@article{JanzWalzCirnuetal.2024, author = {Janz, Anna and Walz, Katharina and Cirnu, Alexandra and Surjanto, Jessica and Urlaub, Daniela and Leskien, Miriam and Kohlhaas, Michael and Nickel, Alexander and Brand, Theresa and Nose, Naoko and W{\"o}rsd{\"o}rfer, Philipp and Wagner, Nicole and Higuchi, Takahiro and Maack, Christoph and Dudek, Jan and Lorenz, Kristina and Klopocki, Eva and Erg{\"u}n, S{\"u}leyman and Duff, Henry J. and Gerull, Brenda}, title = {Mutations in DNAJC19 cause altered mitochondrial structure and increased mitochondrial respiration in human iPSC-derived cardiomyocytes}, series = {Molecular Metabolism}, volume = {79}, journal = {Molecular Metabolism}, issn = {2212-8778}, doi = {10.1016/j.molmet.2023.101859}, url = {http://nbn-resolving.de/urn:nbn:de:bvb:20-opus-350393}, year = {2024}, abstract = {Highlights • Loss of DNAJC19's DnaJ domain disrupts cardiac mitochondrial structure, leading to abnormal cristae formation in iPSC-CMs. • Impaired mitochondrial structures lead to an increased mitochondrial respiration, ROS and an elevated membrane potential. • Mutant iPSC-CMs show sarcomere dysfunction and a trend to more arrhythmias, resembling DCMA-associated cardiomyopathy. Background Dilated cardiomyopathy with ataxia (DCMA) is an autosomal recessive disorder arising from truncating mutations in DNAJC19, which encodes an inner mitochondrial membrane protein. Clinical features include an early onset, often life-threatening, cardiomyopathy associated with other metabolic features. Here, we aim to understand the metabolic and pathophysiological mechanisms of mutant DNAJC19 for the development of cardiomyopathy. Methods We generated induced pluripotent stem cell-derived cardiomyocytes (iPSC-CMs) of two affected siblings with DCMA and a gene-edited truncation variant (tv) of DNAJC19 which all lack the conserved DnaJ interaction domain. The mutant iPSC-CMs and their respective control cells were subjected to various analyses, including assessments of morphology, metabolic function, and physiological consequences such as Ca\(^{2+}\) kinetics, contractility, and arrhythmic potential. Validation of respiration analysis was done in a gene-edited HeLa cell line (DNAJC19tv\(_{HeLa}\)). Results Structural analyses revealed mitochondrial fragmentation and abnormal cristae formation associated with an overall reduced mitochondrial protein expression in mutant iPSC-CMs. Morphological alterations were associated with higher oxygen consumption rates (OCRs) in all three mutant iPSC-CMs, indicating higher electron transport chain activity to meet cellular ATP demands. Additionally, increased extracellular acidification rates suggested an increase in overall metabolic flux, while radioactive tracer uptake studies revealed decreased fatty acid uptake and utilization of glucose. Mutant iPSC-CMs also showed increased reactive oxygen species (ROS) and an elevated mitochondrial membrane potential. Increased mitochondrial respiration with pyruvate and malate as substrates was observed in mutant DNAJC19tv HeLa cells in addition to an upregulation of respiratory chain complexes, while cellular ATP-levels remain the same. Moreover, mitochondrial alterations were associated with increased beating frequencies, elevated diastolic Ca\(^{2+}\) concentrations, reduced sarcomere shortening and an increased beat-to-beat rate variability in mutant cell lines in response to β-adrenergic stimulation. Conclusions Loss of the DnaJ domain disturbs cardiac mitochondrial structure with abnormal cristae formation and leads to mitochondrial dysfunction, suggesting that DNAJC19 plays an essential role in mitochondrial morphogenesis and biogenesis. Moreover, increased mitochondrial respiration, altered substrate utilization, increased ROS production and abnormal Ca\(^{2+}\) kinetics provide insights into the pathogenesis of DCMA-related cardiomyopathy.}, language = {en} } @phdthesis{Rainer2024, author = {Rainer, Johannes}, title = {Vaskulartoxische Wirkung von Taxanen bei fortgeschrittenen Tumorerkrankungen}, doi = {10.25972/OPUS-35072}, url = {http://nbn-resolving.de/urn:nbn:de:bvb:20-opus-350722}, school = {Universit{\"a}t W{\"u}rzburg}, year = {2024}, abstract = {Taxane (wie Paclitaxel oder Cabazitaxel) sind bew{\"a}hrte Arzneimittel in den systemischen Therapieschemata vieler b{\"o}sartiger Erkrankungen, einschließlich Brust- und Eierstockkrebs. Sie f{\"o}rdern die Stabilisierung der Mikrotubuli, was zu einem Stillstand des Zellzyklus w{\"a}hrend der Mitose f{\"u}hrt, auf den die Apoptose folgt. Neben dieser antimitotischen Wirkung von Taxanen ist seit einiger Zeit auch eine gef{\"a}ßver{\"a}ndernde Wirkung von Taxanen bekannt. K{\"u}rzlich wurde gezeigt, dass Taxane tats{\"a}chlich St{\"o}rungen in der Gef{\"a}ßarchitektur verursachen, indem sie den Kalziumeinstrom {\"u}ber TRPC6, einen unselektiven Kationenkanal, ausl{\"o}sen. Der erh{\"o}hte intrazellul{\"a}re Ca2+-Spiegel bewirkt eine Rundung der Endothelzellen, was zu einer St{\"o}rung des endothelialen Monolayers, Serumausfluss und Gef{\"a}ßkollaps f{\"u}hrt. In dieser Arbeit konzentrierten wir uns auf die Gef{\"a}ßbetten von peripheren Organen wie dem Herzen oder der Niere in Abh{\"a}ngigkeit vom Tumorstadium und der Taxol-Behandlung. Die Organe wurden mit immunhistochemischen Techniken angef{\"a}rbt, um Ver{\"a}nderungen in der Architektur und Morphologie der Blutgef{\"a}ße zu untersuchen. Wir fanden Ver{\"a}nderungen in der Morphologie der Kapillaren des Herzens und dar{\"u}ber hinaus Ver{\"a}nderungen in der Expression endothelialer Antigene in Abh{\"a}ngigkeit vom Tumorstadium, insbesondere eine zunehmende endotheliale Expression von TRPC6 in Abh{\"a}ngigkeit vom Tumorstadium. Diese Ergebnisse liefern neue Erkenntnisse f{\"u}r das Verst{\"a}ndnis der systemischen Auswirkungen maligner Erkrankungen und tragen dazu bei, Folgeerkrankungen bei Patienten mit fortgeschrittenem Krebs zu verhindern.}, subject = {Taxane}, language = {de} } @phdthesis{Mueller2023, author = {M{\"u}ller, Jonathan}, title = {Charakterisierung von CEACAM1 in der Retina und Choroidea am Mausmodell}, doi = {10.25972/OPUS-30554}, url = {http://nbn-resolving.de/urn:nbn:de:bvb:20-opus-305546}, school = {Universit{\"a}t W{\"u}rzburg}, year = {2023}, abstract = {Carcinoembryonic antigen-related cell adhesion molecule 1 (CEACAM1) ist ein multifunktionales Zell-Zell Adh{\"a}sionsprotein, das in eine Vielzahl an zellul{\"a}ren Prozessen involviert ist, wie zum Beispiel der Differenzierung von Geweben, der Tumorsuppression, Metastasierung, Angiogenese und Apoptose. Außerdem hat es modulierende Eigenschaften auf die angeborene und erworbene Immunantwort. In der vorliegenden Arbeit charakterisierte ich initial die Lokalisation und die CEACAM1-exprimierenden Zelltypen im Auge und bestimmte quantitativ die Expression von Ceacam1 in der Retina und Choroidea zu unterschiedlichen Zeitpunkten. Es zeigte sich hierbei, dass Ceacam1 zu allen untersuchten Zeitpunkten, sowohl w{\"a}hrend der Entwicklung als auch im adulten retinalen und choroidalen Gewebe nachweisbar war. Mittels Immunhistochemie konnte die Expression von CEACAM1 im Corneaepithel, den Gef{\"a}ßen der Iris und des Ziliark{\"o}rpers, im nicht-pigmentierten Epithel des Ziliark{\"o}rpers, sowie in den retinalen und choroidalen Gef{\"a}ßen nachgewiesen werden. Durch Doppelf{\"a}rbung mit Kollagen IV konnte die endotheliale Expression von CEACAM1 in den Endothelzellen der Gef{\"a}ße best{\"a}tigt werden. Im zweiten Teil meiner Arbeit untersuchte ich die Funktion von CEACAM1 im Auge und verglich dazu wildtypische Retinae mit Cc1-/--Retinae. Es zeigten sich keine offensichtlichen morphologischen Ver{\"a}nderungen der retinalen Schichten und die anschließend durchgef{\"u}hrten morphometrischen Analysen der Schichtdicken der retinalen Neurone zeigte keine Anzeichen einer Neurodegeneration. Allerdings waren in Cc1-/--Retinae kleine Zysten und IBA1 positive, phagozytisch aktive Zellen im subneuroretinalen Raum, also dem Bereich zwischen RPE und den Außensegmenten der Photorezeptoren zu erkennen. Die anschließend durchgef{\"u}hrten Expressionsanalysen immunmodulierender Faktoren und von Mitgliedern des TGF-β-Signalwegs in retinalen und choroidealen Proben wildtypischer und Cc1-/--M{\"a}usen zeigten keine ver{\"a}nderte Expression f{\"u}r Iba1, Ccl2 sowie Tnf-α. Jedoch konnten signifikant erh{\"o}hte Werte f{\"u}r TGF-β1 in der Gruppe der 2-4 als auch der Gruppe der 9 Monate alten Cc1-/--Retinae im Vergleich zu wildtypischen Retinae nachgewiesen werden. Basierend auf den Daten der vorliegenden Arbeit kann geschlussfolgert werden, dass die Deletion von CEACAM1 unter physiologischen Bedingungen die Struktur der Retina und Choroidea nicht offensichtlich beeinflusst. Allerdings f{\"u}hrt die Deletion zu erh{\"o}hten Tgfβ1 Spiegeln in der Retina und zur Aktivierung und Akkumulation von IBA1 positiven Zellen im subneuroretinalen Raum.}, subject = {Angiogenese}, language = {de} } @phdthesis{Ruf2023, author = {Ruf, Theresa}, title = {Immunhistologische Analyse der Effekte einer Kombinationstherapie im Brustkrebsmodell: Inhibition der Kollagensynthese durch PLOD-2-Blockade und Inhibierung des PD-1/PD-L1 Checkpoints}, doi = {10.25972/OPUS-32038}, url = {http://nbn-resolving.de/urn:nbn:de:bvb:20-opus-320380}, school = {Universit{\"a}t W{\"u}rzburg}, year = {2023}, abstract = {In dieser Arbeit wurden die histologischen und immunhistologischen Auswirkungen der Kombination aus Inhibition des PD-1/PD-1L-Checkpoints und PLOD-2-Blockade untersucht. Es konnte festgestellt werden, dass die Immuntherapie anschl{\"a}gt und dabei als Monotherapie die st{\"a}rksten Tumornekrosen induzierte. Das Ansprechen auf die Immuntherapie mit BMS-1166 war jedoch sehr unterschiedlich. In der Kombination mit dem PLOD-2-Inhibitor Minoxidil wurden hingegen einheitlichere, aber auch geringere Nekroseanteile festgestellt. Dabei muss jedoch beachtet werden, dass die Kombinationsbehandlung die st{\"a}rkste Auswirkung auf das Tumorwachstum hatte. So waren diese Tumore die kleinsten und leichtesten, was in Zusammenhang mit dem ausgepr{\"a}gten kollagenen Netzwerk dieser Gruppe stehen k{\"o}nnte. Die Kombination zeigte keine Auswirkung auf die Tumorvaskularisierung und die Zellteilungsaktivit{\"a}t, sowie auch keine Auff{\"a}lligkeiten bez{\"u}glich der Infiltration mit Immunzellen. Lungenmetastasen kamen in allen Behandlungsgruppen vor. Bei der Kombinationsbehandlung waren jedoch die durchschnittlich gr{\"o}ßten Lungenmetastasen festzustellen. In dieser Arbeit konnte keine klare signifikante Verbesserung der Brustkrebstherapie durch die Kombination von Inhibition der Kollagensynthese durch PLOD-2-Blockade und Inhibierung des PD-1/PD-1L-Checkpoints aufgezeigt werden. Das kollagene Netzwerk war auff{\"a}llig und sollte genauer untersucht werden. Es lohnt sich weiter an Kombinationen aus Immuntherapeutikum und EZM-Destabilisierung zu arbeiten. Die TME muss dabei weiterhin Ansatzpunkt der Forschung bleiben, um eine erleichterte Penetration der Medikamente in den Tumor zu erzielen. Hier ist der Austausch des Medikaments zur EZM-Destabilisierung empfehlenswert. Die LOX-Inhibierung hat sich bereits in Kombination mit Chemotherapie als vorteilhaft erwiesen (Rossow et al., 2018) und sollte nachfolgend in einem {\"a}hnlichen Versuchsaufbau mit dem Immuntherapeutikum BMS-1166 ausprobiert werden.}, subject = {Immunhistochemie}, language = {de} } @phdthesis{Michelbach2023, author = {Michelbach, Peter}, title = {Struktur und 3D-Organisation der Kapillarwand-assoziierten Zellen im murinen Myokard}, doi = {10.25972/OPUS-32763}, url = {http://nbn-resolving.de/urn:nbn:de:bvb:20-opus-327634}, school = {Universit{\"a}t W{\"u}rzburg}, year = {2023}, abstract = {Herzkreislauferkrankungen sind weit verbreitet und nicht nur eine große Belastung f{\"u}r die Betroffenen, sondern auch f{\"u}r das Gesundheitssystem. Die Folgen von Herzkreislauferkrankungen wie z.B. Myokardinfarkt und koronare Herzkrankheit stellen weltweit die h{\"a}ufigste Todesursache dar. Pr{\"a}vention, fr{\"u}hzeitige Erkennung und konsequente Behandlung sind daher von großer Bedeutung. Um das Verst{\"a}ndnis f{\"u}r die Pathophysiologie zu f{\"o}rdern und ferner Therapieans{\"a}tze ausfindig zu machen, ist es notwendig, nicht nur die Herzmuskelzellen im Blick zu haben, sondern auch die Komponenten des Herzmuskelstromas, die deren Funktion beeinflussen k{\"o}nnen. Das Verst{\"a}ndnis und die Rekonstruktion des kardialen Gewebes auf ultrastruktureller Ebene, sowie die Charakterisierung und Wechselwirkungen der verschiedenen Zellen des Herzens haben deshalb das Interesse vieler Forschergruppen geweckt. Das Ziel dieser Arbeit war die detaillierte ultrastrukturelle Analyse kardialer Perizyten, Endothelzellen sowie Kapillarwand-assoziierter Zellen und deren Kontakte im Arbeitsmyokard der Maus mittels verschiedener elektronenmikroskopischer Methoden. Zu Beginn der Arbeit wurde die transmissionselektronenmikroskopische Probenaufbereitung optimiert und ein modifiziertes Protokoll zur hervorragenden Kontrastierung der biologischen Membranen und zum bestm{\"o}glichen Erhalt der Ultrastruktur etabliert. Die optimierte Probenaufbereitung bot dann die ideale Grundlage f{\"u}r die Generierung elektronenmikroskopischer Datens{\"a}tze mittels serieller Block-Face Rasterelektronenmikroskopie (SBF-SEM) und anschließender Erzeugung dreidimensionaler Modelle der Mikrovaskulatur des Arbeitsmyokards der Maus. Die detaillierte ultrastrukturelle Analyse in drei Dimensionen offenbart neue morphologische Merkmale der kardialen Mikrovaskulatur und zeigt, dass die kardialen Perizyten vereinzelt Forts{\"a}tze abgeben, die mit den Endothelzellen assoziiert sind. Dadurch entsteht nicht nur eine perizyt{\"a}re-endotheliale Einheit, die von derselben Basallamina umschlossen wird. Die Rekonstruktion zeigt ebenfalls, dass die Kapillarwand-assoziierten Zellen sehr groß und weit verzweigt sind und nicht von der die Perizyten und Endothelzellen umgebenden Basallamina umschlossen werden. Sie stehen an vereinzelten Stellen in direktem Kontakt mit den Endothelzellen. Immunelektronenmikroskopische Analysen zeigen, dass die Kapillarwand-assoziierten Zellen sowohl CD34-positiv als auch CD44-positiv sind. Gr{\"o}ßer angelegte Studien zur weiteren dreidimensionalen Analyse z.B. in der Intima einer Arteriole k{\"o}nnten zur weiteren Charakterisierung der Perizyten und der Kapillarwand-assoziierten Zellen beitragen und sogar eine Einteilung m{\"o}glich machen. Eine Beteiligung von Perizyten im Rahmen des kardialen Remodeling nach einem Myokardinfarkt wurde bereits nachgewiesen. Außerdem spielen die Membranproteine CD34 und CD44 eine wichtige Rolle in der H{\"a}matopoese und auch der Angiogenese. In Zukunft k{\"o}nnten sich auch daraus interessante neue Ans{\"a}tze f{\"u}r gezielte Therapien nach einem Myokardinfarkt ergeben.}, subject = {Perizyt}, language = {de} } @article{GottschlichDrenckhahnMeierottetal.2023, author = {Gottschlich, G{\"u}nter and Drenckhahn, Detlev and Meierott, Lenz and Zonneveld, Ben}, title = {Hieracium maculatum subsp. pseudogougetianum, eine neue Unterart aus dem Mainfr{\"a}nkischen Muschelkalkgebiet}, series = {Forum Geobotanicum}, volume = {11}, journal = {Forum Geobotanicum}, issn = {1867-9315}, doi = {10.3264/FG.2023.0923}, url = {http://nbn-resolving.de/urn:nbn:de:bvb:20-opus-327601}, pages = {15-20}, year = {2023}, abstract = {In Lower Franconia/Northern Bavaria, a well-defined subspecies of the Hieracium maculatum group was detected. This subspecies is restricted to the slopes of the Main valley between W{\"u}rzburg and Hasloch with a hot spot (>90\% of total population) between the villages Th{\"u}ngersheim and Retzbach. Due to some similarities with H. glaucinum subsp. prasiophaeum (synonym: subsp. gougetianum) the subspecies is named H. maculatum subsp. pseudogougetianum. This subspecies grows preferentially on shell-bearing limestone gravels and begins flowering as early as mid-April. Head involucra are whitish hairy mixed with dark stalked glands. The basal leaf rosette consists of ovate to elliptic, toothed to serrate, dark spotted leaves, glabrous, glaucous above. Stems bear 1-3(4) stalked stem leaves and usually form long lateral flowering branches from the leaf axils. Like some other H. maculatum subspecies, H. maculatum subsp. pseudogougetianum is tetraploid with a mean genome weight (2C value) of 14.5 pg, distinguishing it from the H. glaucinum group, whose studied taxa are invariably triploid (mean 10.1 pg).}, subject = {Korbbl{\"u}tler}, language = {de} } @phdthesis{Hoehn2023, author = {H{\"o}hn, Marie}, title = {Ver{\"a}nderung der Tumorimmunumgebung muriner Mamma-Karzinome durch Inhibierung der Kollagensynthese}, doi = {10.25972/OPUS-28492}, url = {http://nbn-resolving.de/urn:nbn:de:bvb:20-opus-284929}, school = {Universit{\"a}t W{\"u}rzburg}, year = {2023}, abstract = {Das Mamma-Karzinom geh{\"o}rt zu den sogenannten desmoplastischen Tumorarten. Hierbei handelt es sich um Tumoren mit erh{\"o}hter Ansammlung von Bindegewebszellen und einer Akkumulation von Extrazellul{\"a}rer Matrix (EZM). Diese verdichtete EZM wirkt sowohl auf mechanischer als auch auf Signalweg-vermittelter Ebene als eine Barriere, welche die therapeutische Wirksamkeit erheblich vermindert. Einer der Hauptbestandteile der EZM ist Kollagen. Durch Anwendung von Pr{\"a}paraten, welche die Kollagensynthese und -reifung inhibieren, kann die rigide Struktur aufgelockert werden. Daraus ergibt sich eine verbesserte Versorgung mit N{\"a}hrstoffen und eine verbesserte Infiltrationsm{\"o}glichkeit f{\"u}r Immunzellen. Dies ist f{\"u}r die Effizienz der Immuntherapie, welche sich in den letzten Jahren als vielversprechende Alternative zu den Grunds{\"a}ulen der Krebstherapie entwickelt hat, unabdinglich. In der vorliegenden Arbeit wurden murine Mamma-Karzinome der 4T1-Linie nach Behandlung mit EZM-destabilisierenden Kollageninhibitoren auf ihre Immunumgebung hin untersucht. Verwendet wurden drei Wirkstoffe, welche an unterschiedlichen Punkten in die Kollagensynthese und -reifung eingreifen: βAPN als LOX(L)-Inhibitor, 1,4-DPCA als P4HA-Inhibitor und Minoxidil als LH-Inhibitor. Die Behandlung f{\"u}hrte zu einem deutlichen Anstieg aller untersuchten Immunzellen und deutet somit auf eine verbesserte Infiltrationsm{\"o}glichkeit hin. Zudem wurde die Expression maligner Signalwege, wie die der Angiogenese, Hypoxie, Metastasierungsneigung, Invasivit{\"a}t und Immunsuppression, verringert und tumorsuppressive Immunantworten verst{\"a}rkt. Die Kollageninhibition hatte zus{\"a}tzlich ein verringertes Tumorwachstum und eine Reduktion der Blutgef{\"a}ßdichte zufolge. Als Fazit gilt es festzuhalten, dass die Verwendung von Kollageninhibitoren in der Immuntherapie eine vielversprechende Option zur Verbesserung der Effizienz dieser Therapeutika darstellt. Diese Erkenntnis gilt es im Rahmen k{\"u}nftiger wissenschaftlicher Untersuchungen weiterzuentwickeln.}, subject = {Brustkrebs}, language = {de} } @article{DrenckhahnGottschlichZonneveld2023, author = {Drenckhahn, Detlev and Gottschlich, G{\"u}nter and Zonneveld, Ben}, title = {Neubeschreibungen und 2C-Werte von Pilosella macranthela subsp. silvae-pici (Spessart-Mausohrhabichtskraut) und Pilosella ottonis (Otto-Mausohrhabichtskraut) mit einer {\"U}bersicht {\"u}ber das Vorkommen von P. macranthela-Taxa in Bayern.}, series = {Forum Geobotanicum}, volume = {11}, journal = {Forum Geobotanicum}, issn = {1867-9315}, doi = {10.3264/FG.2023.1114}, url = {http://nbn-resolving.de/urn:nbn:de:bvb:20-opus-344797}, pages = {21-34}, year = {2023}, abstract = {A pentaploid taxon from the Pilosella macranthela group was discovered in Lower Franconia/Bavaria which is newly described here as P. macranthela subsp. sylvae-pici. It grows mainly in the Bavarian Bunter Spessart and occurs with three small, isolated stands also in the limestone area of the Main valley and Tauber area. Its habit of growth is intermediate between the furcata and laxicephala types of Pilosellae with epigeal and partly hypogeal stolons and a tendency to form clusters. The upper and lower surfaces of the rosette leaves have stellate hairs. The peduncles and the phyllaries are densely covered with dark glandular hairs with yellowish glandular heads. The lateral teeth of the ligules often are separated by incisions. A tetra- und pentaploid transitional taxon between P. macranthela subsp. sylvae-pici and P. officinarum is described as P. ottonis. P. ottonis is tetra- and pentaploid with up to 7 capitula. It is densely covered with dark stalk glands on phyllaries and resembles forms of P. acutifolia in habit. The phyllaries of P. ottonis are covered with numerous epidermal papillae with a diameter of about 10-20 μm and in this aspect resemble P. macranthela subsp. sylvae-pici and P. glomerata. However, in P. officinarum epidermal papillae are absent. Plants of genetically heterogeneous tetra- and heptaploid P. macranthela have been found outside the Spessart as spontaneous hybrids between P. glomerata and P. officinarum and also occur sporadically without P. glomerata in the vicinity.}, subject = {Habichtskraut}, language = {de} } @article{WatermannMeyerWagneretal.2023, author = {Watermann, Christoph and Meyer, Malin Tordis and Wagner, Steffen and Wittekindt, Claus and Klussmann, Jens Peter and Erguen, Sueleyman and Baumgart-Vogt, Eveline and Karnati, Srikanth}, title = {Peroxisomes are highly abundant and heterogeneous in human parotid glands}, series = {International Journal of Molecular Sciences}, volume = {24}, journal = {International Journal of Molecular Sciences}, number = {5}, issn = {1422-0067}, doi = {10.3390/ijms24054783}, url = {http://nbn-resolving.de/urn:nbn:de:bvb:20-opus-311079}, year = {2023}, abstract = {The parotid gland is one of the major salivary glands producing a serous secretion, and it plays an essential role in the digestive and immune systems. Knowledge of peroxisomes in the human parotid gland is minimal; furthermore, the peroxisomal compartment and its enzyme composition in the different cell types of the human parotid gland have never been subjected to a detailed investigation. Therefore, we performed a comprehensive analysis of peroxisomes in the human parotid gland's striated duct and acinar cells. We combined biochemical techniques with various light and electron microscopy techniques to determine the localization of parotid secretory proteins and different peroxisomal marker proteins in parotid gland tissue. Moreover, we analyzed the mRNA of numerous gene encoding proteins localized in peroxisomes using real-time quantitative PCR. The results confirm the presence of peroxisomes in all striated duct and acinar cells of the human parotid gland. Immunofluorescence analyses for various peroxisomal proteins showed a higher abundance and more intense staining in striated duct cells compared to acinar cells. Moreover, human parotid glands comprise high quantities of catalase and other antioxidative enzymes in discrete subcellular regions, suggesting their role in protection against oxidative stress. This study provides the first thorough description of parotid peroxisomes in different parotid cell types of healthy human tissue.}, language = {en} } @article{Haveman2023, author = {Haveman, Rense}, title = {Phytosociological notes on hedges in South Ayrshire, Scotland}, series = {Forum Geobotanicum}, volume = {11}, journal = {Forum Geobotanicum}, issn = {1867-9315}, doi = {10.3264/FG.2023.0420}, url = {http://nbn-resolving.de/urn:nbn:de:bvb:20-opus-312634}, pages = {1-7}, year = {2023}, abstract = {On the basis of four relev{\´e}es of hedges around Straiton en Dailly in South Ayrshire, Scotland, some features of hedges are discussed. On the basis of the brambles, the vegetation of these hedges can be assigned to the Pruno-Rubion sprengelii, which comprises the bramble scrubs of circumneutral and nutrient rich soils in West Europe (Haveman et al. 2017, Haveman \& de Ronde 2019). Until now, this alliance was thought to be restricted to the northwestern edge of the European continent, but based on these relev{\´e}es and the known distribution area of Rubus nemoralis and Rubus polyanthemus, both characteristic for the Pruno-Rubion sprengelii, large parts of North England and Scotland have to be included in the distribution area of the alliance. The Pruno-Rubion sprengelii is optimally developed in rather narrow structures, like hedges, which are pruned every year. Here, brambles and herbs alike can grow with and under the shrubs, facilitated by the light that reaches large parts of the ground. Where the economic base of hedges perishes, they are not longer maintained, and the shrubs can grow out to their natural proportions. This changes the amount of light reaching the surface in the inner parts of the thicket, changing the competition between the species. The brambles as well as the herbs are displaced to the outer edges of the scrub, and the vegetation "dissociates" in a high-growing scrub, a fore-mantle ("cuff") with brambles, and a fringe with perennial herbs. These elements can hardly ever be assigned to the Pruno-Rubion anymore. The Pruno-Rubion sprengelii in optima forma is a scrub in which the three elements (shrubs, brambles, and herbs) grow closely intertwined. This is rarely found in natural landscapes, and thus the alliance is a typical element of the old farmer landscape. What is more: the typical species of the alliance, like Rubus nemoralis and R. polyanthemus, could only evolve after the landscape was opened by farmers in the last six millennia (Matzke-Hajek 1997), giving way to Rubus ulmifolius to expand its distribution area. This caused an explosion of hybrids which stabilised through apomixis into the wealth of Rubus species inhibiting the West European landscape nowadays (Sochor et al. 2015). Many of these species have their original home in a man-made landscape. Therefore, the Pruno-Rubion sprengelii can be characterised as a "farmers alliance" pur sang.}, subject = {Brombeere}, language = {en} } @article{Gottschlich2023, author = {Gottschlich, G{\"u}nter}, title = {Hieracium rotundatum subsp. silvae-bavaricae, eine neue Hieracium-Sippe aus dem Bayerischen Wald (Deutschland)}, series = {Forum Geobotanicum}, volume = {11}, journal = {Forum Geobotanicum}, issn = {1867-9315}, doi = {10.3264/FG.2023.0912}, url = {http://nbn-resolving.de/urn:nbn:de:bvb:20-opus-327145}, pages = {8-14}, year = {2023}, abstract = {Hieracium rotundatum subsp. silvae-bavaricae is described as new for science and illustrated. The new subspecies belongs to a group of species (H. rotundatum, H. transylvanicum) whose main distribution is in the Balkans. The changeful nomenclatural history of the species name is described. Diagnostic features to distinguish the growth habit-similar species H. murorum, H. rotundatum and H. transylvanicum are discussed. Particular attention is drawn to the importance of the development of the basic leaf cycle. Contrary to previous knowledge, the northwestern distribution limit of H.rotundatum extends now to southeastern Bavaria. During the search for H. rotundatum a morphologically conspicuous subspecies of H. rotundatum could be detected, which is described here as new.}, subject = {Habichtskraut}, language = {de} } @article{KirchmeierMeierottJung2023, author = {Kirchmeier, Peter and Meierott, Lenz and Jung, Klaus}, title = {Taraxacum sect. Borealia Hand.-Mazz. in den Alpen}, series = {Forum Geobotanicum}, volume = {11}, journal = {Forum Geobotanicum}, issn = {1867-9315}, doi = {10.3264/FG.2023.1230}, url = {http://nbn-resolving.de/urn:nbn:de:bvb:20-opus-347512}, pages = {35-56}, year = {2023}, abstract = {The presence of Taraxacum microspecies of the section Borealia in the European Alps has been known from France, Suisse, Austria, Italy and Slowenia. The five known species are Taraxacum gallicum, T. handelii, T. kraettlii, T. mazzettii and T. melzerianum. From 2004 up to 2014 these localities have been visited. Detailed examinations of many collections make it possible to add characteristics and precise the descriptions and correct mistakes, eliminate ambiguities and fill gaps in the original descriptions. Numerous photos, drawings and a new determination key will make the access to the section Borealia easier. A new species of section Borealia, T. cimae-gallinae, from the mountain H{\"u}hnerspiel near Sterzing (Italy, South Tyrol) is described. The habitats of the Borealia in the alpine level are mostly gravel floors on wind-swept ridges or on summit levelings. The environment of Borealia-species is threatened by ski tourism or by the changes from global warming.