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Abstract
The known Schiff base compound, (E)1‐benzyl‐3‐((4‐methoxyphenyl)imino)‐5‐methylindolin‐2‐one, was prepared as before by reacting 1‐benzyl‐5‐methylindoline‐2,3‐dione with 4‐methoxyaniline. The product was unambiguously characterized using elemental analysis, 1H and 13C‐NMR spectroscopy, and its new single‐crystal X‐ray structural analysis. Molecular orbital calculations were conducted in order to investigate the structures and relative stabilities of the (E) and (Z) isomers of 1‐benzyl‐3‐([4 methoxyphenyl]‐imino)‐5‐methylindolin‐2‐one. Specific attention was paid to the (E) isomer. The available crystallographic experimental data for the latter ensured also validation of the model structures computationally derived at the theoretical B3LYP/6‐31G(d,p) level.
Trotz der kontinuierlichen Weiterentwicklung der präventiven, diagnostischen und therapeutischen Maßnahmen stellt die koronare Herzkrankheit (KHK) und deren Folgen auch zukünftig eine große Herausforderung in der Medizin dar. Es besteht ein kritischer Bedarf für einen nicht-invasiven Biomarker, um Patienten mit dem größten Risiko für ein akutes pathologisches Ereignis bereits vor der klinischen Manifestation zu identifizieren. Ein solcher Biomarker wird auch für die schnelle Diagnosestellung bei symptomatisch auffälligen Patienten benötigt, bei denen die bisher etablierten Marker (Troponin, CK - Kreatinkinase) noch nicht nachweisbar sind. Als potenzieller Kandidat für einen solchen Biomarker könnten zirkulierende Endothelzellen (cEC) in Frage kommen. Obwohl die Rolle der cEC im akuten Myokardinfarkt (MI) noch nicht vollständig geklärt ist, legen die bisherigen Untersuchungen nahe, dass cEC eine differentialdiagnostische Möglichkeit für die Myokardinfarktpatienten in der so genannten ''Twighlight Zone'' - die Zeit zwischen ischämischem Ereignis und Nachweis von Troponin im Blut - sein können. Ziel dieser Untersuchung war daher, eine Einsicht in die komplexe methodische Quantifizierung der cEC und deren potentielle diagnostischen und prognostischen Rolle bei Patienten mit MI zu liefern.
In der vorliegenden Arbeit wurden 31 Patienten mit Myokardinfarkt eingeschlossen, 14 mit ST-Hebungsinfarkt (ST-elevation myocardial infarction/STEMI) und 17 mit nicht ST-Hebungsinfarkt (non ST-elevation myocardial infarction/NSTEMI). Als Kontrolle wurden 12 gesunden Probanden (Blutspender) ohne das Vorliegen einer KHK in der Anamnese gewonnen. Mittels Durchflusszytometrie wurden cEC der Patienten und der Kontrollgruppe bestimmt.
Die Ergebnisse zeigen einen Anstieg der cEC bei Infarktpatienten im Vergleich zur Kontrollgruppe. Dieser Unterschied war zwischen STEMI- und NSTEMI-Gruppe sowie STEMI- und Kontrollgruppe von statistischer Signifikanz. Des Weiteren wurde in Übereinstimmung mit anderen Untersuchungen keine
Korrelation zwischen cEC-Anzahl und den kardialen Enzymen (Troponin und CK) nachgewiesen. Bezüglich der prognostischen Bedeutung der cEC konnte in dieser Arbeit eine signifikante Korrelation der cEC mit dem GRACE-Score (in hospital risk of death and 6-month probability of death) bei den Patienten adjustiert für Alter, BMI und CRP nachgewiesen werden. Diese Studie gibt erste Hinweise darauf, dass cEC als additive Biomarker bei der Diagnose des Myokardinfarktes eingesetzt werden könnten, wobei weitere klinische Studien klären sollten, inwieweit cEC für diagnostische als auch prognostische Zwecke dienen könnten.
Humans are exposed to a plethora of microorganisms that reside on outer and inner body surfaces. These are collectively referred to as the human microbiome. The evolutionary relationship between humans and their microbiome is very complex. It is now widely accepted that these microorganisms are not just passive spectators but play an important role in health. The presence or absence of certain microbes is also linked to various diseases, including inflammatory bowel disease, cardiovascular disease, obesity, cancer, and allergies.
