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Item 10 Jahre Nationales Referenzzentrum für Hepatitis-B-Viren und Hepatitis-D-Viren in Gießen: Tätigkeiten und Erfahrungen(2022) Glebe, Dieter; Lehmann, Felix; Goldmann, Nora; Giese, Annika; Hida, Yassine; Gerlich, Wolfram H.; Ziebuhr, John; Slanina, Heiko; Schüttler, Christian G.Item 10 Jahre Screening auf Frühgeborenenretinopathie (2009–2019)(2023) Busik, Valentina; Lorenz, Birgit; Mais, Christine; Jäger, Melanie; Friedburg, Christoph; Andrassi-Darida, Monika; Ehrhardt, Harald; Hubert, MechthildHintergrund: Die Frühgeborenenretinopathie („retinopathy of prematurity“ [ROP]) zählt in Deutschland und weltweit zu den häufigsten Erblindungsursachen im Kindesalter; ein adäquates Screening ist essenziell. Der telemedizinische Ansatz mit objektiver Dokumentation des Netzhautbefundes eröffnet die Möglichkeit, untersucherunabhängig alle ROP-Stadien sicher zu diagnostizieren, wenn die Befundung durch ein ROP-geschultes Augenärzteteam erfolgt. Ziel der Arbeit: 10-Jahres-Vergleich des ROP-Screenings an 2 deutschen Level-1-Perinatalzentren: universitär und vor Ort vs. nichtuniversitär und telemedizinisch. Material und Methoden: Retrospektive Analyse der Screeninguntersuchungen stratifiziert nach Gestationsalter (GA), Geburtsgewicht (GG), Geschlecht, Mehrlingsgeburten, ROP-Zeitpunkt, Behandlungsbedürftigkeit, -zeitpunkt und Untersuchungshäufigkeit. Ergebnisse: Von 2009 bis 2019 wurden 1191 Frühgeborene gescreent, davon 29 Externe. Bei den verbleibenden 1162 wurden 3713 Netzhautuntersuchungen durchgeführt; 34 % hatten eine ROP, 5,4 % wurden behandelt (Gießen 3,7 %, Siegen 7,2 %). Mittleres GA war 28,9 SSW (± 2,5), mittleres GG 1155 g (± 417,5). Die Zahl der ROP-Diagnosen stieg in Gießen um 227,3 % und in Siegen um 111,1 % aufgrund zunehmender Zahlen der betreuten Kinder. Diskussion: Die vergleichende Analyse bestätigte national und international steigende ROP-Screening-Untersuchungen und Kinder mit akuter ROP. Das telemedizinische Screening war dem Vor-Ort-Screening gleichwertig und sicher. Mit beiden Screeningmethoden wurden alle Behandlungsbedürftigen zeitgerecht erkannt. Kein Kind mit einem GA > 29 SSW benötigte eine Behandlung, analog zu Ergebnissen des schwedischen Registers. Im deutschen ROP-Register waren aber einzelne Frühgeborene mit einem GA ≥ 30 SSW behandlungsbedürftig.Item 12-week combined strength and endurance exercise attenuates CD8+ T-cell differentiation and affects the kynurenine pathway in the elderly: a randomized controlled trial(2023) Boßlau, Tim Konstantin; Wasserfurth, Paulina; Reichel, Thomas; Weyh, Christopher; Palmowski, Jana; Nebl, Josefine; Joisten, Niklas; Belen, Sergen; Schenk, Alexander; Hahn, Andreas; Zimmer, Philipp; Krüger, KarstenBackground: Age-related accumulation of highly differentiated CD8+ effector memory re-expressing CD45RA (EMRA) T-cells and disruption of the kynurenine (KYN) pathway are associated with chronic inflammation and the development of insulin resistance. In this study the aim was to investigate the effects of 12-week combined strength and endurance exercise on CD8+ T-cell differentiation and KYN pathway metabolites. Ninety-six elderly subjects (f/m, aged 50—70) were randomized to a control (CON) or exercise (EX) group. The EX group completed combined strength and endurance training twice weekly for one hour each time at an intensity of 60% of the one-repetition maximum for strength exercises and a perceived exertion of 15/20 for endurance exercises. The EX group was also randomly subdivided into two groups with or without a concomitant balanced diet intervention in order to examine additional effects besides exercise alone. Before and after the intervention phase, the proportions of CD8+ T-cell subsets and levels of KYN pathway metabolites in peripheral blood were determined. Results: The CD8+ EMRA T-cell subsets increased in the CON group but remained almost unchanged in the EX group (p = .02). Plasma levels of kynurenic acid (KA) increased in the EX group and decreased in the CON group (p = .03). Concomitant nutritional intervention resulted in lower levels of quinolinic acid (QA) compared with exercise alone (p = .03). Overall, there was a