}, subject = {Pflanzen}, language = {de} } @article{GredicKarnatiRuppertetal.2023, author = {Gredic, Marija and Karnati, Srikanth and Ruppert, Clemens and Guenther, Andreas and Avdeev, Sergey N. and Kosanovic, Djuro}, title = {Combined pulmonary fibrosis and emphysema: when Scylla and Charybdis ally}, series = {Cells}, volume = {12}, journal = {Cells}, number = {9}, issn = {2073-4409}, doi = {10.3390/cells12091278}, url = {http://nbn-resolving.de/urn:nbn:de:bvb:20-opus-313571}, year = {2023}, abstract = {Combined pulmonary fibrosis and emphysema (CPFE) is a recently recognized syndrome that, as its name indicates, involves the existence of both interstitial lung fibrosis and emphysema in one individual, and is often accompanied by pulmonary hypertension. This debilitating, progressive condition is most often encountered in males with an extensive smoking history, and is presented by dyspnea, preserved lung volumes, and contrastingly impaired gas exchange capacity. The diagnosis of the disease is based on computed tomography imaging, demonstrating the coexistence of emphysema and interstitial fibrosis in the lungs, which might be of various types and extents, in different areas of the lung and several relative positions to each other. CPFE bears high mortality and to date, specific and efficient treatment options do not exist. In this review, we will summarize current knowledge about the clinical attributes and manifestations of CPFE. Moreover, we will focus on pathophysiological and pathohistological lung phenomena and suspected etiological factors of this disease. Finally, since there is a paucity of preclinical research performed for this particular lung pathology, we will review existing animal studies and provide suggestions for the development of additional in vivo models of CPFE syndrome.}, language = {en} } @article{HuflageGrunzPatzeretal.2023, author = {Huflage, Henner and Grunz, Jan-Peter and Patzer, Theresa Sophie and Pannenbecker, Pauline and Feldle, Philipp and Sauer, Stephanie Tina and Petritsch, Bernhard and Erg{\"u}n, S{\"u}leyman and Bley, Thorsten Alexander and Kunz, Andreas Steven}, title = {Potential of unenhanced ultra-low-dose abdominal photon-counting CT with tin filtration: a cadaveric study}, series = {Diagnostics}, volume = {13}, journal = {Diagnostics}, number = {4}, issn = {2075-4418}, doi = {10.3390/diagnostics13040603}, url = {http://nbn-resolving.de/urn:nbn:de:bvb:20-opus-304122}, year = {2023}, abstract = {Objectives: This study investigated the feasibility and image quality of ultra-low-dose unenhanced abdominal CT using photon-counting detector technology and tin prefiltration. Materials and Methods: Employing a first-generation photon-counting CT scanner, eight cadaveric specimens were examined both with tin prefiltration (Sn 100 kVp) and polychromatic (120 kVp) scan protocols matched for radiation dose at three different levels: standard-dose (3 mGy), low-dose (1 mGy) and ultra-low-dose (0.5 mGy). Image quality was evaluated quantitatively by means of contrast-to-noise-ratios (CNR) with regions of interest placed in the renal cortex and subcutaneous fat. Additionally, three independent radiologists performed subjective evaluation of image quality. The intraclass correlation coefficient was calculated as a measure of interrater reliability. Results: Irrespective of scan mode, CNR in the renal cortex decreased with lower radiation dose. Despite similar mean energy of the applied x-ray spectrum, CNR was superior for Sn 100 kVp over 120 kVp at standard-dose (17.75 ± 3.51 vs. 14.13 ± 4.02), low-dose (13.99 ± 2.6 vs. 10.68 ± 2.17) and ultra-low-dose levels (8.88 ± 2.01 vs. 11.06 ± 1.74) (all p ≤ 0.05). Subjective image quality was highest for both standard-dose protocols (score 5; interquartile range 5-5). While no difference was ascertained between Sn 100 kVp and 120 kVp examinations at standard and low-dose levels, the subjective image quality of tin-filtered scans was superior to 120 kVp with ultra-low radiation dose (p < 0.05). An intraclass correlation coefficient of 0.844 (95\% confidence interval 0.763-0.906; p < 0.001) indicated good interrater reliability. Conclusions: Photon-counting detector CT permits excellent image quality in unenhanced abdominal CT with very low radiation dose. Employment of tin prefiltration at 100 kVp instead of polychromatic imaging at 120 kVp increases the image quality even further in the ultra-low-dose range of 0.5 mGy.}, language = {en} } @article{PatzerKunzHuflageetal.2023, author = {Patzer, Theresa Sophie and Kunz, Andreas Steven and Huflage, Henner and Conrads, Nora and Luetkens, Karsten Sebastian and Pannenbecker, Pauline and Paul, Mila Marie and Erg{\"u}n, S{\"u}leyman and Bley, Thorsten Alexander and Grunz, Jan-Peter}, title = {Ultrahigh-resolution photon-counting CT in cadaveric fracture models: spatial frequency is not everything}, series = {Diagnostics}, volume = {13}, journal = {Diagnostics}, number = {10}, issn = {2075-4418}, doi = {10.3390/diagnostics13101677}, url = {http://nbn-resolving.de/urn:nbn:de:bvb:20-opus-319281}, year = {2023}, abstract = {In this study, the impact of reconstruction sharpness on the visualization of the appendicular skeleton in ultrahigh-resolution (UHR) photon-counting detector (PCD) CT was investigated. Sixteen cadaveric extremities (eight fractured) were examined with a standardized 120 kVp scan protocol (CTDI\(_{vol}\) 10 mGy). Images were reconstructed with the sharpest non-UHR kernel (Br76) and all available UHR kernels (Br80 to Br96). Seven radiologists evaluated image quality and fracture assessability. Interrater agreement was assessed with the intraclass correlation coefficient. For quantitative comparisons, signal-to-noise-ratios (SNRs) were calculated. Subjective image quality was best for Br84 (median 1, interquartile range 1-3; p ≤ 0.003). Regarding fracture assessability, no significant difference was ascertained between Br76, Br80 and Br84 (p > 0.999), with inferior ratings for all sharper kernels (p < 0.001). Interrater agreement for image quality (0.795, 0.732-0.848; p < 0.001) and fracture assessability (0.880; 0.842-0.911; p < 0.001) was good. SNR was highest for Br76 (3.4, 3.0-3.9) with no significant difference to Br80 and Br84 (p > 0.999). Br76 and Br80 produced higher SNRs than all kernels sharper than Br84 (p ≤ 0.026). In conclusion, PCD-CT reconstructions with a moderate UHR kernel offer superior image quality for visualizing the appendicular skeleton. Fracture assessability benefits from sharp non-UHR and moderate UHR kernels, while ultra-sharp reconstructions incur augmented image noise.}, language = {en} } @article{WoersdoerferErguen2023, author = {W{\"o}rsd{\"o}rfer, Philipp and Erg{\"u}n, S{\"u}leyman}, title = {"Organoids": insights from the first issues}, series = {Organoids}, volume = {2}, journal = {Organoids}, number = {2}, issn = {2674-1172}, doi = {10.3390/organoids2020006}, url = {http://nbn-resolving.de/urn:nbn:de:bvb:20-opus-313694}, pages = {79 -- 81}, year = {2023}, abstract = {No abstract available}, language = {en} } @article{RockelWagnerSpengeretal.2023, author = {Rockel, Anna F. and Wagner, Nicole and Spenger, Peter and Erg{\"u}n, S{\"u}leyman and W{\"o}rsd{\"o}rfer, Philipp}, title = {Neuro-mesodermal assembloids recapitulate aspects of peripheral nervous system development \(in\) \(vitro\)}, series = {Stem Cell Reports}, volume = {18}, journal = {Stem Cell Reports}, number = {5}, issn = {2213-6711}, doi = {10.1016/j.stemcr.2023.03.012}, url = {http://nbn-resolving.de/urn:nbn:de:bvb:20-opus-349925}, pages = {1155-1165}, year = {2023}, abstract = {Summary Here we describe a novel neuro-mesodermal assembloid model that recapitulates aspects of peripheral nervous system (PNS) development such as neural crest cell (NCC) induction, migration, and sensory as well as sympathetic ganglion formation. The ganglia send projections to the mesodermal as well as neural compartment. Axons in the mesodermal part are associated with Schwann cells. In addition, peripheral ganglia and nerve fibers interact with a co-developing vascular plexus, forming a neurovascular niche. Finally, developing sensory ganglia show response to capsaicin indicating their functionality. The presented assembloid model could help to uncover mechanisms of human NCC induction, delamination, migration, and PNS development. Moreover, the model could be used for toxicity screenings or drug testing. The co-development of mesodermal and neuroectodermal tissues and a vascular plexus along with a PNS allows us to investigate the crosstalk between neuroectoderm and mesoderm and between peripheral neurons/neuroblasts and endothelial cells. Highlights •Novel neuro-mesodermal assembloid model of peripheral nervous system development •Model covers neural crest cell induction, migration, and ganglion formation •Ganglia send projections to the mesodermal as well as neural compartment •Peripheral ganglia and nerve fibers interact with a co-developing vascular plexus}, language = {en} } @phdthesis{Vix2022, author = {Vix, Patrick}, title = {Die Rolle des Zelladh{\"a}sionsmolek{\"u}ls Carcinoembryonic antigen-related cell adhesion molecule-1 (CEACAM1) bei der lymphogenen Metastasierung des Prostatakarzinoms (PCa)}, doi = {10.25972/OPUS-29002}, url = {http://nbn-resolving.de/urn:nbn:de:bvb:20-opus-290021}, school = {Universit{\"a}t W{\"u}rzburg}, year = {2022}, abstract = {Der epithelialen Pr{\"a}senz des Carcinoembryonic antigen-related cell adhesion molecule-1 (CEACAM1) in Prostatadr{\"u}sen wird eine tumorsupprimierende Funktion zugeschrieben. Maligne Ver{\"a}nderungen des Prostatadr{\"u}senepithels bei einem PCa f{\"u}hren zu einer Abnahme der epithelialen CEACAM1-Expression, zu einem Verlust der Zellpolarit{\"a}t und zu einer erh{\"o}hten Zellproliferation (prostatische intraepitheliale Neoplasie (PIN)). W{\"a}hrend des PIN-Stadiums exprimieren benachbarte Blut- und Lymphgef{\"a}ße CEACAM1. CEACAM1 selbst wirkt pro-angiogen und stimuliert die Gef{\"a}ßneubildung und auch die Neubildung von Lymphgef{\"a}ßen, Lymphangiogenese. Seine Rolle in der Tumor-Lymphangiogenese und dadurch bedingten Metastasierung von Tumoren wurde bisher nicht ausreichend gekl{\"a}rt. Ziel dieser Arbeit war es, die Rolle von CEACAM1 bei der lymphogenen PCa-Metastasierung anhand von immunhistochemischen (IHC) Analysen am humanem PCa-Prostata- und Lymphknoten-(LN)-Gewebe, sowie im Mausmodell zu analysieren. Laut den Immunfluoreszenzanalysen traten in den PIN-Arealen signifikant mehr CEACAM1-positive Blut- und Lymphgef{\"a}ße auf, als in den darauffolgenden Tumorstadien. Weiter wurde eine CEACAM1-Expression in LN-Sinusgewebe bereits bei Niedrig-Risiko-Patienten (pN0) detektiert. Diese fr{\"u}he CEACAM1-Expression trat auch in den LN im PCa-Mausmodell auf. Weiter wurde im LN-Gewebe von „Hoch-Risiko"-Patienten (pN1) eine luminale CEACAM1-Expression innerhalb der aus Tumorzellen bestehenden Dr{\"u}sen beobachtet, die mit der CEACAM1-Expression in nativen Prostatadr{\"u}sen vergleichbar ist. Auch das angiogen-aktivierte Gef{\"a}ßendothel von pN0- und pN1-LN war CEACAM1-positiv. Bei Hoch-Risiko-Patienten (pN1) nahmen die CEACAM1-positiven Blut- und Lymphgef{\"a}ße im Tumorstroma mit zunehmender Dedifferenzierung des Gewebes ab. Die CEACAM1/PSA-Doppelimmun-fluoreszenzanalysen ergaben eine heterogene Expression der beiden Marker bei Intermediate-risk-Patienten und mit zunehmender Dedifferenzierung des Tumorgewebes einen epithelialen Verlust der CEACAM1-Expression in den PSA-positiven G3-Tumordr{\"u}sen. Das Fehlen von PSA in pN0-LN und die nachweisbare Expression von PSA in pN1-LN best{\"a}tigten PSA als geeigneten PCa-Zellmarker in LN. In pN1-LN ohne Dr{\"u}senbildung traten Zellansammlungen mit einer nach außen gerichteten CEACAM1-positiven Front und einem im Zentrum liegenden PSA-positiven Bereich auf. Diese Befunde belegen einen Zusammenhang zwischen der endothelialen CEACAM1-Expression im Sinus und der Mikrometastasierungswahrscheinlichkeit im pN0-LN-Gewebe von PCa-Patienten. Potentiell l{\"a}sst sich daher im Niedrig-Risiko-PCa-Patientenkollektiv {\"u}ber eine CEACAM1-Bestimmung in LN das Risiko f{\"u}r eine Metastasierung fr{\"u}hzeitig erkennen.}, language = {de} } @phdthesis{Bail2022, author = {Bail, Kathrin}, title = {Der Effekt von Fingolimod auf die B-Zell-Distribution und -Aggregation in einem chronischen Mausmodell der Multiplen Sklerose}, doi = {10.25972/OPUS-26928}, url = {http://nbn-resolving.de/urn:nbn:de:bvb:20-opus-269282}, school = {Universit{\"a}t W{\"u}rzburg}, year = {2022}, abstract = {Hintergrund - Die Multiple Sklerose (MS) ist bis heute eine nur teilweise verstandene Autoimmunerkrankung des zentralen Nervensystems (ZNS). Das Tiermodell der experimentellen autoimmunen Enzephalomyelitis (EAE) erm{\"o}glicht die Erforschung von Teilaspekten der Pathogenese der MS und kann zur Etablierung von Therapeutika herangezogen werden. Die MP4-abh{\"a}ngige EAE erm{\"o}glicht als Mausmodell die gezielte Erforschung der Rolle der B-Zelle als Akteur in der Pathogenese der MS. Diese Dissertation untersuchte den Effekt des S1P1-Rezeptor-Modulators FTY720 (Fingolimod) auf die Immunantwort der autoreaktiven B-Zellen in der Peripherie sowie im ZNS. Methoden - MP4-immunisierte M{\"a}use erhielten 50 Tage nach dem EAE-Krankheitsbeginn oral appliziertes FTY720 {\"u}ber einen Zeitraum von 30 Tagen. Die Tiere wurden nach dem Auftreten der Krankheitssymptome t{\"a}glich klinisch evaluiert. Die MP4-spezifische B-Zell-Immunantwort und die MP4-spezifische humorale Immunreaktion wurden mittels ELISPOT und ELISA ausgewertet. Die Verteilung der T- und B-Zell-Anteile im peripheren Blut der M{\"a}use sowie die Aufteilung der B-Zell-Subsets in der Milz wurden mittels Durchflusszytometrie quantifiziert. Mittels Immunhistochemie wurden die B- und T-Zell-Ansammlungen im ZNS der M{\"a}use hinsichtlich ihrer Entwicklung in terti{\"a}r lymphatische Organe (TLOs) untersucht. Ergebnisse - In diesem Versuchsaufbau zeigte FTY720 keine signifikante Verbesserung des klinischen Krankheitsverlaufes der Tiere. Der Anteil von T-Zellen im peripheren Blut der M{\"a}use war unter der Therapie mit FTY720 signifikant reduziert, w{\"a}hrend die Anzahl an B-Zellen nicht-signifikant beeinflusst wurde. Bei der Untersuchung der B-Zell-Subtypen in der Milz fiel zun{\"a}chst ein erh{\"o}hter Anteil an B220+-B-Zellen auf, w{\"a}hrend die Verteilung der weiteren Subsets nicht-signifikant ver{\"a}ndert war. Unter der Therapie mit FTY720 zeigte sich keine Reduktion bereits etablierter B-Zell-Aggregate im ZNS, allerdings ist eine inhibierte Entwicklung in TLOs zu diskutieren. Zusammenfassung - Diese Arbeit impliziert unterschiedliche Effekte von FTY720 auf die B-Zellen in einem B-Zell-abh{\"a}ngigen chronischen Mausmodell der MS.}, subject = {Multiple Sklerose}, language = {de} } @article{DrenckhahnZonneveld2022, author = {Drenckhahn, Detlev and Zonneveld, Ben}, title = {Rubus admirabilis Drenckhahn, eine neue Brombeerart aus dem Formenkreis der Serie Vestiti an der Westk{\"u}ste von Schleswig-Holstein, Deutschland}, series = {Forum Geobotanicum}, volume = {10}, journal = {Forum Geobotanicum}, issn = {1867-9315}, doi = {10.3264/FG.2022.1228}, url = {http://nbn-resolving.de/urn:nbn:de:bvb:20-opus-298658}, pages = {38-44}, year = {2022}, abstract = {Rubus admirabilis Drenckhahn is a tetraploid new species of the Rubus section Rubus, series Vestiti. Stem leaves are 5-foliolate, digitate to weakly pedate with elongated, obovate acuminate terminal leaflets, adpressed hairy upper side and light green shimmering, softly hairy under side. Stems are arching (up to 2 m) partly climbing, obtuse-angled, densely hairy and glandular, gray green to dull brown, armed with 10(3-21) /5cm straight slender prickles, mostly 30-45º declining, 4.6(3-7)mm long. Pedicles of inflorescence are densely hairy (patent and partly adpressed), armed with 2-4 (per cm) slender patent to slightly curved prickles (1-2 mm long) and studded with numerous stalked glands (0.3-0.5 mm long) and some bristles. The species tolerates shadow and prefers moist soil. The type locality is probably the species' site of introduction or genesis. It is located west of the town Garding on the North Frisian peninsula of Eiderstedt (several hundred shrubs), where several non-native Rubus species were probably introduced in the course of reforestation in 1970. Rubus admirabilis spreads south to the town of Heide in Dithmarschen and north to the island of Amrum (maximal range diameter of 70 km) and seems to be in an expansive phase.}, subject = {Brombeere}, language = {de} } @phdthesis{LauerSchmaltz2022, author = {Lauer-Schmaltz, Sandra}, title = {Durchflusszytometrische Analyse CEACAM1-exprimierender Immunzellen bei Patienten mit Multipler Sklerose}, doi = {10.25972/OPUS-28913}, url = {http://nbn-resolving.de/urn:nbn:de:bvb:20-opus-289138}, school = {Universit{\"a}t W{\"u}rzburg}, year = {2022}, abstract = {Da die Pathogenese der Multiplen Sklerose (MS) bis heute nicht vollst{\"a}ndig gekl{\"a}rt ist, befassten wir uns mit der Rolle CEACAM1-exprimierender Immunzellen bei Patienten mit MS und untersuchten diese mittels durchflusszytometrischer Untersuchung. Bei CEACAM1 (Carcinoembryonic-antigen-related cell adhesion molecule) handelt es sich um ein Zelladh{\"a}sionsmolek{\"u}l, das sowohl an inter- als auch intrazellul{\"a}ren Signalmechanismen modulatorisch beteiligt ist. Anhand unserer Ergebnisse scheint CEACAM1 keine zentrale Rolle in der Pathogenese der MS zu spielen. Es ließ sich jedoch eine signifikante Erh{\"o}hung CD56+dim NK-Zellen (nat{\"u}rliche Killerzellen) im peripheren Blut von Patienten mit schubf{\"o}rmig remittierender MS feststellen. Dies st{\"u}tzt die These, dass die „dim"-Subpopulation der NK-Zellen eine proinflammatorische Rolle in der Pathogenese der MS einnehmen k{\"o}nnte. Demnach sollte in Zukunft hinsichtlich der Entwicklung neuer Biomarker in der MS der Fokus auf NK-Zellen und Monozyten sowie deren Subpopulationen gerichtet werden.}, subject = {Durchflusscytometrie}, language = {de} } @phdthesis{Kern2022, author = {Kern, Anna}, title = {Vaskularisierung von humanen neuralen Organoiden mit mesodermalen Progenitorzellen}, doi = {10.25972/OPUS-29111}, url = {http://nbn-resolving.de/urn:nbn:de:bvb:20-opus-291116}, school = {Universit{\"a}t W{\"u}rzburg}, year = {2022}, abstract = {Viele Organoide sind bisher nur stark vereinfachte Modelle der Originalgewebe, da sie nur aus dem Gewebsparenchym bestehen. Um neurale Organoide n{\"a}her an das Originalgewebe zu bringen, ist ein wichtiger Schritt mesenchymale Anteile zu integrieren. In dieser Arbeit war die wichtige Fragenstellung, ob neurale Organoide sich mit mesodermalen Progenitorzellen zu einem gemeinsamen Gewebe vereinigen lassen. Um die Generierung von neuro-mesenchymalen Organoiden zu erreichen, wurden geeignete Differenzierungsprotokolle zur Erzeugung neuroepithelialer und mesodermaler Aggregate aus humanen induzierten pluripotenten Stammzellen etabliert. Anschließend wurden die Sph{\"a}roide vereinigt und eingehend histologisch charakterisiert. Dar{\"u}ber hinaus wurde die Organoidentwicklung unter dem Einfluss von Hypoxie analysiert. Um die Organoide anschaulich mit der tats{\"a}chlichen Embryogenese vergleichen zu k{\"o}nnen, wurden Schnitte von H{\"u}hnerembryonen angefertigt. Die neuro-mesenchymalen Organoide wurden insgesamt 280 Tage kultiviert und an verschieden Zeitpunkten untersucht. Die hier pr{\"a}sentierten Daten zeigen, dass die erzeugten neuro-mesenchymalen Organoide viele Aspekte der nat{\"u}rlichen Embryogenese in Zellkultur nachahmen k{\"o}nnen. So wurde die Ausbildung neuralrohr{\"a}hnlicher Strukturen, die von einem perineuralen Gef{\"a}ßplexus umgeben sind, gezeigt. Des Weiteren wurde eine Interaktion von Astrozyten/radiale Gliazellen mit dem entstehenden Gef{\"a}ßnetz beobachtet. Schließlich zeigten sich das Einwandern von mikrogliaartigen Zellen aus dem mesenchymalen Organoidteil in das Nervengewebe. Diese Arbeit bildet die Basis f{\"u}r die Generierung neuro-mesenchymaler Organoide als realistisches Modellsystem f{\"u}r die Entwicklung des Nervensystems. Solche Modellsysteme k{\"o}nnen f{\"u}r die Erforschung von Krankheiten, Toxizit{\"a}tsstudien sowie Medikamententests verwendet werden.}, subject = {Organoid}, language = {de} } @misc{DunkelDrenckhahnZonneveld2022, author = {Dunkel, Franz G. and Drenckhahn, Detlev and Zonneveld, Ben}, title = {Forum Geobotanicum 10 (2021/2022)}, volume = {10(2021/2022)}, editor = {Meierott, Lenz and Drenckhahn, Detlev and Dunkel, Franz G. and Ewald, J{\"o}rg and Fleischmann, Andreas}, issn = {1867-9315}, url = {http://nbn-resolving.de/urn:nbn:de:bvb:20-opus-298926}, year = {2022}, abstract = {Forum Geobotanicum is an electronic journal devoted to disseminate information concerning geographical distribution, ecology, morphology, taxonomy and conservation of vascular plants in the European Union with a main focus on middle Europe. It covers from molecular biology to environmental aspects. The focus is to publish original papers, reviews and announcements for the educated generalist as well as the specialist in this broad field. Forum Geobotanicum does not aim to supplant existing paper journals, but will be much more flexible in format, publication time and world-wide distribution than paper journals. Many important studies are being currently published in local journals and booklets and some of them are published privately. Hence, these studies will become aware to only a limited readership. Forum Geobotanicum will encourage authors of such papers to submit them as special issues of the journal. Moreover, the journal is planning to build up an E-mail-address section to support communication between geobotanists in Europe. The editors are optimistic that this electronic journal will develop to a widely used communication forum that will help to stimulate activities in the entire field of geobotany in middle Europe. To overcome problems of long term archivation and effective taxonomic publication of articles published electronically in Forum Geobotanicum, print versions of each volume of the journal and appropriate digital storage devices will be delivered freely to selected university libraries and state libraries in middle Europe.}, subject = {Geobotanik}, language = {de} } @phdthesis{Rohde2022, author = {Rohde, J{\"o}rn}, title = {Identifizierung von Schl{\"u}sselgenen, die an der Bildung von terti{\"a}r lymphatischen Organen im zentralen Nervensystem in einem B-Zell-abh{\"a}ngigen Mausmodell der Multiplen Sklerose beteiligt sind}, doi = {10.25972/OPUS-26766}, url = {http://nbn-resolving.de/urn:nbn:de:bvb:20-opus-267669}, school = {Universit{\"a}t W{\"u}rzburg}, year = {2022}, abstract = {Bei Patienten, die an einer speziellen Form der MS erkrankten, konnten Entz{\"u}ndungsinfiltrate in den Meningen nachgewiesen werden, die in ihrem Aufbau lymphoidem Gewebe {\"a}hnelten. Das Auftreten dieser Infiltrate war mit einem schwereren Krankheitsverlauf assoziiert. Das Mausmodell der B-Zell-abh{\"a}ngigen MP4-induzierten experimentellen autoimmunen Enzephalomyelitis (EAE) zeigt in den Kleinhirnen der M{\"a}use Infiltrate, die den Infiltraten beim Menschen {\"a}hneln. Wir nutzten die MP4-induzierte EAE, um die Mechanismen der Erkrankungsentstehung und Progression besser zu verstehen. Ziel dieser Arbeit war es intakte und stabile Ribonukleins{\"a}uren (RNA) aus den Infiltraten der M{\"a}use zu isolieren. Sowie Identifizierung von Gene, die w{\"a}hrend verschiedener Stadien der zerebralen Entz{\"u}ndungsreaktion hochreguliert waren. Wir verglichen die M{\"o}glichkeiten der RNA-Isolation bei Paraffin-eingebettetem Gewebe und kryofixiertem Gewebe. Um die Vergleichbarkeit der Qualit{\"a}ts- und Quantit{\"a}tsanalyse zu gew{\"a}hrleisten, wurde f{\"u}r jede Probe eine RNA Integrit{\"a}tsnummer (RIN) ermittelt. Wir f{\"u}hrten eine Laser Capture Microdissection (LCM) und anschließende Gensequenzierung der zerebralen Infiltrate bei M{\"a}usen durch, bei denen eine MP4-abh{\"a}ngige EAE ausgel{\"o}st wurde. Des Weiteren verglichen wir die Ergebnisse mit den Expressionsprofilen von sekund{\"a}r lymphatischen Organen (SLOs). Insgesamt konnten wir 43 Gene herausfiltern, die im Vergleich zu den Kontrollgruppen hochreguliert waren. Die Entwicklung von ektopen lymphatischen Strukturen (ELS) im zentralen Nervensystem (ZNS) ist ein komplexer und bisher wenig verstandener Prozess. In dieser Studie beobachteten wir 43 Gene, die w{\"a}hrend der Entwicklung von B- Zell-Infiltraten in den Kleinhirnen von MP4-immunisierten M{\"a}usen signifikant hochreguliert waren. Von diesen Genen wurden bereits 14 im Zusammenhang mit der MS erw{\"a}hnt und sind zum Teil Gegenstand aktiver Forschung.}, subject = {Multiple Sklerose}, language = {de} } @article{JordanBroeerFischeretal.2022, author = {Jordan, Martin C. and Br{\"o}er, David and Fischer, Christian and Heilig, Philipp and Gilbert, Fabian and H{\"o}lscher-Doht, Stefanie and Kalogirou, Charis and Popp, Kevin and Grunz, Jan-Peter and Huflage, Henner and Jakubietz, Rafael G. and Erg{\"u}n, S{\"u}leyman and Meffert, Rainer H.}, title = {Development and preclinical evaluation of a cable-clamp fixation device for a disrupted pubic symphysis}, series = {Communications Medicine}, volume = {2}, journal = {Communications Medicine}, number = {1}, doi = {10.1038/s43856-022-00227-z}, url = {http://nbn-resolving.de/urn:nbn:de:bvb:20-opus-299800}, year = {2022}, abstract = {Background Traumatic separation of the pubic symphysis can destabilize the pelvis and require surgical fixation to reduce symphyseal gapping. The traditional approach involves open reduction and the implantation of a steel symphyseal plate (SP) on the pubic bone to hold the reposition. Despite its widespread use, SP-fixation is often associated with implant failure caused by screw loosening or breakage. Methods To address the need for a more reliable surgical intervention, we developed and tested two titanium cable-clamp implants. The cable served as tensioning device while the clamp secured the cable to the bone. The first implant design included a steel cable anterior to the pubic symphysis to simplify its placement outside the pelvis, and the second design included a cable encircling the pubic symphysis to stabilize the anterior pelvic ring. Using highly reproducible synthetic bone models and a limited number of cadaver specimens, we performed a comprehensive biomechanical study of implant stability and evaluated surgical feasibility. Results We were able to demonstrate that the cable-clamp implants provide stability equivalent to that of a traditional SP-fixation but without the same risks of implant failure. We also provide detailed ex vivo evaluations of the safety and feasibility of a trans-obturator surgical approach required for those kind of fixation. Conclusion We propose that the developed cable-clamp fixation devices may be of clinical value in treating pubic symphysis separation.