Large-scale validation of miRNAs by disease association, evolutionary conservation and pathway activity.

The validation of microRNAs (miRNAs) identified by next generation sequencing involves amplification-free and hybridization-based detection of transcripts as criteria for confirming valid miRNAs. Since respective validation is frequently not performed, miRNA repositories likely still contain a substantial fraction of false positive candidates while true miRNAs are not stored in the repositories yet. Especially if downstream analyses are performed with these candidates (e.g. target or pathway prediction), the results may be misleading. In the present study, we evaluated 558 mature miRNAs from miRBase and 1,709 miRNA candidates from next generation sequencing experiments by amplification-free hybridization and investigated their distributions in patients with various disease conditions. Notably, the most significant miRNAs in diseases are often not contained in the miRBase. However, these candidates are evolutionary highly conserved. From the expression patterns, target gene and pathway analyses and evolutionary conservation analyses, we were able to shed light on the complexity of miRNAs in humans. Our data also highlight that a more thorough validation of miRNAs identified by next generation sequencing is required. The results are available in miRCarta ( https://mircarta.cs.uni-saarland.de )

Amplified Host Defense by Toll-Like Receptor-Mediated Downregulation of the Glucocorticoid-Induced Leucine Zipper (GILZ) in Macrophages

Activation of toll-like receptors (TLRs) plays a pivotal role in the host defense against bacteria and results in the activation of NF-κB-mediated transcription of proinflammatory mediators. Glucocorticoid-induced leucine zipper (GILZ) is an anti-inflammatory mediator, which inhibits NF-κB activity in macrophages. Thus, we aimed to investigate the regulation and role of GILZ expression in primary human and murine macrophages upon TLR activation. Treatment with TLR agonists, e.g., Pam3CSK4 (TLR1/2) or LPS (TLR4) rapidly decreased GILZ mRNA and protein levels. In consequence, GILZ downregulation led to enhanced induction of pro-inflammatory mediators, increased phagocytic activity, and a higher capacity to kill intracellular bacteria (Salmonella enterica serovar typhimurium), as shown in GILZ knockout macrophages. Treatment with the TLR3 ligand polyinosinic: polycytidylic acid [Poly(I:C)] did not affect GILZ mRNA levels, although GILZ protein expression was decreased. This effect was paralleled by sensitization toward TLR1/2- and TLR4-agonists. A bioinformatics approach implicated more than 250 miRNAs as potential GILZ regulators. Microarray analysis revealed that the expression of several potentially GILZ-targeting miRNAs was increased after Poly(I:C) treatment in primary human macrophages. We tested the ability of 11 of these miRNAs to target GILZ by luciferase reporter gene assays. Within this small set, four miRNAs (hsa-miR-34b*,−222,−320d,−484) were confirmed as GILZ regulators, suggesting that GILZ downregulation upon TLR3 activation is a consequence of the synergistic actions of multiple miRNAs. In summary, our data show that GILZ downregulation promotes macrophage activation. GILZ downregulation occurs both via MyD88-dependent and -independent mechanisms and can involve decreased mRNA or protein stability and an attenuated translation

Ocean current connectivity propelling the secondary spread of a marine invasive comb jelly across western Eurasia

