Refine
Year of publication
Document Type
- Article (31128)
- Part of Periodical (11550)
- Book (8266)
- Doctoral Thesis (5713)
- Part of a Book (3967)
- Working Paper (3385)
- Review (2939)
- Contribution to a Periodical (2368)
- Preprint (2084)
- Report (1560)
Language
- German (42718)
- English (29217)
- French (1060)
- Portuguese (840)
- Spanish (309)
- Croatian (302)
- Multiple languages (258)
- Italian (198)
- mis (174)
- Turkish (168)
Has Fulltext
- yes (75568) (remove)
Keywords
- Deutsch (1076)
- Literatur (868)
- taxonomy (760)
- Deutschland (553)
- Rezension (511)
- new species (449)
- Rezeption (354)
- Frankfurt <Main> / Universität (341)
- Übersetzung (326)
- Geschichte (300)
Institute
- Medizin (7691)
- Präsidium (5170)
- Physik (4464)
- Extern (2738)
- Wirtschaftswissenschaften (2686)
- Gesellschaftswissenschaften (2369)
- Biowissenschaften (2184)
- Biochemie und Chemie (1975)
- Frankfurt Institute for Advanced Studies (FIAS) (1686)
- Center for Financial Studies (CFS) (1628)
Ausdrucksphänomene fluktuieren zwischen den Sphären von Körperlichkeit und Bewegung sowie Sprachlichkeit und Kommunikation. Um Ausdruck in seiner eigentümlichen Mobilität und Wandlungsfähigkeit zu erfassen, erkunden die Beiträge des vorliegenden Bands seine ästhetischen, sozialen und (inter-)subjektiven Dimensionen und deren Überschneidungen sowohl im Rahmen historischer Entwicklungen als auch vor dem Hintergrund aktueller Erkenntnisse und Debatten der Psychologie, Philosophie, Hirnforschung und Linguistik. Aus den Themenbereichen Körper, Sprache und Künste, aber gleichfalls aus übergreifenden Fragestellungen der Anthropologie heraus werden kommunikative und körperliche Ausdrucksformen, auch in ihrem Zusammenspiel, in systematischer Weise analysiert und auf ihre situative und historische Spezifik hin untersucht. Nicht nur sprachliche Äußerungen, sondern ebenso Gesten und Gebärden, Tanz, Musik, Mienenspiel und Malerei stoßen dabei an Grenzen der Verständlichkeit und verweisen auf Ausdrucksqualitäten jenseits von Verbalität und Körperlichkeit.
The development of the atrioventricular (AV) canal and the cardiac valves is tightly linked and a critically regulated process. Anomalies in components of the involved pathways can lead to congenital valve malformations, a leading cause of morbidity and mortality in neonates. Myocardial Bmp as well as endocardial Notch and Wnt signaling have been identified as critical factors for the induction of EMT during the formation of the endocardial cushions and cardiac valves. Of these, canonical Wnt signaling positively regulates endocardial proliferation and EMT but negatively regulates endocardial differentiation. Further, elevated Wnt signaling leads to the ectopic expression of myocardial Bmp ligands suggesting a high level of integration of the involved pathways and crosstalk amongst the different cardiac tissues.
Here we have identified a novel role for Id4 as a mediator between Bmp and Wnt signaling. Id4 belongs to the Id family of proteins and is known to be involved in bone and nervous system development. We found that in zebrafish, id4 is expressed in the endocardium of the AV canal at embryonic stages and throughout the atrial chamber in addition to AV canal, in adults. Using transcription activator-like effector nucleases (TALENs) we established an id4 mutant allele. Our analysis shows that id4 mutant larvae are susceptible to retrograde blood flow, and show aberrant expression of developmental valvular markers. These include expanded expression domains of markers like bmp4, cspg2a and Alcam. In contrast, valve maturation as assessed by the expression of spp1 is considerably reduced in id4 mutants. Using conditional transgenic systems, along with elegant in vivo imaging of transgenic reporter lines, we further found that id4 is a transcriptional target of Bmp signaling, and it is capable of dose dependently restricting Wnt signaling in the endocardium of the Atrioventricular Canal.
Taken together, our data identifies Id4 as a novel player in Atrioventricular Canal and valve development. We show that Id4 function is important in valve development acting downstream of Bmp signaling by restricting endocardial Wnt to allow valve maturation
Purpose: To analyze the protein profile of human vitreous of untreated patients with retinal vein occlusion (RVO).
Methods: Sixty-eight vitreous humor (VH) samples (44 from patients with treatment naïve RVO, 24 controls with idiopathic floaters) were analyzed in this clinical-experimental study using capillary electrophoresis coupled to mass spectrometer and tandem mass spectrometry. To define potential candidate protein markers of RVO, proteomic analysis was performed on RVO patients (n = 30) and compared with controls (n = 16). To determine validity of potential biomarker candidates in RVO, receiver operating characteristic (ROC) was performed by using proteome data of independent RVO (n = 14) and control samples (n = 8).