Allergies are several conditions caused by a misguided immune response to foreign antigens that are typically harmless. Common allergic diseases include atopic dermatitis (AD), allergic asthma, hay fever, and anaphylaxis. The incidences of allergic diseases are continuously rising, with up to 40% of the human population thought to be sensitised to environmental antigens. This increased incidence is not simply the result of societies becoming more aware and better at diagnosing these diseases. It is believed that the increases in allergies and sensitisation have environmental causes and are related to Western lifestyles. It is known that the rate of allergies is less frequent in developing countries. They are also more likely to occur in urban than rural areas. The prevailing view of the involvement of bacteria in allergies is described by the hygiene hypothesis. The hypothesis claims that decreased exposure to diverse microbial communities early in life increases the risk of developing allergic diseases. There are numerous examples to support this claim. For example, children born and raised in close contact to farm animals or in the presence of pets, and who are thus in direct and constant contact with a complex microbial environment, are protected from allergic diseases. On the other hand, colonisation or infection with certain bacteria increases allergic disease risks. This seems to contradict the hygiene hypothesis.
It appears that the members of the microbiome have different effects on allergy, and the hygiene hypothesis may not apply to every player in the complex microbial diversity that humans are in contact with. Therefore, a better understanding of the host bacterial interaction is required on the level of bacterial species.
This work studies the interplay between bacteria and the immune system to identify and characterise bacterial components with allergenic properties. In this quest, Staphylococcus aureus (S. aureus) and Staphylococcus epidermidis (S. epidermidis) were investigated for their allergenic properties and involvement in different allergic diseases. In the case of S. aureus, evidence is presented on allergic implications for two different components; serine protease-like proteins (Spls) and superantigens (SAg). Furthermore, experimental support is provided on the allergenic properties of the extracellular serine protease (Esp) from S. epidermidis. We argue that stimulating allergic reactions by staphylococci is an immune evasion mechanism that increases the survival chances of the bacteria within the host.
In chapter 1, an introduction is given to both S. aureus and S. epidermidis and their interactions with the immune system. Also, the bacterial components with allergenic properties and allergic diseases with known bacterial involvement are presented. Finally, the question of why bacteria cause allergy is discussed.
Chapter 2 describes allergic reactions to the Spls of S. aureus in a cohort of cystic fibrosis patients. Chapter 3 focuses on the SAgs of S. aureus. SAgs were discovered more than 30 years ago, but their physiological function is still under discussion. In this chapter, the allergenic properties of SAgs and their possible immunological mechanisms are reviewed, and a possible link between SAgs and allergic diseases is discussed. In chapter 4, the focus shifts to S. epidermidis and its involvement in AD. The human immune response to the Esp from S. epidermidis is characterised in healthy and AD individuals. The allergenic properties of Esp imply a detrimental role of S. epidermidis in AD. Finally, chapter 5 summarises and discusses the results of this thesis. In this section, the pieces are put together, and attention is brought back to the question of why bacteria cause allergies.
Abstract
We investigated four subaerial (paleo)lacustrine landforms at the north‐eastern shoreline of Schweriner See, north‐eastern Germany. These included two beach ridges, one subaerial nearshore bar and a silting up sequence located close to a fossil cliff, which marks the former maximum extent of Schweriner See. We used luminescence profiling with a SUERC portable OSL device (POSL) on all four sediment sequences in combination with sedimentological investigations such as grain size, loss‐on‐ignition and magnetic susceptibility to provide information on the various formations in a lacustrine depositional environment. The POSL reader was used on pre‐treated polymineral samples to gain an insight into luminescence distribution within the individual sediment sequences, but also among the four sequences. POSL proved valuable to understand depositional processes, which were not visible in lithology or sedimentological parameters. With somewhat larger uncertainty this method provides relative chronologies of the sediment sequences. Additionally, we carried out radiocarbon dating and full optical stimulated luminescence (OSL) dating to establish a chronological framework. OSL ages proved to be more reliable to date beach ridges in this setting than radiocarbon samples, which were severely influenced by sediment reworking. This combined approach of sedimentological analyses, luminescence profiling and absolute age determinations revealed details in depositional processes at Schweriner See which otherwise would have remained undetected. Furthermore, it helped to set these subaerial (paleo)lacustrine landforms in a chronological framework.
Modulation der biologischen Wirkung von MikroRNA-1 mittels chemischer Modifikation des RNA-Moleküls
(2021)
In vielen Malignitäten kann eine Dysregulation von miR beobachtet werden. Die reduzierte Expression dieser kleinen, nicht-codierenden RNA resultiert in verlängertem Tumorüberleben, einer gesteigerten Proliferationsrate, verbesserter Angiogenese und Metastasierung von Tumorzellen [1, 2]. Da miR-1 in ihrer Funktion als Tumorsuppressor mehrere onkogene Signal- und Effektorkaskaden supprimieren kann, ist die Erforschung ihres therapeutischen Potenzials von gesteigertem Interesse [1]. Dies zu realisieren eignen sich Prostatakarzinomzellen in besonderem Maße, da dort endogene miR-1 enorm herunterreguliert ist [157].