Publication history: Accepted - 15 February 2018; Published - 16 May 2018.Aim: Invasive species are of increasing global concern. Nevertheless, the mechanisms driving further distribution after the initial establishment of non-native species remain largely unresolved, especially in marine systems. Ocean currents can be a major driver governing range occupancy, but this has not been accounted for in most invasion ecology studies so far. We investigate how well initial establishment areas are interconnected to later occupancy regions to test for the potential role of ocean currents driving secondary spread dynamics in order to infer invasion corridors and the source–sink dynamics of a non-native holoplanktonic biological probe species on a continental scale. Location: Western Eurasia. Time period: 1980s–2016. Major taxa studied: ‘Comb jelly’ Mnemiopsis leidyi. Methods: Based on 12,400 geo-referenced occurrence data, we reconstruct the invasion history of M. leidyi in western Eurasia. We model ocean currents and calculate their stability to match the temporal and spatial spread dynamics with large-scale connectivity patterns via ocean currents. Additionally, genetic markers are used to test the predicted connectivity between subpopulations. Results: Ocean currents can explain secondary spread dynamics, matching observed range expansions and the timing of first occurrence of our holoplanktonic non-native biological probe species, leading to invasion corridors in western Eurasia. In northern Europe, regional extinctions after cold winters were followed by rapid recolonizations at a speed of up to 2,000 km per season. Source areas hosting year-round populations in highly interconnected regions can re-seed genotypes over large distances after local extinctions. Main conclusions: Although the release of ballast water from container ships may contribute to the dispersal of non-native species, our results highlight the importance of ocean currents driving secondary spread dynamics. Highly interconnected areas hosting invasive species are crucial for secondary spread dynamics on a continental scale. Invasion risk assessments should consider large-scale connectivity patterns and the potential source regions of non-native marine species.Danish Council for Independent Research; Grant/Award Number: DFF-1325-00102B; FP7 People: Marie-Curie Actions, Grant/Award Number: MOBILEX, DFF - 1325-00025; EU, BONUS, BMBF, Grant/ Award Number: 03F0682; Excellence Cluster “Future Ocean”, Grant/Award Number: CP153

Notärztliche Einschätzung der Verletzungsschwere am Unfallort - Diagnostischer Wert technischer Parameter

Im klinischen Alltag werden im Rahmen der Übergabe zwischen Notarzt und Klinik nach Verkehrsunfällen immer wieder technische Parameter vom Unfallort genannt. Ob diese Informationen zur verbesserten Einschätzung der Verletzungsschwere nützen, ist bisher nicht ausreichend untersucht worden. Um dieser Frage nachzugehen, wurden Daten von 100 realen Fällen aus der Datenbank der Unfallforschung Greifswald in einem mehrstufigen Experiment erfahrenen Notärzten in Fragebogenform präsentiert. Dabei wurden zunächst einfache Routineparameter, dann erweiterte Parameter und schließlich Fotos der Unfälle dargeboten. Gefordert war eine Einschätzung der Verletzungsschwere eines beteiligten PKW-Fahrers in den vier am ISS-Wert orientierten Kategorien „leicht verletzt“, „schwer verletzt“, „lebensgefährlich verletzt“ und „tot“. Zur Auswertung erfolgte eine Dichotomisierung in „leicht und schwer verletzt“ (ISS≤15) versus „lebensgefährlich verletzt und tot“ (ISS 16-75). Berechnet wurden die Übereinstimmung der Teilnehmer im Hinblick auf die Verletzungsschwere jenseits des Zufalls (kappa-Statistik) sowie die diagnostische Testgüte (Sensitivität, Spezifität, Fläche unter der ROC-Kurve, Likelihood Ratios) technischer Unfallparameter. Die Beobachter-Übereinstimmung der Verletzungsschwere unter Kenntnis einfacher oder erweiterter technischer Parameter sowie Bildparameter lag bei kappa-Werten von 0,42, 0,65 und 0,61. Die Sensitivität schwankte zwischen den Beobachtern und je nach unterschiedlicher Informationsmenge zwischen 18 und 80%, die Spezifität zwischen 41 und 89%. Durch die Präsentation von Fotos vom Unfallort ließ sich eine Steigerung der Sensitivität erzielen. Die Verschiebung der Vortest-Wahrscheinlichkeit von 50% für eine lebensbedrohliche Verletzung betrug im Falle negativer technischer Befunde maximal 40%, im positiven Fall 67%. Im Rahmen dieser umfangreichen Untersuchung unter Nutzung realer Unfälle und erfahrener Notärzte konnte erstmals gezeigt werden, dass technische Unfallparameter isoliert keine sichere Vorhersage der Verletzungsschwere zulassen. Ob technische Parameter zusammen mit medizinischen Parametern eine verbesserte erste Einschätzung ermöglichen, muss Ziel weiterer Untersuchungen sein.Technical parameters of road traffic crashes are routinely reported during transfer of patients from emergency medical services to the hospital. It is unknown whether this information contributes to an improvement in the estimation of injury severity. To clarify this matter, questionnaires containing data of 100 cases from the database of the in-depth accident research project in Greifswald were presented to experienced emergency physicians. The technical parameters presented were grouped according to complexity of information, ranging from routine parameters to photo documentation of the crash scene. Three emergency physicians were asked to appraise the injury severity of vehicle drivers according to four ISS-based categories (“moderate”, “severe”, “life-threatening” and “lethal”). For analysis data was dichotomized in “moderate and severe” (ISS ≤ 15) versus “life-threatening and lethal” (ISS 16-75). Statistical analysis comprised inter-rater agreement beyond chance (kappa values) and indicators of diagnostic test accuracy (sensitivity, specificity, area under the ROC-curves, likelihood ratios). The inter-rater agreement of injury severity based on technical crash information was moderate to substantial (kappa 0.42–0.66). Amongst the three observers and various amounts of technical data, sensitivity ranged between 18 and 80%, and specificity ranged between 41 and 89% in predicting the presence of major trauma. Presentation of photographs of the crash scene in¬creased diagnostic accuracy. Still, the pre¬sented information led to a shift from a 50% prior probability of life-threatening injuries to a maximum of 40% in the negative and 67% in the positive case. With this extensive study utilizing real-crash data and experienced emergency physicians it could be demonstrated that technical crash parameters itself do not allow for estimation of a vehicle driver’s injury severity. Whether a combination of medical and technical parameters facilitates a better estimation should be subject of further research