}, language = {en} } @article{KleefeldtUpcinBoemmeletal.2022, author = {Kleefeldt, Florian and Upcin, Berin and B{\"o}mmel, Heike and Schulz, Christian and Eckner, Georg and Allmanritter, Jan and Bauer, Jochen and Braunger, Barbara and Rueckschloss, Uwe and Erg{\"u}n, S{\"u}leyman}, title = {Bone marrow-independent adventitial macrophage progenitor cells contribute to angiogenesis}, series = {Cell Death \& Disease}, volume = {13}, journal = {Cell Death \& Disease}, number = {3}, doi = {10.1038/s41419-022-04605-2}, url = {http://nbn-resolving.de/urn:nbn:de:bvb:20-opus-299724}, year = {2022}, abstract = {Pathological angiogenesis promotes tumor growth, metastasis, and atherosclerotic plaque rupture. Macrophages are key players in these processes. However, whether these macrophages differentiate from bone marrow-derived monocytes or from local vascular wall-resident stem and progenitor cells (VW-SCs) is an unresolved issue of angiogenesis. To answer this question, we analyzed vascular sprouting and alterations in aortic cell populations in mouse aortic ring assays (ARA). ARA culture leads to the generation of large numbers of macrophages, especially within the aortic adventitia. Using immunohistochemical fate-mapping and genetic in vivo-labeling approaches we show that 60\% of these macrophages differentiate from bone marrow-independent Ly6c\(^{+}\)/Sca-1\(^{+}\) adventitial progenitor cells. Analysis of the NCX\(^{-/-}\) mouse model that genetically lacks embryonic circulation and yolk sac perfusion indicates that at least some of those progenitor cells arise yolk sac-independent. Macrophages represent the main source of VEGF in ARA that vice versa promotes the generation of additional macrophages thereby creating a pro-angiogenetic feedforward loop. Additionally, macrophage-derived VEGF activates CD34\(^{+}\) progenitor cells within the adventitial vasculogenic zone to differentiate into CD31\(^{+}\) endothelial cells. Consequently, depletion of macrophages and VEGFR2 antagonism drastically reduce vascular sprouting activity in ARA. In summary, we show that angiogenic activation induces differentiation of macrophages from bone marrow-derived as well as from bone marrow-independent VW-SCs. The latter ones are at least partially yolk sac-independent, too. Those VW-SC-derived macrophages critically contribute to angiogenesis, making them an attractive target to interfere with pathological angiogenesis in cancer and atherosclerosis as well as with regenerative angiogenesis in ischemic cardiovascular disorders.}, language = {en} } @article{BielmeierSchmittKleefeldtetal.2022, author = {Bielmeier, Christina B. and Schmitt, Sabrina I. and Kleefeldt, Nikolai and Boneva, Stefaniya K. and Schlecht, Anja and Vallon, Mario and Tamm, Ernst R. and Hillenkamp, Jost and Erg{\"u}n, S{\"u}leyman and Neueder, Andreas and Braunger, Barbara M.}, title = {Deficiency in retinal TGFβ signaling aggravates neurodegeneration by modulating pro-apoptotic and MAP kinase pathways}, series = {International Journal of Molecular Sciences}, volume = {23}, journal = {International Journal of Molecular Sciences}, number = {5}, issn = {1422-0067}, doi = {10.3390/ijms23052626}, url = {http://nbn-resolving.de/urn:nbn:de:bvb:20-opus-283971}, year = {2022}, abstract = {Transforming growth factor β (TGFβ) signaling has manifold functions such as regulation of cell growth, differentiation, migration, and apoptosis. Moreover, there is increasing evidence that it also acts in a neuroprotective manner. We recently showed that TGFβ receptor type 2 (Tgfbr2) is upregulated in retinal neurons and M{\"u}ller cells during retinal degeneration. In this study we investigated if this upregulation of TGFβ signaling would have functional consequences in protecting retinal neurons. To this end, we analyzed the impact of TGFβ signaling on photoreceptor viability using mice with cell type-specific deletion of Tgfbr2 in retinal neurons and M{\"u}ller cells (Tgfbr2\(_{ΔOC}\)) in combination with a genetic model of photoreceptor degeneration (VPP). We examined retinal morphology and the degree of photoreceptor degeneration, as well as alterations of the retinal transcriptome. In summary, retinal morphology was not altered due to TGFβ signaling deficiency. In contrast, VPP-induced photoreceptor degeneration was drastically exacerbated in double mutant mice (Tgfbr2\(_{ΔOC}\); VPP) by induction of pro-apoptotic genes and dysregulation of the MAP kinase pathway. Therefore, TGFβ signaling in retinal neurons and M{\"u}ller cells exhibits a neuroprotective effect and might pose promising therapeutic options to attenuate photoreceptor degeneration in humans.}, language = {en} } @article{SchlechtWolfBonevaetal.2022, author = {Schlecht, Anja and Wolf, Julian and Boneva, Stefaniya and Prinz, Gabriele and Braunger, Barbara M. and Wieghofer, Peter and Agostini, Hansj{\"u}rgen and Schlunck, G{\"u}nther and Lange, Clemens}, title = {Transcriptional and distributional profiling of microglia in retinal angiomatous proliferation}, series = {International Journal of Molecular Sciences}, volume = {23}, journal = {International Journal of Molecular Sciences}, number = {7}, issn = {1422-0067}, doi = {10.3390/ijms23073443}, url = {http://nbn-resolving.de/urn:nbn:de:bvb:20-opus-284072}, year = {2022}, abstract = {Macular neovascularization type 3, formerly known as retinal angiomatous proliferation (RAP), is a hallmark of age-related macular degeneration and is associated with an accumulation of myeloid cells, such as microglia (MG) and infiltrating blood-derived macrophages (MAC). However, the contribution of MG and MAC to the myeloid cell pool at RAP sites and their exact functions remain unknown. In this study, we combined a microglia-specific reporter mouse line with a mouse model for RAP to identify the contribution of MG and MAC to myeloid cell accumulation at RAP and determined the transcriptional profile of MG using RNA sequencing. We found that MG are the most abundant myeloid cell population around RAP, whereas MAC are rarely, if ever, associated with late stages of RAP. RNA sequencing of RAP-associated MG showed that differentially expressed genes mainly contribute to immune-associated processes, including chemotaxis and migration in early RAP and proliferative capacity in late RAP, which was confirmed by immunohistochemistry. Interestingly, MG upregulated only a few angiomodulatory factors, suggesting a rather low angiogenic potential. In summary, we showed that MG are the dominant myeloid cell population at RAP sites. Moreover, MG significantly altered their transcriptional profile during RAP formation, activating immune-associated processes and exhibiting enhanced proliferation, however, without showing substantial upregulation of angiomodulatory factors.}, language = {en} } @phdthesis{Upcin2022, author = {Upcin, Berin}, title = {Contribution of vascular adventitia-resident progenitor cells to new vessel formation in \(ex\) \(vivo\) 3D models}, doi = {10.25972/OPUS-25507}, url = {http://nbn-resolving.de/urn:nbn:de:bvb:20-opus-255070}, school = {Universit{\"a}t W{\"u}rzburg}, year = {2022}, abstract = {Ongoing research to fight cancer, one of the dominant diseases of the 21st century has led to big progress especially when it comes to understanding the tumor growth and metastasis. This includes the discovery of the molecular mechanisms of tumor vascularization, which is critically required for establishment of tumor metastasis. Formation of new blood vessels is the first step in tumor vascularization. Therefore, understanding the molecular and cellular basis of tumor vascularization attracted a significant effort studying in biomedical research. The blood vessels for supplying tumor can be formed by sprouting from pre-existing vessels, a process called angiogenesis, or by vasculogenesis, that is de novo formation of blood vessels from not fully differentiated progenitor cell populations. Vasculogenic endothelial progenitor cells (EPCs) can either be activated from populations in the bone marrow reaching the pathological region via the circulation or they can be recruited from local reservoirs. Neovessel formation influences tumor progression, hence therapeutic response model systems of angiogenesis/vasculogenesis are necessary to study the underlying mechanisms. Although, initially the research in this area focused more on angiogenesis, it is now well understood that both angiogenesis and postnatal vasculogenesis contribute to neovessel formation in adult under both most pathological as well as physiological conditions. Studies in the last two decades demonstrate that in addition to the intimal layer of fully differentiated mature endothelial cells (ECs) and various smaller supplying vessels (vasa vasorum) that can serve as a source for new vessels by angiogenesis, especially the adventitia of large and medium size blood vessels harbors various vascular wall-resident stem and progenitor cells (VW-SPCs) populations that serve as a source for new vessels by postnatal vasculogenesis. However, little is known about the potential role of VW-SPCs in tumor vascularization. To this end, the present work started first to establish a modified aortic ring assay (ARA) using mouse aorta in order to study the contribution of vascular adventitia-resident VW-SPCs to neovascularization in general and in presence of tumor cells. ARA is already established an ex vivo model for neovascularization allows to study the morphogenetic events of complex new vessel formation that includes all layers of mature blood vessels, a significant advantage over the assays that employ monolayer endothelial cell cultures. Moreover, in contrast to assays employing endothelial cells monocultures, both angiogenic and vasculogenic events take place during new vessel formation in ARA although the exact contribution of these two processes to new vessel formation cannot be easily distinguished in conventional ARA. Thus, in this study, a modified protocol for the ARA (mdARA) was established by either removing or keeping the aortic adventitia in place. The mdARA allows to distinguish the role of VW-SPCs from those of other aortic layers. The present data show that angiogenic sprouting from mature aortic endothelium was markedly delayed when the adventitial layer was removed. Furthermore, the network between the capillary-like sprouts was significantly reduced in absence of aortic adventitia. Moreover, the stabilization of new sprouts by assembling the NG2+ pericyte-like cells that enwrapped the endothelial sprouts from the outside was improved when the adventitial layer remained in place. Next, mimicking the tumor-vessel adventitia-interaction, multicellular tumor spheroids (MCTS) and aortic rings (ARs) with or without adventitia of C57BL/6-Tg (UBC-GFP) mice were confronted within the collagen gel and cultured ex vivo. This 3D model enabled analysis of the mobilization, migration and capillary-like sprouts formation by VW-SPCs within tumor-vessel wall-interface in comparison to tumor-free side of the ARs. Interestingly, while MCTS preferred the uptake of single vascular adventitia-derived cells, neural spheroids were directly penetrated by capillary-like structures that were sprouted from the aortic adventitia. In summary, the model established in this work allows to study new vessel formation by both postnatal vasculogenesis and angiogenesis under same conditions. It can be applied in various mouse models including reporter mouse models, e.g. Cxcr1 CreER+/mTmG+/- mice, in which GFP-marked macrophages of the vessel wall were directly observed as they mobilized from their niche and migrated into collagen gel. Another benefit of the model is that it can be used for testing different factors such as small molecules, growth factors, cytokines, and drugs with both pro- and anti-angiogenic/vasculogenic effects.}, language = {en} } @phdthesis{Elsner2022, author = {Elsner, Clara Dorothea}, title = {Ultrastructural analysis of biogenesis and release of endothelial extracellular vesicles}, doi = {10.25972/OPUS-28852}, url = {http://nbn-resolving.de/urn:nbn:de:bvb:20-opus-288526}, school = {Universit{\"a}t W{\"u}rzburg}, year = {2022}, abstract = {Extracellular vesicle (EV)-mediated intercellular communication through exosomes, microvesicles (MVs) and apoptotic bodies has been shown to be implicated in various physiological as well as pathological processes such as the development and progression of atherosclerosis. While the cellular machinery controlling EV formation and composition has been studied extensively, little is known about the underlying morphological processes. This study focuses on a detailed ultrastructural analysis of the different steps of EV formation and release in Myocardial Endothelial (MyEnd) and Aortic Endothelial (AoEnd) cells cultured under serum starvation and inflammatory stimulation with TNF-α. Detailed morphological analyses were conducted applying and comparing different high- resolution light and electron microscopic methods. In this study, we could depict all steps of MV biogenesis named in literature. However, during the study of exosome biogenesis, we discovered a yet undescribed process: Instead of a direct fusion with the plasma membrane, multivesicular bodies were incorporated into a new distinct cellular compartment bound by fenestrated endothelium first. This may present a novel step in exosome biogenesis and warrants further study. Regarding the conditions of cell cultivation, we observed that the commonly used serum starvation causes MyEnd cells, but not AoEnd cells, to enter apoptosis after 48 hours. When preparing functional EV studies, we therefore recommend assessing the morphological condition of the serum-starved cells at different cultivation points first. When evaluating MV production, a statistical analysis showed that the more time AoEnd cells spent in cultivation under serum starvation, the higher the percentage of MV producing cells. However, additional TNF-α stimulation induced a significantly higher MV production than serum starvation alone. Lastly, our results show that TNF-α stimulation of AoEnd cells in vitro leads to the upregulation of CD44, an adhesion molecule critical in the early stages of atherosclerosis. CD44 was then depicted on the surface of generated MVs and exosomes. We conclude that under inflammatory conditions, EVs can mediate the transfer of CD44 from endothelial cells to target cells. This could be a novel mechanism by which MVs contribute to the development and progression of atherosclerotic disease and should be clarified by further studies.}, subject = {Vesikel}, language = {en} } @article{GergsJahnSchulzetal.2022, author = {Gergs, Ulrich and Jahn, Tina and Schulz, Nico and Großmann, Claudia and Rueckschloss, Uwe and Demus, Uta and Buchwalow, Igor B. and Neumann, Joachim}, title = {Protein phosphatase 2A improves cardiac functional response to ischemia and sepsis}, series = {International Journal of Molecular Sciences}, volume = {23}, journal = {International Journal of Molecular Sciences}, number = {9}, issn = {1422-0067}, doi = {10.3390/ijms23094688}, url = {http://nbn-resolving.de/urn:nbn:de:bvb:20-opus-284035}, year = {2022}, abstract = {Reversible protein phosphorylation is a posttranslational modification of regulatory proteins involved in cardiac signaling pathways. Here, we focus on the role of protein phosphatase 2A (PP2A) for cardiac gene expression and stress response using a transgenic mouse model with cardiac myocyte-specific overexpression of the catalytic subunit of PP2A (PP2A-TG). Gene and protein expression were assessed under basal conditions by gene chip analysis and Western blotting. Some cardiac genes related to the cell metabolism and to protein phosphorylation such as kinases and phosphatases were altered in PP2A-TG compared to wild type mice (WT). As cardiac stressors, a lipopolysaccharide (LPS)-induced sepsis in vivo and a global cardiac ischemia in vitro (stop-flow isolated perfused heart model) were examined. Whereas the basal cardiac function was reduced in PP2A-TG as studied by echocardiography or as studied in the isolated work-performing heart, the acute LPS- or ischemia-induced cardiac dysfunction deteriorated less in PP2A-TG compared to WT. From the data, we conclude that increased PP2A activity may influence the acute stress tolerance of cardiac myocytes.}, language = {en} } @article{BeheraJainGangulietal.2022, author = {Behera, Ananyaashree and Jain, Preeti and Ganguli, Geetanjali and Biswas, Mainak and Padhi, Avinash and Pattanaik, Kali Prasad and Nayak, Barsa and Erg{\"u}n, S{\"u}leyman and Hagens, Kristine and Redinger, Natalja and Saqib, Mohd and Mishra, Bibhuti B. and Schaible, Ulrich E. and Karnati, Srikanth and Sonawane, Avinash}, title = {Mycobacterium tuberculosis acetyltransferase suppresses oxidative stress by inducing peroxisome formation in macrophages}, series = {International Journal of Molecular Sciences}, volume = {23}, journal = {International Journal of Molecular Sciences}, number = {5}, issn = {1422-0067}, doi = {10.3390/ijms23052584}, url = {http://nbn-resolving.de/urn:nbn:de:bvb:20-opus-284080}, year = {2022}, abstract = {Mycobacterium tuberculosis (Mtb) inhibits host oxidative stress responses facilitating its survival in macrophages; however, the underlying molecular mechanisms are poorly understood. Here, we identified a Mtb acetyltransferase (Rv3034c) as a novel counter actor of macrophage oxidative stress responses by inducing peroxisome formation. An inducible Rv3034c deletion mutant of Mtb failed to induce peroxisome biogenesis, expression of the peroxisomal β-oxidation pathway intermediates (ACOX1, ACAA1, MFP2) in macrophages, resulting in reduced intracellular survival compared to the parental strain. This reduced virulence phenotype was rescued by repletion of Rv3034c. Peroxisome induction depended on the interaction between Rv3034c and the macrophage mannose receptor (MR). Interaction between Rv3034c and MR induced expression of the peroxisomal biogenesis proteins PEX5p, PEX13p, PEX14p, PEX11β, PEX19p, the peroxisomal membrane lipid transporter ABCD3, and catalase. Expression of PEX14p and ABCD3 was also enhanced in lungs from Mtb aerosol-infected mice. This is the first report that peroxisome-mediated control of ROS balance is essential for innate immune responses to Mtb but can be counteracted by the mycobacterial acetyltransferase Rv3034c. Thus, peroxisomes represent interesting targets for host-directed therapeutics to tuberculosis.}, language = {en} } @article{WangKarnatiMadhusudhan2022, author = {Wang, Hongjie and Karnati, Srikanth and Madhusudhan, Thati}, title = {Regulation of the homeostatic unfolded protein response in diabetic nephropathy}, series = {Pharmaceuticals}, volume = {15}, journal = {Pharmaceuticals}, number = {4}, issn = {1424-8247}, doi = {10.3390/ph15040401}, url = {http://nbn-resolving.de/urn:nbn:de:bvb:20-opus-267143}, year = {2022}, abstract = {A growing body of scientific evidence indicates that protein homeostasis, also designated as proteostasis, is causatively linked to chronic diabetic nephropathy (DN). Experimental studies have demonstrated that the insulin signaling in podocytes maintain the homeostatic unfolded protein response (UPR). Insulin signaling via the insulin receptor non-canonically activates the spliced X-box binding protein-1 (sXBP1), a highly conserved endoplasmic reticulum (ER) transcription factor, which regulates the expression of genes that control proteostasis. Defective insulin signaling in mouse models of diabetes or the genetic disruption of the insulin signaling pathway in podocytes propagates hyperglycemia induced maladaptive UPR and DN. Insulin resistance in podocytes specifically promotes activating transcription factor 6 (ATF6) dependent pathogenic UPR. Akin to insulin, recent studies have identified that the cytoprotective effect of anticoagulant serine protease-activated protein C (aPC) in DN is mediated by sXBP1. In mouse models of DN, treatment with chemical chaperones that improve protein folding provides an additional benefit on top of currently used ACE inhibitors. Understanding the molecular mechanisms that transmute renal cell specific adaptive responses and that deteriorate renal function in diabetes will enable researchers to develop new therapeutic regimens for DN. Within this review, we focus on the current understanding of homeostatic mechanisms by which UPR is regulated in DN.}, language = {en} } @article{KarnatiGuntasRajendranetal.2022, author = {Karnati, Srikanth and Guntas, Gulcan and Rajendran, Ranjithkumar and Shityakov, Sergey and H{\"o}ring, Marcus and Liebisch, Gerhard and Kosanovic, Djuro and Erg{\"u}n, S{\"u}leyman and Nagai, Michiaki and F{\"o}rster, Carola Y.}, title = {Quantitative lipidomic analysis of Takotsubo syndrome patients' serum}, series = {Frontiers in Cardiovascular Medicine}, volume = {9}, journal = {Frontiers in Cardiovascular Medicine}, number = {797154}, issn = {2297-055X}, doi = {10.3389/fcvm.2022.797154}, url = {http://nbn-resolving.de/urn:nbn:de:bvb:20-opus-270832}, year = {2022}, abstract = {Takotsubo syndrome (TTS), also known as the transient left ventricular apical ballooning syndrome, is in contemporary times known as novel acute cardiac syndrome. It is characterized by transient left ventricular apical akinesis and hyperkinesis of the basal left ventricular portions. Although the precise etiology of TTS is unknown, events like the sudden release of stress hormones, such as the catecholamines and the increased inflammatory status might be plausible causes leading to the cardiovascular pathologies. Recent studies have highlighted that an imbalance in lipid accumulation might promote a deviant immune response as observed in TTS. However, there is no information on comprehensive profiling of serum lipids of TTS patients. Therefore, we investigated a detailed quantitative lipid analysis of TTS patients using ES-MSI. Our results showed significant differences in the majority of lipid species composition in the TTS patients compared to the control group. Furthermore, the computational analyses presented was able to link the altered lipids to the pro-inflammatory cytokines and disseminate possible mechanistic pathways involving TNFα and IL-6. Taken together, our study provides an extensive quantitative lipidome of TTS patients, which may provide a valuable Pre-diagnostic tool. This would facilitate the elucidation of the underlying mechanisms of the disease and to prevent the development of TTS in the future.}, language = {en} } @article{KustiatiErguenKarnatietal.2022, author = {Kustiati, Ulayatul and Erg{\"u}n, Suleyman and Karnati, Srikanth and Nugrahaningsih, Dwi Aris Agung and Kusindarta, Dwi Liliek and Wihadmadyatami, Hevi}, title = {Ethanolic extract of Ocimum sanctum Linn. Inhibits cell migration of human lung adenocarcinoma cells (A549) by downregulation of integrin αvβ3, α5β1, and VEGF}, series = {Scientia Pharmaceutica}, volume = {90}, journal = {Scientia Pharmaceutica}, number = {4}, issn = {2218-0532}, doi = {10.3390/scipharm90040069}, url = {http://nbn-resolving.de/urn:nbn:de:bvb:20-opus-290540}, year = {2022}, abstract = {Adenocarcinoma lung cancer is a type of non-small cell lung carcinoma (NSCLC), which accounts for 85\% of lung cancer incidence globally. The therapies that are being applied, both conventional therapies and antibody-based treatments, are still found to have side effects. Several previous studies have demonstrated the ability of the ethanolic extract of Ocimum sanctum Linn. (EEOS) as an ethnomedicine with anti-tumor properties. The aim of this study was to determine the effect of Ocimum sanctum Linn. ethanolic extract in inhibiting the proliferation, angiogenesis, and migration of A549 cells (NSCLC). The adhesion as well as the migration assay was performed. Furthermore, enzyme-linked immunosorbent assay (ELISA) was used to measure the expression of αvβ3 integrins, α5β1 integrins, and VEGF. The cells were divided into the following treatment groups: control (non-treated/NT), positive control (AP3/inhibitor β3 80 µg/mL), cisplatin (9 µg/mL), and EEOS at concentrations of 50, 70, 100, and 200 µg/mL. The results showed that EEOS inhibits the adhesion ability and migration of A549 cells, with an optimal concentration of 200 µg/mL. ELISA testing showed that the group of A549 cells given EEOS 200 µg/mL presented a decrease in the optimal expression of integrin α5β1, integrin αvβ3, and VEGF.}, language = {en} } @article{ShityakovNagaiErguenetal.2022, author = {Shityakov, Sergey and Nagai, Michiaki and Erg{\"u}n, S{\"u}leyman and Braunger, Barbara M. and F{\"o}rster, Carola Y.}, title = {The protective effects of neurotrophins and microRNA in diabetic retinopathy, nephropathy and heart failure via regulating endothelial function}, series = {Biomolecules}, volume = {12}, journal = {Biomolecules}, number = {8}, issn = {2218-273X}, doi = {10.3390/biom12081113}, url = {http://nbn-resolving.de/urn:nbn:de:bvb:20-opus-285966}, year = {2022}, abstract = {Diabetes mellitus is a common disease affecting more than 537 million adults worldwide. The microvascular complications that occur during the course of the disease are widespread and affect a variety of organ systems in the body. Diabetic retinopathy is one of the most common long-term complications, which include, amongst others, endothelial dysfunction, and thus, alterations in the blood-retinal barrier (BRB). This particularly restrictive physiological barrier is important for maintaining the neuroretina as a privileged site in the body by controlling the inflow and outflow of fluid, nutrients, metabolic end products, ions, and proteins. In addition, people with diabetic retinopathy (DR) have been shown to be at increased risk for systemic vascular complications, including subclinical and clinical stroke, coronary heart disease, heart failure, and nephropathy. DR is, therefore, considered an independent predictor of heart failure. In the present review, the effects of diabetes on the retina, heart, and kidneys are described. In addition, a putative common microRNA signature in diabetic retinopathy, nephropathy, and heart failure is discussed, which may be used in the future as a biomarker to better monitor disease progression. Finally, the use of miRNA, targeted neurotrophin delivery, and nanoparticles as novel therapeutic strategies is highlighted.}, language = {en} } @article{SchmidtAltDeoghareetal.2022, author = {Schmidt, Sven and Alt, Yvonne and Deoghare, Nikita and Kr{\"u}ger, Sarah and Kern, Anna and Rockel, Anna Frederike and Wagner, Nicole and Erg{\"u}n, S{\"u}leyman and W{\"o}rsd{\"o}rfer, Philipp}, title = {A blood vessel organoid model recapitulating aspects of vasculogenesis, angiogenesis and vessel wall maturation}, series = {Organoids}, volume = {1}, journal = {Organoids}, number = {1}, issn = {2674-1172}, doi = {10.3390/organoids1010005}, url = {http://nbn-resolving.de/urn:nbn:de:bvb:20-opus-284043}, pages = {41 -- 53}, year = {2022}, abstract = {Blood vessel organoids are an important in vitro model to understand the underlying mechanisms of human blood vessel development and for toxicity testing or high throughput drug screening. Here we present a novel, cost-effective, and easy to manufacture vascular organoid model. To engineer the organoids, a defined number of human induced pluripotent stem cells are seeded in non-adhesive agarose coated wells of a 96-well plate and directed towards a lateral plate mesoderm fate by activation of Wnt and BMP4 signaling. We observe the formation of a circular layer of angioblasts around days 5-6. Induced by VEGF application, CD31\(^+\) vascular endothelial cells appear within this vasculogenic zone at approximately day 7 of organoid culture. These cells arrange to form a primitive vascular plexus from which angiogenic sprouting is observed after 10 days of culture. The differentiation outcome is highly reproducible, and the size of organoids is scalable depending on the number of starting cells. We observe that the initial vascular ring forms at the interface between two cell populations. The inner cellular compartment can be distinguished from the outer by the expression of GATA6, a marker of lateral plate mesoderm. Finally, 14-days-old organoids were transplanted on the chorioallantois membrane of chicken embryos resulting in a functional connection of the human vascular network to the chicken circulation. Perfusion of the vessels leads to vessel wall maturation and remodeling as indicated by the formation of a continuous layer of smooth muscle actin expressing cells enwrapping the endothelium. In summary, our organoid model recapitulates human vasculogenesis, angiogenesis as well as vessel wall maturation and therefore represents an easy and cost-effective tool to study all steps of blood vessel development and maturation directly in the human setting without animal experimentation.