Results: Ninety-four different proteins (736 tryptic peptides) could be identified. Sixteen proteins were found to be significant when comparing RVO and control samples (P = 1.43E-05 to 4.48E-02). Five proteins (Clusterin, Complement C3, Ig lambda-like polypeptide 5 (IGLL5), Opticin and Vitronectin), remained significant after using correction for multiple testing. These five proteins were also detected significant when comparing subgroups of RVO (central RVO, hemi-central RVO, branch RVO) to controls. Using independent samples ROC-Area under the curve was determined proving the validity of the results: Clusterin 0.884, Complement C3 0.955, IGLL5 1.000, Opticin 0.741, Vitronectin 0.786. In addition, validation through ELISA measurements was performed.
Conclusion: The results of the study reveal that the proteomic composition of VH differed significantly between the patients with RVO and the controls. The proteins identified may serve as potential biomarkers for pathogenesis induced by RVO.
Deciduous plants avoid the costs of maintaining leaves in the unfavourable season, but carry the costs of constructing new leaves every year. Deciduousness is therefore expected in ecological situations with pronounced seasonality and low costs of leaf construction. In our study system, a seasonally dry tropical savanna, many trees are deciduous, suggesting that leaf construction costs must be low. Previous studies have, however, shown that nitrogen is limiting in this system, suggesting that leaf construction costs are high. Here we examine this conundrum using a time series of soil moisture availability, leaf phenology and nitrogen distribution in the tree canopy to illustrate how trees resorb nitrogen before leaf abscission and use stored reserves of nitrogen and carbon to construct new leaves at the onset of the growing season. Our results show that trees deployed leaves shortly before and in anticipation of the first rains with its associated pulse of nitrogen mineralisation. Our results also show that trees rapidly constructed a full canopy of leaves within two weeks of the first rains. We detected an increase in leaf nitrogen content that corresponded with the first rains and with the movement of nitrogen to more distal branches, suggesting that stored nitrogen reserves are used to construct leaves. Furthermore the stable carbon isotope ratios (δ13C) of these leaves suggest the use of stored carbon for leaf construction. Our findings suggest that the early deployment of leaves using stored nitrogen and carbon reserves is a strategy that is integrally linked with the onset of the first rains. This strategy may confer a competitive advantage over species that deploy leaves at or after the onset of the rains.
Background: Hypothermia has been discussed as playing a role in improving the early phase of systemic inflammation. However, information on the impact of hypothermia on the local inflammatory response is sparse. We therefore investigated the kinetics of local and systemic inflammation in the late posttraumatic phase after induction of hypothermia in an established porcine long-term model of combined trauma.
Materials & Methods: Male pigs (35 ± 5kg) were mechanically ventilated and monitored over the study period of 48 h. Combined trauma included tibia fracture, lung contusion, liver laceration and pressure-controlled hemorrhagic shock (MAP < 30 ± 5 mmHg for 90 min). After resuscitation, hypothermia (33°C) was induced for a period of 12 h (HT-T group) with subsequent re-warming over a period of 10 h. The NT-T group was kept normothermic. Systemic and local (fracture hematoma) cytokine levels (IL-6, -8, -10) and alarmins (HMGB1, HSP70) were measured via ELISA.
Results: Severe signs of shock as well as systemic and local increases of pro-inflammatory mediators were observed in both trauma groups. In general the local increase of pro- and anti-inflammatory mediator levels was significantly higher and prolonged compared to systemic concentrations. Induction of hypothermia resulted in a significantly prolonged elevation of both systemic and local HMGB1 levels at 48 h compared to the NT-T group. Correspondingly, local IL-6 levels demonstrated a significantly prolonged increase in the HT-T group at 48 h.
Conclusion: A prolonged inflammatory response might reduce the well-described protective effects on organ and immune function observed in the early phase after hypothermia induction. Furthermore, local immune response also seems to be affected. Future studies should aim to investigate the use of therapeutic hypothermia at different degrees and duration of application.
Background: Common ECG criteria such as ST-segment changes are of limited value in patients with suspected acute myocardial infarction (AMI) and bundle branch block or wide QRS complex. A large proportion of these patients do not suffer from an AMI, whereas those with ST-elevation myocardial infarction (STEMI) equivalent AMI benefit from an aggressive treatment. Aim of the present study was to evaluate the diagnostic information of cardiac troponin I (cTnI) in hemodynamically stable patients with wide QRS complex and suspected AMI.
Methods: In 417 out of 1818 patients presenting consecutively between 01/2007 and 12/2008 in a prospective multicenter observational study with suspected AMI a prolonged QRS duration was observed. Of these, n = 117 showed significant obstructive coronary artery disease (CAD) used as diagnostic outcome variable. cTnI was determined at admission.
