Die Bedeutung der oberen Atemwege bei zystischer Fibrose – Pathomechanismen, Monitoring und Therapie

Zusammenfassung der Habilitationsschrift von Dr. rer. nat. Julia Hentschel mit dem Titel: „Die Bedeutung der oberen Atemwege bei zystischer Fibrose – Pathomechanismen, Monitoring und Therapie“, Universität Leipzig Lange Jahre standen rezidivierende Infekte der Lunge und Gedeihstörung durch Maldigestion als lebensbegrenzende Symptome eines Patienten mit Mukoviszidose (Cystische Fibrose/CF) im Vordergrund. Das Hauptaugenmerk in der Diagnostik und Therapie von CF-Patienten lag daher zumeist auf den unteren Atemwegen bzw. dem Verdauungstrakt. Verbesserte Therapiemöglichkeiten und eine erhebliche Steigerung der Lebenserwartung brachten andere Symptome in den klinischen und wissenschaftlichen Fokus. Seit 2005 untersuchen wir systematisch die Rolle der oberen Atemwege bei Patienten mit CF, indem wir bei jeder Routinevorstellung Material der oberen Atemwege nicht-invasiv gewinnen, analysieren und asservieren. Wir konnten Methoden etablieren, um nasale Lavagen mit wenig Aufwand auch für große Studien einfach und schnell zu gewinnen und zu asservieren. Wir konnten zeigen, dass sich diese nasalen Lavagen eignen, um eine Besiedlung mit Pathogenen zu erfassen und die resultierende inflammatorische Wirtsantwort zu untersuchen. Wir konnten erstmals beweisen, dass die oberen Atemwege ein Keimreservoir darstellen und von dort aus Keime die Lunge (wieder) besiedeln können. Dies fand Eingang in aktuelle Leitlinien zur Mukoviszidosebetreuung und Lungentransplantation und hat Einfluss auf den Erfolg dieses ohnehin schon stark risikobehafteten Eingriffs. CF ist eine Erkrankung, bei der häufig eine überschießende Entzündungsreaktion beobachtet wird. Unsere Studien legten dabei den Schluss nahe, dass in den oberen Atemwegen andere Imbalancen eine Rolle spielen als in den unteren Atemwegen und sich daraus auch unterschiedliche therapeutische Ansätze ergeben könnten. Weiterhin konnten wir verdeutlichen, dass sich auch der Einfluss therapeutischer Interventionen in den Inflammationsmediatoren der oberen Atemwege widerspiegelt. Eine wichtige Erkenntnis daraus war, dass v.a. systemische Antibiosen deutlich langsamer und/oder schlechter in den Nasennebenhöhlen und den oberen Atemwegen wirken als auf die unteren und dass hier ein Umdenken in der antibiotischen Therapie der oberen Atemwege erfolgen muss. Unsere Studien beweisen einmal mehr das Kontinuum der Atemwege von der Nasenspitze über die paranasalen Sinus bis in die distalen Alveolen und dass ein gutes Therapiekonzept immer das große Ganze im Blick haben sollte. Schlussendlich stellt die CF hier ein Modellsystem dar. Viele unserer Erkenntnisse können auch Nicht-CF-Patienten mit Erkrankungen der oberen Atemwege zugutekommen

DNA damage-induced translocation of S100A11 into the nucleus regulates cell proliferation

<p>Abstract</p> <p>Background</p> <p>Proteins are able to react in response to distinct stress stimuli by alteration of their subcellular distribution. The stress-responsive protein S100A11 belongs to the family of multifunctional S100 proteins which have been implicated in several key biological processes. Previously, we have shown that S100A11 is directly involved in DNA repair processes at damaged chromatin in the nucleus. To gain further insight into the underlying mechanism subcellular trafficking of S100A11 in response to DNA damage was analyzed.</p> <p>Results</p> <p>We show that DNA damage induces a nucleolin-mediated translocation of S100A11 from the cytoplasm into the nucleus. This translocation is impeded by inhibition of the phosphorylation activity of PKCα. Translocation of S100A11 into the nucleus correlates with an increased cellular p21 protein level. Depletion of nucleolin by siRNA severely impairs translocation of S100A11 into the nucleus resulting in a decreased p21 protein level. Additionally, cells lacking nucleolin showed a reduced colony forming capacity.</p> <p>Conclusions</p> <p>These observations suggest that regulation of the subcellular distribution of S100A11 plays an important role in the DNA damage response and p21-mediated cell cycle control.</p

Bdellovibrio and Like Organisms Are Predictors of Microbiome Diversity in Distinct Host Groups