}, language = {en} } @article{ErguenWoersdoerfer2022, author = {Erg{\"u}n, S{\"u}leyman and W{\"o}rsd{\"o}rfer, Philipp}, title = {Organoids, assembloids and embryoids: New avenues for developmental biology, disease modeling, drug testing and toxicity assessment without animal experimentation}, series = {Organoids}, volume = {1}, journal = {Organoids}, number = {1}, issn = {2674-1172}, doi = {10.3390/organoids1010004}, url = {http://nbn-resolving.de/urn:nbn:de:bvb:20-opus-284101}, pages = {37 -- 40}, year = {2022}, abstract = {No abstract available}, language = {en} } @article{FeldheimWendLaueretal.2022, author = {Feldheim, Jonas and Wend, David and Lauer, Mara J. and Monoranu, Camelia M. and Glas, Martin and Kleinschnitz, Christoph and Ernestus, Ralf-Ingo and Braunger, Barbara M. and Meybohm, Patrick and Hagemann, Carsten and Burek, Malgorzata}, title = {Protocadherin Gamma C3 (PCDHGC3) is strongly expressed in glioblastoma and its high expression is associated with longer progression-free survival of patients}, series = {International Journal of Molecular Sciences}, volume = {23}, journal = {International Journal of Molecular Sciences}, number = {15}, issn = {1422-0067}, doi = {10.3390/ijms23158101}, url = {http://nbn-resolving.de/urn:nbn:de:bvb:20-opus-284433}, year = {2022}, abstract = {Protocadherins (PCDHs) belong to the cadherin superfamily and represent the largest subgroup of calcium-dependent adhesion molecules. In the genome, most PCDHs are arranged in three clusters, α, β, and γ on chromosome 5q31. PCDHs are highly expressed in the central nervous system (CNS). Several PCDHs have tumor suppressor functions, but their individual role in primary brain tumors has not yet been elucidated. Here, we examined the mRNA expression of PCDHGC3, a member of the PCDHγ cluster, in non-cancerous brain tissue and in gliomas of different World Health Organization (WHO) grades and correlated it with the clinical data of the patients. We generated a PCDHGC3 knockout U343 cell line and examined its growth rate and migration in a wound healing assay. We showed that PCDHGC3 mRNA and protein were significantly overexpressed in glioma tissue compared to a non-cancerous brain specimen. This could be confirmed in glioma cell lines. High PCDHGC3 mRNA expression correlated with longer progression-free survival (PFS) in glioma patients. PCDHGC3 knockout in U343 resulted in a slower growth rate but a significantly faster migration rate in the wound healing assay and decreased the expression of several genes involved in WNT signaling. PCDHGC3 expression should therefore be further investigated as a PFS-marker in gliomas. However, more studies are needed to elucidate the molecular mechanisms underlying the PCDHGC3 effects.}, language = {en} } @article{RajendranRajendranGuptaetal.2022, author = {Rajendran, Ranjithkumar and Rajendran, Vinothkumar and Gupta, Liza and Shirvanchi, Kian and Schunin, Darja and Karnati, Srikanth and Giraldo-Vel{\´a}squez, Mario and Berghoff, Martin}, title = {Interferon beta-1a versus combined interferon beta-1a and oligodendrocyte-specific FGFR1 deletion in experimental autoimmune encephalomyelitis}, series = {International Journal of Molecular Sciences}, volume = {23}, journal = {International Journal of Molecular Sciences}, number = {20}, issn = {1422-0067}, doi = {10.3390/ijms232012183}, url = {http://nbn-resolving.de/urn:nbn:de:bvb:20-opus-290401}, year = {2022}, abstract = {Recombinant beta interferons-1 (IFNβ-1) are used as first line therapies in patients with relapsing multiple sclerosis (MS), a chronic inflammatory and neurodegenerative disease of the CNS. IFNβ-1a/b has moderate effects on the prevention of relapses and slowing of disease progression. Fibroblast growth factors (FGFs) and FGF receptors (FGFRs) are known to play a key role in the pathology of MS and its model EAE. To investigate the effects of short-term treatment with s.c. IFNβ-1a versus the combined application of s.c. IFNβ-1a and oligodendrocyte-specific deletion of FGFR1 (Fgfr1\(^{ind-/-}\) mice) in MOG\(_{35-55}\)-induced EAE. IFNβ-1a (30 mg/kg) was applied s.c. from days 0-7 p.i. of EAE in controls and Fgfr1\(^{ind-/-}\) mice. FGFR signaling proteins associated with inflammation/degeneration in MS/EAE were analyzed by western blot in the spinal cord. Further, FGFR1 in Oli-neu oligodendrocytes were inhibited by PD166866 and treated with IFNβ-1a (400 ng/mL). Application of IFNβ-1a over 8 days resulted in less symptoms only at the peak of disease (days 9-11) compared to controls. Application of IFNβ-1a in Fgfr1\(^{ind-/-}\) mice resulted in less symptoms primarily in the chronic phase of EAE. Fgfr1\(^{ind-/-}\) mice treated with IFNβ-1a showed increased expression of pERK and BDNF. In Oli-neu oligodendrocytes, treatment with PD166866 and IFNβ-1a also showed an increased expression of pERK and BDNF/TrkB. These data suggest that the beneficial effects in the chronic phase of EAE and on signaling molecules associated with ERK and BDNF expression are caused by the modulation of FGFR1 and not by interferon beta-1a. FGFR may be a potential target for therapy in MS.}, language = {en} } @article{ReschkeSalvadorSchlegeletal.2022, author = {Reschke, Moritz and Salvador, Ellaine and Schlegel, Nicolas and Burek, Malgorzata and Karnati, Srikanth and Wunder, Christian and F{\"o}rster, Carola Y.}, title = {Isosteviol sodium (STVNA) reduces pro-inflammatory cytokine IL-6 and GM-CSF in an in vitro murine stroke model of the blood-brain barrier (BBB)}, series = {Pharmaceutics}, volume = {14}, journal = {Pharmaceutics}, number = {9}, issn = {1999-4923}, doi = {10.3390/pharmaceutics14091753}, url = {http://nbn-resolving.de/urn:nbn:de:bvb:20-opus-286275}, year = {2022}, abstract = {Early treatment with glucocorticoids could help reduce both cytotoxic and vasogenic edema, leading to improved clinical outcome after stroke. In our previous study, isosteviol sodium (STVNA) demonstrated neuroprotective effects in an in vitro stroke model, which utilizes oxygen-glucose deprivation (OGD). Herein, we tested the hypothesis that STVNA can activate glucocorticoid receptor (GR) transcriptional activity in brain microvascular endothelial cells (BMECs) as previously published for T cells. STVNA exhibited no effects on transcriptional activation of the glucocorticoid receptor, contrary to previous reports in Jurkat cells. However, similar to dexamethasone, STVNA inhibited inflammatory marker IL-6 as well as granulocyte-macrophage colony-stimulating factor (GM-CSF) secretion. Based on these results, STVNA proves to be beneficial as a possible prevention and treatment modality for brain ischemia-reperfusion injury-induced blood-brain barrier (BBB) dysfunction.}, language = {en} } @article{Drenckhahn2021, author = {Drenckhahn, Detlev}, title = {Zur Vegetation der Seedeiche der Nordseek{\"u}ste Schleswig-Holsteins - Implikationen f{\"u}r die Umsetzung des Generalplans K{\"u}stenschutz}, series = {Forum Geobotanicum}, volume = {10}, journal = {Forum Geobotanicum}, issn = {1867-9315}, doi = {10.3264/FG.2021.0825a}, url = {http://nbn-resolving.de/urn:nbn:de:bvb:20-opus-243795}, pages = {28-37}, year = {2021}, abstract = {Bis zum Jahr 2100 prognostiziert der Weltklimarat (IPCC 2021) einen Anstieg des Meeresspiegels von bis zu 63-101 cm gegen{\"u}ber heutigen Wasserst{\"a}nden. Im Rahmen des Generalplans K{\"u}stenschutz Schleswig-Holstein(GKSH) soll als Klimafolgeanpassung eine Erh{\"o}hung und Profil{\"a}nderung der meisten Nordseedeiche und Elbedeiche erfolgen (zusammen 363,3 km mit einer Vegetationsfl{\"a}che von 3.500 ha). Diese Maßnahmen werden mit einem vollst{\"a}ndigen Verlust der alten Deichvegetation einhergehen und zur Freisetzung von großen Mengen an CO₂ aus dem Bodenkohlenstoff f{\"u}hren. Die Seedeiche der Nordseek{\"u}ste (262 km) z{\"a}hlen zu den artenreichen, semi-nat{\"u}rlichen und von Schafen beweideten Grasl{\"a}ndern (Fl{\"a}che von 2600 ha) in Schleswig-Holstein mit bis zu 18 Gras- und 64 zweikeim-bl{\"a}ttrigen Bl{\"u}tenpflanzen und an die Vegetation gebundene 800-1000 Arten von Invertebraten (darunter 200 K{\"a}ferarten). Auf die Außenb{\"o}schung dringen Pflanzen der Salzwiesengesellschaften vor. Die steileren, w{\"a}rmeexponierten ({\"u}berwiegend nach Osten und S{\"u}den ausgerichtet) und durch Vertritt l{\"u}ckigen Innenb{\"o}schungen der Seedeiche sind wertvolle Refugien w{\"a}rmeliebender, konkurrenzschwacher Arten von Magerstandorten und Trittgesellschaften wie die folgenden mediterran-subatlantischen Arten: Knotenklettenkerbel (Torilis nodosa), Zwergklee/Armbl{\"u}tiger Klee (Trifolium micranthum) und Vogelfußklee (Trifolium ornithopodioides). F{\"u}r die Erhaltung beider Kleearten (die aktuelle Verbreitung wird dokumentiert) besitzt Schleswig-Holstein eine nationale und nordwest-europ{\"a}isch-kontinentale Verantwortlichkeit. Folgende Maßnahmen zum Schutz der reichhaltigen Deichvegetation und Teilen seiner Invertebratenfauna bei der Deichverst{\"a}rkung im Rahmen des GKSH werden vorgeschlagen: 1. Abheben der Grasnarbe mit Wurzelraum und zeitnahe Wiederverlegung der alten Grasnarbe (Soden) auf das neue Deichprofil; das ist auch wichtig zum Erhalt des Bodenkohlenstoffs (Klimaschutz). 2. Einsaat von neuen Deichprofilen mit Saatgut von artenreichen Deichabschnitten. 3. Aufnahme substanzieller Forschungsprogramme/Forschungsf{\"o}rderung zur {\"O}kologie der Seedeiche. Weiterhin sollte auf den Einsatz von Herbiziden auf Deichen zur Bek{\"a}mpfung von Disteln verzichtet werden.}, subject = {Seedeich}, language = {de} } @phdthesis{Dambacher2021, author = {Dambacher, Helena}, title = {Die Etablierung des CRISPR/Cas9-Systems in humanen induzierten pluripotenten Stammzellen zur Untersuchung der Funktion des Kanalproteins Connexin 43 in der Embryonalentwicklung}, doi = {10.25972/OPUS-24015}, url = {http://nbn-resolving.de/urn:nbn:de:bvb:20-opus-240152}, school = {Universit{\"a}t W{\"u}rzburg}, year = {2021}, abstract = {Die Rolle von Connexinen und Gap Junction-vermittelter Kommunikation in pluripotenten Stammzellen sowie der fr{\"u}hen Embryonalentwicklung sind bis heute nicht vollst{\"a}ndig aufgekl{\"a}rt. Mutationen in humanen Connexinen verursachen eine Vielzahl von Krankheiten. Connexin-defiziente iPS Zellen stellen eine gute Basis f{\"u}r die Erforschung der Rolle von Connexinen w{\"a}hrend der Embryonalentwicklung und bei der Krankheitsentstehung dar. Das Ziel der vorliegenden Arbeit war es, das CRISPR/Cas9-System in pluripotenten Stammzellen erfolgreich anzuwenden und ein Protokoll zur Erstellung verschiedener Cx43-Defektmutanten zu entwerfen. Nach der Etablierung der CRSIPR/Cas9-Methode in HEK293T-Zellen konnte in der vorliegenden Arbeit dar{\"u}ber hinaus erfolgreich eine Cx43-Defizienz in FSiPS-Zellen erzeugt werden. Weiterhin wurden mehrere Cx43-Mutanten geschaffen und initial auf Pluripotenzmarker und ihr Differenzierungspotential untersucht. Diese Arbeit bildet die Basis f{\"u}r weitere Untersuchungen des Cx43 in iPS-Zellklonen und davon abgeleiteten Zelltypen sowie artifiziellen 3D-Gewebekulturen. Dar{\"u}ber hinaus bildet sie die Grundlage f{\"u}r die Bildung weiterer Connexin-Defektmutanten sowie von iPS-Zellen mit krankheitsrelevanten Mutationen.}, subject = {CRISPR/Cas-Methode}, language = {de} } @article{Dunkel2021, author = {Dunkel, Franz G.}, title = {Sieben neue Arten aus dem Schweizer Ranunculus auricomus-Komplex}, series = {Forum Geobotanicum}, volume = {10}, journal = {Forum Geobotanicum}, issn = {1867-9315}, doi = {10.3264/FG.2021.0825}, url = {http://nbn-resolving.de/urn:nbn:de:bvb:20-opus-243782}, pages = {1-27}, year = {2021}, abstract = {Die Arten des Schweizer Ranunculus-auricomus-Komplexes sind nur zu einem Teil bekannt. Zur vollst{\"a}ndigeren Erfassung des Komplexes wurden Exkursionen in die s{\"u}dwestliche und {\"o}stliche Schweiz unternommen. Es wurden sieben neue Arten entdeckt, die hier beschrieben und abgebildet sind. Ihre Taxonomie und Gef{\"a}hrdung wird diskutiert. R. chalarocarpus W. Koch ex Dunkel ist bereits bei Koch provisorisch erw{\"a}hnt, R. clavicornis Dunkel wird nun g{\"u}ltig beschrieben. Beide Arten sind aufgrund ihres Vorkommens in Auw{\"a}ldern und feuchten Laubw{\"a}ldern stark gef{\"a}hrdet, R. clavicornis sogar fast ausgestorben. Der neu beschriebene R. thurgoviae kommt im Osten der Schweiz vor (Kanton Thurgau). Die bislang bekannte Verbreitung von R. allobrogorum Dunkel, R. crenulatus Dunkel, R. genevensis Dunkel und R. lineatus ist fast vollst{\"a}ndig auf den Kanton Genf beschr{\"a}nkt. Die Arten des Ranunculus auricomus-Komplexes sind ein sensibler Indikator f{\"u}r Ver{\"a}nderungen der Vegetation und Umwelt und sollten diesbez{\"u}glich deutlich mehr Gewicht bekommen.}, subject = {Ranunculus}, language = {de} } @phdthesis{Loew2021, author = {L{\"o}w, Kornelia}, title = {Identifizierung und Charakterisierung koronarer Gef{\"a}ßwand-residenter Stammzellen}, doi = {10.25972/OPUS-22340}, url = {http://nbn-resolving.de/urn:nbn:de:bvb:20-opus-223402}, school = {Universit{\"a}t W{\"u}rzburg}, year = {2021}, abstract = {In der vorliegenden Arbeit gelang es die Bedeutung sowie die Aktivierung und Mobilisierung der koronaren Gef{\"a}ßwand-residenten Stammzellen bei der Angiogenese des Herzgewebes mittels Cardiac Angiogenesis Assay pr{\"a}zise zu charakterisieren und beeinflussende Faktoren zu identifizieren.}, subject = {Vorl{\"a}uferzelle}, language = {de} } @phdthesis{Endlich2021, author = {Endlich, Alexander Dominic}, title = {Die Rolle der Dsg3-Depletion in der Pathogenese des Pemphigus vulgaris}, doi = {10.25972/OPUS-22557}, url = {http://nbn-resolving.de/urn:nbn:de:bvb:20-opus-225573}, school = {Universit{\"a}t W{\"u}rzburg}, year = {2021}, abstract = {Pemphigus vulgaris (PV) ist eine blasenbildende Autoimmunerkrankung, die durch Autoantik{\"o}rper gegen Dsg1 und Dsg3 gekennzeichnet ist. Der genaue Pathomechanismus, der zu einem PV-IgG vermittelten Verlust der interzellul{\"a}ren Adh{\"a}sion f{\"u}hrt, ist noch unklar. Die Dsg3-Depletion und die Modulation von Signalkaskaden stellen hierbei kennzeichnende Merkmale der Erkrankung dar. Mit den Ergebnissen der vorliegenden Arbeit ist eine bessere Einordnung der Dsg3-Depletion in den pathogenetischen Kontext von Pemphigus vulgaris m{\"o}glich. Die Experimente zeigen, dass die Dsg3-Depletion von Differenzierungsprozessen abh{\"a}ngig ist und mit einem Adh{\"a}sionsverlust einhergehen kann. Die Hemmung der PKC verhindert hierbei sowohl die PV-IgG vermittelten Effekte in der Zellkultur als auch die Blasenbildung im Mausmodell in vivo und in humaner Haut ex vivo. Des Weiteren liefert die Arbeit neue Erkenntnisse, welche f{\"u}r die suprabasale Lokalisation der Blasenbildung bedeutsam sein k{\"o}nnten.}, subject = {Zelladh{\"a}sion}, language = {de} } @article{RajendranBoettigerDentzienetal.2021, author = {Rajendran, Ranjithkumar and B{\"o}ttiger, Gregor and Dentzien, Niklas and Rajendran, Vinothkumar and Sharifi, Bischand and Erg{\"u}n, S{\"u}leyman and Stadelmann, Christine and Karnati, Srikanth and Berghoff, Martin}, title = {Effects of FGFR tyrosine kinase inhibition in OLN-93 oligodendrocytes}, series = {Cells}, volume = {10}, journal = {Cells}, number = {6}, issn = {2073-4409}, doi = {10.3390/cells10061318}, url = {http://nbn-resolving.de/urn:nbn:de:bvb:20-opus-239600}, year = {2021}, abstract = {Fibroblast growth factor (FGF) signaling is involved in the pathogenesis of multiple sclerosis (MS). Data from neuropathology studies suggest that FGF signaling contributes to the failure of remyelination in MS. In MOG\(_{35-55}\)-induced EAE, oligodendrocyte-specific deletion of FGFR1 and FGFR2 resulted in a less severe disease course, reduced inflammation, myelin and axon degeneration and changed FGF/FGFR and BDNF/TrkB signaling. Since signaling cascades in oligodendrocytes could not be investigated in the EAE studies, we here aimed to characterize FGFR-dependent oligodendrocyte-specific signaling in vitro. FGFR inhibition was achieved by application of the multi-kinase-inhibitor dovitinib and the FGFR1/2/3-inhibitor AZD4547. Both substances are potent inhibitors of FGF signaling; they are effective in experimental tumor models and patients with malignancies. Effects of FGFR inhibition in oligodendrocytes were studied by immunofluorescence microscopy, protein and gene analyses. Application of the tyrosine kinase inhibitors reduced FGFR1, phosphorylated ERK and Akt expression, and it enhanced BDNF and TrkB expression. Furthermore, the myelin proteins CNPase and PLP were upregulated by FGFR inhibition. In summary, inhibition of FGFR signaling in oligodendrocytes can be achieved by application of tyrosine kinase inhibitors. Decreased phosphorylation of ERK and Akt is associated with an upregulation of BDNF/TrkB signaling, which may be responsible for the increased production of myelin proteins. Furthermore, these data suggest that application of FGFR inhibitors may have the potential to promote remyelination in the CNS.}, language = {en} } @article{HorderGuazaLasherasGrummeletal.2021, author = {Horder, Hannes and Guaza Lasheras, Mar and Grummel, Nadine and Nadernezhad, Ali and Herbig, Johannes and Erg{\"u}n, S{\"u}leyman and Teßmar, J{\"o}rg and Groll, J{\"u}rgen and Fabry, Ben and Bauer-Kreisel, Petra and Blunk, Torsten}, title = {Bioprinting and differentiation of adipose-derived stromal cell spheroids for a 3D breast cancer-adipose tissue model}, series = {Cells}, volume = {10}, journal = {Cells}, number = {4}, doi = {10.3390/cells10040803}, url = {http://nbn-resolving.de/urn:nbn:de:bvb:20-opus-236496}, year = {2021}, abstract = {Biofabrication, including printing technologies, has emerged as a powerful approach to the design of disease models, such as in cancer research. In breast cancer, adipose tissue has been acknowledged as an important part of the tumor microenvironment favoring tumor progression. Therefore, in this study, a 3D-printed breast cancer model for facilitating investigations into cancer cell-adipocyte interaction was developed. First, we focused on the printability of human adipose-derived stromal cell (ASC) spheroids in an extrusion-based bioprinting setup and the adipogenic differentiation within printed spheroids into adipose microtissues. The printing process was optimized in terms of spheroid viability and homogeneous spheroid distribution in a hyaluronic acid-based bioink. Adipogenic differentiation after printing was demonstrated by lipid accumulation, expression of adipogenic marker genes, and an adipogenic ECM profile. Subsequently, a breast cancer cell (MDA-MB-231) compartment was printed onto the adipose tissue constructs. After nine days of co-culture, we observed a cancer cell-induced reduction of the lipid content and a remodeling of the ECM within the adipose tissues, with increased fibronectin, collagen I and collagen VI expression. Together, our data demonstrate that 3D-printed breast cancer-adipose tissue models can recapitulate important aspects of the complex cell-cell and cell-matrix interplay within the tumor-stroma microenvironment}, language = {en} } @article{JordanJaeckleScheidtetal.2021, author = {Jordan, Martin C. and J{\"a}ckle, Veronika and Scheidt, Sebastian and Gilbert, Fabian and H{\"o}lscher-Doht, Stefanie and Erg{\"u}n, S{\"u}leyman and Meffert, Rainer H. and Heintel, Timo M.}, title = {Trans-obturator cable fixation of open book pelvic injuries}, series = {Scientific Reports}, volume = {11}, journal = {Scientific Reports}, number = {1}, doi = {10.1038/s41598-021-92755-2}, url = {http://nbn-resolving.de/urn:nbn:de:bvb:20-opus-261212}, year = {2021}, abstract = {Operative treatment of ruptured pubic symphysis by plating is often accompanied by complications. Trans-obturator cable fixation might be a more reliable technique; however, have not yet been tested for stabilization of ruptured pubic symphysis. This study compares symphyseal trans-obturator cable fixation versus plating through biomechanical testing and evaluates safety in a cadaver experiment. APC type II injuries were generated in synthetic pelvic models and subsequently separated into three different groups. The anterior pelvic ring was fixed using a four-hole steel plate in Group A, a stainless steel cable in Group B, and a titan band in Group C. Biomechanical testing was conducted by a single-leg-stance model using a material testing machine under physiological load levels. A cadaver study was carried out to analyze the trans-obturator surgical approach. Peak-to-peak displacement, total displacement, plastic deformation and stiffness revealed a tendency for higher stability for trans-obturator cable/band fixation but no statistical difference to plating was detected. The cadaver study revealed a safe zone for cable passage with sufficient distance to the obturator canal. Trans-obturator cable fixation has the potential to become an alternative for symphyseal fixation with less complications.}, language = {en} } @phdthesis{Andreska2021, author = {Andreska, Thomas}, title = {Effects of dopamine on BDNF / TrkB mediated signaling and plasticity on cortico-striatal synapses}, doi = {10.25972/OPUS-17431}, url = {http://nbn-resolving.de/urn:nbn:de:bvb:20-opus-174317}, school = {Universit{\"a}t W{\"u}rzburg}, year = {2021}, abstract = {Progressive loss of voluntary movement control is the central symptom of Parkinson's disease (PD). Even today, we are not yet able to cure PD. This is mainly due to a lack of understanding the mechanisms of movement control, network activity and plasticity in motor circuits, in particular between the cerebral cortex and the striatum. Brain-derived neurotrophic factor (BDNF) has emerged as one of the most important factors for the development and survival of neurons, as well as for synaptic plasticity. It is thus an important target for the development of new therapeutic strategies against neurodegenerative diseases. Together with its receptor, the Tropomyosin receptor kinase B (TrkB), it is critically involved in development and function of the striatum. Nevertheless, little is known about the localization of BDNF within presynaptic terminals in the striatum, as well as the types of neurons that produce BDNF in the cerebral cortex. Furthermore, the influence of midbrain derived dopamine on the control of BDNF / TrkB interaction in striatal medium spiny neurons (MSNs) remains elusive so far. Dopamine, however, appears to play an important role, as its absence leads to drastic changes in striatal synaptic plasticity. This suggests that dopamine could regulate synaptic activity in the striatum via modulation of BDNF / TrkB function. To answer these questions, we have developed a sensitive and reliable protocol for the immunohistochemical detection of endogenous BDNF. We find that the majority of striatal BDNF is provided by glutamatergic, cortex derived afferents and not dopaminergic inputs from the midbrain. In fact, we found BDNF in cell bodies of neurons in layers II-III and V of the primary and secondary motor cortex as well as layer V of the somatosensory cortex. These are the brain areas that send dense projections to the dorsolateral striatum for control of voluntary movement. Furthermore, we could show that these projection neurons significantly downregulate the expression of BDNF during the juvenile development of mice between 3 and 12 weeks. In parallel, we found a modulatory effect of dopamine on the translocation of TrkB to the cell surface in postsynaptic striatal Medium Spiny Neurons (MSNs). In MSNs of the direct pathway (dMSNs), which express dopamine receptor 1 (DRD1), we observed the formation of TrkB aggregates in the 6-hydroxydopamine (6-OHDA) model of PD. This suggests that DRD1 activity controls TrkB surface expression in these neurons. In contrast, we found that DRD2 activation has opposite effects in MSNs of the indirect pathway (iMSNs). Activation of DRD2 promotes a rapid decrease in TrkB surface expression which was reversible and depended on cAMP. In parallel, stimulation of DRD2 led to induction of phospho-TrkB (pTrkB). This effect was significantly slower than the effect on TrkB surface expression and indicates that TrkB is transactivated by DRD2. Together, our data provide evidence that dopamine triggers dual modes of plasticity on striatal MSNs by acting on TrkB surface expression in DRD1 and DRD2 expressing MSNs. This surface expression of the receptor is crucial for the binding of BDNF, which is released from corticostriatal afferents. This leads to the induction of TrkB-mediated downstream signal transduction cascades and long-term potentiation (LTP). Therefore, the dopamine-mediated translocation of TrkB could be a mediator that modulates the balance between dopaminergic and glutamatergic signaling to allow synaptic plasticity in a spatiotemporal manner. This information and the fact that TrkB is segregated to persistent aggregates in PD could help to improve our understanding of voluntary movement control and to develop new therapeutic strategies beyond those focusing on dopaminergic supply.}, subject = {Brain-derived neurotrophic factor}, language = {en} } @article{KarnatiSeimetzKleefeldtetal.2021, author = {Karnati, Srikanth and Seimetz, Michael and Kleefeldt, Florian and Sonawane, Avinash and Madhusudhan, Thati and Bachhuka, Akash and Kosanovic, Djuro and Weissmann, Norbert and Kr{\"u}ger, Karsten and Erg{\"u}n, S{\"u}leyman}, title = {Chronic Obstructive Pulmonary Disease and the Cardiovascular System: Vascular Repair and Regeneration as a Therapeutic Target}, series = {Frontiers in Cardiovascular Medicine}, volume = {8}, journal = {Frontiers in Cardiovascular Medicine}, issn = {2297-055X}, doi = {10.3389/fcvm.2021.649512}, url = {http://nbn-resolving.de/urn:nbn:de:bvb:20-opus-235631}, year = {2021}, abstract = {Chronic obstructive pulmonary disease (COPD) is a major cause of morbidity and mortality worldwide and encompasses chronic bronchitis and emphysema. It has been shown that vascular wall remodeling and pulmonary hypertension (PH) can occur not only in patients with COPD but also in smokers with normal lung function, suggesting a causal role for vascular alterations in the development of emphysema. Mechanistically, abnormalities in the vasculature, such as inflammation, endothelial dysfunction, imbalances in cellular apoptosis/proliferation, and increased oxidative/nitrosative stress promote development of PH, cor pulmonale, and most probably pulmonary emphysema. Hypoxemia in the pulmonary chamber modulates the activation of key transcription factors and signaling cascades, which propagates inflammation and infiltration of neutrophils, resulting in vascular remodeling. Endothelial progenitor cells have angiogenesis capabilities, resulting in transdifferentiation of the smooth muscle cells via aberrant activation of several cytokines, growth factors, and chemokines. The vascular endothelium influences the balance between vaso-constriction and -dilation in the heart. Targeting key players affecting the vasculature might help in the development of new treatment strategies for both PH and COPD. The present review aims to summarize current knowledge about vascular alterations and production of reactive oxygen species in COPD. The present review emphasizes on the importance of the vasculature for the usually parenchyma-focused view of the pathobiology of COPD.}, language = {en} } @article{DoganScheuringWagneretal.2021, author = {Dogan, Leyla and Scheuring, Ruben and Wagner, Nicole and Ueda, Yuichiro and Schmidt, Sven and W{\"o}rsd{\"o}rfer, Philipp and Groll, J{\"u}rgen and Erg{\"u}n, S{\"u}leyman}, title = {Human iPSC-derived mesodermal progenitor cells preserve their vasculogenesis potential after extrusion and form hierarchically organized blood vessels}, series = {Biofabrication}, volume = {13}, journal = {Biofabrication}, number = {4}, doi = {10.1088/1758-5090/ac26ac}, url = {http://nbn-resolving.de/urn:nbn:de:bvb:20-opus-254046}, year = {2021}, abstract = {Post-fabrication formation of a proper vasculature remains an unresolved challenge in bioprinting. Established strategies focus on the supply of the fabricated structure with nutrients and oxygen and either rely on the mere formation of a channel system using fugitive inks or additionally use mature endothelial cells and/or peri-endothelial cells such as smooth muscle cells for the formation of blood vessels in vitro. Functional vessels, however, exhibit a hierarchical organization and multilayered wall structure that is important for their function. Human induced pluripotent stem cell-derived mesodermal progenitor cells (hiMPCs) have been shown to possess the capacity to form blood vessels in vitro, but have so far not been assessed for their applicability in bioprinting processes. Here, we demonstrate that hiMPCs, after formulation into an alginate/collagen type I bioink and subsequent extrusion, retain their ability to give rise to the formation of complex vessels that display a hierarchical network in a process that mimics the embryonic steps of vessel formation during vasculogenesis. Histological evaluations at different time points of extrusion revealed the initial formation of spheres, followed by lumen formation and further structural maturation as evidenced by building a multilayered vessel wall and a vascular network. These findings are supported by immunostainings for endothelial and peri-endothelial cell markers as well as electron microscopic analyses at the ultrastructural level. Moreover, endothelial cells in capillary-like vessel structures deposited a basement membrane-like matrix at the basal side between the vessel wall and the alginate-collagen matrix. After transplantation of the printed constructs into the chicken chorioallantoic membrane (CAM) the printed vessels connected to the CAM blood vessels and get perfused in vivo. These results evidence the applicability and great potential of hiMPCs for the bioprinting of vascular structures mimicking the basic morphogenetic steps of de novo vessel formation during embryogenesis.