Digital Competencies for Science Teaching : Adapting the DiKoLAN Framework to Teacher Education in Switzerland

Prospective teachers need both general and subject-specific digital competencies. However, available competency frameworks usually address only non-subject-specific general digital competencies. Although the joint framework curriculum of the German-speaking cantons of Switzerland not only provides a comprehensive definition of expected digital student competencies, but also suggestions for linking them to and implementing them in subject lessons, there is still no integral planning and structuring for subject-specific teacher training in the canton of Thurgau. For a curricular design of the promotion of subject-specific digital competencies, three competency frameworks for the study programs Primary level, Secondary level 1, and Secondary level 2 at the Thurgau University of Education were derived in expert interviews based on the DiKoLAN framework (Digital Competencies for Teaching in Science Education). While there are significant overlaps, the original reference framework clearly needs to be adapted. Furthermore, there are important commonalities between the newly formed individual competency frameworks. The methodological approach has proven to be fruitful and is recommended to follow-up studies and studies with similar research questions for imitation.publishe

Implementation of the World Health Organization Trauma Care Checklist Program in 11 centers across multiple economic strata: effect on care process measures

BACKGROUND: Trauma contributes more than ten percent of the global burden of disease. Initial assessment and resuscitation of trauma patients often requires rapid diagnosis and management of multiple concurrent complex conditions, and errors are common. We investigated whether implementing a trauma care checklist would improve care for injured patients in low-, middle-, and high-income countries. METHODS: From 2010 to 2012, the impact of the World Health Organization (WHO) Trauma Care Checklist program was assessed in 11 hospitals using a stepped wedge pre- and post-intervention comparison with randomly assigned intervention start dates. Study sites represented nine countries with diverse economic and geographic contexts. Primary end points were adherence to process of care measures; secondary data on morbidity and mortality were also collected. Multilevel logistic regression models examined differences in measures pre- versus post-intervention, accounting for patient age, gender, injury severity, and center-specific variability. RESULTS: Data were collected on 1641 patients before and 1781 after program implementation. Patient age (mean 34 ± 18 vs. 34 ± 18), sex (21 vs. 22 % female), and the proportion of patients with injury severity scores (ISS) ≥ 25 (10 vs. 10 %) were similar before and after checklist implementation (p \u3e 0.05). Improvement was found for 18 of 19 process measures, including greater odds of having abdominal examination (OR 3.26), chest auscultation (OR 2.68), and distal pulse examination (OR 2.33) (all p \u3c 0.05). These changes were robust to several sensitivity analyses. CONCLUSIONS: Implementation of the WHO Trauma Care Checklist was associated with substantial improvements in patient care process measures among a cohort of patients in diverse settings