Results: Patients with significant CAD had higher cTnI levels compared to individuals without (median 250ng/L vs. 11ng/L; p<0.01). To identify patients needing a coronary intervention, cTnI yielded an area under the receiver operator characteristics curve of 0.849. Optimized cut-offs with respect to a sensitivity driven rule-out and specificity driven rule-in strategy were established (40ng/L/96ng/L). Application of the specificity optimized cut-off value led to a positive predictive value of 71% compared to 59% if using the 99th percentile cut-off. The sensitivity optimized cut-off value was associated with a negative predictive value of 93% compared to 89% provided by application of the 99th percentile threshold.
Conclusion: cTnI determined in hemodynamically stable patients with suspected AMI and wide QRS complex using optimized diagnostic thresholds improves rule-in and rule-out with respect to presence of a significant obstructive CAD.
Background: Advanced liver diseases are associated with profound alterations of the coagulation system increasing the risk not only of bleeding, but also of thromboembolic complications. A recent milestone study has shown that prophylactic anticoagulation in liver cirrhosis patients results in a reduced frequency of hepatic decompensation. Yet, INR measurement, one of the most widely applied tests to assess liver function, only inaccurately predicts the risk of hepatic decompensation related to alterations of the coagulation system. To assess the relationship between selected coagulation factors / natural anticoagulants with INR, MELD score, and hepatic decompensation, we performed the present pilot study. A total number of 92 patients with various stages of liver cirrhosis were included and prospectively followed for at least 6 months. We found that important natural anticoagulants, namely antithrombin and protein C, as well as factor XI (which may also serve as an anticoagulant) decreased earlier and by a larger magnitude than one would expect from classical coagulation test results. The correlation between these factors and INR was only moderate. Importantly, reduced plasma activities of natural anticoagulants but not INR or MELD score were independent predictors of hepatic encephalopathy (P = 0.013 and 0.003 for antithrombin and protein C, respectively).
Conclusion: In patients with liver cirrhosis plasma activities of several natural anticoagulants are earlier and stronger affected than routine coagulation tests. Reduced activities of natural anticoagulants may be predictive for the development of hepatic encephalopathy.
Thromboembolic events are one of the world’s leading causes of death among patients. Embolus or clot formations have several etiologies including paraneoplastic, post-surgery, cauterization, transplantation, or extracorporeal circuits. Despite its medical significance, little progress has been made in early embolus detection, screening and control. The aim of our study is to test the utility of the in vivo photoacoustic (PA) flow cytometry (PAFC) technique for non-invasive embolus detection in real-time. Using in vivo PAFC, emboli were non-invasively monitored in the bloodstream of two different mouse models. The tumor-free mouse model consisted of two groups, one in which the limbs were clamped to produce vessel stasis (7 procedures), and one where the mice underwent surgery (7 procedures). The melanoma-bearing mouse model also consisted of two groups, one in which the implanted tumor underwent compression (8 procedures), and one where a surgical excision of the implanted tumor was performed (8 procedures). We demonstrated that the PAFC can detect a single embolus, and has the ability to distinguish between erythrocyte–rich (red) and leukocyte/platelet-rich (white) emboli in small vessels. We show that, in tumor-bearing mice, the level of circulating emboli was increased compared to tumor-free mice (p = 0.0013). The number of circulating emboli temporarily increased in the tumor-free control mice during vessel stasis (p = 0.033) and after surgical excisions (signed-rank p = 0.031). Similar observations were noted during tumor compression (p = 0.013) and after tumor excisions (p = 0.012). For the first time, it was possible to detect unlabeled emboli in vivo non-invasively, and to confirm the presence of pigmented tumor cells within circulating emboli. The insight on embolus dynamics during cancer progression and medical procedures highlight the clinical potential of PAFC for early detection of cancer and surgery-induced emboli to prevent the fatal thromboembolic complications by well-timed therapy.
Leptomeningeal dissemination of a primary brain tumor is a condition which is challenging to treat, as it often occurs in rather late disease stages in highly pretreated patients. Its prognosis is dismal and there is still no accepted standard of care. We report here a good clinical effect with a partial response in three out of nine patients and a stable disease with improvement on symptoms in two more patients following systemic anti-angiogenic treatment with bevacizumab (BEV) alone or in combination with chemo- and/or radiotherapy in a series of patients with leptomeningeal dissemination from primary brain tumors (diffuse astrocytoma WHO°II, anaplastic astrocytoma WHO°III, anaplastic oligodendroglioma WHO°III, primitive neuroectodermal tumor and glioblastoma, both WHO°IV). This translated into effective symptom control in five out of nine patients, but only moderate progression-free and overall survival times were reached. Partial responses as assessed by RANO criteria were observed in three patients (each one with anaplastic oligodendroglioma, primitive neuroectodermal tumor and glioblastoma). In these patients progression-free survival (PFS) intervals of 17, 10 and 20 weeks were achieved. In three patients (each one with diffuse astrocytoma, anaplastic astrocytoma and primitive neuroectodermal tumor) stable disease was observed with PFS of 13, 30 and 8 weeks. Another three patients (all with glioblastoma) were primary non-responders and deteriorated rapidly with PFS of 3 to 4 weeks. No severe adverse events were seen. These experiences suggest that the combination of BEV with more conventional therapy schemes with chemo- and/or radiotherapy may be a palliative treatment option for patients with leptomeningeal dissemination of brain tumors.
