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Function of Peripheral Blood Eosinophils in Melanoma (2023)
Wendlinger, Simone Alice
Despite accounting for only a small proportion of all skin cancers, malignant melanoma displays a serious health risk with increasing incidence and high mortality rate. Fortunately, advances in the treatment of malignant melanoma now prolong survival and enhance response and treatment efficacy. Established biomarkers help evaluate disease progression and facilitate choosing appropriate and individual treatment options. However, the need for easily accessible and reliable biomarkers is rising to predict patient-specific clinical outcome. Eosinophil infiltration into the tumor and high peripheral eosinophil counts prior and during treatment have been associated with better response in patients for various cancer entities, including melanoma. An analysis of a heterogeneous study cohort reported high serum ECP levels in non-responders. Hence, eosinophil frequency and serum ECP as a soluble eosinophil-secreted mediator were suggested as prognostic biomarkers in melanoma. We examined whether melanoma patients treated with first-line targeted therapy could also benefit from the effects of eosinophils. In total, 243 blood and serum samples from patients with advanced melanoma were prospectively and retrospectively collected before and after drug initiation. To link eosinophil function to improved clinical outcome, soluble serum markers and peripheral blood counts were used for correlative studies using a homogeneous study cohort. In addition, functional and phenotypical characterizations provided insights into the expression profile and activity of freshly isolated eosinophils, including comparisons between patients and healthy donors. Our data showed a significant correlation between high pre-treatment blood eosinophil counts and improved response to targeted therapy and by trend to combinatorial immunotherapy in patients with metastatic melanoma. In accordance with previous studies our results links eosinophil blood counts to better response in melanoma patients. High pre-treatment ECP serum concentration correlated with response to immunotherapy but not to targeted therapy. Eosinophils from healthy donors and patients showed functional and phenotypical similarities. Functional assays revealed a strong cytotoxic potential of blood eosinophils towards melanoma cells in vitro, inducing apoptosis and necrosis. In addition, in vitro cytotoxicity was an active process of peripheral eosinophils and melanoma cells with bidirectional features and required close cell-cell interaction. The extent of cytotoxicity was dose-dependent and showed susceptibility to changes in physical factors like adherence. Importantly, we provide evidence of an additive tumoricidal function of eosinophils and combinatorial targeted therapy in vitro. In summary, we give valuable insights into the complex and treatment-dependent role of eosinophils in melanoma. As a result, our data support the suggestion of eosinophils and their secreted mediators as potential prognostic biomarkers. It will take additional studies to examine the molecular mechanisms that underlie our findings.
Plasticity of Functional MAOA Gene Methylation in Acrophobia (2018)
Schiele, Miriam A. ; Ziegler, Christiane ; Kollert, Leonie ; Katzorke, Andrea ; Schartner, Christoph ; Busch, Yasmin ; Gromer, Daniel ; Reif, Andreas ; Pauli, Paul ; Deckert, Jürgen ; Herrmann, Martin J. ; Domschke, Katharina
Epigenetic mechanisms have been proposed to mediate fear extinction in animal models. Here, MAOA methylation was analyzed via direct sequencing of sodium bisulfite-treated DNA extracted from blood cells before and after a 2-week exposure therapy in a sample of n = 28 female patients with acrophobia as well as in n = 28 matched healthy female controls. Clinical response was measured using the Acrophobia Questionnaire and the Attitude Towards Heights Questionnaire. The functional relevance of altered MAOA methylation was investigated by luciferase-based reporter gene assays. MAOA methylation was found to be significantly decreased in patients with acrophobia compared with healthy controls. Furthermore, MAOA methylation levels were shown to significantly increase after treatment and correlate with treatment response as reflected by decreasing Acrophobia Questionnaire/Attitude Towards Heights Questionnaire scores. Functional analyses revealed decreased reporter gene activity in presence of methylated compared with unmethylated pCpGfree_MAOA reporter gene vector constructs. The present proof-of-concept psychotherapy-epigenetic study for the first time suggests functional MAOA methylation changes as a potential epigenetic correlate of treatment response in acrophobia and fosters further investigation into the notion of epigenetic mechanisms underlying fear extinction.
On Lennard-Jones systems with finite range interactions and their asymptotic analysis (2018)
Schäffner, M. ; Schlömerkemper, A.