}, language = {en} } @article{LuetkensErguenHuflageetal.2021, author = {Luetkens, Karsten Sebastian and Erg{\"u}n, S{\"u}leyman and Huflage, Henner and Kunz, Andreas Steven and Gietzen, Carsten Herbert and Conrads, Nora and Pennig, Lenhard and Goertz, Lukas and Bley, Thorsten Alexander and Gassenmaier, Tobias and Grunz, Jan-Peter}, title = {Dose reduction potential in cone-beam CT imaging of upper extremity joints with a twin robotic x-ray system}, series = {Scientific Reports}, volume = {11}, journal = {Scientific Reports}, number = {1}, doi = {10.1038/s41598-021-99748-1}, url = {http://nbn-resolving.de/urn:nbn:de:bvb:20-opus-270429}, year = {2021}, abstract = {Cone-beam computed tomography is a powerful tool for 3D imaging of the appendicular skeleton, facilitating detailed visualization of bone microarchitecture. This study evaluated various combinations of acquisition and reconstruction parameters for the cone-beam CT mode of a twin robotic x-ray system in cadaveric wrist and elbow scans, aiming to define the best possible trade-off between image quality and radiation dose. Images were acquired with different combinations of tube voltage and tube current-time product, resulting in five scan protocols with varying volume CT dose indices: full-dose (FD; 17.4 mGy), low-dose (LD; 4.5 mGy), ultra-low-dose (ULD; 1.15 mGy), modulated low-dose (mLD; 0.6 mGy) and modulated ultra-low-dose (mULD; 0.29 mGy). Each set of projection data was reconstructed with three convolution kernels (very sharp [Ur77], sharp [Br69], intermediate [Br62]). Five radiologists subjectively assessed the image quality of cortical bone, cancellous bone and soft tissue using seven-point scales. Irrespective of the reconstruction kernel, overall image quality of every FD, LD and ULD scan was deemed suitable for diagnostic use in contrast to mLD (very sharp/sharp/intermediate: 60/55/70\%) and mULD (0/3/5\%). Superior depiction of cortical and cancellous bone was achieved in FD\(_{Ur77}\) and LD\(_{Ur77}\) examinations (p < 0.001) with LD\(_{Ur77}\) scans also providing favorable bone visualization compared to FD\(_{Br69}\) and FD\(_{Br62}\) (p < 0.001). Fleiss' kappa was 0.618 (0.594-0.641; p < 0.001), indicating substantial interrater reliability. In this study, we demonstrate that considerable dose reduction can be realized while maintaining diagnostic image quality in upper extremity joint scans with the cone-beam CT mode of a twin robotic x-ray system. Application of sharper convolution kernels for image reconstruction facilitates superior display of bone microarchitecture.}, language = {en} } @phdthesis{Ipek2021, author = {Ipek, Rojda}, title = {Einfluss der murinen Anti-CD52-Antik{\"o}rper-Therapie auf die Neurodegeneration in der chronischen MP4-induzierten experimentellen autoimmunen Enzephalomyelitis als Mausmodell der Multiplen Sklerose}, doi = {10.25972/OPUS-22576}, url = {http://nbn-resolving.de/urn:nbn:de:bvb:20-opus-225766}, school = {Universit{\"a}t W{\"u}rzburg}, year = {2021}, abstract = {Die Multiple Sklerose (MS) ist eine chronisch-entz{\"u}ndliche Autoimmunerkrankung des zentralen Nervensystems (ZNS) und stellt die h{\"a}ufigste Ursache fr{\"u}hzeitiger Behinderung junger Erwachsener dar. Kennzeichnend sind multifokale ZNS-L{\"a}sionen, die durch Inflammation, Demyelinisierung und Axonsch{\"a}den gepr{\"a}gt sind und zu multiplen neurologischen Defiziten f{\"u}hren. Derzeit ist es mithilfe der verlaufsmodifizierenden Therapie m{\"o}glich, die Immunantwort abzuschw{\"a}chen und damit die Krankheitsprogression zu verz{\"o}gern. Geheilt werden kann die Erkrankung jedoch bislang nicht. Dabei ist nicht hinreichend gekl{\"a}rt, ob die neuen Therapieoptionen {\"u}ber die Immunmodulation/-suppression hinaus einen anhaltenden Schutz vor der langfristigen Neurodegeneration bieten. Basierend auf den vielversprechenden Ergebnissen klinischer Studien zur Therapie der schubf{\"o}rmig-remittierenden MS mit dem Anti-CD52-Antik{\"o}rper Alemtuzumab, der zu einer Depletion CD52-exprimierender Immunzellen f{\"u}hrt, wurden diesbez{\"u}glich Analysen in MS-Tiermodellen durchgef{\"u}hrt. Da die Untersuchung der zugrunde liegenden Patho- und Effektormechanismen am Menschen kaum m{\"o}glich ist, ist die MS-Forschung f{\"u}r ein tiefergehendes Verst{\"a}ndnis auf Tiermodelle angewiesen. Die experimentelle autoimmune Enzephalomyelitis (EAE) ist hierbei das am weitesten verbreitete Modell der MS, wof{\"u}r vor allem der C57BL/6 (B6) -Mausstamm verwendet wird, da auf diesem Hintergrund die meisten genmodifizierten M{\"a}use gez{\"u}chtet werden. Jene tierexperimentellen Studien, in denen ein muriner Anti-CD52-Antik{\"o}rper im fr{\"u}hen Krankheitsstadium der EAE (Auftreten erster paralytischer Symptome) verabreicht wurde, erbrachten den Hinweis einer neuroprotektiven und scheinbar regenerativen Wirkung des Antik{\"o}rpers. {\"U}ber einen neuroprotektiven Effekt von Alemtuzumab im schwer behandelbaren chronisch-progredienten Stadium der MS ist jedoch wenig bekannt. Die vorliegende Arbeit ist die erste detaillierte Untersuchung zum Einfluss des murinen Anti-CD52-Antik{\"o}rpers auf die Demyelinisierung, den Axonschaden und die Hirnatrophie in der MP4-induzierten EAE der B6-Maus im chronischen Verlauf der Erkrankung (ab stabilem Plateau der klinischen Symptomatik). MP4 ist ein Myelinfusionsprotein aus MBP (Myelin-Basisches-Protein) und PLP (Proteolipidprotein), welches in B6-M{\"a}usen durch aktive Immunisierung eine EAE induziert, die chronisch verl{\"a}uft und als eines von wenigen Modellen neben der T-Zell-Abh{\"a}ngigkeit die an Bedeutung zunehmende B-Zell-Komponente der MS darstellt. Histopathologisch finden sich in der chronischen MP4-induzierten EAE eine ausgepr{\"a}gte R{\"u}ckenmarks- und Kleinhirnsch{\"a}digung, die vor allem im Kleinhirn durch eine B-Zell-Aggregation charakterisiert ist. Nachdem die MP4-immunisierten M{\"a}use im chronischen Stadium der EAE an f{\"u}nf aufeinanderfolgenden Tagen mit 10 mg/kg K{\"o}rpergewicht murinem Anti-CD52-spezifischem IgG2a-Isotypantik{\"o}rper bzw. murinem unspezifischem IgG2a-Isotyp-Kontroll-Antik{\"o}rper behandelt worden waren, wurde die Lymphozytendepletion im peripheren Blut durchflusszytometrisch ermittelt und deren Einfluss auf MP4-spezifische Antik{\"o}rper anhand eines indirekten Enzyme-linked Immunosorbent Assays (ELISAs) untersucht. Als Marker f{\"u}r Axonsch{\"a}den wurde im Serum vorhandenes phosphoryliertes Neurofilament-Heavy (pNF-H) mithilfe eines indirekten Sandwich-ELISAs quantitativ bestimmt. R{\"u}ckenmark und Kleinhirn wurden ultrastrukturell auf Ver{\"a}nderungen der Myelinisierung (mittels g-Ratio: Axondurchmesser geteilt durch Gesamtdurchmesser der Nervenfaser) und auf Axonpathologien (verringerter Abstand benachbarter Neurofilamente, axolytische Axone, axonaler Verlust) untersucht. Die Hirnatrophie wurde MRT-basiert gemessen und der klinische Verlauf t{\"a}glich evaluiert. Durch die Anti-CD52-Antik{\"o}rperbehandlung wurde die T- und B-Zellzahl zwar drastisch vermindert, die MP4-spezifische Antik{\"o}rperproduktion blieb davon jedoch unbeeintr{\"a}chtigt. Ein g{\"u}nstiger Effekt auf die De- und Remyelinisierung war nicht festzustellen. Das Hirnvolumen und die klinische Pr{\"a}sentation der M{\"a}use blieben ebenfalls unver{\"a}ndert. W{\"a}hrend kein Unterschied der pNF-H-Konzentration zu erkennen war, konnte ultrastrukturell jedoch ein geringerer Axonschaden nachgewiesen werden. Insgesamt legen diese Ergebnisse nahe, dass der Anti-CD52-Antik{\"o}rper im chronischen Verlauf der EAE/MS wenig Einfluss auf die neurodegenerativen Prozesse nimmt und die Regeneration nicht f{\"o}rdern kann. Die Ursache liegt vermutlich in der Undurchl{\"a}ssigkeit der Bluthirnschranke f{\"u}r Antik{\"o}rper sowie dem limitierten Verst{\"a}ndnis der Antik{\"o}rperwirkung im ZNS. Die vorliegende Studie regt somit zur Etablierung von ZNS-wirksamen Antik{\"o}rpern an und unterstreicht die Bedeutung der Entwicklung von selektiveren neuroprotektiven und remyelinisierungsf{\"o}rdernden Behandlungsans{\"a}tzen, die eine wertvolle Erg{\"a}nzung zur verlaufsmodifizierenden Therapie darstellen k{\"o}nnten.}, subject = {Multiple Sklerose}, language = {de} } @phdthesis{Reeh2021, author = {Reeh, Laurens}, title = {Immunmodulatorische Effekte CD44-positiver Gef{\"a}ßwand-residenter Stamm- und Vorl{\"a}uferzellen im myokardialen Gewebe}, doi = {10.25972/OPUS-25102}, url = {http://nbn-resolving.de/urn:nbn:de:bvb:20-opus-251020}, school = {Universit{\"a}t W{\"u}rzburg}, year = {2021}, abstract = {Die Identifizierung endogener Stammzellen mit kardiogenem Potenzial und die M{\"o}glichkeit, deren Differenzierung zu steuern, w{\"u}rde einen Meilenstein in der kardioregenerativen Therapie darstellen. Innerhalb der Gef{\"a}ßwand konnten unterschiedliche Stamm- und Vorl{\"a}uferzellen identifiziert werden, die sog. Gef{\"a}ßwand-residenten Stammzellen (VW-SCs). Zuletzt konnten aus CD34(+) VW-SCs, ohne genetische Manipulation, Kardiomyozyten generiert werden. Zus{\"a}tzlich fungiert die Gef{\"a}ßwand als Quelle inflammatorischer Zellen, die essenziell f{\"u}r die kardiogene Differenzierung der VW-SCs zu sein scheinen. Ziel dieser Arbeit war es, das Verhalten von CD44(+) VW-SCs zu untersuchen, um herauszufinden, inwieweit dieser Stammzelltyp eine endogene Generierung von Kardiomyozyten unterst{\"u}tzen k{\"o}nnte. Dabei wurde mit infarzierten M{\"a}useherzen, dem Aortenringassay (ARA) und dem kardialen Angiogeneseassay (CAA) gearbeitet. Sowohl in vivo in isch{\"a}mischen Arealen infarzierter M{\"a}useherzen als auch ex vivo im CAA kam es zu einem signifikanten Anstieg von CD44(+) Zellen. Mittels F{\"a}rbungen auf CD44 und Ki-67 konnte die Teilungsf{\"a}higkeit dieser Zellen demonstriert werden. Ex vivo ließen sich aus CD44(+) Zellen F4/80(+) Makrophagen generieren. Die CD44(+) VW-SCs k{\"o}nnen sich dabei sowohl zu pro-inflammatorischen iNOS(+) M1- als auch zu anti-inflammatorischen IL-10(+) M2-Makrophagen differenzieren. Eine Modulation der kardialen Inflammation k{\"o}nnte einen entscheidenden Einfluss auf die Kardiomyogenese haben. Unter VEGF-A kam es im CAA zu einer deutlichen Zunahme von CD44(+) Zellen. Unter Lenvatinib blieb das kardiale Sprouting g{\"a}nzlich aus, die Anzahl der CD44(+) Zellen stagnierte und die VW-SCs verblieben in ihren physiologischen Nischen innerhalb der Gef{\"a}ßwand. Warum es nach einem MI kaum zu einer funktionellen Herzmuskelregeneration kommt, ist weiterhin unklar. Die therapeutische Beeinflussung koronaradventitieller CD44(+) VW-SCs und inflammatorischer Prozesse k{\"o}nnte dabei zuk{\"u}nftig eine wichtige therapeutische Option darstellen.}, subject = {Antigen CD44}, language = {de} } @article{MeyerWatermannDreyeretal.2021, author = {Meyer, Malin Tordis and Watermann, Christoph and Dreyer, Thomas and Wagner, Steffen and Wittekindt, Claus and Klussmann, Jens Peter and Erg{\"u}n, S{\"u}leyman and Baumgart-Vogt, Eveline and Karnati, Srikanth}, title = {Differential expression of peroxisomal proteins in distinct types of parotid gland tumors}, series = {International Journal of Molecular Sciences}, volume = {22}, journal = {International Journal of Molecular Sciences}, number = {15}, issn = {1422-0067}, doi = {10.3390/ijms22157872}, url = {http://nbn-resolving.de/urn:nbn:de:bvb:20-opus-261047}, year = {2021}, abstract = {Salivary gland cancers are rare but aggressive tumors that have poor prognosis and lack effective cure. Of those, parotid tumors constitute the majority. Functioning as metabolic machinery contributing to cellular redox balance, peroxisomes have emerged as crucial players in tumorigenesis. Studies on murine and human cells have examined the role of peroxisomes in carcinogenesis with conflicting results. These studies either examined the consequences of altered peroxisomal proliferators or compared their expression in healthy and neoplastic tissues. None, however, examined such differences exclusively in human parotid tissue or extended comparison to peroxisomal proteins and their associated gene expressions. Therefore, we examined differences in peroxisomal dynamics in parotid tumors of different morphologies. Using immunofluorescence and quantitative PCR, we compared the expression levels of key peroxisomal enzymes and proliferators in healthy and neoplastic parotid tissue samples. Three parotid tumor subtypes were examined: pleomorphic adenoma, mucoepidermoid carcinoma and acinic cell carcinoma. We observed higher expression of peroxisomal matrix proteins in neoplastic samples with exceptional down regulation of certain enzymes; however, the degree of expression varied between tumor subtypes. Our findings confirm previous experimental results on other organ tissues and suggest peroxisomes as possible therapeutic targets or markers in all or certain subtypes of parotid neoplasms.}, language = {en} } @article{SchlechtVallonWagneretal.2021, author = {Schlecht, Anja and Vallon, Mario and Wagner, Nicole and Erg{\"u}n, S{\"u}leyman and Braunger, Barbara M.}, title = {TGFβ-Neurotrophin Interactions in Heart, Retina, and Brain}, series = {Biomolecules}, volume = {11}, journal = {Biomolecules}, number = {9}, issn = {2218-273X}, doi = {10.3390/biom11091360}, url = {http://nbn-resolving.de/urn:nbn:de:bvb:20-opus-246159}, year = {2021}, abstract = {Ischemic insults to the heart and brain, i.e., myocardial and cerebral infarction, respectively, are amongst the leading causes of death worldwide. While there are therapeutic options to allow reperfusion of ischemic myocardial and brain tissue by reopening obstructed vessels, mitigating primary tissue damage, post-infarction inflammation and tissue remodeling can lead to secondary tissue damage. Similarly, ischemia in retinal tissue is the driving force in the progression of neovascular eye diseases such as diabetic retinopathy (DR) and age-related macular degeneration (AMD), which eventually lead to functional blindness, if left untreated. Intriguingly, the easily observable retinal blood vessels can be used as a window to the heart and brain to allow judgement of microvascular damages in diseases such as diabetes or hypertension. The complex neuronal and endocrine interactions between heart, retina and brain have also been appreciated in myocardial infarction, ischemic stroke, and retinal diseases. To describe the intimate relationship between the individual tissues, we use the terms heart-brain and brain-retina axis in this review and focus on the role of transforming growth factor β (TGFβ) and neurotrophins in regulation of these axes under physiologic and pathologic conditions. Moreover, we particularly discuss their roles in inflammation and repair following ischemic/neovascular insults. As there is evidence that TGFβ signaling has the potential to regulate expression of neurotrophins, it is tempting to speculate, and is discussed here, that cross-talk between TGFβ and neurotrophin signaling protects cells from harmful and/or damaging events in the heart, retina, and brain.}, language = {en} } @article{SchlechtThienWolfetal.2021, author = {Schlecht, Anja and Thien, Adrian and Wolf, Julian and Prinz, Gabriele and Agostini, Hansj{\"u}rgen and Schlunck, G{\"u}nther and Wieghofer, Peter and Boneva, Stefaniya and Lange, Clemens}, title = {Immunosenescence in choroidal neovascularization (CNV) — Transcriptional profiling of na{\"i}ve and CNV-associated retinal myeloid cells during aging}, series = {International Journal of Molecular Sciences}, volume = {22}, journal = {International Journal of Molecular Sciences}, number = {24}, issn = {1422-0067}, doi = {10.3390/ijms222413318}, url = {http://nbn-resolving.de/urn:nbn:de:bvb:20-opus-284342}, year = {2021}, abstract = {Immunosenescence is considered a possible factor in the development of age-related macular degeneration and choroidal neovascularization (CNV). However, age-related changes of myeloid cells (MCs), such as microglia and macrophages, in the healthy retina or during CNV formation are ill-defined. In this study, Cx3cr1-positive MCs were isolated by fluorescence-activated cell sorting from six-week (young) and two-year-old (old) Cx3cr1\(^{GFP/+}\) mice, both during physiological aging and laser-induced CNV development. High-throughput RNA-sequencing was performed to define the age-dependent transcriptional differences in MCs during physiological aging and CNV development, complemented by immunohistochemical characterization and the quantification of MCs, as well as CNV size measurements. These analyses revealed that myeloid cells change their transcriptional profile during both aging and CNV development. In the steady state, senescent MCs demonstrated an upregulation of factors contributing to cell proliferation and chemotaxis, such as Cxcl13 and Cxcl14, as well as the downregulation of microglial signature genes. During CNV formation, aged myeloid cells revealed a significant upregulation of angiogenic factors such as Arg1 and Lrg1 concomitant with significantly enlarged CNV and an increased accumulation of MCs in aged mice in comparison to young mice. Future studies need to clarify whether this observation is an epiphenomenon or a causal relationship to determine the role of immunosenescence in CNV formation.}, language = {en} } @article{LiuHanBlairetal.2021, author = {Liu, Fengming and Han, Kun and Blair, Robert and Kenst, Kornelia and Qin, Zhongnan and Upcin, Berin and W{\"o}rsd{\"o}rfer, Philipp and Midkiff, Cecily C. and Mudd, Joseph and Belyaeva, Elizaveta and Milligan, Nicholas S. and Rorison, Tyler D. and Wagner, Nicole and Bodem, Jochen and D{\"o}lken, Lars and Aktas, Bertal H. and Vander Heide, Richard S. and Yin, Xiao-Ming and Kolls, Jay K. and Roy, Chad J. and Rappaport, Jay and Erg{\"u}n, S{\"u}leyman and Qin, Xuebin}, title = {SARS-CoV-2 Infects Endothelial Cells In Vivo and In Vitro}, series = {Frontiers in Cellular and Infection Microbiology}, volume = {11}, journal = {Frontiers in Cellular and Infection Microbiology}, issn = {2235-2988}, doi = {10.3389/fcimb.2021.701278}, url = {http://nbn-resolving.de/urn:nbn:de:bvb:20-opus-241948}, year = {2021}, abstract = {SARS-CoV-2 infection can cause fatal inflammatory lung pathology, including thrombosis and increased pulmonary vascular permeability leading to edema and hemorrhage. In addition to the lung, cytokine storm-induced inflammatory cascade also affects other organs. SARS-CoV-2 infection-related vascular inflammation is characterized by endotheliopathy in the lung and other organs. Whether SARS-CoV-2 causes endotheliopathy by directly infecting endothelial cells is not known and is the focus of the present study. We observed 1) the co-localization of SARS-CoV-2 with the endothelial cell marker CD31 in the lungs of SARS-CoV-2-infected mice expressing hACE2 in the lung by intranasal delivery of adenovirus 5-hACE2 (Ad5-hACE2 mice) and non-human primates at both the protein and RNA levels, and 2) SARS-CoV-2 proteins in endothelial cells by immunogold labeling and electron microscopic analysis. We also detected the co-localization of SARS-CoV-2 with CD31 in autopsied lung tissue obtained from patients who died from severe COVID-19. Comparative analysis of RNA sequencing data of the lungs of infected Ad5-hACE2 and Ad5-empty (control) mice revealed upregulated KRAS signaling pathway, a well-known pathway for cellular activation and dysfunction. Further, we showed that SARS-CoV-2 directly infects mature mouse aortic endothelial cells (AoECs) that were activated by performing an aortic sprouting assay prior to exposure to SARS-CoV-2. This was demonstrated by co-localization of SARS-CoV-2 and CD34 by immunostaining and detection of viral particles in electron microscopic studies. Moreover, the activated AoECs became positive for ACE-2 but not quiescent AoECs. Together, our results indicate that in addition to pneumocytes, SARS-CoV-2 also directly infects mature vascular endothelial cells in vivo and ex vivo, which may contribute to cardiovascular complications in SARS-CoV-2 infection, including multipleorgan failure.}, language = {en} } @phdthesis{Kuhn2021, author = {Kuhn, Anja}, title = {Rekrutierung von Stromazellen aus gef{\"a}ßwandresidenten Vorl{\"a}uferzellen w{\"a}hrend der Tumorgenese}, doi = {10.25972/OPUS-22431}, url = {http://nbn-resolving.de/urn:nbn:de:bvb:20-opus-224315}, school = {Universit{\"a}t W{\"u}rzburg}, year = {2021}, abstract = {Tumore bestehen nicht nur aus malignen Zellen, sondern ebenfalls aus einer Vielzahl an nicht tumorigenen Zellen, die den Tumor auf vielf{\"a}ltige Weise unterst{\"u}tzen und den Tumor vor therapeutischen Maßnahmen sch{\"u}tzen. Die Frage der Herkunft dieser Zellen insbesondere in einem nicht vaskularisierten Tumor ist daher auch f{\"u}r die Entwicklung zuk{\"u}nftiger Therapeutika relevant. In dieser Arbeit wurde eine Methode etabliert, die im dreidimensionalen Raum die Untersuchung des Einflusses von Tumorzellen auf die vaskul{\"a}re Adventitia am Model der Mausaorta erm{\"o}glicht. Dazu erfolgte die Einbettung von Alginatbeads aus verschiedenen Tumorzelllinien in eine gemeinsame Kollagenmatrix mit murinen Aortenringen. W{\"a}hrend des zehnt{\"a}gigem Versuchszeitraums wurde die Aussprossung von Zellen aus den Aortenringen beobachtet und quantifiziert. Es wurde festgestellt, dass die Auswanderung w{\"a}hrend des Versuchszeitraums zunimmt und dass die Konfrontation mit der Zytokinmischung der Tumorzellen zu einer st{\"a}rkeren Aussprossung f{\"u}hrt, als die Stimulation mit VEGF oder keine Stimulation. Eine gerichtete Auswanderung der Zellen in Richtung der Tumorbeads konnte nicht nachgewiesen bzw. best{\"a}tigt werden. Kapill{\"a}re Aussprossungen waren nur in geringem Ausmaß zu beobachten. Bei Charakterisierung der ausgewanderten Zellen mittels immunhistochemischer F{\"a}rbungen waren keine F4/80-positiven und nur einzelne CD34-positive Zellen zu finden. CD31-positive Endothelzellen stellten die Mehrheit der ausgewanderten Zellen bei Tumorzellkonfrontation. Perizyten, die mit dem Marker NG2 gef{\"a}rbt wurden, stellten eine Mehrheit der migrierten Zellen bei allen Bedingungen. Die in dieser Arbeit etablierte Methode des Aortenring-Bead-Konfrontationsassays erm{\"o}glicht es, in Echtzeit den Einfluss von Tumorzellen auf die Gef{\"a}ßwand im dreidimensionalen Raum zu beobachten. Der Aortenring-Bead-Konfrontationsassay bietet eine Vielzahl an Variationsm{\"o}glichkeiten und stellt daher eine vielversprechende M{\"o}glichkeit dar, die L{\"u}cke zwischen zweidimensionalen in vitro-Experimenten und kostenintensiven in vivo-Versuchen zu schließen.}, subject = {Stroma}, language = {de} } @phdthesis{Kempf2021, author = {Kempf, Bettina}, title = {Interaktion ausgew{\"a}hlter Mechanismen der Pemphigus-Pathogenese}, doi = {10.25972/OPUS-22048}, url = {http://nbn-resolving.de/urn:nbn:de:bvb:20-opus-220480}, school = {Universit{\"a}t W{\"u}rzburg}, year = {2021}, abstract = {Bei der Autoimmunerkrankung Pemphigus vulgaris f{\"u}hren Antik{\"o}rper zur charakteristischen suprabasalen Akantholyse und Blasenbildung der Epidermis, indem sie an spezifische Antigene, Dsg3 (Desmoglein 3) und Dsg1 (Desmoglein 1), auf der Zelloberfl{\"a}che der Keratinozyten binden. Die Art und Weise, wie die multiplen zellul{\"a}ren Pathomechanismen zusammenwirken und das potenziell t{\"o}dliche Krankheitsbild hervorrufen, ist jedoch bislang noch weitgehend unklar. In der vorliegenden Arbeit wurden entscheidende, durch die Autoantik{\"o}rper hervorgerufene, pathologische intrazellul{\"a}re Prozesse genauer untersucht und deren Stellenwert beleuchtet.}, subject = {pemphigus}, language = {de} } @article{RajendranRajendranGiraldoVelasquezetal.2021, author = {Rajendran, Ranjithkumar and Rajendran, Vinothkumar and Giraldo-Velasquez, Mario and Megalofonou, Fevronia-Foivi and Gurski, Fynn and Stadelmann, Christine and Karnati, Srikanth and Berghoff, Martin}, title = {Oligodendrocyte-specific deletion of FGFR1 reduces cerebellar inflammation and neurodegeneration in MOG\(_{35-55}\)-induced EAE}, series = {International Journal of Molecular Sciences}, volume = {22}, journal = {International Journal of Molecular Sciences}, number = {17}, issn = {1422-0067}, doi = {10.3390/ijms22179495}, url = {http://nbn-resolving.de/urn:nbn:de:bvb:20-opus-284296}, year = {2021}, abstract = {Multiple sclerosis (MS) is a chronic inflammatory and degenerative disease of the central nervous system (CNS). MS commonly affects the cerebellum causing acute and chronic symptoms. Cerebellar signs significantly contribute to clinical disability, and symptoms such as tremor, ataxia, and dysarthria are difficult to treat. Fibroblast growth factors (FGFs) and their receptors (FGFRs) are involved in demyelinating pathologies such as MS. In autopsy tissue from patients with MS, increased expression of FGF1, FGF2, FGF9, and FGFR1 was found in lesion areas. Recent research using mouse models has focused on regions such as the spinal cord, and data on the expression of FGF/FGFR in the cerebellum are not available. In recent EAE studies, we detected that oligodendrocyte-specific deletion of FGFRs results in a milder disease course, less cellular infiltrates, and reduced neurodegeneration in the spinal cord. The objective of this study was to characterize the role of FGFR1 in oligodendrocytes in the cerebellum. Conditional deletion of FGFR1 in oligodendrocytes (Fgfr1\(^{ind-/-}\) was achieved by tamoxifen application, EAE was induced using the MOG\(_{35-55}\) peptide. The cerebellum was analyzed by histology, immunohistochemistry, and western blot. At day 62 p.i., Fgfr1\(^{ind-/-}\) mice showed less myelin and axonal degeneration compared to FGFR1-competent mice. Infiltration of CD3(+) T cells, Mac3(+) cells, B220(+) B cells and IgG(+) plasma cells in cerebellar white matter lesions (WML) was less in Fgfr1\(^{ind-/-}\)mice. There were no effects on the number of OPC or mature oligodendrocytes in white matter lesion (WML). Expression of FGF2 and FGF9 associated with less myelin and axonal degeneration, and of the pro-inflammatory cytokines IL-1β, IL-6, and CD200 was downregulated in Fgfr1\(^{ind-/-}\) mice. The FGF/FGFR signaling protein pAkt, BDNF, and TrkB were increased in Fgfr1\(^{ind-/-}\) mice. These data suggest that cell-specific deletion of FGFR1 in oligodendrocytes has anti-inflammatory and neuroprotective effects in the cerebellum in the EAE disease model of MS.