Triple therapy of chronic hepatitis C virus (HCV) infection with boceprevir (BOC) or telaprevir (TVR) leads to virologic failure in many patients which is often associated with the selection of resistance-associated variants (RAVs). These resistance profiles are of importance for the selection of potential rescue treatment options. In this study, we sequenced baseline NS3 RAVs population-based and investigated the sensitivity of NS3 phenotypes in an HCV replicon assay together with clinical factors for a prediction of treatment response in a cohort of 165 German and Swiss patients treated with a BOC or TVR-based triple therapy. Overall, the prevalence of baseline RAVs was low, although the frequency of RAVs was higher in patients with virologic failure compared to those who achieved a sustained virologic response (SVR) (7% versus 1%, P = 0.06). The occurrence of RAVs was associated with a resistant NS3 quasispecies phenotype (P<0.001), but the sensitivity of phenotypes was not associated with treatment outcome (P = 0.2). The majority of single viral and host predictors of SVR was only weakly associated with treatment response. In multivariate analyses, low AST levels, female sex and an IFNL4 CC genotype were independently associated with SVR. However, a combined analysis of negative predictors revealed a significantly lower overall number of negative predictors in patients with SVR in comparison to individuals with virologic failure (P<0.0001) and the presence of 2 or less negative predictors was indicative for SVR. These results demonstrate that most single baseline viral and host parameters have a weak influence on the response to triple therapy, whereas the overall number of negative predictors has a high predictive value for SVR.
Background: Expected growth in the demand for health services has generated interest in the more effective deployment of health care assistants. Programs encouraging German general practitioners (GPs) to share responsibility for care with specially qualified health care assistants in the family practice (VERAHs) have existed for several years. But no studies have been conducted on the tasks German GPs are willing to rely on specially qualified personnel to perform, what they are prepared to delegate to all non-physician practice staff and what they prefer to do themselves.
Methods: As part of an evaluation study on the deployment of VERAHs in GP-centered health care, we used a questionnaire to ask about task delegation within the practice team. From a list of tasks that VERAHs are specifically trained to carry out, GPs were asked to indicate which they actually delegate. We also asked GPs why they had employed a VERAH in their practice and for their opinions on the benefits and limitations of assigning tasks to VERAHs. The aim of the study was to find out which tasks GPs delegate to their specially qualified personnel, which they permit all HCAs to carry out, and which tasks they do not delegate at all.
Results: The survey was filled in and returned by 245 GPs (83%). Some tasks were exclusively delegated to VERAHs (e.g. home visits), while others were delegated to all HCAs (e.g. vaccinations). About half the GPs rated the assessment of mental health, as part of the comprehensive assessment of a patient’s condition, as the sole responsibility of a GP.
The possibility to delegate more complex tasks was the main reason given for employing a VERAH. Doctors said the delegation of home visits provided them with the greatest relief.
Conclusions: In Germany, where GPs are solely accountable for the health care provided in their practices, experience with the transfer of responsibility to other non-physician health care personnel is still very limited. When HCAs have undergone special training, GPs seem to be prepared to delegate tasks that demand a substantial degree of know-how, such as home visits and case management. This “new” role allocation within the practice may signal a shift in the provision of health care by family practice teams in Germany.
Objective: Mucoactive drugs should increase the ability to expectorate sputum and, ideally, have anti-inflammatory properties. The aim of the study was to evaluate the mucolytic activity of Tyloxapol compared to saline (0.9%) in COPD.
Design: A randomized, placebo-controlled, double-blinded crossover, clinical trial was carried out. Patients were randomly assigned to either inhale 5 ml Tyloxapol 1% or saline 0.9% solution three times daily for 3 weeks and vice versa for another 3 weeks. 28 patients (18 male, 10 female, 47 to 73 years old, median age 63.50) were screened, 21 were treated and 19 patients completed the study per protocol.
Results: A comparison of the two treatment phases showed that the primary endpoint sputum weight was statistically significant higher when patients inhaled Tyloxapol (mean 4.03 g, 95% CI: 2.34–5.73 g at week 3) compared to saline (mean 2.63 g, 95% CI: 1.73–3.53 g at week 3). The p-value at three weeks of treatment was 0.041 between treatment arms. Sputum cells decreased during the Tyloxapol treatment after 3 weeks, indicating that Tyloxapol might have some anti-neutrophilic properties. Lung function parameters (FVC, FEV1, RV, and RV/TLC) remained stable during the study, and no treatment effect was shown. Interestingly, there was a mean increase in all inflammatory cytokines (IL-1β, IL-6, and IL-8) during the saline treatment from day 1 to week 3, whereas during the Tyloxapol treatment, all cytokines decreased. Due to the small sample size and the large individual variation in sputum cytokines, these differences were not significant. However, analyses confirmed that Tyloxapol has significant anti-inflammatory properties in vitro. Despite the high number of inhalations (more than 1000), only 27 adverse events (20 during the Tyloxapol and seven during saline) were recorded. Eleven patients experienced AEs under Tyloxapol and six under saline treatment, which indicates that inhalation of saline or Tyloxapol is a very safe procedure.