Biodiversity is generally believed to be a main determinant of ecosystem functioning. This principle also applies to the microbiome and could consequently contribute to host health. According to ecological theory, communities are shaped by top predators whose direct and indirect interactions with community members cause stability and diversity. Bdellovibrio and like organisms (BALOs) are a neglected group of predatory bacteria that feed on Gram-negative bacteria and can thereby influence microbiome composition. We asked whether BALOs can predict biodiversity levels in microbiomes from distinct host groups and environments. We demonstrate that genetic signatures of BALOs are commonly found within the 16S rRNA reads from diverse host taxa. In many cases, their presence, abundance, and especially richness are positively correlated with overall microbiome diversity. Our findings suggest that BALOs can act as drivers of microbial alpha-diversity and should therefore be considered candidates for the restoration of microbiomes and the prevention of dysbiosis

Reliability of electrical and hydraulic pitch systems in wind turbines based on field-data analysis

The pitch system is notably one of the critical subsystems of a wind turbine, supporting its effective control towards maximising wind capture and at the same time protecting its integrity in cases of excessive loads. A pitching mechanism is also responsible for operational downtime, hence its reliability performance needs to be carefully evaluated so as to ensure operational availability. This study aims to derive failure rates of two configurations of pitch systems, namely the electrical and hydraulic, based on statistical analysis of a large population of onshore assets, followed by a classification of findings by turbine rating, effect of seasonality, and reliability performance of different manufacturers. The data sets underlying the present analysis are based on maintenance reports and comprise 1847 operational years of wind turbines with electrical and 848 operational years of turbines with hydraulic pitch system. Results of this study show high failure rates in pitch systems of both types, with hydraulic systems performing slightly better than electrical (0.54 vs. 0.56 failures per turbine per year), a significant variation between turbines of different manufacturers, and a tendency for higher failure rates for larger turbines

Bi-allelic loss of function variants in SLC30A5 as cause of perinatal lethal cardiomyopathy

Perinatal mortality is a heavy burden for both affected parents and physicians. However, the underlying genetic causes have not been sufficiently investigated and most cases remain without diagnosis. This impedes appropriate counseling or therapy. We describe four affected children of two unrelated families with cardiomyopathy, hydrops fetalis, or cystic hygroma that all deceased perinatally. In the four patients, we found the following homozygous loss of function (LoF) variants in SLC30A5 NM_022902.4:c.832_836del p.(Ile278Phefs*33) and NM_022902.4:c.1981_1982del p.(His661Tyrfs*10). Knockout of SLC30A5 has previously been shown a cardiac phenotype in mouse models and no homozygous LoF variants in SLC30A5 are currently described in gnomAD. Taken together, we present SLC30A5 as a new gene for a severe and perinatally lethal form of cardiomyopathy

Ivacaftor Reduces Inflammatory Mediators in Upper Airway Lining Fluid From Cystic Fibrosis Patients With a G551D Mutation: Serial Non- Invasive Home-Based Collection of Upper Airway Lining Fluid

In cystic fibrosis (CF) therapy, the recent approval of CF-transmembrane conductance regulator (CFTR) channel modulators is considered to be the major breakthrough. However, the current first-line approach based mainly on pulmonary function to measure effects of the novel therapy, tested by forced expiratory volumes in one second (FEV1), provides restricted sensitivity to detect early structural damages. Accordingly, there is a need for new sensitive surrogate parameters. Most interestingly, these should quantify inflammation that precedes a decline of pulmonary function. We present a novel method assessing inflammatory markers in the upper airways’ epithelial lining fluid (ELF) obtained by nasal lavage (NL). In contrast to broncho-alveolar lavage, ELF sampling by NL is an attractive method due to its limited invasiveness which allows repeated analyses, even performed in a home-based setting. In a longitudinal cohort study (ClinicalTrials.gov, Identifier: NCT02311140), we assessed changes of inflammatory mediators in 259 serially obtained nasal lavages taken up to every second day before and during therapy with ivacaftor from ten CF patients carrying a G551D mutation. Patients were trained to sample NL-fluid at home, to immediately freeze and transfer chilled secretions to centers. Neutrophil Elastase, Interleukins IL-1b, IL-6 and IL-8 in NL were quantified. During 8-12 weeks of ivacaftor-treatment, median values of IL-1b and IL-6 significantly declined 2.29-fold (2.97!1.30 pg/mL), and 1.13-fold (6.48!5.72 pg/mL), respectively. In parallel, sweat tests and pulmonary function improved considerably. This is the first study assessing changes of airway inflammation on a day-to-day basis in CF patients receiving a newly administered CFTR-modulator therapy. It proves a decline of airway inflammation during ivacaftor-therapy