The aim of this work is to provide further insight into the qualitative behavior of mechanical systems that are well described by Lennard-Jones type interactions on an atomistic scale. By means of Gamma-convergence techniques, we study the continuum limit of one-dimensional chains of atoms with finite range interactions of Lennard-Jones type, including the classical Lennard-Jones potentials. So far, explicit formula for the continuum limit were only available for the case of nearest and next-to-nearest neighbour interactions. In this work, we provide an explicit expression for the continuum limit in the case of finite range interactions. The obtained homogenization formula is given by the convexification of a Cauchy-Born energy density. Furthermore, we study rescaled energies in which bulk and surface contributions scale in the same way. The related discrete-to-continuum limit yields a rigorous derivation of a one-dimensional version of Griffith' fracture energy and thus generalizes earlier derivations for nearest and next-to-nearest neighbors to the case of finite range interactions. A crucial ingredient to our proofs is a novel decomposition of the energy that allows for re fined estimates.
Entwicklung eines dreidimensionalen Zellkulturmodells zur Untersuchung der Apoptoseinduktion von kolorektalen Tumorzellen durch periphere Blutlymphozyten (2023)
Scherg, Florian Alexander Rudolf
Das kolorektale Karzinom zählt zu den häufigsten Tumoren in den westlichen Nationen. Für die Heilung ist neben der frühen Diagnose und der korrekten Klassifikation in UICC-Stadien, die Auswahl einer effektiven individuellen Therapiestrategie von Bedeutung. Der Schlüssel für die erfolgreiche Verbesserung dieser und weiterer Schritte in der Behandlung eines kolorektalen Karzinom ist die Analyse der Interaktion zwischen kolorektalen Karzinomzellen und Immunzellen in einem 3D-Zellkulturmodell, das es ermöglicht, die in vivo Situation möglichst realitätsnah zu imitieren. Im Vergleich zwischen einem 2D- und 3D-Zellkulturmodell konnten in dieser Arbeit bedeutende und reproduzierbare Unterschiede sowohl im Wachstumsverhalten und in der Zellmorphologie von Kolonkarzinomzellen (SW480-Zellen), Fibroblasten und Immunzellen (PBMC) als auch in deren Interaktion untereinander aufgezeigt werden. PBMC induzieren im 2D-System sowohl bei der MK der SW480-Zellen als auch bei der KK mit Fibroblasten im Vergleich zum 3D-System deutlich mehr Apoptose. So gibt es bei der MK mit 1, 2 und 3 Millionen PBMC eine durchschnittliche Apoptosezunahme von 88 % nach 24 h und von 95 % nach 48 h. Bei der KK mit ebenfalls 1, 2 und 3 Millionen PBMC gibt es mit durchschnittlich 89 % nach 24 h und 92 % nach 48 h vergleichbare Apoptosezunahmen. Zellkulturen auf azellularisierter Darmmatrix bieten den Raum für die bereits erwähnten wichtigen 3D-Zell-ECM- und Zell-Zellinteraktionen, realitätsnahe extrazelluläre Matrizen und soluble Substanzen, die eine sehr große Bedeutung für die Ausbildung tumorspezifischer Charakteristika haben wie z. B. Karzinogenese, Gewebsdifferenzierung Proliferationsgeschwindigkeit, Metastasierungspotential, Malignität, Angiogeneseeigenschaften und Mikroumgebungsbedingungen wie sie im System Mensch vorkommen. Sie besitzen durch ihre viel komplexere histologische Struktur im Vergleich zu den zweidimensionalen Monolayer-Systemen eine höhere Aussagekraft, bieten eine bessere Vergleichbarkeit mit Tumorsystemen in lebenden Organismen und mindern die experimentelle Fehlinterpretationsraten im Vergleich zu 2D-Modellen. So ist es auch nicht verwunderlich, dass die Ergebnisse bezüglich der Apoptoserate der SW480-Zellen in beiden Zellkulturmodellen so unterschiedlich ausfallen. Durch eine gute strategische Planung der Durchführung weiterer Experimente und der Quantifizierung von weiteren möglichen Zytokinen könnte der Pool an möglichen Wegen hin zur Apoptose der SW480-Zelle eingeengt werden. Unsere Ergebnisse zeigen, wie wichtig es ist, auch zukünftig weiter an dieser Thematik zu forschen, um so die Therapie des kolorektalen Karzinoms entscheidend zu verbessern. So wäre es durchaus vorstellbar, innerhalb dieses 3D-Modells durch Experimente, die beispielsweise die Tumorzellumgebung oder die Immunzelleigenschaften verändern, dem tumorzellbiologischen Verhalten und der immunologischen Tumor-Wirtsbeziehung in vivo entscheidend näher zu kommen.
