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Item type: Item , The hemodynamic influence of the intrapelvic bleeding volume measured with three dimensional (3D) CT Volumetry by mechanic partially stable or instable pelvic ring fractures(2025) Georgievski, GoranIntroduction: Pelvic fractures, often resulting from high-energy trauma, pose significant challenges in emergency medicine and trauma surgery, particularly due to their association with substantial hemorrhage. The severity and complexity of these injuries necessitate accurate and timely assessment to guide management strategies. Traditionally, the emphasis on evaluating bleeding volume in pelvic fractures has focused on intrabdominal bleeding, with less attention given to intramuscular bleeding and fracture hematomas. Given the dynamic nature of hemorrhagic complications in trauma settings, this study explores the utility of 3D Slicer Volumetry in measuring bleeding volumes associated with unstable pelvic fractures, targeting primarily patients with Type B and Type C fractures. Goal: The primary aim of this research is to leverage advanced imaging techniques through 3D Slicer Volumetry to quantify bleeding volumes in patients with partially and completely unstable pelvic fractures. The investigation specifically seeks to: (1) determine differences in bleeding volume between pelvic fractures classified as Type B versus Type C by the AO Classification system; (2) evaluate the efficacy of 3D Slicer Volumetry as a viable clinical tool for defining bleeding volume; and (3) assess whether the quantified intracavitary bleeding correlates with overall blood loss in these traumatic injuries. Material and Methods: This retrospective study was carried out using data from 70 patients who sustained Type B or Type C pelvic fractures at the University Hospital of Giessen between 2016 and 2021. Inclusion criteria encompassed patients aged 14 years or older, experiencing stable clinical conditions upon arrival, and having undergone a full-body CT scan with contrast. Following data collection, 3D Slicer software was employed for the volumetric analysis of bleeding volumes, allowing interactive segmentation and comprehensive assessment of hematomas. Statistical analysis utilized non-parametric methods, particularly the Mann-Whitney U test, to compare bleeding volumes and clinical parameters across different fracture types. Key variables included age, gender, mechanism of injury, and laboratory findings at admission. Results: The findings indicate a predominance of pelvic fractures due to traffic accidents, encompassing 57.14% of cases. The average age of the patients was 49.98 years, with a male-to-female ratio of approximately 3:1. The mean volume of bleeding recorded was 591.9 ml (±455.4 ml). Notably, 42.86% of patients sustained Type B fractures, while the remainder had Type C fractures. Results demonstrated statistically significant differences in bleeding volumes between the two fracture types, with Type C fractures presenting greater hemorrhagic volumes. Systolic blood pressure upon admission was primarily above 89 mmHg in 90% of patients, indicating a relatively stable hemodynamic state despite significant injuries. The average duration of hospitalization was 33.55 days, reflecting the complexity of treatments necessitated by unstable pelvic fractures. Discussion: The results underscore the clinical relevance of accurately assessing bleeding volumes in pelvic fractures, which can significantly influence treatment pathways in trauma settings. The application of 3D Slicer Volumetry proved effective in providing precise measurements of bleeding volume, thus enhancing injury characterization. The observed trend of increased hemorrhage in Type C pelvic fractures reinforces the need for swift intervention and potential surgical exploration in cases of severe instability. Additionally, the study emphasizes the variable responses of patients to trauma based on age and sex, highlighting the need for tailored assessment protocols. While the results reveal promising data regarding the quantification of bleeding volumes, further research is warranted to correlate volumetric data with clinical outcomes and resource allocation in trauma care. Conclusion: This study demonstrates the feasibility and utility of using 3D Slicer Volumetry to evaluate bleeding volumes in patients with unstable pelvic fractures. The findings reveal critical differences in hemorrhagic profiles between fracture types, enhancing the understanding and management