Conclusion: Our study demonstrated that inhalation of Tyloxapol by patients with COPD is safe and superior to saline and has some anti-inflammatory effects.
We integrated recent improvements within the floating catchment area (FCA) method family into an integrated ‘iFCA`method. Within this method we focused on the distance decay function and its parameter. So far only distance decay functions with constant parameters have been applied. Therefore, we developed a variable distance decay function to be used within the FCA method. We were able to replace the impedance coefficient β by readily available distribution parameter (i.e. median and standard deviation (SD)) within a logistic based distance decay function. Hence, the function is shaped individually for every single population location by the median and SD of all population-to-provider distances within a global catchment size. Theoretical application of the variable distance decay function showed conceptually sound results. Furthermore, the existence of effective variable catchment sizes defined by the asymptotic approach to zero of the distance decay function was revealed, satisfying the need for variable catchment sizes. The application of the iFCA method within an urban case study in Berlin (Germany) confirmed the theoretical fit of the suggested method. In summary, we introduced for the first time, a variable distance decay function within an integrated FCA method. This function accounts for individual travel behaviors determined by the distribution of providers. Additionally, the function inherits effective variable catchment sizes and therefore obviates the need for determining variable catchment sizes separately.
Background: Dengue virus infection is the most rapidly spreading vector-borne disease in the world. Essential research on dengue virus transmission and its prevention requires community participation. Therefore, it is crucial to understand the factors that are associated with the willingness of communities in high prevalence areas to participate in dengue research. The aim of this study was to explore factors associated with the willingness of healthy community members in Aceh province, Indonesia, to participate in dengue research that would require phlebotomy.
Methodology/Principal Findings: A community-based cross-sectional study was carried out in nine regencies and municipalities of Aceh from November 2014 to March 2015. Interviews using a set of validated questionnaires were conducted to collect data on demography, history of dengue infection, socioeconomic status, and knowledge, attitude and practice regarding dengue fever. Two-step logistic regression and Spearman’s rank correlation (rs) analysis were used to assess the influence of independent variables on dependent variables. Among 535 participants, less than 20% had a good willingness to participate in the dengue study. The factors associated with good willingness to participate were being female, working as a civil servant, private employee or entrepreneur, having a high socioeconomic status and good knowledge, attitude and practice regarding dengue. Good knowledge and attitude regarding dengue were positive independent predictors of willingness to participate (OR: 2.30 [95% CI: 1.36–3.90] and 3.73 [95% CI: 2.24–6.21], respectively).
Conclusion/Significance: The willingness to participate in dengue research is very low among community members in Aceh, and the two most important associated factors are knowledge and attitude regarding dengue. To increase participation rate, efforts to improve the knowledge and attitude of community members regarding dengue fever and dengue-related research is required before such studies are launched.
1. Objective: Chronic hepatitis C virus infections (HCV) cause a significant public health burden. Introduction of telaprevir (TVR) and boceprevir (BOC) has increased sustained virologic response rates (SVR) in genotype 1 patients but were accompanied by higher treatment costs and more side effects. Aim of the study was to assess outcomes and costs of treating HCV with TVR or BOC in routine care.
2. Material and Methods: Data was obtained from a non-interventional study. This analysis relates on a subset of 1,786 patients for whom resource utilisation was documented. Sociodemografic and clinical parameters as well as resource utilisation were collected using a web-based data recording system. Costs were calculated using official remuneration schemes.
3. Results: Mean age of patients was 49.2 years, 58.6% were male. In treatment-naive patients SVR-rates of 62.2% and 55.7% for TVR and BOC were observed (prior relapser: 68.5% for TVR and 63.5% for BOC; prior nonresponder: 45.6% for TVR and 39.1% for BOC). Treatment costs are dominated by costs for pharmaceuticals and range between €39,081 and €53,491. We calculated average costs per SVR of €81,347 (TVR) and €70,163 (BOC) in treatment-naive patients (prior relapser: 78,089 €/SVR for TVR and 82,077 €/SVR for BOC; prior non-responder: 116,509 €/SVR for TVR and 110,156 €/SVR for BOC). Quality of life data showed a considerable decrease during treatment.
4. Conclusion: Our study is one of few investigating both, outcomes and costs, of treating HCV in a real-life setting. Data can serve as a reference in the discussion of increasing costs in recently introduced agents
Mediation in der Türkei : Betrachtung ausgewählter Aspekte im Vergleich zur Mediation in Deutschland
(2016)
Angesichts der vergleichsweise noch sehr jungen Entwicklung der Mediation in der Türkei mag man es auf den ersten Blickerstaunlich finden, dass in der Türkei zeitgleich mit Deutschland ein Mediationsgesetz geschaffen wurde. Die Mediation als außergerichtliches Vermittlungsverfahren gründet darauf, dass Streitparteien freiwillig und selbstbestimmt ihren Konflikt mit Unterstützung eines Mediators einer gemeinsam entwickelten Lösung zuführen. Dies sind die Grundprinzipien der Mediation, die sowohl dem deutschen als auch dem türkischen Mediationsgesetz als Basis dienen.