}, language = {en} } @article{WagnerMottUpcinetal.2021, author = {Wagner, Nicole and Mott, Kristina and Upcin, Berin and Stegner, David and Schulze, Harald and Erg{\"u}n, S{\"u}leyman}, title = {CXCL12-abundant reticular (CAR) cells direct megakaryocyte protrusions across the bone marrow sinusoid wall}, series = {Cells}, volume = {10}, journal = {Cells}, number = {4}, issn = {2073-4409}, doi = {10.3390/cells10040722}, url = {http://nbn-resolving.de/urn:nbn:de:bvb:20-opus-234180}, year = {2021}, abstract = {Megakaryocytes (MKs) release platelets into the lumen of bone marrow (BM) sinusoids while remaining to reside within the BM. The morphogenetic events of this complex process are still not fully understood. We combined confocal laser scanning microscopy with transmission and serial block-face scanning electron microscopy followed by 3D-reconstruction on mouse BM tissue sections. These analyses revealed that MKs in close vicinity to BM sinusoid (BMS) wall first induce the lateral retraction of CXCL12-abundant reticular (CAR) cells (CAR), followed by basal lamina (BL) degradation enabling direct MK-sinusoidal endothelial cells (SECs) interaction. Subsequently, an endothelial engulfment starts that contains a large MK protrusion. Then, MK protrusions penetrate the SEC, transmigrate into the BMS lumen and form proplatelets that are in direct contact to the SEC surface. Furthermore, such processes are induced on several sites, as observed by 3D reconstructions. Our data demonstrate that MKs in interaction with CAR-cells actively induce BMS wall alterations, including CAR-cell retraction, BL degradation, and SEC engulfment containing a large MK protrusion. This results in SEC penetration enabling the migration of MK protrusion into the BMS lumen where proplatelets that are adherent to the luminal SEC surface are formed and contribute to platelet release into the blood circulation.}, language = {en} } @article{KoenigerBellMifkaetal.2021, author = {Koeniger, Tobias and Bell, Luisa and Mifka, Anika and Enders, Michael and Hautmann, Valentin and Mekala, Subba Rao and Kirchner, Philipp and Ekici, Arif B. and Schulz, Christian and W{\"o}rsd{\"o}rfer, Philipp and Mencl, Stine and Kleinschnitz, Christoph and Erg{\"u}n, S{\"u}leyman and Kuerten, Stefanie}, title = {Bone marrow-derived myeloid progenitors in the leptomeninges of adult mice}, series = {Stem Cells}, volume = {39}, journal = {Stem Cells}, number = {2}, doi = {10.1002/stem.3311}, url = {http://nbn-resolving.de/urn:nbn:de:bvb:20-opus-224452}, pages = {227 -- 239}, year = {2021}, abstract = {Although the bone marrow contains most hematopoietic activity during adulthood, hematopoietic stem and progenitor cells can be recovered from various extramedullary sites. Cells with hematopoietic progenitor properties have even been reported in the adult brain under steady-state conditions, but their nature and localization remain insufficiently defined. Here, we describe a heterogeneous population of myeloid progenitors in the leptomeninges of adult C57BL/6 mice. This cell pool included common myeloid, granulocyte/macrophage, and megakaryocyte/erythrocyte progenitors. Accordingly, it gave rise to all major myelo-erythroid lineages in clonogenic culture assays. Brain-associated progenitors persisted after tissue perfusion and were partially inaccessible to intravenous antibodies, suggesting their localization behind continuous blood vessel endothelium such as the blood-arachnoid barrier. Flt3\(^{Cre}\) lineage tracing and bone marrow transplantation showed that the precursors were derived from adult hematopoietic stem cells and were most likely continuously replaced via cell trafficking. Importantly, their occurrence was tied to the immunologic state of the central nervous system (CNS) and was diminished in the context of neuroinflammation and ischemic stroke. Our findings confirm the presence of myeloid progenitors at the meningeal border of the brain and lay the foundation to unravel their possible functions in CNS surveillance and local immune cell production.}, language = {en} } @article{MeyerWatermannDreyeretal.2021, author = {Meyer, Malin Tordis and Watermann, Christoph and Dreyer, Thomas and Erg{\"u}n, S{\"u}leyman and Karnati, Srikanth}, title = {2021 update on diagnostic markers and translocation in salivary gland tumors}, series = {International Journal of Molecular Sciences}, volume = {22}, journal = {International Journal of Molecular Sciences}, number = {13}, issn = {1422-0067}, doi = {10.3390/ijms22136771}, url = {http://nbn-resolving.de/urn:nbn:de:bvb:20-opus-261057}, year = {2021}, abstract = {Salivary gland tumors are a rare tumor entity within malignant tumors of all tissues. The most common are malignant mucoepidermoid carcinoma, adenoid cystic carcinoma, and acinic cell carcinoma. Pleomorphic adenoma is the most recurrent form of benign salivary gland tumor. Due to their low incidence rates and complex histological patterns, they are difficult to diagnose accurately. Malignant tumors of the salivary glands are challenging in terms of differentiation because of their variability in histochemistry and translocations. Therefore, the primary goal of the study was to review the current literature to identify the recent developments in histochemical diagnostics and translocations for differentiating salivary gland tumors.}, language = {en} } @article{RajendranBoettigerStadelmannetal.2021, author = {Rajendran, Ranjithkumar and B{\"o}ttiger, Gregor and Stadelmann, Christine and Karnati, Srikanth and Berghoff, Martin}, title = {FGF/FGFR pathways in multiple sclerosis and in its disease models}, series = {Cells}, volume = {10}, journal = {Cells}, number = {4}, issn = {2073-4409}, doi = {10.3390/cells10040884}, url = {http://nbn-resolving.de/urn:nbn:de:bvb:20-opus-236594}, year = {2021}, abstract = {Multiple sclerosis (MS) is a chronic inflammatory and neurodegenerative disease of the central nervous system (CNS) affecting more than two million people worldwide. In MS, oligodendrocytes and myelin sheaths are destroyed by autoimmune-mediated inflammation, while remyelination is impaired. Recent investigations of post-mortem tissue suggest that Fibroblast growth factor (FGF) signaling may regulate inflammation and myelination in MS. FGF2 expression seems to correlate positively with macrophages/microglia and negatively with myelination; FGF1 was suggested to promote remyelination. In myelin oligodendrocyte glycoprotein (MOG)\(_{35-55}\)-induced experimental autoimmune encephalomyelitis (EAE), systemic deletion of FGF2 suggested that FGF2 may promote remyelination. Specific deletion of FGF receptors (FGFRs) in oligodendrocytes in this EAE model resulted in a decrease of lymphocyte and macrophage/microglia infiltration as well as myelin and axon degeneration. These effects were mediated by ERK/Akt phosphorylation, a brain-derived neurotrophic factor, and downregulation of inhibitors of remyelination. In the first part of this review, the most important pharmacotherapeutic principles for MS will be illustrated, and then we will review recent advances made on FGF signaling in MS. Thus, we will suggest application of FGFR inhibitors, which are currently used in Phase II and III cancer trials, as a therapeutic option to reduce inflammation and induce remyelination in EAE and eventually MS.}, language = {en} } @article{KamaliRajendranStadelmannetal.2021, author = {Kamali, Salar and Rajendran, Ranjithkumar and Stadelmann, Christine and Karnati, Srikanth and Rajendran, Vinothkumar and Giraldo-Velasquez, Mario and Berghoff, Martin}, title = {Oligodendrocyte-specific deletion of FGFR2 ameliorates MOG\(_{35-55}\)-induced EAE through ERK and Akt signalling}, series = {Brain Pathology}, volume = {31}, journal = {Brain Pathology}, doi = {10.1111/bpa.12916}, url = {http://nbn-resolving.de/urn:nbn:de:bvb:20-opus-224354}, pages = {297 -- 311}, year = {2021}, abstract = {Fibroblast growth factors (FGFs) and their receptors (FGFRs) are involved in demyelinating pathologies including multiple sclerosis (MS). In our recent study, oligodendrocyte-specific deletion of FGFR1 resulted in a milder disease course, less inflammation, reduced myelin and axon damage in EAE. The objective of this study was to elucidate the role of oligodendroglial FGFR2 in MOG\(_{35-55}\)-induced EAE. Oligodendrocyte-specific knockout of FGFR2 (Fgfr2\(^{ind-/-}\)) was achieved by application of tamoxifen; EAE was induced using the MOG\(_{35-55}\) peptide. EAE symptoms were monitored over 62 days. Spinal cord tissue was analysed by histology, immunohistochemistry and western blot. Fgfr2\(^{ind-/-}\) mice revealed a milder disease course, less myelin damage and enhanced axonal density. The number of oligodendrocytes was not affected in demyelinated areas. However, protein expression of FGFR2, FGF2 and FGF9 was downregulated in Fgfr2\(^{ind-/-}\) mice. FGF/FGFR dependent signalling proteins were differentially regulated; pAkt was upregulated and pERK was downregulated in Fgfr2\(^{ind-/-}\) mice. The number of CD3(+) T cells, Mac3(+) cells and B220(+) B cells was less in demyelinated lesions of Fgfr2\(^{ind-/-}\) mice. Furthermore, expression of IL-1β, TNF-α and CD200 was less in Fgfr2\(^{ind-/-}\) mice than controls. Fgfr2ind-/- mice showed an upregulation of PLP and downregulation of the remyelination inhibitors SEMA3A and TGF-β expression. These data suggest that cell-specific deletion of FGFR2 in oligodendrocytes has anti-inflammatory and neuroprotective effects accompanied by changes in FGF/FGFR dependent signalling, inflammatory cytokines and expression of remyelination inhibitors. Thus, FGFRs in oligodendrocytes may represent potential targets for the treatment of inflammatory and demyelinating diseases including MS.}, language = {en} } @article{JanzZinkCirnuetal.2021, author = {Janz, Anna and Zink, Miriam and Cirnu, Alexandra and Hartleb, Annika and Albrecht, Christina and Rost, Simone and Klopocki, Eva and G{\"u}nther, Katharina and Edenhofer, Frank and Erg{\"u}n, S{\"u}leyman and Gerull, Brenda}, title = {CRISPR/Cas9-edited PKP2 knock-out (JMUi001-A-2) and DSG2 knock-out (JMUi001-A-3) iPSC lines as an isogenic human model system for arrhythmogenic cardiomyopathy (ACM)}, series = {Stem Cell Research}, volume = {53}, journal = {Stem Cell Research}, doi = {10.1016/j.scr.2021.102256}, url = {http://nbn-resolving.de/urn:nbn:de:bvb:20-opus-259846}, pages = {102256}, year = {2021}, abstract = {Arrhythmogenic cardiomyopathy (ACM) is characterized by fibro-fatty replacement of the myocardium, heart failure and life-threatening ventricular arrhythmias. Causal mutations were identified in genes encoding for proteins of the desmosomes, predominantly plakophilin-2 (PKP2) and desmoglein-2 (DSG2). We generated gene-edited knock-out iPSC lines for PKP2 (JMUi001-A-2) and DSG2 (JMUi001-A-3) using the CRISPR/Cas9 system in a healthy control iPSC background (JMUi001A). Stem cell-like morphology, robust expression of pluripotency markers, embryoid body formation and normal karyotypes confirmed the generation of high quality iPSCs to provide a novel isogenic human in vitro model system mimicking ACM when differentiated into cardiomyocytes.}, language = {en} } @article{UpcinHenkeKleefeldtetal.2021, author = {Upcin, Berin and Henke, Erik and Kleefeldt, Florian and Hoffmann, Helene and Rosenwald, Andreas and Irmak-Sav, Ster and Aktas, Huseyin Bertal and R{\"u}ckschloß, Uwe and Erg{\"u}n, S{\"u}leyman}, title = {Contribution of adventitia-derived stem and progenitor cells to new vessel formation in tumors}, series = {Cells}, volume = {10}, journal = {Cells}, number = {7}, doi = {10.3390/cells10071719}, url = {http://nbn-resolving.de/urn:nbn:de:bvb:20-opus-242577}, year = {2021}, abstract = {Blocking tumor vascularization has not yet come to fruition to the extent it was hoped for, as angiogenesis inhibitors have shown only partial success in the clinic. We hypothesized that under- appreciated vascular wall-resident stem and progenitor cells (VW-SPCs) might be involved in tumor vascularization and influence effectiveness of anti-angiogenic therapy. Indeed, in patient samples, we observed that vascular adventitia-resident CD34\(^+\) VW-SPCs are recruited to tumors in situ from co-opted vessels. To elucidate this in detail, we established an ex vivo model using concomitant embedding of multi-cellular tumor spheroids (MCTS) and mouse aortic rings (ARs) into collagen gels, similar to the so-called aortic ring assay (ARA). Moreover, ARA was modified by removing the ARs' adventitia that harbors VW-SPCs. Thus, this model enabled distinguishing the contribution of VW-SPCs from that of mature endothelial cells (ECs) to new vessel formation. Our results show that the formation of capillary-like sprouts is considerably delayed, and their number and network formation were significantly reduced by removing the adventitia. Substituting iPSC-derived neural spheroids for MCTS resulted in distinct sprouting patterns that were also strongly influenced by the presence or absence of VW-SPCs, also underlying the involvement of these cells in non-pathological vascularization. Our data suggest that more comprehensive approaches are needed in order to block all of the mechanisms contributing to tumor vascularization.}, language = {en} } @article{BielmeierRothSchmittetal.2021, author = {Bielmeier, Christina B. and Roth, Saskia and Schmitt, Sabrina I. and Boneva, Stefaniya K. and Schlecht, Anja and Vallon, Mario and Tamm, Ernst R. and Erg{\"u}n, S{\"u}leyman and Neueder, Andreas and Braunger, Barbara M.}, title = {Transcriptional profiling identifies upregulation of neuroprotective pathways in retinitis pigmentosa}, series = {International Journal of Molecular Sciences}, volume = {22}, journal = {International Journal of Molecular Sciences}, number = {12}, issn = {1422-0067}, doi = {10.3390/ijms22126307}, url = {http://nbn-resolving.de/urn:nbn:de:bvb:20-opus-260769}, year = {2021}, abstract = {Hereditary retinal degenerations like retinitis pigmentosa (RP) are among the leading causes of blindness in younger patients. To enable in vivo investigation of cellular and molecular mechanisms responsible for photoreceptor cell death and to allow testing of therapeutic strategies that could prevent retinal degeneration, animal models have been created. In this study, we deeply characterized the transcriptional profile of mice carrying the transgene rhodopsin V20G/P23H/P27L (VPP), which is a model for autosomal dominant RP. We examined the degree of photoreceptor degeneration and studied the impact of the VPP transgene-induced retinal degeneration on the transcriptome level of the retina using next generation RNA sequencing (RNASeq) analyses followed by weighted correlation network analysis (WGCNA). We furthermore identified cellular subpopulations responsible for some of the observed dysregulations using in situ hybridizations, immunofluorescence staining, and 3D reconstruction. Using RNASeq analysis, we identified 9256 dysregulated genes and six significantly associated gene modules in the subsequently performed WGCNA. Gene ontology enrichment showed, among others, dysregulation of genes involved in TGF-β regulated extracellular matrix organization, the (ocular) immune system/response, and cellular homeostasis. Moreover, heatmaps confirmed clustering of significantly dysregulated genes coding for components of the TGF-β, G-protein activated, and VEGF signaling pathway. 3D reconstructions of immunostained/in situ hybridized sections revealed retinal neurons and M{\"u}ller cells as the major cellular population expressing representative components of these signaling pathways. The predominant effect of VPP-induced photoreceptor degeneration pointed towards induction of neuroinflammation and the upregulation of neuroprotective pathways like TGF-β, G-protein activated, and VEGF signaling. Thus, modulation of these processes and signaling pathways might represent new therapeutic options to delay the degeneration of photoreceptors in diseases like RP.}, language = {en} } @phdthesis{Huebner2020, author = {H{\"u}bner, Lisa-Christina}, title = {Bedeutung von neuroendokrinen Zellen f{\"u}r die Signal{\"u}bertragung an sensorischen Nervenfasern in den Atemwegen}, doi = {10.25972/OPUS-20751}, url = {http://nbn-resolving.de/urn:nbn:de:bvb:20-opus-207517}, school = {Universit{\"a}t W{\"u}rzburg}, year = {2020}, abstract = {Die vorliegende Arbeit hatte zum Ziel, neuroendokrine Zellen in den Atemwegen bei M{\"a}usen zu untersuchen, welche Kontakt zu sensorischen Nervenfasern ausbilden. In vorangegangenen Versuchen konnte bereits die Menge des ausgesch{\"u}tteten CGRPs nach Stimulation mit Bitterstoffen bestimmt werden. Die Methode zur Messung der Freisetzung von CGRP aus verschiedenen Organen wurde von Prof. Reeh und seiner Arbeitsgruppe etabliert. Ziel der vorliegenden Arbeit war es, zu untersuchen, woher das ausgesch{\"u}ttete CGRP kommt und ob die Stimulation von B{\"u}rstenzellen mit Bitterstoffen zur Aussch{\"u}ttung von CGRP aus den neuroendokrinen Zellen f{\"u}hrt. Anhand der elektronenmikroskopischen Auswertung und der dreidimensionalen Rekonstruktion konnte gezeigt werden, dass es Kontakt zwischen den neuroendokrinen Zellen im Epithel der Trachea und sensorischen Nervenfasern gibt. Die immunhistochemischen Versuche zeigten, dass es nach Stimulation mit Denatonium h{\"o}chstwahrscheinlich zur Aussch{\"u}ttung von CGRP durch die intraepithelialen Fasern gekommen ist. Diese Annahme spiegelt sich in der ver{\"a}nderten Morphologie sowie der geringeren Quantit{\"a}t der intraepithelialen Fasern nach Stimulation mit Denatonium deutlich wider. Dass es weder bei der Anzahl der neuroendokrinen Zellen, noch bei der Erscheinung und Anzahl der extraepithelialen Fasern nach Denatoniumstimulation zu einer Ver{\"a}nderung gekommen ist, unterst{\"u}tzt diese Annahme ebenfalls. Im Hinblick auf die durchgef{\"u}hrten Versuche mit den TRPM5-gendefizienten M{\"a}usen zeigte sich, dass die Stimulation mit Denatonium keine Auswirkungen auf die Anzahl der neuroendokrinen Zellen hatte. Dieses Ergebnis unterst{\"u}tzt die Erkenntnisse der vorangegangenen Untersuchungen, welche gezeigt haben, dass das CGRP nicht von den neuroendokrinen Zellen ausgesch{\"u}ttet wurde. Des Weiteren l{\"a}sst das Ergebnis darauf schließen, dass die Aussch{\"u}ttung von CGRP nicht abh{\"a}ngig von der Anwesenheit von B{\"u}rstenzellen ist. Insgesamt zeigen die Untersuchungen, dass es nach Stimulation mit Bittersubstanzen zu einer CGRP-Aussch{\"u}ttung durch die intraepithelialen Fasern gekommen ist. Interessant w{\"a}re es weiterhin zu kl{\"a}ren, welche Effekte diese Aussch{\"u}ttung bewirkt und welche Bedeutung der Freisetzung von Substanz P in diesem Zusammenhang zukommt.}, subject = {Luftr{\"o}hre}, language = {de} } @phdthesis{Eckner2020, author = {Eckner, Georg Ludwig}, title = {Der Einfluss vaskul{\"a}r-adventitieller Makrophagen auf Gef{\"a}ßwand-residente Stammzellen}, doi = {10.25972/OPUS-20608}, url = {http://nbn-resolving.de/urn:nbn:de:bvb:20-opus-206080}, school = {Universit{\"a}t W{\"u}rzburg}, year = {2020}, abstract = {In dieser Dissertation wurden murine Aorta-Explantate im experimentellen Ansatz des "Aortic Ring Assay" {\"u}ber elf Tage kultiviert, erstmalig die Differenzierung zu F4/80(+)-Makrophagen gezeigt und eine nahezu vollst{\"a}ndige Depletion dieser Zellen durch Clodronat-Liposomen bewirkt. Diese Adventitia-generierten Makrophagen wurden als die Hauptquelle des lokal gebildeten VEGF nachgewiesen. Die daraus resultierende Depletion des parakrin wirkenden VEGF resultierte in einer teilweisen Konservierung der CD34(+) „Vaskulogenen Zone" der aortalen Adventitia. Zu der Best{\"a}tigung dieser Ergebnisse wurde experimentell {\"u}ber den VEGF-Rezeptor-Blocker E7080 in das VEGF-VEGFR-2 System eingegriffen. Die Versuchsans{\"a}tze mit diesem Rezeptorblocker resultierten in einem {\"a}hnlichen Ergebnis wie die Versuche unter Makrophagen-Depletion.}, subject = {Gef{\"a}ßwand}, language = {de} } @phdthesis{Burow2020, author = {Burow, Wera Tamara}, title = {Die Rolle des CEACAM1-Molek{\"u}ls bei der Entstehung von neurogener Entz{\"u}ndung in den Atemwegen}, doi = {10.25972/OPUS-20933}, url = {http://nbn-resolving.de/urn:nbn:de:bvb:20-opus-209331}, school = {Universit{\"a}t W{\"u}rzburg}, year = {2020}, abstract = {Neurogene Entz{\"u}ndung ist charakterisiert durch Vasodilatation, Plasmaextravasation und Leukozytenmigration. Im Zuge dieser Dissertationsarbeit konnte ein in vivo Versuchsmodell zur Quantifizierung neurogener Entz{\"u}ndungsreaktionen in den Atemwegen etabliert werden. Der bakterielle Bitterstoff Cycloheximid ist in der Lage, eine Erh{\"o}hung der Plasmaextravasation und Migration neutrophiler Granulozyten zu bewirken. Somit kann Cycloheximid nicht nur protektive Schutzreflexe ausl{\"o}sen, sondern f{\"u}hrt auch lokal zu einer neurogenen Entz{\"u}ndungsreaktion. Das carcinoembryonic antigen-related cell adhesion molecule-1 (CEACAM1) ist an der Regulierung der endothelialen Barrierefunktion beteiligt. Die Versuche zeigen bei CC1-/--M{\"a}usen eine Verminderung der basalen Permeabilit{\"a}t in trachealen postkapill{\"a}ren Venolen. Nach Stimulation mit Cycloheximid zeigen CC1-/--M{\"a}use im Vergleich mit WT-M{\"a}usen eine verminderte Plasmaextravasation in bronchialen postkapill{\"a}ren Venolen. Auch die Permeabilit{\"a}t des Endothels f{\"u}r neutrophile Granulozyten scheint durch CEACAM1-Defizienz in trachealen und bronchialen Venolen herabgesetzt zu werden. Die Anwesenheit des CEACAM1-Molek{\"u}ls verursacht offenbar eine verminderte Stabilit{\"a}t der endothelialen Barriere in postkapill{\"a}ren Venolen der Atemwege. Diese Ergebnisse zeigen eine gegenteilige Funktion von CEACAM1 in postkapill{\"a}ren Venolen der Atemwege im Vergleich mit großen, herznahen Blutgef{\"a}ßen. Des Weiteren scheint sich die Rolle von CEACAM1 in der Entstehung von akuten und chronischen Entz{\"u}ndungsreaktionen zu unterscheiden. Das in dieser Arbeit etablierte Versuchsmodell stellt eine M{\"o}glichkeit dar, neurogene Entz{\"u}ndungsreaktionen als Reaktion auf verschiedene gustatorische Stimulanzien zu testen und zu quantifizieren.}, subject = {Entz{\"u}ndung}, language = {de} } @article{Gottschlich2020, author = {Gottschlich, G{\"u}nter}, title = {Synopse der f{\"u}r Deutschland nachgewiesenen Arten und Unterarten der Gattung Hieracium s. l. (Hieracium s. str. und Pilosella), aufgeschl{\"u}sselt nach Vorkommen in den einzelnen Bundesl{\"a}ndern}, series = {Forum geobotanicum}, volume = {9}, journal = {Forum geobotanicum}, issn = {1867-9315}, doi = {10.3264/FG.2020.0114}, url = {http://nbn-resolving.de/urn:nbn:de:bvb:20-opus-198175}, pages = {1-59}, year = {2020}, abstract = {Eine Liste der 205 Arten und 1561 Unterarten der Gattung Hieracium s. l. , die in Deutschland, aufgeschl{\"u}sselt nach Bundesl{\"a}ndern vorkommen, wird vorgestellt. Da die meisten infraspezifischen Namen unter Hieracium publiziert wurden und um die Zahl der invaliden Namen unter Pilosella in der Liste zu minimieren, wird auf eine Aufteilung in Hieracium und Pilosella verzichtet. Durch Farbmarkierungen wird gekennzeichnet, welche Unterart urspr{\"u}nglich aus einem Bundesland beschrieben wurde bzw. ob ein Syntypus aus einem Bundesland stammt.}, subject = {Habichtskraut}, language = {de} } @article{Dunkel2020, author = {Dunkel, Franz G.}, title = {Ranunculus sarntheinianus Dunkel, spec. nova, eine neue Art aus dem Ranunculus-auricomus-Komplex - seit 135 Jahren im Oberen Inntal bei Innsbruck}, series = {Forum geobotanicum}, volume = {9}, journal = {Forum geobotanicum}, issn = {1867-9315}, doi = {10.3264/FG.2020.0115}, url = {http://nbn-resolving.de/urn:nbn:de:bvb:20-opus-198183}, pages = {60-65}, year = {2020}, abstract = {F{\"u}r das Vorkommen einer Ranunculus auricomus-Sippe im Oberen Inntal bei Innnsbuck, Nordtirol, {\"O}sterreich, existieren belegte Nachweise seit 135 Jahren. Diese Sippe sammelte L. Sarnthein am 08.06.1884, J. Murr 1887 bei Flaurling. Von beiden Aufsammlungen befinden sich Belege im Tiroler Landesmuseum (Herbarium Ferdinandeum Innsbruck; IBF). Trotz Trockenlegung der Sumpfwiesen und Eutrophierung des Biotops kommt die Art noch aktuell in einem kleinen Nasswiesenrest vor. Sie wird hier als R. sarntheinianus Dunkel beschrieben, abgebildet und ihre Taxonomie wird diskutiert. Der mutmaßlich letzte Wuchsort ist vom Aussterben bedroht. Eine graphische Darstellung soll bei Artbestimmung und Auffinden neuer Wuchsorte behilflich sein.}, subject = {Hahnenfuß}, language = {de} } @article{DrenckhahnWeber2020, author = {Drenckhahn, Detlev and Weber, Heinrich E.}, title = {Die Nordfriesische Brombeere, Rubus boreofrisicus Drenckhahn \& H. E. Weber, eine endemische Rubus-Art der Westk{\"u}ste von Schleswig-Holstein, Deutschland}, series = {Forum geobotanicum}, volume = {9}, journal = {Forum geobotanicum}, issn = {1867-9315}, doi = {10.3264/FG.2020.0116}, url = {http://nbn-resolving.de/urn:nbn:de:bvb:20-opus-198194}, pages = {66-69}, year = {2020}, abstract = {Rubus boreofrisicus Drenckhahn \& H. E.Weber ist eine bisher unbeschriebene Rubus-Art der Untergattung Rubus, Serie Discolores. Die Endbl{\"a}ttchen der 5-z{\"a}hligen, handf{\"o}rmigen Sch{\"o}sslingsbl{\"a}tter sind breit eif{\"o}rmig bis ann{\"a}hernd rund, unterseits grau-weißlich, oberseits dunkelgr{\"u}n und schwach behaart. Die Sch{\"o}sslinge sind braunrot, kantig bis schwach gefurcht, etwas behaart mit geraden bis schwach gekr{\"u}mmten, 5-8 mm langen, braunroten Stacheln mit heller Spitzenh{\"a}lfte. Die Bl{\"u}tenstiele besitzen Stieldr{\"u}sen. Rubus boreofrisicus kommt h{\"a}ufig im Waldg{\"u}rtel und in der angrenzenden D{\"u}nenheide der nordfriesischen Insel Amrum vor und ist auch 50 km s{\"u}dlich von Amrum in St. Peter-Ording im Westen der Halbinsel Eiderstedt vertreten.}, subject = {Rubus}, language = {de} } @article{Gallo2020, author = {Gallo, Lorenzo}, title = {Nomenclatural adjustments and typifications in the genus Phedimus (Crassulaceae)}, series = {Forum Geobotanicum}, volume = {9}, journal = {Forum Geobotanicum}, issn = {1867-9315}, doi = {10.3264/FG.2020.0616}, url = {http://nbn-resolving.de/urn:nbn:de:bvb:20-opus-206347}, pages = {70-73}, year = {2020}, abstract = {This paper deals with the taxonomical position and the nomenclature of two taxa belonging to the genus Sedum (Crassulaceae), today treated as Phedimus, namely Sedum middendorffianum Maxim var. diffusum Praeger and Sedum oppositifolium Sims. The correct taxonomical application of names is based on the nomenclatural types designated here.