Fluorescence in situ hybridization (FISH) and cell sorting of living bacteria

Despite the development of several cultivation methods, the rate of discovery of microorganisms that are yet-to-be cultivated outpaces the rate of isolating and cultivating novel species in the laboratory. Furthermore, no current cultivation technique is capable of selectively isolating and cultivating specific bacterial taxa or phylogenetic groups independently of morphological or physiological properties. Here, we developed a new method to isolate living bacteria solely based on their 16S rRNA gene sequence. We showed that bacteria can survive a modified version of the standard fluorescence in situ hybridization (FISH) procedure, in which fixation is omitted and other factors, such as centrifugation and buffers, are optimized. We also demonstrated that labelled DNA probes can be introduced into living bacterial cells by means of chemical transformation and that specific hybridization occurs. This new method, which we call live-FISH, was then combined with fluorescence-activated cell sorting (FACS) to sort specific taxonomic groups of bacteria from a mock and natural bacterial communities and subsequently culture them. Live-FISH represents the first attempt to systematically optimize conditions known to affect cell viability during FISH and then to sort bacterial cells surviving the procedure. No sophisticated probe design is required, making live-FISH a straightforward method to be potentially used in combination with other single-cell techniques and for the isolation and cultivation of new microorganisms

Divergent dynamics of inflammatory mediators and multiplex PCRs during airway infection in cystic fibrosis patients and healthy controls: Serial upper airway sampling by nasal lavage

Background In cystic fibrosis (CF), acute respiratory exacerbations critically enhance pulmonary destruction. Since these mainly occur outside regular appointments, they remain unexplored. We previously elaborated a protocol for home-based upper airway (UAW) sampling obtaining nasal-lavage fluid (NLF), which, in contrast to sputum, does not require immediate processing. The aim of this study was to compare UAW inflammation and pathogen colonization during stable phases and exacerbations in CF patients and healthy controls. Methods Initially, we obtained NLF by rinsing 10 ml of isotonic saline/nostril during stable phases. During exacerbations, subjects regularly collected NLF at home. CF patients directly submitted one aliquot for microbiological cultures. The remaining samples were immediately frozen until transfer on ice to our clinic, where PCR analyses were performed and interleukin (IL)-1β/IL-6/IL-8, neutrophil elastase (NE), matrix metalloproteinase (MMP)-9, and tissue inhibitor of metalloproteinase (TIMP)-1 were assessed. Results Altogether, 49 CF patients and 38 healthy controls (HCs) completed the study, and 214 NLF samples were analyzed. Of the 49 CF patients, 20 were at least intermittently colonized with P. aeruginosa and received azithromycin and/or inhaled antibiotics as standard therapy. At baseline, IL-6 and IL-8 tended to be elevated in CF compared to controls. During infection, inflammatory mediators increased in both cohorts, reaching significance only for IL-6 in controls (p=0.047). Inflammatory responses tended to be higher in controls [1.6-fold (NE) to 4.4-fold (MMP-9)], while in CF, mediators increased only moderately [1.2-1.5-fold (IL-6/IL-8/NE/TIMP-1/MMP-9)]. Patients receiving inhalative antibiotics or azithromycin (n=20 and n=15, respectively) revealed lower levels of IL-1β/IL-6/IL-8 and NE during exacerbation compared to CF patients not receiving those antibiotics. In addition, CF patients receiving azithromycin showed MMP-9 levels significantly lower than CF patients not receiving azithromycin at stable phase and exacerbation. Altogether, rhinoviruses were the most frequently detected virus, detected at least once in n=24 (49.0%) of the 49 included pwCF and in n=26 (68.4%) of the 38 healthy controls over the 13-month duration of the study. Remarkably, during exacerbation, rhinovirus detection rates were significantly higher in the HC group compared to those in CF patients (65.8% vs. 22.4%; p<0.0001). Conclusion Non-invasive and partially home-based UAW sampling opens new windows for the assessment of inflammation and pathogen colonization in the unified airway system

Goos-Haenchen shift and localization of optical modes in deformed microcavities

Recently, an interesting phenomenon of spatial localization of optical modes along periodic ray trajectories near avoided resonance crossings has been observed [J. Wiersig, Phys. Rev. Lett. 97, 253901 (2006)]. For the case of a microdisk cavity with elliptical cross section we use the Husimi function to analyse this localization in phase space. Moreover, we present a semiclassical explanation of this phenomenon in terms of the Goos-Haenchen shift which works very well even deep in the wave regime. This semiclassical correction to the ray dynamics modifies the phase space structure such that modes can localize either on stable islands or along unstable periodic ray trajectories.Comment: 9 pages, 14 figures in reduced qualit