Trotz vieler Ähnlichkeiten haben die kulturellen Besonderheiten beider Länder Einfluss auf die rechtliche Ausgestaltung dieses Einigungsverfahrens sowie dessen Umsetzung in der Praxis .Ziel des vorliegenden Arbeitspapiers ist es, dem Leser einen Einblick in die Unterschiede und Gemeinsamkeiten der Mediation in der Türkei und Deutschland zu vermitteln und dabei vergleichend zu untersuchen , ob und inwieweit landestypischen Spezifika in der Entstehungsgeschichte, den Grundlagen und der Praxis der Mediation erkennbar und durch gesellschaftliche und kulturelle Faktoren erklärbar sind.
Die vorliegende Studie vermittelt einen epidemiologischen Überblick über das mit Haut- und Nagelläsionen assoziierte Pilzspektrum im Westen Panamas. Hierzu wurden Proben von vermutlich durch Pilzinfektionen verursachten Haut- sowie Nagelläsionen gesammelt und zum Anlegen von Kulturen verwendet. Die isolierten Pilze wurden basierend auf dem D-H-S System (Rieth), anhand morphologischer Merkmale, rDNA Sequenzdaten sowie phylogenetischen Analysen klassifiziert und mit Hilfe von Literaturdaten sowie physiologischen Eigenschaften als saprotrophe, opportunistische oder pathogene Organismen beurteilt. In Panama wurden 52 Proben von 51 Personen gesammelt, wobei das Material von 42 Haut- und Nagelläsionen der Füße, vier Läsionen der Fingernägel, zwei Chromomykosen, einer Tinea nigra und drei sonstigen Hautläsionen stammt. Bei 75 Prozent (n = 39) der Proben konnten Pilze kultiviert und insgesamt 201 Pilzstämme isoliert und subkultiviert werden. Hiervon wurden 50 Isolate (24,9 %) als Dermatophyten, 24 Stämme (11,9 %) als Hefen und 127 Isolate (63,2 %) als Schimmelpilze klassifiziert. Bei 19 Probanden (48,7 %) konnten Dermatophyten isoliert werden, wobei aus dem Probenmaterial von 12 Personen (63,2 %) ebenfalls andere Pilzarten nachgewiesen wurden. Von zwei Läsionen (5,1 %) wurden nur Hefen isoliert, wobei einmal eine Schwarze Hefe kultiviert wurde. In dem Material acht weiterer Proben (20,5 %) wurden Schimmelpilze und Hefestämme nachgewiesen und bei zehn Probanden (25,6 %) konnten aus dem Probenmaterial nur Schimmelpilze kultiviert werden. 172 Isolate wurden taxonomisch klassifiziert und 44 Arten aus 25 Gattungen, 17 Familien, 15 Ordnungen, sechs Klassen sowie den Abteilungen Ascomycota oder Basidiomycota zugeordnet. Die Ascomyceten stellen mit 164 Stämmen 40 verschiedener Arten aus 23 Gattungen, 15 Familien, 11 Ordnungen und vier Klassen die am häufigsten isolierte und vielfältigste Gruppe dar, während die Basidiomycota nur mit acht Isolaten vier verschiedener Arten zwei unterschiedlicher Gattungen, Familien, Ordnungen und Klassen nachgewiesen wurden. Im Rahmen dieser Arbeit wurden in Panama die anthropophilen Dermatophyten Trichophyton rubrum und T. interdigitale dokumentiert, wobei T. rubrum die am häufigsten isolierte Art darstellt. Kultivierte Hefen waren Candida albicans, C. duobushaemulonii, C. tropicalis, Hortaea werneckii, Sporobolomyces sp., Trichosporon asahii, T. japonicum und T. montevideense. Die Schimmelpilze stellen die größte und ökologisch diverseste Organismengruppe der kultivierten Pilze dar. So wurden von den untersuchten Läsionen sowohl humanpathogene Erreger, als auch opportunistische Arten und rein saprotrophe Pilze sowie mehrere Vertreter wahrscheinlich bisher nicht wissenschaftlich beschriebener Arten bzw. Gattungen nachgewiesen. Aus dem Probenmaterial wurden die Pilze Acremonium collariferum, Aspergillus awamori, A. clavatus, A. flavus, A. giganteus, A. heteromorphus, A. niger, A. ochraceus, A. sclerotiorum, A. versicolor, Chaetomium globosum, Chrysosporium tuberculatum, Cladosporium sphaerospermum, C. tenuissimum, Curvularia geniculata, C. lunata, Fonsecaea pedrosoi, Fusarium oxysporum, F. solani, Lophotrichus bartlettii, Microascus cinereus, Neoscytalidium dimidiatum, Penicillium commune, Scolecobasidium sp., Scopulariopsis carbonaria, S. croci, Verticillium cf. epiphytum und Wardomycopsis litoralis isoliert. Zudem wurden vier Isolate von zwei vermutlich neuen Arten der Gattung Acremonium (Bionectriaceae, Hypocreales), zwei Stämme mit einer genetischen Affinität zu der Gattung Cryptendoxyla (Cephalothecaceae, Sordariales) und jeweils ein mit den Gattungen Fusicladium (Venturiaceae, Venturiales), Knufia (Trichomeriaceae, Chaetothyriales) bzw. Rhexothecium (Eremomycetaceae, Dothideomycetidae) assoziierter Stamm kultiviert. Im Rahmen dieser Studie wurden