Der Hygieniker und Medizinhistoriker Georg Sticker (1860-1960) - Leben und Werk (2023)
Maier-Unverfehrt, Kristin
Georg Sticker war Arzt, Hochschullehrer, Hygieniker und Medizinhistoriker. Das Themenspektrum seiner Arbeiten ist sehr umfangreich. Der Schwerpunkt dieser Arbeit liegt auf seinen Hauptforschungsgebieten, der Medizingeschichte und hier vor allem auf der Seuchengeschichte zu Pest, Lepra und Syphilis und der Bedeutung der Geschichte der Epidemien für die Epidemiologie; sowie seinen großen ärztlichen Vorbildern Hippokrates und Paracelsus. Auch über Naturheilkunst und Gesundheit und Erziehung berichtet er mit großem Engagement.Nach seiner Berufung an die Universität Würzburg beendet er seine ärztliche Tätigkeit, gründet hier im Jahr 1921 das Institut für Geschichte der Medizin und widmet sich ganz der Medizingeschichte
Mapping membrane receptor distribution on resting platelets combining Expansion Microscopy and fluorescence confocal microscopy (2023)
Maier, Sophia Edith
Stroke and myocardial infarction are the most prominent and severe consequences of pathological thrombus formation. For prevention and/or treatment of thrombotic events there is a variety of anti-coagulation and antiplatelet medication that all have one side effect in common: the increased risk of bleeding. To design drugs that only intervene in the unwanted aggregation process but do not disturb general hemostasis, it is crucial to decipher the exact clotting pathway which has not been fully understood yet. Platelet membrane receptors play a vital role in the clotting pathway and, thus, the aim of this work is to establish a method to elucidate the interactions, clustering, and reorganization of involved membrane receptors such as GPIIb/IIIa and GPIX as part of the GPIb-IX-V complex. The special challenges regarding visualizing membrane receptor interactions on blood platelets are the high abundancy of the first and the small size of the latter (1—3µm of diameter). The resolution limit of conventional fluorescence microscopy and even super-resolution approaches prevents the successful differentiation of densely packed receptors from one another. Here, this issue is approached with the combination of a recently developed technique called Expansion Microscopy (ExM). The image resolution of a conventional fluorescence microscope is enhanced by simply enlarging the sample physically and thus pulling the receptors apart from each other. This method requires a complex sample preparation and holds lots of obstacles such as variable or anisotropic expansion and low images contrast. To increase ExM accuracy and sensitivity for interrogating blood platelets, it needs optimized sample preparation as well as image analysis pipelines which are the main part of this thesis. The colocalization results show that either fourfold or tenfold expanded, resting platelets allow a clear distinction between dependent, clustered, and independent receptor organizations compared to unexpanded platelets.Combining dual-color Expansion and confocal fluorescence microscopy enables to image in the nanometer range identifying GPIIb/IIIa clustering in resting platelets – a pattern that may play a key role in the clotting pathway
Plastin 3 rescues defective cell surface translocation and activation of TrkB in mouse models for spinal muscular atrophy (2023)
Hennlein, Luisa
Spinal muscular atrophy (SMA) is a genetic pediatric condition that affects lower motoneurons leading to their degeneration and muscle weakness. It is caused by homozygous loss or mutations in the Survival Motor Neuron 1 (SMN1) gene; however, the pathomechanism leading to motoneuron degeneration is not fully resolved. Cultured embryonic SMA motoneurons display axon elongation and differentiation defects accompanied by collapsed growth cones with a disturbed actin cytoskeleton. Intriguingly, motoneurons cultured from mice deficient for the Tropomyosin-kinase receptor B (TrkB), exhibit similar pathological features. Thus, the question arises whether SMA motoneurons suffer from defective Brain-derived