slight increase in the QA/KA ratio in the CON group, whereas it decreased in the EX group (p > .05). Conclusions: Combined strength and endurance training seems to be a suitable approach to attenuate CD8+ T-cell differentiation in the elderly and to redirect the KYN pathway towards KA. The clinical relevance of these effects needs further investigation.Item 3D In Vitro Models: Novel Insights into Idiopathic Pulmonary Fibrosis Pathophysiology and Drug Screening(2022) Vazquez-Armendariz, Ana Ivonne; Barroso, Margarida Maria; El Agha, Elie; Herold, SusanneItem 3D-analysis of unwanted tooth movements despite bonded orthodontic retainers: a pilot study(2020) Klaus, Katharina; Xirouchaki, Faidra; Ruf, SabineItem 8p23.1 duplication syndrome: narrowing of critical interval to 1.80 Mbp(2014) Weber, Axel; Köhler, Angelika; Hahn, Andreas; Müller, UlrichBACKGROUND: A 3.68 Mbp duplication of 8p23.1 defines the 8p23.1 duplication syndrome. The main features of this syndrome are developmental delay and/or learning problems. RESULTS: Here we present a patient with a 1.80 Mbp duplication in 8p23.1 and characteristic signs and symptoms of the syndrome, including delay of motor and speech development and intellectual disability. DISCUSSION: The case indicates that genes within this interval, in particular dosage sensitive genes SOX7 and TNKS1, and possibly MIR124-1 and MIR598 as well suffice to cause the pathognomonic features of the 8p23.1 duplication syndrome.Item A 20:1 synergetic mixture of cafedrine\/theodrenaline accelerates particle transport velocity in murine tracheal epithelium via IP3 receptor-associated calcium release(2023) Schmidt, Götz; Rienas, Gerrit; Müller, Sabrina; Edinger, Fabian; Sander, Michael; Koch, Christian; Henrich, MichaelBackground: Mucociliary clearance is a pivotal physiological mechanism that protects the lung by ridding the lower airways of pollution and colonization by pathogens, thereby preventing infections. The fixed 20:1 combination of cafedrine and theodrenaline has been used to treat perioperative hypotension or hypotensive states due to emergency situations since the 1960s. Because mucociliary clearance is impaired during mechanical ventilation and critical illness, the present study aimed to evaluate the influence of cafedrine/theodrenaline on mucociliary clearance. Material and Methods: The particle transport velocity (PTV) of murine trachea preparations was measured as a surrogate for mucociliary clearance under the influence of cafedrine/theodrenaline, cafedrine alone, and theodrenaline alone. Inhibitory substances were applied to elucidate relevant signal transduction cascades. Results: All three applications of the combination of cafedrine/theodrenaline, cafedrine alone, or theodrenaline alone induced a sharp increase in PTV in a concentration-dependent manner with median effective concentrations of 0.46 µM (consisting of 9.6 µM cafedrine and 0.46 µM theodrenaline), 408 and 4 μM, respectively. The signal transduction cascades were similar for the effects of both cafedrine and theodrenaline at the murine respiratory epithelium. While PTV remained at its baseline value after non-selective inhibition of β-adrenergic receptors and selective inhibition of β1 receptors, cafedrine/theodrenaline, cafedrine alone, or theodrenaline alone increased PTV despite the inhibition of the protein kinase A. However, IP3 receptor activation was found to be the pivotal mechanism leading to the increase in murine PTV, which was abolished when IP3 receptors were inhibited. Depleting intracellular calcium stores with caffeine confirmed calcium as another crucial messenger altering the PTV after the application of cafedrine/theodrenaline. Discussion: Cafedrine/theodrenaline, cafedrine alone, and theodrenaline alone exert their effects via IP3 receptor-associated calcium release that is ultimately triggered by β1-adrenergic receptor stimulation. Synergistic effects at the β1-adrenergic receptor are highly relevant to alter the PTV of the respiratory epithelium at clinically relevant concentrations. Further investigations are needed to assess the value of cafedrine/theodrenaline-mediated alterations in mucociliary function in clinical