of pelvic injuries in trauma patients. Integration of advanced imaging techniques may facilitate improved outcomes through better-informed clinical decision-making in emergency trauma care. Future investigations should focus on longitudinal outcomes related to volumetric assessment and treatment effectiveness in pelvic fracture management.Item type: Item , Biotechnological Control Strategies for Managing Drosophila suzukii(2025) Abdelhafiz, Ibrahim AmeedDrosophila suzukii (D. suzukii) is an invasive pest native to East Asia that has established itself across much of the world. Unlike most Drosophila species, which prefer decaying organic material for reproduction, D. suzukii targets fresh and ripening fruits, causing significant agricultural damage. Larval feeding and subsequent secondary infections can result in complete crop loss. Current control strategies rely heavily on chemical insecticides, which pose environmental risks and affect non-target organisms. This underscores the urgent need for safer and more sustainable alternatives. This dissertation investigates three biotechnological control strategies against D. suzukii. The first approach enhances the Sterile Insect Technique (SIT), a method that reduces pest populations through the release of sterilized males. Three key advancements are presented: (1) development of a non-destructive sexing method based on pupal weight differences, (2) identification of optimal X-ray sterilization conditions (90 kV/40 Gy), and (3) implementation of a temperature-based sterilization technique utilizing the natural thermosensitivity of D. suzukii males. The second strategy focuses on characterizing La Jolla virus (LJV), a candidate for virus-based biocontrol. The study examines natural transmission routes, including airborne, venereal, oral, and fecal, and investigates the virus’s pathology. LJV infection was shown to affect feeding behavior, nutrient absorption, fecundity, and egg-to-adult viability, offering insight into its potential as a biological control agent. The third approach explores RNA interference (RNAi) as a control tool in two contexts: enhancing SIT by generating sterile male-only populations through gene silencing, and deploying RNAi as a biopesticide by targeting essential genes. However, under the tested conditions, RNAi did not yield significant effects in either application. Collectively, the findings provide valuable contributions to the development of targeted, environmentally friendly control methods for D. suzukii, highlighting both promising advances and existing limitations in the field of biological pest management.Item type: Item , Development of SNAP tag and horseradish peroxidase-based nanobodies as secondary antibody mimics for indirect immunoassays(2025) Sheng, WenjieImmunoassays are widely used in diagnosis and biomedical research for the detecting and quantifying specific biomolecules. Based on antigen-antibody interaction system, immunoassays enable the localization, qualitative analysis and quantification of target proteins. Among these, indirect immunoassays offer enhanced signal amplification and flexibility by using fluorescence-conjugated secondary antibodies. However, conventional secondary antibodies, predominantly immunoglobulin (IgG), present challenges due to their large size (150 kDa). Moreover, most animal-derived antibodies raise ethical concerns and exhibit batch-to batch variability. In contrast, small antibody fragments such as nanobodies (Nbs), which are derived from camelids and consist of only a single variable domain, are significantly smaller size (15 kDa) and can be efficiently produced using mammalian cell expression system. In this study, five previously established anti-mouse and anti-rabbit IgG secondary Nbs were selected and incorporated with a self-labeling SNAP-tag. The SNAP-tag (20 kDa) catalyzes the covalent, site-specific attachment of O6-benzylguanine (BG)-modified fluorophores to recombinant Nbs anti-IgG-SNAP proteins. These Nbs anti-IgG-SNAP were expressed in HEK293T cells. Following a rapid and straightforward conjugation protocol involving the SNAP-tag and BG modified Alexa Fluor dyes, the specific detection capability of Nbs anti-IgG-SNAP for mouse- or rabbit-derived primary antibodies was validated using flow cytometry and multi-color fluorescence microscopy. Additionally, these secondary nanobodies were further developed to be combined with horseradish peroxidase (HRP) and the recombinant Nbs anti-IgG-HRP proteins were expressed in HEK293T cells. Their