neurotrophic factor (BDNF)/TrkB signaling and whether there is a link to the disturbed actin cytoskeleton. In the recent years, modifier genes such as Plastin 3 (PLS3) were shown to beneficially interfere with SMA pathology. Nevertheless, the mechanism of how the actin-bundler PLS3 counteracts SMN deficiency is not well understood. In this study, we investigated TrkB localization and its activation in cultured SMA motoneurons and neuromuscular junctions (NMJs). While TrkB levels are only mildly affected locally in axon terminals, BDNF-mediated TrkB phosphorylation was massively disturbed. The activity-dependent TrkB translocation to the cell surface and its activation via BDNF were shown to be Pls3-dependent processes, that can be abolished by knockdown of Pls3. In contrast, PLS3 overexpression in SMA motoneurons rescued the defects on morphological and functional level. In particular, the relocation of TrkB to the cell surface after BDNF-induced internalization is disturbed in SMA, which is based on an actin-dependent TrkB translocation defect from intracellular stores. Lastly, AAV9-mediated PLS3 overexpression in vivo in neonatal SMA mice provided further evidence for the capacity of PLS3 to modulate actin dynamics necessary for accurate BDNF/TrkB signaling. In conclusion, we provide a novel role for PLS3 in mediating proper alignment of transmembrane proteins as prerequisite for their appropriate functioning. Hence, PLS3 is required for a key process indispensable for the development and function of motoneurons even beyond the context of SMA.
Analyse der Schusstodesfälle aus dem Sektionsgut des Instituts für Rechtsmedizin der Universität Würzburg von 2002-2017 (2023)
Hensel, Jan
Die vorliegende Arbeit ist eine retrospektive Analyse der Schusstodesfälle aus dem Sektionsgut des Instituts für Rechtsmedizin der Universität Würzburg aus dem Zeitraum 2002-2017. Bei 173 Schusstodesfällen kam es zu 209 registrierten Einschüssen. Hierbei entfielen 83,2 % der Schusstoten auf Suizide, 10,4 % auf Homizide und 3,5 % auf Unfälle. Rund 80 % der Opfer waren 30-79 Jahre alt und zu 91,9 % männlich, zu 8,1 % weiblich, ein Verhältnis von 11:1. Schusstreffer fanden sich zu 75,1 % in der Kopfregion gefolgt von Brust (12,9 %) und Extremitäten (4,8 %). Zu über ¾ ist die Todesursache ein zentrales Regulationsversagen infolge eines Kopfschusses. Der Schusswaffensuizident ist zu 98,6 % männlichen Geschlechts. Der Schusstod wird zu 95,9 % durch einen Nahschuss (meist absoluter Nahschuss) herbeigeführt. In 91,3 % ist die Einschussregion der Kopf in 8,7 % die Brust. Der Suizident ist zu 70,8 % 40-79 Jahre alt. Psychische Erkrankungen stellen das führende Tatmotiv dar. Die Tat findet zumeist im Frühjahr oder Sommer statt. Wenn die Blutalkoholkonzentration des Opfers erhoben wurde, war diese zu rund 50 % positiv. In 61,1 % aller Homizide waren Frauen das Opfer. Die Schussabgabe erfolgte in 90,9 % seitens eines Familienmitgliedes, meist dem Partner bzw. Ex-Partner. Klammert man die Tötungen im Polizeieinsatz (4 Fälle) aus, waren 92,9 % aller Homizide Beziehungstaten, auf die eine schusswaffenbedingte Selbsttötung des Täters folgte. Das Homizidopfer war zu 77,8 % 5-49 Jahre alt. Bei den weiblichen Homiziden überwog zu 63,6 % der Nahschuss, bei den männlichen Opfern der Tötung von fremder Hand lag in den meisten Fällen ein Fernschuss vor. In 26,1 % bestand nachweislich ein beruflich- oder freizeitbedingter Waffenbezug. Zu 64,2 % fand die Schussabgabe im häuslichen Umfeld statt. In 68,2 % aller Fälle wurde eine Kurzwaffe, meist Pistole (59,3 %) benutzt, in 32,8 % war das verwendete Munitionskaliber 9 mm/.357. 85,0 % aller abgegebenen Schüsse waren Nahschüsse, 63,2 % aller Schüsse waren Durchschüsse. In 89,0 % wurde ein einzelner Schuss abgegeben. Die erhobenen Ergebnisse dieser Arbeit weißen viele Parallelen mit bereits bestehenden Publikationen, insbesondere aus dem deutschsprachigen Raum und dem nordeuropäischen Ausland auf, grenzen sich aber vor allem in Hinsicht auf den Todesumstand von anderen Ländern teils deutlich ab.