Die vorliegende Arbeit untersuchte die miR-1 Re-Expressionstherapie zur Hemmung der Tumorprogression auf molekularer Ebene an Prostatakarzinomzellen mit synthetischen miR-1 Molekülen. Darüber hinaus wurde die synthetische miR-1 durch unterschiedliche Substituenten modifiziert, um zu evaluieren ob diese Modifikationen Einfluss auf die biologische Wirkung auf die untersuchten Prostatakarzinomzelllinien haben. Ein weiterer Fokus lag dabei auf der Fragestellung, ob die Position und die chemischen Eigenschaften der Substituenten selbst einen Einfluss auf die miR-1 Effekte ausüben.
Es konnte gezeigt werden, dass die Wiederherstellung der intrazellulären miR-1 Spiegel durch die unmodifizierte, synthetische miR-1 bereits einen hemmenden Effekt auf das Tumorwachstum hatte. Ausserdem deuten die Ergebnisse darauf hin, dass die chemisch synthetisierte und modifizierte miR-1, der unveränderten, synthetischen miR-1 in ihrer tumorsupprimierenden Funktion überlegen sein kann.
Die antiproliferative Wirksamkeit der modifizierten miR-1 Moleküle hing jedoch eher von der Position, als von der Art des modifizierten Nukleotids ab. Es konnte gezeigt werden, dass, unabhängig vom Substituenten, Modifikationen an vermutlich kritischen Positionen sogar ein gesteigertes Tumorwachstum zur Folge hatten. Substituenten an ungünstigen Positionen konnten zu Reduktion der antiproliferativen Effekte von miR-1-Molekülen und somit sogar zu vermehrten Tumorwachstum führen. Dies bietet die Möglichkeit, durch die Auswahl gut charakterisierter miR mit hohem antiproliferativem Potenzial maßgeschneiderte, synthetisch modifizierte miR für die zukünftige, individualisierte Therapie verschiedener Krebsentitäten zu entwickeln.
Background: COVID-19 lead to the adoption of containment measures including temporary closure of dental clinics. Despite the risk of infection transmission, dental emergencies have not ceased during this pandemic and had to be managed also in the lockdown period.
Aim: To analyze the profiles and offered management options of pediatric patients presenting with dental emergencies during a COVID-19 lockdown.
Design: Retrospective analysis of patient records of children seeking emergency dental treatment during a 7-week lockdown period in 2020 in a university pedodontics clinic in Germany, compared to a similar cohort from 2019. Data on patient level, tooth level, and session level were collected. An analysis of the digital records after 6 months follow-up was performed for the patients who received Non-Aerosol Generating Procedures (NAGP) as management for dental emergency in the lockdown period in 2020.
Results: The 2020 cohort consisted of 83 patients, while the 2019 cohort included 46 patients showing 45% higher necessity for emergency treatment in 2020. Most common chief complaint was oral mucosal conditions in 2020 (26.4%), and irreversible pulpitis in 2019 (25.5%). Dental caries (without spontaneous pain) was the second most chief complaint in both cohorts (20.7% and 23.4% respectively). Most interventions in 2020 were Minimally Invasive Treatments such as the hall technique and silver diammine fluoride (20.3%), which were in 2019 not considered, followed by pharmacological treatment (16.9%), which were in 2019 also highly used (35.9%). The 6 months follow up for the NAGP revealed benefit in management of the acute dental problem, by either direct treatment or by postponing the treatment need to a later time period.
Conclusion: The COVID-19 pandemic led to increase in emergency pediatric dental visits and shifted treatment options towards less invasive procedures.
In challenging situations, where aerosols increase the risk of infection transmission, NAPD are a viable option in the management of dental emergencies, especially in pediatric dentistry.
Störungen im Calcium- und Phosphatstoffwechsel bei chronisch Nierenkranken führen zur sogenannten „Chronic Kidney Disease - Mineral and Bone Disorder“ (CKD-MBD). Die CKD- MBD umfasst neben laborchemischen Veränderungen und Knochenerkrankungen vor allem Gefäßverkalkungen. Letztere sind aktive vaskuläre Umbauprozesse und finden sich insbesondere in der Tunica media arterieller Gefäße, mit einer Assoziation zu einer erhöhten kardiovaskulären Mortalität.