practice.Item A comparative clinical study on the transfer accuracy of conventional and digital implant impressions using a new reference key-based method(2021) Schmidt, Alexander; Rein, Peter Engelbert; Wöstmann, Bernd; Schlenz, Maximiliane AmelieItem A comparative retrospective study on the prevalence and therapeutic treatment of dental agenesis between healthy children and children with systemic disease or congenital malformation(2023) Nadolinski, Mirja; Schlenz, Maximiliane Amelie; Rahman, Alexander; Krämer, Norbert; Schulz-Weidner, NellyBackground: Dental agenesis (DA) in the permanent dentition is one of the most common dental anomalies, with a prevalence up to 2–10%. Therefore, the aim of this retrospective study was to investigate the prevalence and therapeutic treatment of DA in healthy children (HC) compared to children with systemic disease or congenital malformation (SD/CM). Methods: Out of 3407 patients treated at the Department of Paediatric Dentistry of the Justus Liebig University Giessen (Germany) between January 2015 and December 2020, a total of 1067 patients (594 female, 473 male) aged between 4.5 and 18 years were included in this study due to DA. Besides the patients’ general medical history and therapeutic treatments, panoramic radiographs were analysed. Results: In contrast to the HC group with 9.7% DA, the SD/CM group showed a significantly higher prevalence of DA (19.8%; p < 0.05). The latter group was further classified into children with ectodermal dysplasia (4.4%), down syndrome (8.2%), cleft lip and palate (4.4%), intellectual disability/developmental delay (16.4%), and other genetic/organic diseases without intellectual disability (45.9%). Regarding therapeutic treatments, the HC group (59.5%) was significantly more often treated with an orthodontic gap opening compared to the SD/CM group (42.6%; p < 0.05), followed by orthodontic gap closing 36.5% in the HC group and 22.9% in the SD/CM group (p < 0.05), whereas no treatment was predominantly performed in the SD/CM group (37.7%) compared to the HC group (4%; p < 0.05). Furthermore, 50% in the SD/CM group required general anaesthesia for therapeutic treatment (vs. 8.1% in the HC group; p < 0.05). Conclusions: Children with SD/CM suffered more often from DA compared to HC that underlines multi- and interdisciplinary treatment of utmost importance. Furthermore, due to intellectual disability, common treatment methods can be complicated by insufficient compliance. This fact underlines the importance of an early attempt to establish the necessary cooperation enabling children with SD/CM to receive therapy.Item A comparative study on the lipidome of normal knee synovial fluid from humans and horses(2021) Kosinska, Marta Krystyna; Eichner, Gerrit; Schmitz, Gerd; Liebisch, Gerhard; Steinmeyer, JürgenItem A cross-sectional survey on knowledge and attitudes of Greek dentists regarding molar incisor hypomineralisation diagnosis and treatment(2022) Seremidi, Kyriaki; Amend, Stefanie; Krämer, Norbert; Gizani, SotiriaBackground: Molar Incisor Hypomineralisation (MIH) remains a challenge for clinicians underlining the gap in the literature regarding the condition. The study aimed to record knowledge and attitudes of Greek dentists regarding diagnosis and treatment of MIH and correlate findings with non-dental characteristics. Methods: It is a cross-sectional study based on a questionnaire consisting of 37 multiple-choice questions. Data regarding diagnosis, aetiopathogenesis, and clinical management of MIH were collected from active members of three Greek Dental Associations. Chi-square and student’s t-test were used to correlate responses with practitioners’ characteristics and odds ratios calculated to evaluate differences on treatment of MIH-affected teeth (p < 0.05). Results: From the 360 participants (response rate = 94%), 185 were general dental practitioners (GDPs) and 175 dental specialists (59 paediatric dentists (PDs), 38 orthodontists and 78 of other specialties).MIH was commonly encountered as a clinical problem, with GDPs reporting genetics and fluoride intake as common aetiological factors at significantly higher percentages as compared to PDs (p < 0.05). Permanent