A. giganteus, C. tenuissimum, L. bartlettii, S. carbonaria, S. croci, V. epiphytum und W. litoralis erstmalig von Mykosen des Menschen dokumentiert und die in der Literatur als Verursacher sowie Besiedler von Haut- und Nagelläsionen beschriebenen Organismen A. clavatus, A. flavus, A. niger, A. ochraceus, C. tropicalis, C. globosum, C. sphaerospermum, C. lunata, F. oxysporum, M. cinereus, P. commune, T. asahii, T. japonicum und T. montevideense wurden das erste Mal in klinischem Probenmaterial aus Panama nachgewiesen. Die Arten A. awamori, A. heteromorphus, C. globosum, C. tenuissimum, L. bartlettii, M. cinereus, P. commune, S. croci, T. asahii, T. japonicum, T. montevideense, V. epiphytum, W. litoralis und die Gattung Scolecobasidium wurden zudem erstmalig für Panama dokumentiert. Die Isolation von W. litoralis ist ebenfalls der erste Nachweis dieses Pilzes außerhalb von Spanien und auf dem amerikanischen Kontinent. Die große Anzahl im Rahmen dieser Arbeit beschriebener, bisher für die Wissenschaft unbekannter bzw. nicht in Panama dokumentierter Pilzarten lässt auf eine große mykologische Biodiversität in Panama schließen und zeigt den Bedarf weiterer Forschung.
Structural characterization of stressosome complexes by single-particle cryo-electron microscopy
(2015)
The stressosome is a Mega Dalton macromolecular complex involved in stress adaptation in bacteria. Stressosomes are considered as stress signaling hubs. They are able to perceive a variety of different stress stimuli and transduce them into one single cellular answer, which is the initialization of a transcriptional up-regulation of hundreds of different genes encoding for universal but also very specific stress response proteins.
The stressosome of Bacillus subtilis became a prime example for this intriguing stress-triggered transcriptional regulation when its architecture was determined by Single-particle cryo-electron microscopy (cryo-EM) in 2008. In Gram-positive Bacillus species, the stressosome complex senses changes in salt concentration, ethanol content, blue-light, heat or acid stress contributing to the general stress response by activation of the alternative σB factor. σB is a transcriptional promoter that initiates the transcription of over 150 general stress genes, e.g., genes that encode osmolyte transporters to counteract osmotic and chill stress. The B. subtilis stressosome (stressosome_Bc) is composed of multiple copies of the 3 proteins: RsbR, RsbS and RsbT. These three Rsb proteins (Regulator of Sigma B) are found clustered in one operon forming the conserved RST module. RsbS and RsbR are scaffold proteins comprising a STAS domain, respectively. Because these domains are dominantly associated to sulfate transporters and anti-sigma antagonist they were named STAS domains, however, they were also identified in other sensor proteins. In the stressosome they form the internal ball-shaped core, while the N-terminal globin-fold sensor domain of RsbR, protruding to the outside, facilitates stress sensing. It is assumed that the stress signal is transduced to the stressosome core via the STAS domain resulting in conformational changes of the core. These changes affect the binding of the third protein, RsbT, a serin-threonine kinase. As a direct consequence of stress sensing the RsbT kinase is released from the complex to start an activation cascade involving the stepwise activation of RsbU, V, W, and X, which are all part of the same operon, and finally of σB. In Bacillus species, several RsbR orthologs were identified varying mainly in the sequence of the N-terminal sensor domains. It is assumed that the stressosome_Bc assembles with a still unknown combination of RsbR orthologs allowing for the broad spectrum of stress stimuli that can be processed in vivo. The pathogenic bacteria Listeria monocytogenes is a close relative of Bacillus. Its potent stress response allows Listeria to survive the harsh environmental conditions during host infection and therefore the stress regulation machinery is contributing heavily to the virulence of this pathogen. In Listeria the Rsb operon is conserved and highly homologous to the Bacillus one. In the frame of this thesis, the in vitro assembly of Listeria innocua stressosomes was shown for the first time by Single-particle (SP) negative stain EM. Moreover, binding of Listeria RsbT to the assembled RsbR-RsbS complex was demonstrated biochemically.