}, subject = {Sedum oppositifolium}, language = {en} } @phdthesis{Kleefeldt2020, author = {Kleefeldt, Florian}, title = {Einfluss von CEACAM1 auf die endotheliale Funktion}, doi = {10.25972/OPUS-20172}, url = {http://nbn-resolving.de/urn:nbn:de:bvb:20-opus-201726}, school = {Universit{\"a}t W{\"u}rzburg}, year = {2020}, abstract = {Dem Endothel, welches die luminale Oberfl{\"a}che aller Blutgef{\"a}ße auskleidet, kommt eine wichtige Barrierefunktion zwischen Blut und Gewebe zu. Nur durch eine bedarfsgerechte Justierung dieser Barriere, die den Durchtritt von Molek{\"u}len und Zellen reguliert, kann die Gewebehom{\"o}ostase aufrechterhalten werden. Dabei ist das Endothel nicht nur passive Barriere, sondern auch an dieser dynamischen Regulation aktiv beteiligt. St{\"o}rungen oder Fehlregulationen dieser Prozesse f{\"u}hren zu Pathologien, z.B. Arteriosklerose. Es ist seit l{\"a}ngerem bekannt, dass Carcinoembryonic antigen-related cell adhesion molecule-1 (CEACAM1), ein Mitglied der Immunglobulin-Superfamilie, die Bildung und Morphogenese neuer Blutgef{\"a}ße beeinflusst. Die spontane Entwicklung kleiner Arteriosklerose-{\"a}hnlicher L{\"a}sionen in CEACAM1 knockout (Cc1-/-) M{\"a}usen zeigt, dass CEACAM1 auch f{\"u}r die Hom{\"o}ostase ausgereifter Blutgef{\"a}ße von Bedeutung ist. Ziel dieser Dissertationsarbeit war daher, den Einfluss von CEACAM1 auf wesentliche Aspekte der Endothelfunktion in Aorten in situ bzw. in Endothelzellkulturen in vitro zu analysieren. Es konnte zun{\"a}chst gezeigt werden, dass CEACAM1-defiziente Endothelzellen im Vergleich zu Wildtyp (WT) Endothelzellen eine rundlichere Zellmorphologie mit meanderf{\"o}rmigen Zellgrenzen und interzellul{\"a}ren L{\"u}cken aufweisen. Diese morphologischen Unterschiede stimmen mit Befunden in situ an Aorten von WT und Cc1-/- M{\"a}usen {\"u}berein. Weiterhin wurde eine Translokation der endothelialen NO-Synthase (eNOS) von der Zellmembran in den peri-nukle{\"a}ren Bereich bei CEACAM1-Defizienz festgestellt. Die erhobenen Daten bieten zwei m{\"o}gliche Erkl{\"a}rungen daf{\"u}r. Einerseits k{\"o}nnte CEACAM1 durch Interaktion mit eNOS als Membrananker fungieren. Daneben wiesen CEACAM1-defiziente Endothelzellen eine erh{\"o}hte Expression des Enzyms APT1 auf, welches eNOS depalmitoyliert. Die daraus resultierende, ebenfalls nachgewiesene geringere Palmitoylierung k{\"o}nnte auch zur verminderten Membran-lokalisation von eNOS beitragen. Zur endothelialen Funktion geh{\"o}rt, die Adh{\"a}sion von Blutzellen an die Gef{\"a}ßwand weitestgehend zu beschr{\"a}nken. CEACAM1-defiziente Endothelzellen zeigten im Vergleich zu WT Endothelzellen eine verst{\"a}rkte Adh{\"a}sivit{\"a}t gegen{\"u}ber murinen und humanen Monozyten. {\"A}hnliche Unterschiede wurden f{\"u}r Aortenexplantate aus WT und Cc1-/- M{\"a}usen festgestellt. Dies ist einerseits mit einer verst{\"a}rkten Expression des Zelladh{\"a}sionsmolek{\"u}ls ICAM-1 bei CEACAM1-Defizienz erkl{\"a}rbar. Dar{\"u}ber hinaus vermittelt die Glykokalyx anti-adh{\"a}sive Eigenschaften. Aus Vorbefunden war bekannt, dass die endotheliale Glykokalyx in der Aorta von Cc1-/- M{\"a}use reduziert ist. Im Rahmen dieser Arbeit konnte dies auf eine verst{\"a}rkte Expression der Glykokalyx-degradierenden Enzyme MMP9, Chondroitinase sowie Hyaluronidase-2 in Cc1-/- Endothelzellen zur{\"u}ckgef{\"u}hrt werden. Eine erh{\"o}hte Permeabilit{\"a}t stellt einen Indikator f{\"u}r ein dysfunktionales Endothel, eines der initialen Schritte in der Pathogenese der Arteriosklerose, dar. Zur Analyse der aortalen Permeabilit{\"a}t wurde ein modifizierter Miles-Assay etabliert. Unter Verwendung etablierter muriner Arteriosklerosemodelle konnte gezeigt werden, dass dieser Assay eine St{\"o}rung der vaskul{\"a}ren Permeabilit{\"a}t bereits vor Auftreten makroskopischer Ver{\"a}nderungen zuverl{\"a}ssig detektiert. Im Rahmen der folgenden Analysen an WT und Cc1-/- M{\"a}usen zeigte sich ein altersabh{\"a}ngiger Effekt von CEACAM1 auf die Gef{\"a}ßpermeabilit{\"a}t: Aorten von 3 Monate alten Cc1-/- M{\"a}use wiesen eine im Vergleich zum WT erh{\"o}hte Gef{\"a}ßpermeabilit{\"a}t auf, welche wahrscheinlich Folge einer verz{\"o}gerten Gef{\"a}ßreifung ist. Im Alter von 9 Monaten zeigte sich dagegen ein entgegengesetztes Bild. Dies wurde auf eine verst{\"a}rkte Expression des die Barriere sch{\"a}digenden Inflammationsmediators TNF-α in 9 Monate alten WT M{\"a}usen zur{\"u}ckgef{\"u}hrt. Außerdem modulierte CEACAM1 die TNF-α-vermittelte Lockerung der endothelialen Barriere, indem es die Phosphorylierung von Adherens Junction Proteinen beeinflusste. Basal stabilisierte CEACAM1 die endotheliale Barriere durch Hemmung der Phosphorylierung von Caveolin-1, welches Adherens Junctions destabilisiert. Unter Einfluss von TNF-α war CEACAM1 verst{\"a}rkt im Bereich von Adherens Junctions lokalisiert und rekrutierte dort Src-Kinase. Src-Kinase wiederum destabilisierte Adherens Junctions durch Phosphorylierung von β-Catenin, was in verst{\"a}rkter Gef{\"a}ßpermeabilit{\"a}t resultierte. Dagegen f{\"u}hrte TNF-α in CEACAM1-defizienten Endothelzellen zu einer Dephosphorylierung von Caveolin-1 und β-Catenin, wodurch Adherens Junctions und damit die endotheliale Barriere stabilisiert wurden. Diese CEACAM1-abh{\"a}ngige differenzielle Regulation der Stabilit{\"a}t von Adherens Junctions unter TNF-α tr{\"a}gt wahrscheinlich maßgeblich zu den Unterschieden der vaskul{\"a}ren Permeabilit{\"a}t in 3 bzw. 9 Monate alten WT und Cc1-/- M{\"a}usen bei. Zusammenfassend konnte im Rahmen dieser Arbeit nachgewiesen werden, dass CEACAM1 zentrale Funktionen des Endothels und hier{\"u}ber die Hom{\"o}ostase reifer Gef{\"a}ße beeinflusst. Da eine Expression von CEACAM1 auch in arteriosklerotischen Plaques nachgewiesen werden konnte, soll in weiteren Untersuchungen auch der Beitrag von CEACAM1 zur arteriosklerotischen Plaquebildung analysiert werden.}, subject = {Endothel}, language = {de} } @phdthesis{Muenster2020, author = {M{\"u}nster, Wienke}, title = {Die Rolle von Acetylcholin und cholinergem Signalling in der Angiogenese}, doi = {10.25972/OPUS-20740}, url = {http://nbn-resolving.de/urn:nbn:de:bvb:20-opus-207408}, school = {Universit{\"a}t W{\"u}rzburg}, year = {2020}, abstract = {Ziel dieser Arbeit war die Untersuchung des Einflusses von ACh und vom cholinergen Signalling in der Angiogenese. Neben der klassischen Rolle des AChs im neuronalen System, konnten es in den letzten Jahren auch in nicht-neuronal innerviertem Gewebe festgestellt werden. In dieser Arbeit konzentrierte man sich auf beobachtete cholinerge Zellen in der Aortenwand einer ChAT-eGFP-Maus und in davon ausgehenden neu gebildeten Gef{\"a}ßen. Nach Durchf{\"u}hrung des Aortenringassays nach Baker konnte zun{\"a}chst nachgewiesen werden, dass es sich bei ACh um einen Angiogenese-Stimulator handelt. Eine Carbacholkonzentration von 1 µM erwies sich als bester Angiogenesestimulator. Der Inhibitorversuch mit Lenvatinib, einem Hemmer des VEGFR, zeigte, dass die cholinerge Wirkung zur Aktivierung {\"a}hnlicher angiogeneseseinhibierenden Mechanismen wie bei einer VEGF-Stimulation kommt. Bei der zeitlichen Weiterverfolgung des Versuchs konnte man feststellen, dass sich die Gef{\"a}ßformation mit Dauer des Versuches {\"a}ndert. Bei der immunhistologischen Analyse der neu gebildeten Gef{\"a}ße wurde am ehesten eine {\"U}berlappung der ChAT-Zellen mit Zellen gefunden, die sich positiv f{\"u}r die Perizytenmarker NG2 und Desmin zeigten. Es ist anzunehmen, dass das ACh, exprimiert von einigen Perizyten, eine Leitschiene f{\"u}r das sich neu bildende Gef{\"a}ß bietet und zu einem gerichteten Wachstum f{\"u}hrt. Vor Kurzem wurde in der Wand erwachsener Blutgef{\"a}ße eine Nische f{\"u}r Stammzellen identifiziert. Um zu untersuchen ob die ChAT-Zellen dort ihren Ursprung haben, wurde mit Stammzellmarkern gef{\"a}rbt. Diese Arbeit hat mit ihren Ergebnissen eine signifikante Grundlage f{\"u}r weiterf{\"u}hrende Studien geliefert.}, subject = {Acetylcholin}, language = {de} } @phdthesis{Kwok2020, author = {Kwok, Chee Keong}, title = {Scaling up production of reprogrammed cells for biomedical applications}, doi = {10.25972/OPUS-19186}, url = {http://nbn-resolving.de/urn:nbn:de:bvb:20-opus-191865}, school = {Universit{\"a}t W{\"u}rzburg}, year = {2020}, abstract = {Induced pluripotent stem cells (iPSCs) have been recognised as a virtually unlimited source of stem cells that can be generated in a patient-specific manner. Due to these cells' potential to give rise to all differentiated cell types of the human body, they have been widely used to derive differentiated cells for drug screening and disease modelling purposes. iPSCs also garner much interest as they can potentially serve as a source for cell replacement therapy. Towards the realisation of these biomedical applications, this thesis aims to address challenges that are associated with scale-up, safety and biofabrication. Firstly, the manufacture of a high number of human iPSCs (hiPSCs) will require standardised procedures for scale-up and the development of a flexible bioprocessing method, since standard adherent hiPSC culture exhibits limited scalability and is labour-intensive. While the quantity of cells that are required for cell therapy depends largely on the tissue and defect that these replacing cells are meant to correct, an estimate of 1 × 10^9 has been suggested to be sufficient for several indications, including myocardial infarction and islet replacement for diabetes. Here, the development of an integrated, microcarrier-free workflow to transition standard adherent hiPSC culture (6-well plates) to scalable stirred suspension culture in bioreactors (1 L working volume, 2.4 L maximum working volume) is presented. The two-phase bioprocess lasts 14 days and generates hiPSC aggregates measuring 198 ± 58 μm in diameter on the harvesting day, yielding close to 2 × 10^9 cells. hiPSCs can be maintained in stirred suspension for at least 7 weeks with weekly passaging, while exhibiting pluripotency-associated markers TRA-1-60, TRA-1-81, SSEA-4, OCT4, and SOX2. These cells retain their ability to differentiate into cells of all the three germ layers in vitro, exemplified by cells positive for AFP, SMA, or TUBB3. Additionally, they maintain a stable karyotype and continue to respond to specification cues, demonstrated by directed differentiation into beating cardiomyocyte-like cells. Therefore, the aim of manufacturing high hiPSC quantities was met using a state-of-the-art scalable suspension bioreactor platform. Secondly, multipotent stem cells such as induced neural stem cells (iNSCs) may represent a safer source of renewable cells compared to pluripotent stem cells. However, pre-conditioning of stem cells prior to transplantation is a delicate issue to ensure not only proper function in the host but also safety. Here, iNSCs which are normally maintained in the presence of factors such as hLIF, CHIR99021, and SB431542 were cultured in basal medium for distinct periods of time. This wash-out procedure results in lower proliferation while maintaining key neural stem cell marker PAX6, suggesting a transient pre-differentiated state. Such pre-treatment may aid transplantation studies to suppress tumourigenesis through transplanted cells, an approach that is being evaluated using a mouse model of experimental focal demyelination and autoimmune encephalomyelitis. Thirdly, biomedical applications of stem cells can benefit from recent advancements in biofabrication, where cells can be arranged in customisable topographical layouts. Employing a 3DDiscovery bioprinter, a bioink consisting of hiPSCs in gelatin-alginate was extruded into disc-shaped moulds or printed in a cross-hatch infill pattern and cross-linked with calcium ions. In both discs and printed patterns, hiPSCs recovered from these bioprints showed viability of around 70\% even after 4 days of culture when loaded into gelatin-alginate solution in aggregate form. They maintained pluripotency-associated markers TRA-1-60 and SSEA-4 and continued to proliferate after re-plating. As further proof-of-principle, printed hiPSC 3D constructs were subjected to targeted neuronal differentiation, developing typical neurite outgrowth and resulting in a widespread network of cells throughout and within the topology of the printed matrix. Staining against TUBB3 confirmed neuronal identity of the differentiated cellular progeny. In conclusion, these data demonstrate that hiPSCs not only survive the 3D-printing process but were able to differentiate along the printed topology in cellular networks.}, subject = {scale-up}, language = {en} } @article{OttoFriedrichMadunićetal.2020, author = {Otto, Christoph and Friedrich, Alexandra and Madunić, Ivana Vrhovac and Baumeier, Christian and Schwenk, Robert W. and Karaica, Dean and Germer, Christoph-Thomas and Sch{\"u}rmann, Annette and Sabolić, Ivan and Koepsell, Hermann, Hermann}, title = {Antidiabetic Effects of a Tripeptide That Decreases Abundance of Na\(^+\)-D-glucose Cotransporter SGLT1 in the Brush-Border Membrane of the Small Intestine}, series = {ACS Omega}, volume = {5}, journal = {ACS Omega}, number = {45}, doi = {10.1021/acsomega.0c03844}, url = {http://nbn-resolving.de/urn:nbn:de:bvb:20-opus-230654}, pages = {29127-29139}, year = {2020}, abstract = {In enterocytes, protein RS1 (RSC1A1) mediates an increase of glucose absorption after ingestion of glucose-rich food via upregulation of Na+-D-glucose cotransporter SGLT1 in the brush-border membrane (BBM). Whereas RS1 decelerates the exocytotic pathway of vesicles containing SGLT1 at low glucose levels between meals, RS1-mediated deceleration is relieved after ingestion of glucose-rich food. Regulation of SGLT1 is mediated by RS1 domain RS1-Reg, in which Gln-Ser-Pro (QSP) is effective. In contrast to QSP and RS1-Reg, Gln-Glu-Pro (QEP) and RS1-Reg with a serine to glutamate exchange in the QSP motif downregulate the abundance of SGLT1 in the BBM at high intracellular glucose concentrations by about 50\%. We investigated whether oral application of QEP improves diabetes in db/db mice and affects the induction of diabetes in New Zealand obese (NZO) mice under glucolipotoxic conditions. After 6-day administration of drinking water containing 5 mM QEP to db/db mice, fasting glucose was decreased, increase of blood glucose in the oral glucose tolerance test was blunted, and insulin sensitivity was increased. When QEP was added for several days to a high fat/high carbohydrate diet that induced diabetes in NZO mice, the increase of random plasma glucose was prevented, accompanied by lower plasma insulin levels. QEP is considered a lead compound for development of new antidiabetic drugs with more rapid cellular uptake. In contrast to SGLT1 inhibitors, QEP-based drugs may be applied in combination with insulin for the treatment of type 1 and type 2 diabetes, decreasing the required insulin amount, and thereby may reduce the risk of hypoglycemia.}, language = {en} } @misc{GottschlichDunkelDrenckhahnetal.2020, author = {Gottschlich, G{\"u}nter and Dunkel, Franz G. and Drenckhahn, Detlev and Weber, Heinrich E. and Gallo, Lorenzo}, title = {Forum Geobotanicum Vol. 9 (2020)}, volume = {9(2020)}, editor = {Meierott, Lenz and Drenckhahn, Detlev and Dunkel, Franz G. and Ewald, J{\"o}rg and Fleischmann, Andreas}, issn = {1867-9315}, url = {http://nbn-resolving.de/urn:nbn:de:bvb:20-opus-298919}, year = {2020}, abstract = {Forum Geobotanicum is an electronic journal devoted to disseminate information concerning geographical distribution, ecology, morphology, taxonomy and conservation of vascular plants in the European Union with a main focus on middle Europe. It covers from molecular biology to environmental aspects. The focus is to publish original papers, reviews and announcements for the educated generalist as well as the specialist in this broad field. Forum Geobotanicum does not aim to supplant existing paper journals, but will be much more flexible in format, publication time and world-wide distribution than paper journals. Many important studies are being currently published in local journals and booklets and some of them are published privately. Hence, these studies will become aware to only a limited readership. Forum Geobotanicum will encourage authors of such papers to submit them as special issues of the journal. Moreover, the journal is planning to build up an E-mail-address section to support communication between geobotanists in Europe. The editors are optimistic that this electronic journal will develop to a widely used communication forum that will help to stimulate activities in the entire field of geobotany in middle Europe. To overcome problems of long term archivation and effective taxonomic publication of articles published electronically in Forum Geobotanicum, print versions of each volume of the journal and appropriate digital storage devices will be delivered freely to selected university libraries and state libraries in middle Europe.}, subject = {Geobotanik}, language = {de} } @article{HenkeNandigamaErguen2020, author = {Henke, Erik and Nandigama, Rajender and Erg{\"u}n, S{\"u}leyman}, title = {Extracellular matrix in the tumor microenvironment and its impact on cancer therapy}, series = {Frontiers in Molecular Biosciences}, volume = {6}, journal = {Frontiers in Molecular Biosciences}, number = {160}, issn = {2296-889X}, doi = {10.3389/fmolb.2019.00160}, url = {http://nbn-resolving.de/urn:nbn:de:bvb:20-opus-199341}, year = {2020}, abstract = {Solid tumors are complex organ-like structures that consist not only of tumor cells but also of vasculature, extracellular matrix (ECM), stromal, and immune cells. Often, this tumor microenvironment (TME) comprises the larger part of the overall tumor mass. Like the other components of the TME, the ECM in solid tumors differs significantly from that in normal organs. Intratumoral signaling, transport mechanisms, metabolisms, oxygenation, and immunogenicity are strongly affected if not controlled by the ECM. Exerting this regulatory control, the ECM does not only influence malignancy and growth of the tumor but also its response toward therapy. Understanding the particularities of the ECM in solid tumor is necessary to develop approaches to interfere with its negative effect. In this review, we will also highlight the current understanding of the physical, cellular, and molecular mechanisms by which the pathological tumor ECM affects the efficiency of radio-, chemo-, and immunotherapy. Finally, we will discuss the various strategies to target and modify the tumor ECM and how they could be utilized to improve response to therapy.}, language = {en} } @article{HollenhorstJurastowNandigamaetal.2020, author = {Hollenhorst, Monika I. and Jurastow, Innokentij and Nandigama, Rajender and Appenzeller, Silke and Li, Lei and Vogel, J{\"o}rg and Wiederhold, Stephanie and Althaus, Mike and Empting, Martin and Altm{\"u}ller, Janine and Hirsch, Anna K. H. and Flockerzi, Veit and Canning, Brendan J. and Saliba, Antoine-Emmanuel and Krasteva-Christ, Gabriela}, title = {Tracheal brush cells release acetylcholine in response to bitter tastants for paracrine and autocrine signaling}, series = {The FASEB Journal}, volume = {34}, journal = {The FASEB Journal}, number = {1}, doi = {10.1096/fj.201901314RR}, url = {http://nbn-resolving.de/urn:nbn:de:bvb:20-opus-213516}, pages = {316 -- 332}, year = {2020}, abstract = {For protection from inhaled pathogens many strategies have evolved in the airways such as mucociliary clearance and cough. We have previously shown that protective respiratory reflexes to locally released bacterial bitter "taste" substances are most probably initiated by tracheal brush cells (BC). Our single-cell RNA-seq analysis of murine BC revealed high expression levels of cholinergic and bitter taste signaling transcripts (Tas2r108, Gnat3, Trpm5). We directly demonstrate the secretion of acetylcholine (ACh) from BC upon stimulation with the Tas2R agonist denatonium. Inhibition of the taste transduction cascade abolished the increase in [Ca\(^{2+}\)]\(_{i}\) in BC and subsequent ACh-release. ACh-release is regulated in an autocrine manner. While the muscarinic ACh-receptors M3R and M1R are activating, M2R is inhibitory. Paracrine effects of ACh released in response to denatonium included increased [Ca\(^{2+}\)]\(_{i}\) in ciliated cells. Stimulation by denatonium or with Pseudomonas quinolone signaling molecules led to an increase in mucociliary clearance in explanted tracheae that was Trpm5- and M3R-mediated. We show that ACh-release from BC via the bitter taste cascade leads to immediate paracrine protective responses that can be boosted in an autocrine manner. This mechanism represents the initial step for the activation of innate immune responses against pathogens in the airways.}, language = {en} } @article{GrunzWenigKunzetal.2020, author = {Grunz, Jan-Peter and Wenig, Andreas Max and Kunz, Andreas Steven and Veyhl-Wichmann, Maike and Schmitt, Rainer and Gietzen, Carsten Herbert and Pennig, Lenhard and Herz, Stefan and Erg{\"u}n, S{\"u}leyman and Bley, Thorsten Alexander and Gassenmaier, Tobias}, title = {3D cone-beam CT with a twin robotic x-ray system in elbow imaging: comparison of image quality to high-resolution multidetector CT}, series = {European Radiology Experimental}, volume = {4}, journal = {European Radiology Experimental}, doi = {10.1186/s41747-020-00177-y}, url = {http://nbn-resolving.de/urn:nbn:de:bvb:20-opus-229877}, year = {2020}, abstract = {Background Elbow imaging is challenging with conventional multidetector computed tomography (MDCT), while cone-beam CT (CBCT) provides superior options. We compared intra-individually CBCT versus MDCT image quality in cadaveric elbows. Methods A twin robotic x-ray system with new CBCT mode and a high-resolution clinical MDCT were compared in 16 cadaveric elbows. Both systems were operated with a dedicated low-dose (LD) protocol (equivalent volume CT dose index [CTDI\(_{vol(16 cm)}\)] = 3.3 mGy) and a regular clinical scan dose (RD) protocol (CTDI\(_{vol(16 cm)}\) = 13.8 mGy). Image quality was evaluated by two radiologists (R1 and R2) on a seven-point Likert scale, and estimation of signal intensity in cancellous bone was conducted. Wilcoxon signed-rank tests and intraclass correlation coefficient (ICC) statistics were used. Results The CBCT prototype provided superior subjective image quality compared to MDCT scans (for RD, p ≤ 0.004; for LD, p ≤ 0.001). Image quality was rated very good or excellent in 100\% of the cases by both readers for RD CBCT, 100\% (R1) and 93.8\% (R2) for LD CBCT, 62.6\% and 43.8\% for RD MDCT, and 0.0\% and 0.0\% for LD MDCT. Single-measure ICC was 0.95 (95\% confidence interval 0.91-0.97; p < 0.001). Software-based assessment supported subjective findings with less "undecided" pixels in CBCT than dose-equivalent MDCT (p < 0.001). No significant difference was found between LD CBCT and RD MDCT. Conclusions In cadaveric elbow studies, the tested cone-beam CT prototype delivered superior image quality compared to high-end multidetector CT and showed a potential for considerable dose reduction.}, language = {en} } @article{Koepsell2020, author = {Koepsell, Hermann}, title = {Glucose transporters in the small intestine in health and disease}, series = {Pfl{\"u}gers Archiv - European Journal of Physiology}, volume = {472}, journal = {Pfl{\"u}gers Archiv - European Journal of Physiology}, issn = {0031-6768}, doi = {10.1007/s00424-020-02439-5}, url = {http://nbn-resolving.de/urn:nbn:de:bvb:20-opus-232552}, pages = {1207-1248}, year = {2020}, abstract = {Absorption of monosaccharides is mainly mediated by Na\(^+\)-d-glucose cotransporter SGLT1 and the facititative transporters GLUT2 and GLUT5. SGLT1 and GLUT2 are relevant for absorption of d-glucose and d-galactose while GLUT5 is relevant for d-fructose absorption. SGLT1 and GLUT5 are constantly localized in the brush border membrane (BBM) of enterocytes, whereas GLUT2 is localized in the basolateral membrane (BLM) or the BBM plus BLM at low and high luminal d-glucose concentrations, respectively. At high luminal d-glucose, the abundance SGLT1 in the BBM is increased. Hence, d-glucose absorption at low luminal glucose is mediated via SGLT1 in the BBM and GLUT2 in the BLM whereas high-capacity d-glucose absorption at high luminal glucose is mediated by SGLT1 plus GLUT2 in the BBM and GLUT2 in the BLM. The review describes functions and regulations of SGLT1, GLUT2, and GLUT5 in the small intestine including diurnal variations and carbohydrate-dependent regulations. Also, the roles of SGLT1 and GLUT2 for secretion of enterohormones are discussed. Furthermore, diseases are described that are caused by malfunctions of small intestinal monosaccharide transporters, such as glucose-galactose malabsorption, Fanconi syndrome, and fructose intolerance. Moreover, it is reported how diabetes, small intestinal inflammation, parental nutrition, bariatric surgery, and metformin treatment affect expression of monosaccharide transporters in the small intestine. Finally, food components that decrease d-glucose absorption and drugs in development that inhibit or downregulate SGLT1 in the small intestine are compiled. Models for regulations and combined functions of glucose transporters, and for interplay between d-fructose transport and metabolism, are discussed.}, language = {en} } @article{Koepsell2020, author = {Koepsell, Hermann}, title = {Glucose transporters in brain in health and disease}, series = {Pfl{\"u}gers Archiv - European Journal of Physiology}, volume = {472}, journal = {Pfl{\"u}gers Archiv - European Journal of Physiology}, issn = {0031-6768}, doi = {10.1007/s00424-020-02441-x}, url = {http://nbn-resolving.de/urn:nbn:de:bvb:20-opus-232746}, pages = {1299-1343}, year = {2020}, abstract = {Energy demand of neurons in brain that is covered by glucose supply from the blood is ensured by glucose transporters incapillaries and brain cells. In brain, the facilitative diffusion glucose transporters GLUT1-6 and GLUT8, and the Na+-D-glucosecotransporters SGLT1 are expressed. The glucose transporters mediate uptake of D-glucose across the blood-brain barrier anddelivery of D-glucose to astrocytes and neurons. They are critically involved in regulatory adaptations to varying energy demandsin response to differing neuronal activities and glucose supply. In this review, a comprehensive overview about verified andproposed roles of cerebral glucose transporters during health and diseases is presented. Our current knowledge is mainly based onexperiments performed in rodents. First, the functional properties of human glucose transporters expressed in brain and theircerebral locations are described. Thereafter, proposed physiological functions of GLUT1, GLUT2, GLUT3, GLUT4, andSGLT1 for energy supply to neurons, glucose sensing, central regulation of glucohomeostasis, and feeding behavior are compiled, and their roles in learning and memory formation are discussed. In addition, diseases are described in which functionalchanges of cerebral glucose transporters are relevant. These are GLUT1 deficiency syndrome (GLUT1-SD), diabetes mellitus, Alzheimer's disease (AD), stroke, and traumatic brain injury (TBI). GLUT1-SD is caused by defect mutations in GLUT1. Diabetes and AD are associated with changed expression of glucose transporters in brain, and transporter-related energy defi-ciency of neurons may contribute to pathogenesis of AD. Stroke and TBI are associated with changes of glucose transporter expression that influence clinical outcome}, language = {en} } @article{CapetianMuellerVolkmannetal.2020, author = {Capetian, Philipp and M{\"u}ller, Lorenz and Volkmann, Jens and Heckmann, Manfred and Erg{\"u}n, S{\"u}leyman and Wagner, Nicole}, title = {Visualizing the synaptic and cellular ultrastructure in neurons differentiated from human induced neural stem cells - an optimized protocol}, series = {International Journal of Molecular Sciences}, volume = {21}, journal = {International Journal of Molecular Sciences}, number = {5}, issn = {1422-0067}, doi = {10.3390/ijms21051708}, url = {http://nbn-resolving.de/urn:nbn:de:bvb:20-opus-236053}, year = {2020}, abstract = {The size of the synaptic subcomponents falls below the limits of visible light microscopy. Despite new developments in advanced microscopy techniques, the resolution of transmission electron microscopy (TEM) remains unsurpassed. The requirements of tissue preservation are very high, and human post mortem material often does not offer adequate quality. However, new reprogramming techniques that generate human neurons in vitro provide samples that can easily fulfill these requirements. The objective of this study was to identify the culture technique with the best ultrastructural preservation in combination with the best embedding and contrasting technique for visualizing neuronal elements. Two induced neural stem cell lines derived from healthy control subjects underwent differentiation either adherent on glass coverslips, embedded in a droplet of highly concentrated Matrigel, or as a compact neurosphere. Afterward, they were fixed using a combination of glutaraldehyde (GA) and paraformaldehyde (PFA) followed by three approaches (standard stain, Ruthenium red stain, high contrast en-bloc stain) using different combinations of membrane enhancing and contrasting steps before ultrathin sectioning and imaging by TEM. The compact free-floating neurospheres exhibited the best ultrastructural preservation. High-contrast en-bloc stain offered particularly sharp staining of membrane structures and the highest quality visualization of neuronal structures. In conclusion, compact neurospheres growing under free-floating conditions in combination with a high contrast en-bloc staining protocol, offer the optimal preservation and contrast with a particular focus on visualizing membrane structures as required for analyzing synaptic structures.