molars and incisors (44%) were the teeth most commonly affected, with yellow/brown demarcated opacities (68%) the most common clinical feature with PDs reporting them in a significantly higher percentage (p < 0.05). Dentists with specialisation, dentists that treat > 10 children per week and children with MIH-affected teeth had a 2–5.5 times greater probability to report difficulty achieving sufficient anaesthesia and hypersensitivity problems (p < 0.001). Agreement between GDPs’ and dental specialists’ views was found on less invasive treatment of anterior lesions. Non-PDs reported bulk-fill restorations and onlays as the ideal treatment of severely-affected posterior teeth, as compared to PDs that preferred preformed metal crowns (p < 0.05). Multivariate logistic regression analysis revealed that the age of the clinician, years of experience and number of children treated per week were the factors significantly associated with the decision for the treatment of only severely-affected posterior MIH teeth. Conclusions: Most participants had encountered MIH-affected teeth in their clinical practice and were able to recognize main aetiological factors and clinical findings related to the condition. Nevertheless, their knowledge regarding treatment is limited.Item A fixed 20:1 combination of cafedrine/theodrenaline increases cytosolic Ca2+ concentration in human tracheal epithelial cells via ryanodine receptor-mediated Ca2+ release(2023) Schmidt, Götz; Rienas, Gerrit; Müller, Sabrina; Richter, Katrin; Sander, Michael; Koch, Christian; Henrich, MichaelMucociliary clearance is a pivotal physiological mechanism that protects the lung by cleaning the airways from pollution and colonization, thereby preventing infection. Ciliary function is influenced by various signal transduction cascades, and Ca2+ represents a key second messenger. A fixed 20:1 combination of cafedrine and theodrenaline has been widely used to treat perioperative hypotension and emergency hypotensive states since the 1960s; however, its effect on the intracellular Ca2+ concentration ([Ca2+]i) of respiratory epithelium remains unknown. Therefore, human tracheal epithelial cells were exposed to the clinically applied 20:1 mixture of cafedrine/theodrenaline and the individual substances separately. [Ca2+]i was assessed by FURA-2 340/380 fluorescence ratio. Pharmacological inhibitors were applied to elucidate relevant signal transduction cascades, and reverse transcription polymerase chain reaction (RT-PCR) was performed on murine tracheal epithelium to analyze ryanodine receptor (RyR) subtype expression. All three pharmacological preparations instantaneously induced a steep increase in [Ca2+]i that quickly returned to its baseline value despite the persistence of each substance. Peak [Ca2+]i following the administration of 20:1 cafedrine/theodrenaline, cafedrine alone, and theodrenaline alone increased in a dose-dependent manner, with median effective concentrations of 0.35 mM (7.32 mM cafedrine and 0.35 mM theodrenaline), 3.14 mM, and 3.45 mM, respectively. When extracellular Ca2+ influx was inhibited using a Ca2+-free buffer solution, the peak [Ca2+]i following the administration of cafedrine alone and theodrenaline alone were reduced but not abolished. No alteration in [Ca2+]i compared with baseline [Ca2+]i was observed during β-adrenergic receptor inhibition. Depletion of caffeine-sensitive stores and inhibition of RyR, but not IP3 receptors, completely abolished any increase in [Ca2+]i. However, [Ca2+]i still increased following the depletion of mitochondrial Ca2+ stores using 2,4-dinitrophenol. RT-PCR revealed RyR-2 and RyR-3 expression on murine tracheal epithelium. Although our experiments showed that cafedrine/theodrenaline, cafedrine alone, or theodrenaline alone release Ca2+ from intracellular stores through mechanisms that are exclusively triggered by β-adrenergic receptor stimulation, which most probably lead to RyR activation, clinical plasma concentrations are considerably lower than those used in our experiments to elicit an increase in [Ca2+]i; therefore, further studies are needed to evaluate the ability of cafedrine/theodrenaline to alter mucociliary clearance in