Application based personalized food choices and health sustainment: scientific background and investigation of biomarkers in human tissue specimens (2023)
Jaud, Tobias Armin
Dietary fatty acids serve as objective biomarkers for the estimation of habitual diet mainly because biomarkers are free of memory bias or inaccuracies of food databases. The aim of the present work encompassed the implementation of a gas chromatographical method coupled with a mass spectrometrical and flame-ionization detector for analysis of fatty acid biomarkers in human biospecimens, their analytical determination and statistical evaluation in two different study populations and different biospecimens as well as the elaboration of adverse reactions to food ingredients with special focus on food allergies and food intolerances in the context of a possible implementation into an application for consumer health. The first aim was the identification of potential influence of fatty acid biomarkers on desaturase and elongase indexes (Δ9DI, Δ6DI, Δ5DI and ELOVLI5), which are factors in type 2 diabetes risk, in breast adipose tissue from healthy women. Influence of further variables on respective indexes was also investigated. 40 samples were investigated and potential variables were either collected by questionnaire or determined. Principle component analysis was applied for fatty acid biomarkers (PCdiet1, PCdiet2 and PCdiet3 representative for the dietary intake of vegetable oils/nuts, fish and partially hydrogenated vegetable oils), endogenous estrogens (PCE1) and oxysterols (PCOxy1). Multiple linear regression models were applied. Δ9DI and Δ6DI were influenced non-significantly and significantly negatively by PCdiet2 supporting a putative beneficial effect of vegetable oils and nuts on type 2 diabetes risk factors. ELOVLI5 and Δ5DI were influenced significantly and non-significantly positively by PCdiet1 supporting a putative beneficial effect of fish consumption on type 2 diabetes risk factors. On the other hand, PCdiet1 also significantly and non-significantly positively influenced Δ9DI and Δ6DI supporting a putative adverse effect of fish biomarkers on type 2 diabetes risk factors. The opposing influences of PCdiet1 suggesting an ambivalent role of dietary intake of fish on investigated indexes. Δ6DI was significantly positively influenced by PCdiet3 and number of pregnancies supporting a putative adverse effect of partially hydrogenated vegetable oils and pregnancies on type 2 diabetes risk factors. Lifestyle factors like smoking significantly and non-significantly influenced Δ9DI and Δ6DI putatively adversely. Δ5DI was influenced significantly positively by estrogen active drugs suggesting a putative beneficial effect on type 2 diabetes risk factors. It must be considered that a variation coefficient of up to 0.44 only explained 44% of variance of the respective indexes, suggesting other influencing factors might play a role. The second aim was the implementation of a gas chromatographical method coupled with a mass spectrometrical and flame-ionization detector for analysis of fatty acid biomarkers in human biospecimens. The method was optimized for separation and detection of 40 fatty acids. Mean recovery for tridecanoic acid was x(tridecanoic acid) = 90.51% and for nonadecanoic acid x(nonadecanoic acid) = 96.21%. Thus, there was no significant loss of fatty acids with shorter and longer carbon chains over the extraction process to be expected. Limit of detections were calculated in adipose tissue samples and ranged from 0.007 to 0.077% of the proportion of the respective fatty acid to total fatty acids. The third aim was the investigation if differentiation between breast glandular and adipose tissue had a relevant impact on the analysis of dietary fatty acid biomarkers or if contamination of breast glandular with breast adipose tissue and vice versa was neglectable for the analysis of dietary fatty acid biomarkers. No statistical significant differences were observed for all investigated fatty acid biomarkers (pentadecanoic-, heptadecanoic-, trans palmitoleic-, eicosapentaenoic-, docosahexaenoic-, linoleic and α-linolenic acid) between breast glandular and adipose tissue. Thus, differentiation between breast glandular and adipose tissue seems not to be necessary for the analysis of fatty acids serving as biomarkers for the intake of specific food groups. Potential influence of mixed breast tissue on fatty acid biomarkers analysis seems to be neglectable. The fourth aim was the determination of fatty acid biomarkers in adipose tissue in another study population from healthy participants. 