}, language = {en} } @article{IUedaWoersdoerferetal.2020, author = {I, Takashi and Ueda, Yuichiro and W{\"o}rsd{\"o}rfer, Philipp and Sumita, Yoshinori and Asahina, Izumi and Erg{\"u}n, S{\"u}leyman}, title = {Resident CD34-positive cells contribute to peri-endothelial cells and vascular morphogenesis in salivary gland after irradiation}, series = {Journal of Neural Transmission}, volume = {127}, journal = {Journal of Neural Transmission}, issn = {0300-9564}, doi = {10.1007/s00702-020-02256-1}, url = {http://nbn-resolving.de/urn:nbn:de:bvb:20-opus-235613}, pages = {1467-1479}, year = {2020}, abstract = {Salivary gland (SG) hypofunction is a common post-radiotherapy complication. Besides the parenchymal damage after irradiation (IR), there are also effects on mesenchymal stem cells (MSCs) which were shown to contribute to regeneration and repair of damaged tissues by differentiating into stromal cell types or releasing vesicles and soluble factors supporting the healing processes. However, there are no adequate reports about their roles during SG damage and regeneration so far. Using an irradiated SG mouse model, we performed certain immunostainings on tissue sections of submandibular glands at different time points after IR. Immunostaining for CD31 revealed that already one day after IR, vascular impairment was induced at the level of capillaries. In addition, the expression of CD44—a marker of acinar cells—diminished gradually after IR and, by 20 weeks, almost disappeared. In contrast, the number of CD34-positive cells significantly increased 4 weeks after IR and some of the CD34-positive cells were found to reside within the adventitia of arteries and veins. Laser confocal microscopic analyses revealed an accumulation of CD34-positive cells within the area of damaged capillaries where they were in close contact to the CD31-positive endothelial cells. At 4 weeks after IR, a fraction of the CD34-positive cells underwent differentiation into α-SMA-positive cells, which suggests that they may contribute to regeneration of smooth muscle cells and/or pericytes covering the small vessels from the outside. In conclusion, SG-resident CD34-positive cells represent a population of progenitors that could contribute to new vessel formation and/or remodeling of the pre-existing vessels after IR and thus, might be an important player during SG tissue healing.}, language = {en} } @article{WoersdoerferIAsahinaetal.2020, author = {W{\"o}rsd{\"o}rfer, Philipp and I, Takashi and Asahina, Izumi and Sumita, Yoshinori and Erg{\"u}n, S{\"u}leyman}, title = {Do not keep it simple: recent advances in the generation of complex organoids}, series = {Journal of Neural Transmission}, volume = {127}, journal = {Journal of Neural Transmission}, issn = {0300-9564}, doi = {10.1007/s00702-020-02198-8}, url = {http://nbn-resolving.de/urn:nbn:de:bvb:20-opus-235628}, pages = {1569-1577}, year = {2020}, abstract = {3D cell culture models which closely resemble real human tissues are of high interest for disease modelling, drug screening as well as a deeper understanding of human developmental biology. Such structures are termed organoids. Within the last years, several human organoid models were described. These are usually stem cell derived, arise by self-organization, mimic mechanisms of normal tissue development, show typical organ morphogenesis and recapitulate at least some organ specific functions. Many tissues have been reproduced in vitro such as gut, liver, lung, kidney and brain. The resulting entities can be either derived from an adult stem cell population, or generated from pluripotent stem cells using a specific differentiation protocol. However, many organoid models only recapitulate the organs parenchyma but are devoid of stromal components such as blood vessels, connective tissue and inflammatory cells. Recent studies show that the incorporation of endothelial and mesenchymal cells into organoids improved their maturation and might be required to create fully functional micro-tissues, which will allow deeper insights into human embryogenesis as well as disease development and progression. In this review article, we will summarize and discuss recent works trying to incorporate stromal components into organoids, with a special focus on neural organoid models.}, language = {en} } @phdthesis{Rossow2019, author = {Rossow, Leonie}, title = {Einfluss der Lysyloxidase-katalysierten Matrix-Quervernetzung auf Tumorwachstum, -metabolismus und -malignit{\"a}t}, doi = {10.25972/OPUS-18034}, url = {http://nbn-resolving.de/urn:nbn:de:bvb:20-opus-180346}, school = {Universit{\"a}t W{\"u}rzburg}, year = {2019}, abstract = {Die EZM bildet ein Netzwerk quervernetzter Proteine, welches alle Zellen im Tumor umgibt. Sie {\"u}bt direkte Effekte auf die Medikamenteneinbringung und -verteilung aus und somit auch auf die therapeutische Effizienz von Chemotherapeutika. Die LOX(L)-Proteinfamilie katalysiert die oxidative Desaminierung von Lysinresten in Elastin und Kollagenfasern und erm{\"o}glicht dadurch eine intra- und intermolekulare Quervernetzung. Diese wird f{\"u}r die Reifung und Stabilisierung der Kollagene in der EZM ben{\"o}tigt. Eine erh{\"o}hte LOX(L)-Expression steigert durch eine verst{\"a}rkte EZM-Quervernetzung die Gewebesteifheit im Tumor und bildet so eine physikalische Diffusionsbarriere. Durch diese Barriere wird die Versorgung mit Sauerstoff und N{\"a}hrstoffen reduziert. Die resultierende Hypoxie im Tumor kann eine fehlgeleitete Angiogenese triggern und zu einer Aktivierung maligner Signalkaskaden f{\"u}hren. In dieser Arbeit wurden durch eine LOX(L)-Inhibierung mittels βAPN einerseits und eine ektopische LOX-/LOXL2-{\"U}berexpression andererseits Auswirkungen solcher Eingriffe auf verschiedene Indikatoren wie Zellproliferation und apoptose, Versorgung mit Sauerstoff und N{\"a}hrstoffen, Angiogenese, Hypoxie, Makrophageninfiltration und die Expression verschiedener Wachstumsfaktoren analysiert. Die Versuche wurden an f{\"u}nf verschiedenen Tumoren (4T1-, E0771- und EMT6-Brustkarzinome, LLC-Lungenkarzinome und MT6-Fibrosarkome) durchgef{\"u}hrt. Die Ergebnisse dieser Arbeit demonstrieren eine direkte Verbindung zwischen der EZM und einer Therapieresistenz. Nach βAPN-Behandlung konnte eine verbesserte Versorgung mit Sauerstoff und N{\"a}hrstoffen beobachtet werden, welches in einer Verringerung maligner Signalkaskaden und folglich auch in einer verbesserten Vaskularisierung resultierte. Als Konsequenz wurde die therapeutische Effizienz von Chemotherapeutika verbessert. Im Gegensatz dazu f{\"u}hrte eine LOX-/LOXL2-{\"U}berexpression zu einer erh{\"o}hten Therapieresistenz. Die vorliegende Studie zeigt, dass die Modifizierung der EZM durch eine Hemmung von LOX(L) das Potenzial birgt, das Ansprechen von Chemotherapeutika in der Behandlung von Krebserkrankungen zu verbessern.}, subject = {Lysyl-Oxidase}, language = {de} } @misc{DrenckhahnDrenckhahnWeberetal.2019, author = {Drenckhahn, Detlev and Drenckhahn, Helga and Weber, Heinrich E. and Jansen, Werner and Weber, Heinrich E. and Zonnenveld, Ben J. M.}, title = {Forum Geobotanicum Vol. 8 (2018/2019)}, volume = {8(2018/2019)}, editor = {Meierott, Lenz and Drenckhahn, Detlev and Dunkel, Franz G. and Ewald, J{\"o}rg and Fleischmann, Andreas}, issn = {1867-9315}, url = {http://nbn-resolving.de/urn:nbn:de:bvb:20-opus-175292}, year = {2019}, abstract = {Forum Geobotanicum is an electronic journal devoted to disseminate information concerning geographical distribution, ecology, morphology, taxonomy and conservation of vascular plants in the European Union with a main focus on middle Europe. It covers from molecular biology to environmental aspects. The focus is to publish original papers, reviews and announcements for the educated generalist as well as the specialist in this broad field. Forum Geobotanicum does not aim to supplant existing paper journals, but will be much more flexible in format, publication time and world-wide distribution than paper journals. Many important studies are being currently published in local journals and booklets and some of them are published privately. Hence, these studies will become aware to only a limited readership. Forum Geobotanicum will encourage authors of such papers to submit them as special issues of the journal. Moreover, the journal is planning to build up an E-mail-address section to support communication between geobotanists in Europe. The editors are optimistic that this electronic journal will develop to a widely used communication forum that will help to stimulate activities in the entire field of geobotany in middle Europe. To overcome problems of long term archivation and effective taxonomic publication of articles published electronically in Forum Geobotanicum, print versions of each volume of the journal and appropriate digital storage devices will be delivered freely to selected university libraries and state libraries in middle Europe.}, subject = {Geobotanik}, language = {de} } @phdthesis{Wunsch2019, author = {Wunsch, Marie}, title = {Das enterische Nervensystem als m{\"o}gliche Zielstruktur der Autoimmunreaktion in der Multiplen Sklerose}, url = {http://nbn-resolving.de/urn:nbn:de:bvb:20-opus-175888}, school = {Universit{\"a}t W{\"u}rzburg}, year = {2019}, abstract = {Bei der Multiplen Sklerose (MS) handelt es sich um eine Autoimmunerkrankung des zentralen Nervensystems (ZNS). Abh{\"a}ngig von der betroffenen ZNS-Region kann es zu vielf{\"a}ltigen Symptomen kommen. Neben neurologischen Symptomen verursacht durch ZNS-L{\"a}sionen leidet ein Großteil der MS-Patienten auch unter gastrointestinalen Funktionsst{\"o}rungen. Diese gastrointestinalen Symptome wurden bisher eher auf L{\"a}sionen im R{\"u}ckenmark zur{\"u}ckgef{\"u}hrt und nicht direkt in Verbindung mit der autoimmunen {\"A}tiologie der Erkrankung gebracht. In dieser Studie wurde das enterische Nervensystem (ENS) in einem B-Zell- und Antik{\"o}rper-abh{\"a}ngigen Mausmodell der MS untersucht. Daf{\"u}r wurde der Autoimmunprozess durch Immunisierung mit MP4, einem Fusionsprotein aus dem Myelin-Basischen-Protein (MBP) und dem Proteolipid-Protein (PLP), ausgel{\"o}st. Das ZNS und ENS wurden in den unterschiedlichen Erkrankungsstadien immunhistochemisch und elektronenmikroskopisch analysiert. Neben der Immunpathologie des ZNS konnte dabei eine Degeneration des ENS schon vor dem Einsetzen der ersten neurologischen Defizite nachgewiesen werden. Die ENS-Pathologie war antik{\"o}rper-mediiert und ging einher mit einer verringerten gastrointestinalen Motilit{\"a}t sowie mit einer Gliose und Neurodegeneration des ENS. Mithilfe von Immunpr{\"a}zipitation und Massenspektrometrie konnten im ENS vier m{\"o}gliche Zielstrukturen des Autoimmunprozesses identifiziert werden, was auf sog. epitope spreading hindeutet. Auch im Plasma von MS-Patienten konnten Antik{\"o}rper gegen drei dieser Antigene nachgewiesen werden. Des Weiteren zeigten sich in Kolon-Resektaten von MS-Patienten erste Ans{\"a}tze einer Neurodegeneration und Gliose des ENS. In dieser Studie wurde zum ersten Mal ein direkter Zusammenhang zwischen der Autoimmunreaktion gegen das ZNS und einer simultanen Reaktion gegen das ENS gezeigt. Dies kann einen Paradigmenwechsel im Verst{\"a}ndnis der Immunpathogenese der MS anstoßen und neue therapeutische und diagnostische Ans{\"a}tze initiieren.}, subject = {Multiple Sklerose}, language = {de} } @phdthesis{Notz2019, author = {Notz, Quirin Julius}, title = {Der Einfluss von Fingolimod auf die autoreaktive B-Zell-Antwort in einem B-Zell-abh{\"a}ngigen Mausmodell der Multiplen Sklerose}, doi = {10.25972/OPUS-19154}, url = {http://nbn-resolving.de/urn:nbn:de:bvb:20-opus-191540}, school = {Universit{\"a}t W{\"u}rzburg}, year = {2019}, abstract = {Die MP4-induzierte experimentelle autoimmune Encephalomyelitis (EAE) erlaubt eine fokussierte Betrachtung von B-Zellen, die eine wichtige Rolle bei der Pathogenese der Multiplen Sklerose (MS) spielen. Es konnte zum Beispiel gezeigt werden, dass das Vorhandensein von B-Zell-Aggregaten im zentralen Nervensystem (ZNS) von MS-Patienten mit einem aggravierten Krankheitsverlauf assoziiert war. Diese Follikel k{\"o}nnten dabei als ektope lymphatische Strukturen den Immunprozess aktiv gestalten und somit ein therapeutisches Ziel darstellen. In der vorliegenden Studie wurde der Effekt des Sphingosin-1-Phosphat-Rezeptor-Modulators Fingolimod (FTY720) auf die autoreaktive B-Zell-Antwort und speziell die Bildung von B-Zell-Aggregaten im Kleinhirn der MP4-EAE-M{\"a}use untersucht.}, subject = {Multiple Sklerose}, language = {de} } @phdthesis{AlZuraiqi2019, author = {Al-Zuraiqi, Yaser}, title = {Die Rolle der residenten Stammzellen der Gef{\"a}ßwand bei der Bildung der Mikroglia und Angiogenese im adulten Gehirn}, doi = {10.25972/OPUS-18832}, url = {http://nbn-resolving.de/urn:nbn:de:bvb:20-opus-188329}, school = {Universit{\"a}t W{\"u}rzburg}, year = {2019}, abstract = {The role of vessel wall-resident stem cells in the generation of microglia and angiogenisis in the adult CNS Das Zentralnervensystem (ZNS) wird kontinuierlich durch ein eigenes Immunsystem {\"u}berwacht. Die Mikroglia sind ein wichtiger Vertreter dieses Immunsystems und ein besonderes Charakteristikum des ZNS. F{\"u}r die Aufrechterhaltung der H{\"a}mostase im ZNS spielen die Mikroglia eine zentrale Rolle. Die Herkunft der Mikroglia war f{\"u}r lange Zeit Gegenstand der kontroversen wissenschaftlichen Diskussion. Zusammengefasst wurde deren Ursprung als h{\"a}matopoetisch, mesodermal und neuroektodermal beschrieben. Allerdings {\"u}berwiegt derzeit die Meinung, dass die Mikroglia von Vorl{\"a}uferzellen geliefert wird, die w{\"a}hrend der Embryonalentwicklung aus der Dottersackwand ins Gehirn migrieren, dort bis zum Erwachsenenalter persistieren und immer wieder zur Erneuerung der Mikroglia herangezogen werden. Wo genau im Hirngewebe derartige oder andere potenzielle Mikrogliavorl{\"a}uferzellen im ZNS residieren, ist bis heute nicht abschließend gekl{\"a}rt. In der vorliegenden Arbeit konnte gezeigt werden, dass bereits die frisch pr{\"a}parierten Hirngef{\"a}ße sowohl CD44+ als auch CD45+ Zellen in ihren W{\"a}nden aufweisen. Außerdem ließ sich beobachten, dass die CD44+ Zellen im BRA nach außen wanderten und sich zu Perizyten-{\"a}hnlichen und glatten Muskelzellen differenzierten. Diese Befunde ließen darauf schließen, dass die CD44+ Zellen mit diesen Eigenschaften das Potenzial haben, zur Gef{\"a}ßneubildung beizutragen. Dar{\"u}ber hinaus konnten CD45+ Zellen in der Adventitia frisch isolierter Hirngef{\"a}ße nachgewiesen werden, die im BRA teilweise f{\"u}r F4/80 und/oder Iba-1 positiv wurden. Dies wiederum l{\"a}sst vermuten, dass aus der Wand der Hirngef{\"a}ße Mikroglia- und Makrophagen-{\"a}hnliche Zellen generiert werden k{\"o}nnen. Es blieb jedoch offen, ob diese CD45+ Vorl{\"a}uferzellen dauerhaft in der Adventitia der Hirngef{\"a}ße residieren oder aber immer wieder durch im Blut zirkulierende Monozyten erneuert werden. Diese Frage zu kl{\"a}ren, ist von klinischer Relevanz, bleibt jedoch zuk{\"u}nftigen Arbeiten {\"u}berlassen. Das hier etablierte BRA k{\"o}nnte auch bei solchen Analysen hilfreich sein.}, subject = {Mikroglia}, language = {de} } @phdthesis{Mekala2019, author = {Mekala, SubbaRao}, title = {Generation of cardiomyocytes from vessel wall-resident stem cells}, url = {http://nbn-resolving.de/urn:nbn:de:bvb:20-opus-146046}, school = {Universit{\"a}t W{\"u}rzburg}, year = {2019}, abstract = {Myocardial infarction (MI) is a major cause of health problems and is among the leading deadly ending diseases. Accordingly, regenerating functional myocardial tissue and/or cardiac repair by stem cells is one of the most desired aims worldwide. Indeed, the human heart serves as an ideal target for regenerative intervention, because the capacity of the adult myocardium to restore itself after injury or infarct is limited. Thus, identifying new sources of tissue resident adult stem or progenitor cells with cardiovascular potential would help to establish more sophisticated therapies in order to either prevent cardiac failure or to achieve a functional repair. Ongoing research worldwide in this field is focusing on a) induced pluripotent stem (iPS) cells, b) embryonic stem (ES) cells and c) adult stem cells (e. g. mesenchymal stem cells) as well as cardiac fibroblasts or myofibroblasts. However, thus far, these efforts did not result in therapeutic strategies that were transferable into the clinical management of MI and heart failure. Hence, identifying endogenous and more cardiac-related sources of stem cells capable of differentiating into mature cardiomyocytes would open promising new therapeutic opportunities. The working hypothesis of this thesis is that the vascular wall serves as a niche for cardiogenic stem cells. In recent years, various groups have identified different types of progenitors or mesenchymal stem cell-like cells in the adventitia and sub-endothelial zone of the adult vessel wall, the so called vessel wall-resident stem cells (VW-SCs). Considering the fact that heart muscle tissue contains blood vessels in very high density, the physiological relevance of VW-SCs for the myocardium can as yet only be assumed. The aim of the present work is to study whether a subset of VW-SCs might have the capacity to differentiate into cardiomyocyte-like cells. This assumption was challenged using adult mouse aorta-derived cells cultivated in different media and treated with selected factors. The presented results reveal the generation of spontaneously beating cardiomyocyte-like cells using specific media conditions without any genetic manipulation. The cells reproducibly started beating at culture days 8-10. Further analyses revealed that in contrast to several publications reporting the Sca-1+ cells as cardiac progenitors the Sca-1- fraction of aortic wall-derived VW-SCs reproducibly delivered beating cells in culture. Similar to mature cardiomyocytes the beating cells developed sarcomeric structures indicated by the typical cross striated staining pattern upon immunofluorescence analysis detecting α-sarcomeric actinin (α-SRA) and electron microscopic analysis. These analyses also showed the formation of sarcoplasmic reticulum which serves as calcium store. Correspondingly, the aortic wall-derived beating cardiomyocyte-like cells (Ao-bCMs) exhibited calcium oscillations. This differentiation seems to be dependent on an inflammatory microenvironment since depletion of VW-SC-derived macrophages by treatment with clodronate liposomes in vitro stopped the generation of Ao bCMs. These locally generated F4/80+ macrophages exhibit high levels of VEGF (vascular endothelial growth factor). To a great majority, VW-SCs were found to be positive for VEGFR-2 and blocking this receptor also stopped the generation VW-SC-derived beating cells in vitro. Furthermore, the treatment of aortic wall-derived cells with the ß-receptor agonist isoproterenol or the antagonist propranolol resulted in a significant increase or decrease of beating frequency. Finally, fluorescently labeled aortic wall-derived cells were implanted into the developing chick embryo heart field where they became positive for α-SRA two days after implantation. The current data strongly suggest that VW-SCs resident in the vascular adventitia deliver both progenitors for an inflammatory microenvironment and beating cells. The present study identifies that the Sca-1- rather than Sca-1+ fraction of mouse aortic wall-derived cells harbors VW-SCs differentiating into cardiomyocyte-like cells and reveals an essential role of VW-SCs-derived inflammatory macrophages and VEGF-signaling in this process. Furthermore, this study demonstrates the cardiogenic capacity of aortic VW-SCs in vivo using a chimeric chick embryonic model.}, subject = {Herzmuskelzelle}, language = {en} } @phdthesis{Groth2019, author = {Groth, Sofie Claire}, title = {Korrelation der Elastizit{\"a}t von R{\"u}ckenmarksgewebe und histologischen Ver{\"a}nderungen in einem Tiermodell der Multiplen Sklerose}, doi = {10.25972/OPUS-17937}, url = {http://nbn-resolving.de/urn:nbn:de:bvb:20-opus-179370}, school = {Universit{\"a}t W{\"u}rzburg}, year = {2019}, abstract = {Multiple Sklerose ist eine der h{\"a}ufigsten und bedeutsamsten entz{\"u}ndlichen Autoimmunerkrankungen bei jungen Erwachsenen. Obwohl die klassischen Kennzeichen der Krankheit wie Infiltration von Immunzellen, Demyelinisierung, Astrogliose und axonale Sch{\"a}digung bekannt sind, sind die genauen Ursachen und die zugrundeliegende Pathophysiologie noch nicht gekl{\"a}rt. In der Fachliteratur wurden bereits biomechanische Ver{\"a}nderungen mit histologischen Ver{\"a}nderungen im ZNS in Verbindung gebracht. Der genaue Zusammenhang und das Ausmaß zwischen den mechanischen Gewebeeigenschaften und den zugrundeliegenden histologischen Ver{\"a}nderungen wurde bis heute jedoch nur wenig erforscht. Die vorliegende Arbeit untersuchte in ihrem methodischen Rahmen den m{\"o}glichen Zusammenhang zwischen den mechanischen Ver{\"a}nderungen des Gewebes und den zugrundeliegenden histologischen Gewebever{\"a}nderungen in den unterschiedlichen Krankheitsstadien der EAE, dem Tiermodell der MS. Die hier dargestellten Experimente konnten demonstrieren, dass das ZNS-Gewebe durch zunehmende Zelldichte steifer wird, w{\"a}hrend es bei fortschreitender Demyelinisierung zur Erweichung des Gewebes kommt. Ferner wurden die mechanischen Gewebeeigenschaften in den unterschiedlichen Krankheitsstadien der EAE durch die Astrogliose und die Mikroglia/Makrophageninfiltration beeinflusst.}, subject = {Multiple Sklerose}, language = {de} } @article{Zonneveld2019, author = {Zonneveld, Ben J. M.}, title = {The DNA weights per nucleus (genome size) of more than 2350 species of the Flora of The Netherlands, of which 1370 are new to science, including the pattern of their DNA peaks}, series = {Forum Geobotanicum}, volume = {8}, journal = {Forum Geobotanicum}, issn = {1867-9315}, doi = {10.3264/FG.2019.1022}, url = {http://nbn-resolving.de/urn:nbn:de:bvb:20-opus-189724}, pages = {24-78}, year = {2019}, abstract = {Besides external characteristics and reading a piece of DNA (barcode), the DNA weight per nucleus (genome size) via flow cytometry is a key value to detect species and hybrids and determine ploidy. In addition, the DNA weight appears to be related to various properties, such as the size of the cell and the nucleus, the duration of mitosis and meiosis and the generation time. Sometimes it is even possible to distinguish between groups or sections, which can lead to new classification of the genera. The variation in DNA weight is also useful to analyze biodiversity, genome evolution and relationships between related taxa. Moreover, it is important to know how large a genome is before one determines the base sequence of the DNA of a plant. Flow cytometry is also important for understanding fundamental processes in plants such as growth and development and recognizing chimeras. In the literature, DNA weight measurements are usually limited to one genus and often only locally (Siljak et al. 2010; Bai et al. 2012). In this study, however, it was decided to investigate all vascular plants from one country. This can also contribute to the protection of rare plants. This study is the first flora in the world whose weight of DNA per nucleus and peak patterns has been determined. More than 6400 plants, representing more than 2350 (sub)species (more than 90\%) have been collected, thanks to the help of almost 100 volunteers of Floristisch Onderzoek Nederland (Floron). Multiple specimens of many species have therefore been measured, preferably from different populations, in some cases more than fifty. For 1370 species, these values were not previously published. Moreover, a good number of the remaining 45\% are new for The Netherlands. In principle, each species has a fixed weight of DNA per nucleus. It has also been found that, especially between the genera, there are strong differences in the number of peaks that determine the DNA weight, from one to five peaks. This indicates that in a plant or organ there are sometimes nuclei with multiples of its standard DNA weight (multiple ploidy levels). It is impossible to show graphs of more than 2350 species. Therefore, we have chosen to show the peak pattern in a new way in a short formula. Within most genera there are clear differences in the DNA weights per nucleus between the species, in some other genera the DNA weight is hardly variable. Based on about twenty genera that were previously measured completely in most cases ('t Hart et al. 2003: Veldkamp and Zonneveld 2011; Soes et al. 2012; Dirkse et al. 2014, 2015; Verloove et al. 2017; Zonneveld [et al.] 2000-2018), it can be noted that even if all species of a genus have the same number of chromosomes, there can still be a difference of up to three times in the weight of the DNA. Therefore, a twice larger DNA weight does not have to indicate four sets of chromosomes. Finally, this research has also found clues to examine further the current taxonomy of a number of species or genera.}, subject = {Pflanzen}, language = {en} } @article{KleefeldtBoemmelBroedeetal.2019, author = {Kleefeldt, Florian and B{\"o}mmel, Heike and Broede, Britta and Thomsen, Michael and Pfeiffer, Verena and W{\"o}rsd{\"o}rfer, Philipp and Karnati, Srikanth and Wagner, Nicole and Rueckschloss, Uwe and Erg{\"u}n, S{\"u}leyman}, title = {Aging-related carcinoembryonic antigen-related cell adhesion molecule 1 signaling promotes vascular dysfunction}, series = {Aging Cell}, volume = {2019}, journal = {Aging Cell}, number = {18}, doi = {10.1111/acel.13025}, url = {http://nbn-resolving.de/urn:nbn:de:bvb:20-opus-201231}, pages = {e13025}, year = {2019}, abstract = {Aging is an independent risk factor for cardiovascular diseases and therefore of particular interest for the prevention of cardiovascular events. However, the mechanisms underlying vascular aging are not well understood. Since carcinoembryonic antigen-related cell adhesion molecule 1 (CEACAM1) is crucially involved in vascular homeostasis, we sought to identify the role of CEACAM1 in vascular aging. Using human internal thoracic artery and murine aorta, we show that CEACAM1 is upregulated in the course of vascular aging. Further analyses demonstrated that TNF-α is CEACAM1-dependently upregulated in the aging vasculature. Vice versa, TNF-α induces CEACAM1 expression. This results in a feed-forward loop in the aging vasculature that maintains a chronic pro-inflammatory milieu. Furthermore, we demonstrate that age-associated vascular alterations, that is, increased oxidative stress and vascular fibrosis, due to increased medial collagen deposition crucially depend on the presence of CEACAM1. Additionally, age-dependent upregulation of vascular CEACAM1 expression contributes to endothelial barrier impairment, putatively via increased VEGF/VEGFR-2 signaling. Consequently, aging-related upregulation of vascular CEACAM1 expression results in endothelial dysfunction that may promote atherosclerotic plaque formation in the presence of additional risk factors. Our data suggest that CEACAM1 might represent an attractive target in order to delay physiological aging and therefore the transition to vascular disorders such as atherosclerosis.}, language = {en} } @article{WoersdoerferDaldaKernetal.2019, author = {W{\"o}rsd{\"o}rfer, Philipp and Dalda, Nahide and Kern, Anna and Kr{\"u}ger, Sarah and Wagner, Nicole and Kwok, Chee Keong and Henke, Erik and Erg{\"u}n, S{\"u}leyman}, title = {Generation of complex human organoid models including vascular networks by incorporation of mesodermal progenitor cells}, series = {Scientific Reports}, volume = {9}, journal = {Scientific Reports}, doi = {10.1038/s41598-019-52204-7}, url = {http://nbn-resolving.de/urn:nbn:de:bvb:20-opus-202681}, pages = {15663}, year = {2019}, abstract = {Organoids derived from human pluripotent stem cells are interesting models to study mechanisms of morphogenesis and promising platforms for disease modeling and drug screening. However, they mostly remain incomplete as they lack stroma, tissue resident immune cells and in particular vasculature, which create important niches during development and disease. We propose, that the directed incorporation of mesodermal progenitor cells (MPCs) into organoids will overcome the aforementioned limitations. In order to demonstrate the feasibility of the method, we generated complex human tumor as well as neural organoids. We show that the formed blood vessels display a hierarchic organization and mural cells are assembled into the vessel wall. Moreover, we demonstrate a typical blood vessel ultrastructure including endothelial cell-cell junctions, a basement membrane as well as luminal caveolae and microvesicles. We observe a high plasticity in the endothelial network, which expands, while the organoids grow and is responsive to anti-angiogenic compounds and pro-angiogenic conditions such as hypoxia. We show that vessels within tumor organoids connect to host vessels following transplantation. Remarkably, MPCs also deliver Iba1\(^+\) cells that infiltrate the neural tissue in a microglia-like manner.}, language = {en} }