Ziel dieser Arbeit war es, vertiefende Einblicke in den Prozess der vaskulären Kalzifikation zu erlangen. Hierbei sollte die Frage geklärt werden, über welche Signalwege das in urämischen Gefäßen überexprimierte TGF-ß1 möglicherweise zur Inhibition vaskulärer Kalzifikation beiträgt und ob eine Beeinflussung möglich ist.
Zur Klärung dieses Sachverhalts wurde ein in vitro Zellkulturmodell (human vascular smooth muscle cells, hVSMCs) durch Zusatz von erhöhtem Calcium- und Phosphat etabliert, ähnlich den laborchemischen Veränderungen bei CKD-MBD. Die kalzifikationsbeeinflussende Rolle von TGF-ß1 sollte geklärt und mit dem SAPK/JNK (stress- activated protein-kinase/ c-Jun N-terminale Kinase) Signalweg eine mögliche Interaktion dieser beiden Signalkaskaden identifiziert werden. Außerdem sollte der Stellenwert der Apoptose sowie der osteogenen Differenzierung im Prozess vaskulärer Kalzifikation analysiert werden.
TGF-ß1 zeigte eine konzentrationsabhängige, kalzifikationsinhibierende Wirkung. TGF-ß1 vermittelte diesen Effekt unter anderem über Phosphorylierung der Proteinkinase SAPK/JNK, die im Zusammenhang mit Apoptose, Zellproliferation und Differenzierungsprozessen steht. Weiterhin konnte bestätigt werden, dass die Apoptose eine wichtige Rolle bei der Initiation der vaskulären Kalzifikation spielt.
Die weiterführende Erforschung der Interaktion des SAPK/JNK Signalwegs mit TGF-ß1 und einem damit verbundenem, potentiellen Therapieansatz bei Patienten mit vaskulärer Kalzifikation sollte in vivo in einem Tiermodell untersucht werden, insbesondere um herauszufinden, ob der Signalweg ohne Beeinträchtigung wichtiger zellulärer Funktionen beeinflusst werden kann. Sowohl TGF-ß1 als auch SAPK/JNK könnten mögliche Targets für neue therapeutische Strategien wie die Entwicklung von sog. „biased ligands“ darstellen, welche selektiv bestimmte Signalwege regulieren können.
Streptococcus pneumoniae has evolved versatile strategies to colonize the nasopharynx of humans. Colonization is facilitated by direct interactions with host cell receptors or via binding to components of the extracellular matrix. In addition, pneumococci hijack host-derived extracellular proteases such as the serine protease plasmin(ogen) for ECM and mucus degradation as well as colonization. S. pneumoniae expresses strain-dependent up to four serine proteases. In this study, we assessed the role of secreted or cell-bound serine proteases HtrA, PrtA, SFP, and CbpG, in adherence assays and in a mouse colonization model. We hypothesized that the redundancy of serine proteases compensates for the deficiency of a single enzyme. Therefore, double and triple mutants were generated in serotype 19F strain EF3030 and serotype 4 strain TIGR4. Strain EF3030 produces only three serine proteases and lacks the SFP encoding gene. In adherence studies using Detroit-562 epithelial cells, we demonstrated that both TIGR4Δcps and 19F mutants without serine proteases or expressing only CbpG, HtrA, or PrtA have a reduced ability to adhere to Detroit-562 cells. Consistent with these results, we show that the mutants of strain 19F, which preferentially colonizes mice, abrogate nasopharyngeal colonization in CD-1 mice after intranasal infection. The bacterial load in the nasopharynx was monitored for 14 days. Importantly, mutants showed significantly lower bacterial numbers in the nasopharynx two days after infection. Similarly, we detected a significantly reduced pneumococcal colonization on days 3, 7, and 14 post-inoculations. To assess the impact of pneumococcal serine proteases on acute infection, we infected mice intranasally with bioluminescent and invasive TIGR4 or isogenic triple mutants expressing only CbpG, HtrA, PrtA, or SFP. We imaged the acute lung infection in real-time and determined the survival of the mice. The TIGR4lux mutant expressing only PrtA showed a significant attenuation and was less virulent in the acute pneumonia model. In conclusion, our results showed that pneumococcal serine proteases contributed significantly to pneumococcal colonization but played only a minor role in pneumonia and invasive diseases. Because colonization is a prerequisite for invasive diseases and transmission, these enzymes could be promising candidates for the development of antimicrobials to reduce pneumococcal transmission.