27 adipose tissue samples were analyzed. Milk and ruminant fat biomarkers exhibited proportions of 0.47% for pentadecanoic acid, 0.34% for heptadecanoic acid and 0.25% for trans palmitoleic acid. Fish fatty acid biomarkers revealed proportions of 0.034% for eicosapentaenoic acid and 0.061% for docosahexaenoic acid. The mean proportion of vegetable oils and nuts biomarkers were 9.58% for linoleic acid and 0.48% for α-linolenic acid in all adipose tissues. Principle component analysis was applied for the fatty acid biomarkers to provide objective markers of habitual diet for this study population. PCdiet1 was mainly characterized by pentadecanoic acid, heptadecanoic acid and trans palmitoleic acid and therefore served as a principle component for the dietary intake of milk and ruminant fat. PCdiet2 and PCdiet3 only exhibited pattern for ω3 and ω6 fatty acids but not for dietary intake of specific food groups and could therefore not used as objective marker. PCdiet1, 2 and 3 explained 82.76% of variance. The last aim of this thesis was the elaboration of adverse reactions to food ingredients with special focus on food allergies and food intolerances in the context of a possible implementation into an application for consumer health. Scientific information on adverse reactions to food ingredients and trigger substances was provided in this thesis and possible implementation strategies were evaluated. For food allergens, which have regulatory requirements in the context of labelling, a strategy was elaborated, where it is necessary to provide information on the list of ingredients, the nexus ’contain’ and the respective food allergen as well as information on the name of the product. For food intolerances, which do not have regulatory requirements, limits were shown in the context of the application. If the elaborated food intolerances shall be implemented into the application, a professional dietary concept has to be developed for every food intolerance because of the complexity of the implementation.
Der Einfluss von Alter und Ängstlichkeit auf die Furchtgeneralisierung und die Aufmerksamkeitslenkung bei gesunden Kindern und Jugendlichen (2023)
Daub, Jonas
Mittels einer klinischen Studie wurden die Furchtgeneralisierung und Aufmerksamkeitslenkung von 44 gesunden Kindern und Jugendlichen im Alter von 9-17 Jahren untersucht. Eine Übergeneralisierung konditionierter Furcht sowie veränderte Aufmerksamkeitsprozesse werden in zahlreichen Arbeiten mit der Entstehung und Aufrechterhaltung von Angsterkrankungen in Verbindung gebracht. Der Hauptteil der Forschung beschränkte sich bislang auf die Untersuchung von erwachsenen Probanden. Da Angsterkrankungen jedoch häufig bereits im Kindes- und Jugendalter entstehen und sich in der Erforschung psychiatrischer Erkrankungen zunehmend eine dimensionale Betrachtungsweise durchsetzt, bestand das Ziel der Studie darin, etwaige Alterseffekte und den Einfluss der Ängstlichkeit auf die genannten Phänomene bei gesunden Probanden zu untersuchen. Darüber hinaus wurde ein potentiell präventiver Ansatz erforscht. Im Ergebnis zeigten sich in den Gruppenvergleichen keine relevanten Differenzen. Interessanterweise deutete sich in der Gruppe der älteren Probanden entgegen der Erwartung eine verstärkte Furchtgeneralisierung an, die womöglich mit einer veränderten Beziehung zu Furcht und Risiko in der Adoleszenz zusammenhängt. Aus den Befunden ergibt sich die Notwendigkeit weiterer, prospektiver Arbeiten, um unser Verständnis der Ätiologie von Angsterkrankungen zu verbessern. Weiterhin ist noch offen, inwiefern es sich bei der Übergeneralisierung und einer veränderten Aufmerksamkeitslenkung um Risikofaktoren für die Entwicklung von Angsterkrankungen oder vielmehr um Epiphänomene handelt, die erst mit Ausbruch der Erkrankung auftreten. Der Einsatz von Methoden der virtuellen Realität erscheint besonders geeignet, diese Prozesse zukünftig noch besser zu erforschen.
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