Despite the conservation of the RST-module the entire Rsb operon is not conserved in the bacterial kingdom suggesting that signal transduction and regulation of gene expression might occur by very different mechanisms in stressosomes of different species. We have focused here on a stressosome type from the Gram-negative pathogen Vibrio vulnificus that is quite distinct from the Bacillus ones with respect to (1) the missing conservation of the Rsb operon, (2) the role of RsbT, (3) the activation of a different transcriptional promoter, and (4) the absence of additional RsbR orthologs. Interestingly, there is only one RsbR protein encoded in the genome. This one contains a Haem-group in its N-terminal domain being oxygen sensitive. It is assumed that the Vibrio stressosome perceive only oxidative stress and that regulation occurs via a diguanylate cyclase with a GAF domain that synthesizes the second messenger c-di-GMP from GTP.
We have started a structure determination of the Vibrio vulnificus stressosome by SP cryo-EM to elucidate the differences in the molecular mechanism of stress sensing in divers stressosome types. A 3D map of the oxidized (activated) Vibrio vulnificus stressosome was determined to 7.6 Å resolution revealing an increased flexibility of both the core and the N-terminal sensor domains in comparison to the Bacillus stressosome suggesting that our structure has trapped for the first time an active state of a stressosome complex. A 3D map of the stressosome core to 7 Å resolution allowed fitting of a homology model of the Vibrio stressosome based on the Bacillus stressosome as template. The conformational changes could be attributed to the entire core, which was confirmed by MD simulations.
Mitochondrial cristae are connected to the inner boundary membrane via crista junctions which are implicated in the regulation of oxidative phosphorylation, apoptosis, and import of lipids and proteins. The MICOS complex determines formation of crista junctions. We performed complexome profiling and identified Mic13, also termed Qil1, as a subunit of the MICOS complex. We show that MIC13 is an inner membrane protein physically interacting with MIC60, a central subunit of the MICOS complex. Using the CRISPR/Cas method we generated the first cell line deleted for MIC13. These knockout cells show a complete loss of crista junctions demonstrating that MIC13 is strictly required for the formation of crista junctions. MIC13 is required for the assembly of MIC10, MIC26, and MIC27 into the MICOS complex. However, it is not needed for the formation of the MIC60/MIC19/MIC25 subcomplex suggesting that the latter is not sufficient for crista junction formation. MIC13 is also dispensable for assembly of respiratory chain complexes and for maintaining mitochondrial network morphology. Still, lack of MIC13 resulted in a moderate reduction of mitochondrial respiration. In summary, we show that MIC13 has a fundamental role in crista junction formation and that assembly of respiratory chain supercomplexes is independent of mitochondrial cristae shape.
Background & Aim: The resistance profile of anti-hepatitis C virus (HCV) agents used in combination is important to guide optimal treatment regimens. We evaluated baseline and treatment-emergent NS3/4A and NS5B amino-acid variants among HCV genotype (GT)-1a and -1b-infected patients treated with faldaprevir (HCV protease inhibitor), deleobuvir (HCV polymerase non-nucleoside inhibitor), and ribavirin in multiple clinical studies.
Methods: HCV NS3/4A and NS5B population sequencing (Sanger method) was performed on all baseline plasma samples (n = 1425 NS3; n = 1556 NS5B) and on post-baseline plasma samples from patients with virologic failure (n = 113 GT-1a; n = 221 GT-1b). Persistence and time to loss of resistance-associated variants (RAVs) was estimated using Kaplan–Meier analysis.
Results: Faldaprevir RAVs (NS3 R155 and D168) and deleobuvir RAVs (NS5B 495 and 496) were rare (<1%) at baseline. Virologic response to faldaprevir/deleobuvir/ribavirin was not compromised by common baseline NS3 polymorphisms (e.g. Q80K in 17.5% of GT-1a) or by NS5B A421V, present in 20% of GT-1a. In GT-1b, alanine at NS5B codon 499 (present in 15% of baseline sequences) was associated with reduced response. Treatment-emergent RAVs consolidated previous findings: NS3 R155 and D168 were key faldaprevir RAVs; NS5B A421 and P495 were key deleobuvir RAVs. Among on-treatment virologic breakthroughs, RAVs emerged in both NS3 and NS5B (>90%). Virologic relapse was associated with RAVs in both NS3 and NS5B (53% GT-1b; 52% GT-1b); some virologic relapses had NS3 RAVs only (47% GT-1a; 17% GT-1b). Median time to loss of GT-1b NS5B P495 RAVs post-treatment (5 months) was less than that of GT-1b NS3 D168 (8.5 months) and GT-1a R155 RAVs (11.5 months).
Conclusion: Faldaprevir and deleobuvir RAVs are more prevalent among virologic failures than at baseline. Treatment response was not compromised by common NS3 polymorphisms; however, alanine at NS5B amino acid 499 at baseline (wild-type in GT-1a, polymorphism in GT-1b) may reduce response to this deleobuvir-based regimen.