 nm), which lays in such a region of strong absorption through the atmosphere, were attached to the animals. The photoluminescent properties of passivated lead sulphide quantum dots were responsible for the emission of light by the tags and provide a superb signal-to noise ratio. We developed prototype markers with a weight of 12.5 mg and a diameter of 5 mm. Furthermore, we developed a short wave infrared detection system which can record and determine the position of an animal in a heterogeneous environment with a delay smaller than 10 ms. With this method we were able to track tagged bumblebees as well as hawk moths in a flight arena that was placed outside on a natural meadow. Conclusion Our new method eliminates the necessity of a constant or predictable environment for many experimental setups. Furthermore, we postulate that the developed matrix-detector mounted to a multicopter will enable tracking of small flying insects, over medium range distances (>1000m) in the near future because: a) the matrix-detector equipped with an 70 mm interchangeable lens weighs less than 380 g, b) it evaluates the position of an animal in real-time and c) it can directly control and communicate with electronic devices. KW - natural environment KW - insect tracking KW - real-time KW - movement ecology KW - heterogeneous background Y1 - 2021 U6 - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-265716 VL - 6 ER -