clinical practice.Item A Microfluidic System for Simultaneous Raman Spectroscopy, Patch-Clamp Electrophysiology, and Live-Cell Imaging to Study Key Cellular Events of Single Living Cells in Response to Acute Hypoxia(2021) Knoepp, Fenja; Wahl, Joel; Andersson, Anders; Kraut, Simone; Sommer, Natascha; Weissmann, Norbert; Ramser, KerstinItem A Nationwide Survey on Patient Empowerment in Pediatric Inflammatory Bowel Disease in Germany(2023) Kaul, Kalina; Schumann, Stefan; Sander, Cornelia; Däbritz, Jan; de Laffolie, JanBackground: The incidence/prevalence of pediatric inflammatory bowel disease (IBD) is increasing. IBD places a significant burden on young patients during important developmental stages and affects their families. Children and adolescents with IBD require increased support from health care services. However, little is known about the additional support needed and how to provide it. Therefore, a large survey was conducted with a focus on patient empowerment. Methods: For the anonymous survey, called CEDNA, a patient questionnaire for adolescents with IBD and a questionnaire for parents of children and adolescents with IBD were made available throughout Germany (distributed n = 2810). Questions covered various aspects of coping with the disease, utilization of care, use and need of information and communication services, and how information should be provided. Results: From October 2021 to April 2022, 1158 (n = 708 parents (61.1%), n = 450 patients (38.9%)) questionnaires were completed. The results show a deficit in pediatric IBD care and revealed a large gap in knowledge about core IBD topics (e.g., 74.8% of patients feel poorly informed about transition, 62.4% know little about patient organizations and self-help groups, and 54.7% have little information about preventive health measures), indicating a low level of health literacy among affected children and adolescents. Conclusion: Pediatric IBD poses significant challenges for patients, their families, and healthcare teams. By empowering patients and families, and providing targeted information and communication tailored to the age of the child or adolescent and the needs of the parents, care can be improved and better adapted to the needs of patients. Actions would include providing quality information online through scientific societies and patient organizations and facilitating self-management in adolescents.Item A New Type of Chronic Wound Infection after Wisdom Tooth Extraction: A Diagnostic Approach with 16S-rRNA Gene Analysis, Next-Generation Sequencing, and Bioinformatics(2020) Böttger, Sebastian; Zechel-Gran, Silke; Streckbein, Philipp; Knitschke, Michael; Hain, Torsten; Weigel, Markus; Wilbrand, Jan-Falco; Domann, Eugen; Howaldt, Hans-Peter; Attia, SamehItem A Novel Insertion in the Hepatitis B Virus Surface Protein Leading to Hyperglycosylation Causes Diagnostic and Immune Escape(2023) Lehmann, Felix; Slanina, Heiko; Roderfeld, Martin; Roeb, Elke; Trebicka, Jonel; Ziebuhr, John; Gerlich, Wolfram H.; Schüttler, Christian G.; Schlevogt, Bernhard; Glebe, DieterChronic hepatitis B virus (HBV) infection is a global health threat. Mutations in the surface antigen of HBV (HBsAg) may alter its antigenicity, infectivity, and transmissibility. A patient positive for HBV DNA and detectable but low-level HBsAg in parallel with anti-HBs suggested the presence of immune and/or diagnostic escape variants. To support this hypothesis, serumderived HBs gene sequences were amplified and cloned for sequencing, which revealed infection with exclusively non-wildtype HBV subgenotype (sgt) D3. Three distinct mutations in the antigenic loop of HBsAg that caused additional N-glycosylation were found in the variant sequences, including a previously undescribed six-nucleotide insertion. Cellular and secreted HBsAg was analyzed for N-glycosylation in Western blot after expression in human hepatoma cells. Secreted HBsAg was also subjected to four widely used, state-of-the-art diagnostic assays, which all failed to detect the hyperglycosylated insertion variant. Additionally, the recognition of mutant HBsAg by vaccine- and natural infection-induced anti-HBs antibodies was severely impaired. Taken