functionality was validated as secondary antibodies in Western blot (WB) and tyramide signal amplification (TSA)-based multiplex immunofluorescence (mIF) assays. The results demonstrated that Nbs anti-IgG-SNAP and Nbs anti-IgG-HRP specifically bound to mouse or rabbit antibodies, exhibiting fluorescence intensities, quantitative validity and specificity comparable to conventional anti-mouse or anti-rabbit secondary antibodies. Moreover, their cost-effectiveness, scalable expression, easy of purification and simple site-specific conjugation procedures present an innovation alternative to traditional animal-derived antibody production, ensuring greater standardization and reproducibility in research applications. Taking together, these findings suggest that recombinant anti-mouse and anti-rabbit IgG secondary nanobodies present a promising and reliable alternative to traditional secondary antibodies in various indirect immunoassays.Item type: Item , Experimental Data for 'Ozonido- and Oxido-TMC Complexes of Iron and Cobalt'(2026) Gerbig, Dennis; Schaub, StefanAnalytical data to accompany 'Ozonido- and Oxido-TMC Complexes of Iron and Cobalt' by Schaub et al. Iron and cobalt complexes of tetramethylcyclam and D12-tetramethylcyclam were ozonized at low temperature and the samples subjected to analysis by cold electrospray mass spectrometry (cold ESI-MS; measured on a UHR-TOF Bruker Daltonik maXis plus instrument) and ultraviolet-visible spectroscopy (UV-Vis; measured on a Jasco V-760 instrument). Data is presented as x,y pairs of values in ASCII encoding and in CSV format. Additional experimental metadata is provided for each data subset in file readme.txt. All files can be read by any standard text or stream editor and imported into spreadsheet or plotting software for visualization. Dataset contents: - coldESIMS.zip (cold ESI-MS data) - UVVis.zip (UV-Vis data)Item type: Item , tRNA-Mimikry in Mengoviren: virale RNA-Elemente, welche die Glycyl-tRNA-Synthetase binden(2025) Droß, FabianPicornaviren sind kleine RNA-Viren, welche aus einer positiven einzelsträngigen RNA bestehen. Diese (+)ssRNA codiert das gesamte virale Polyprotein. Der codierende Bereich des Polyproteins ist am 5´- und 3´-Ende von untranslatierten Regionen (UTRs) mit komplexer Sekundärstruktur flankiert. Die virale RNA ist am 5´-Ende anstatt mit einem Cap-Nukleotid mit einem viralen Protein (VPg) versehen, das 3´-Ende endet mit einem Poly(A)-Schwanz. Aufgrund der fehlenden 5´-Cap-Struktur wird die virale Translation über eine "internal ribosome entry site" (IRES) in der 5´-UTR initiiert. Der Zusammenbau des Ribosoms an der viralen RNA findet mithilfe von Teilen des humanen 48S-Inititationskomplexes, eukaryotischen Initiationsfaktoren (eIFs) und weiteren Proteinen, die an die virale IRES binden, statt. Diese IRES bindenden Proteine werden als IRES trans acting factors (ITAFs) bezeichnet. Zu den ITAFs gehören Proteine wie PTB, hnRNP K, UNRIP und PCBP 1 und 2. Auch für die Glycyl-tRNA Synthetase (GARS) konnte bereits eine Funktion als ITAF in Polioviren gezeigt werden. Hier bindet GARS an die Domäne V der 5´-UTR. Es erkennt die tRNA ähnliche Struktur der UTR und bindet an eine (einzelsträngige) 5´-CCA-3´ Sequenz eines Loops. Diese Bindung wird aber nicht nur in Enteroviren vermutet. Aufgrund der Bedingungen der Bindung von GARS an die virale RNA finden sich viele weitere theoretische GARS-Bindestellen (GBE) in den verschiedenen Picornaviren, so auch im Mengovirus (MV), einem Isolat des Encephalomyocarditis Virus (EMCV) aus der Gruppe der Cardioviren. Das Mengovirus stammt aus der Region Mengo in Uganda und infiziert in erster Linie Nager und nicht-humane Primaten, in welchen es dann eine schwere Myokarditis auslöst, aber auch der Mensch kann infiziert werden. In dieser Arbeit wurden die gefundenen GBEs in der MV 5´- und 3´-UTR untersucht. Hierfür wurden Reporter-RNAs mit Mutationen der beiden UTRs in HeLa Zellen transfiziert. Durch Optimierung der verwendeten Pull-Down-Methode konnte die Bindung von GARS an die MV RNA nachgewiesen werden. Mittels Massenspektrometrie wurde die Auswirkung der GARS-Bindung untersucht. Hierbei wurde beobachtet, dass durch die Bindung von GARS die Bindung einer Vielzahl weiterer Proteine gefördert wird. Diese Proteine gehören unter anderem zu den Translationis-Initiationsfaktoren, Elongationsfaktoren oder zu Teilen der ribosomalen Untereinheiten. Es konnte die Bindung von GARS an die virale RNA nachgewiesen werden, und die daraus folgende positive Wirkung wird in dieser Arbeit diskutiert.