together, these data suggest that the novel six-nucleotide insertion as well as two other previously described mutations causing hyperglycosylation in combination with immune escape mutations have a critical impact on in vitro diagnostics and likely increase the risk of breakthrough infection by evasion of vaccineinduced immunity.Item A novel model of a biomechanically induced osteoarthritis-like cartilage for pharmacological in vitro studies(2021) Sauerland, Katrin; Wolf, Amela; Schudok, Manfred; Steinmeyer, JürgenItem A Novel Model of Venovenous Extracorporeal Membrane Oxygenation in Rats with Femoral Cannulation and Insights into Hemodynamic Changes(2024) Edinger, Fabian; Zajonz, Thomas; Mayer, Nico; Schmidt, Götz; Schneck, Emmanuel; Sander, Michael; Koch, ChristianThe application of venovenous (VV) extracorporeal membrane oxygenation (ECMO) has gained wide acceptance for the treatment of acute severe respiratory failure. Since no rat model of VV ECMO therapy with femoral drainage has yet been described, although this cannulation strategy is commonly used in humans, this study aimed to establish such a model. Twenty male Lewis rats were randomly assigned to receive a sham procedure or VV ECMO therapy. After the inhalative induction of anesthesia, animals were intubated and the vascular accesses were placed surgically. While venous drainage was achieved through a modified multi-orifice 18 G cannula that was placed in the inferior vena cava through the femoral vein over a guide wire with an ultra-flexible tip, the venous return was realized via a shortened 20 G cannula into the jugular vein. Hemodynamic data were obtained from a tail artery and left ventricular pressure–volume catheter. Repetitive blood gas analyses were carried out, and systemic inflammation was measured using an enzyme-linked immunosorbent assay. While animals in the ECMO group showed adequate oxygenation and decarboxylation, there was no evidence of recirculation. VV ECMO therapy increased stroke volume (SV), cardiac output (CO), and left ventricular end-diastolic volume (LVEDV). ECMO-induced inflammation was reflected in increased levels of tumor necrosis factor alpha. However, no differences in interleukins 6 and 10 were seen. This study describes a frequently used cannulation strategy in humans for a rat model of VV ECMO. Despite successful oxygenation and decarboxylation, the oxygenated blood may reduce pulmonary vascular resistance and lead to an increased LVEDV, which is associated with increased SV and CO. This model allows us to answer research questions about topics such as intestinal microcirculation in further studies.Item A Novel Rat Model of Mild Pulmonary Hypertension Associated with Pulmonary Venous Congestion Induced by Left Pulmonary Vein Banding(2024) Münks, Jonas; Yogeswaran, Athiththan; Antoine, Tobiah Kevin; Blumrich, Leonhard Anton; Dorfmüller, Peter; Ghofrani, Hossein Ardeschir; Assmus, Birgit; Schermuly, Ralph Theo; Sydykov, AkylbekPulmonary hypertension (PH) associated with left heart disease (PH-LHD) is the most common form of PH. In PH-LHD, changes in the pulmonary vasculature are assumed to be mainly caused by pulmonary venous congestion. However, the underlying mechanisms of this form of PH are poorly understood. We aimed to establish a model of PH associated with pulmonary venous congestion. Wistar–Kyoto rats underwent partial occlusion of the left pulmonary vein to induce pulmonary venous congestion or sham surgery and were assessed at various time points post-surgery (3, 6, 9, 12 weeks). In vivo cardiopulmonary phenotyping was performed by using echocardiography along with heart catheterization. Histomorphometry methods were used to assess pulmonary vascular remodeling (e.g., wall thickness, degree of muscularization). Left pulmonary vein banding (PVB) resulted in mildly elevated right ventricular systolic pressure and moderate right ventricular hypertrophy. In PVB rats, small- and medium-sized pulmonary vessels in the left lung were characterized by increased wall thickness and muscularization. Taken together, our data demonstrate that left PVB-induced pulmonary venous congestion is associated with pulmonary vascular remodeling and mild PH.