Autoantikörpernachweis mittels indirekter Immunfluoreszenz an HEp-2-Zellen

Systemic autoimmune diseases are characterized by the presence of antinuclear autoantibodies (ANAs). Diluted patient sera are typically used to screen for the presence of ANAs by immunofluorescence microscopy with fixed HEp-2 cells. Despite high quality test kits, reports of different laboratories frequently present controversial results. This study presents a recommendation for a unified processing and interpretation of HEp-2 based screening for autoantibodies. We provide suggestions for selection of high quality test kits, optimized processing, and diagnostic procedures. For good laboratory practice, in addition to a relevant clinical diagnosis and an experienced laboratory specialist, the following procedure is highly recommended: initial HEp-2 based screening by indirect immunofluorescence, starting with a 1:80 serum dilution and evaluation in a bright fluorescence microscope, pathological values from a titer of 1:160, internal quality checks, and unified interpretation. We aim to improve diagnostics and care for patients with autoimmune diseases as a central focus of the European Autoimmunity Standardization Initiative (EASI)

Potential value of automated daily screening of cardiac resynchronization therapy defibrillator diagnostics for prediction of major cardiovascular events: results from Home-CARE (Home Monitoring in Cardiac Resynchronization Therapy) study

Aim To investigate whether diagnostic data from implanted cardiac resynchronization therapy defibrillators (CRT-Ds) retrieved automatically at 24 h intervals via a Home Monitoring function can enable dynamic prediction of cardiovascular hospitalization and death. Methods and results Three hundred and seventy-seven heart failure patients received CRT-Ds with Home Monitoring option. Data on all deaths and hospitalizations due to cardiovascular reasons and Home Monitoring data were collected prospectively during 1-year follow-up to develop a predictive algorithm with a predefined specificity of 99.5%. Seven parameters were included in the algorithm: mean heart rate over 24 h, heart rate at rest, patient activity, frequency of ventricular extrasystoles, atrial–atrial intervals (heart rate variability), right ventricular pacing impedance, and painless shock impedance. The algorithm was developed using a 25-day monitoring window ending 3 days before hospitalization or death. While the retrospective sensitivities of the individual parameters ranged from 23.6 to 50.0%, the combination of all parameters was 65.4% sensitive in detecting cardiovascular hospitalizations and deaths with 99.5% specificity (corresponding to 1.83 false-positive detections per patient-year of follow-up). The estimated relative risk of an event was 7.15-fold higher after a positive predictor finding than after a negative predictor finding. Conclusion We developed an automated algorithm for dynamic prediction of cardiovascular events in patients treated with CRT-D devices capable of daily transmission of their diagnostic data via Home Monitoring. This tool may increase patients’ quality of life and reduce morbidity, mortality, and health economic burden, it now warrants prospective studies

Large expert-curated database for benchmarking document similarity detection in biomedical literature search

Document recommendation systems for locating relevant literature have mostly relied on methods developed a decade ago. This is largely due to the lack of a large offline gold-standard benchmark of relevant documents that cover a variety of research fields such that newly developed literature search techniques can be compared, improved and translated into practice. To overcome this bottleneck, we have established the RElevant LIterature SearcH consortium consisting of more than 1500 scientists from 84 countries, who have collectively annotated the relevance of over 180 000 PubMed-listed articles with regard to their respective seed (input) article/s. The majority of annotations were contributed by highly experienced, original authors of the seed articles. The collected data cover 76% of all unique PubMed Medical Subject Headings descriptors. No systematic biases were observed across different experience levels, research fields or time spent on annotations. More importantly, annotations of the same document pairs contributed by different scientists were highly concordant. We further show that the three representative baseline methods used to generate recommended articles for evaluation (Okapi Best Matching 25, Term Frequency-Inverse Document Frequency and PubMed Related Articles) had similar overall performances. Additionally, we found that these methods each tend to produce distinct collections of recommended articles, suggesting that a hybrid method may be required to completely capture all relevant articles. The established database server located at https://relishdb.ict.griffith.edu.au is freely available for the downloading of annotation data and the blind testing of new methods. We expect that this benchmark will be useful for stimulating the development of new powerful techniques for title and title/abstract-based search engines for relevant articles in biomedical research.Peer reviewe

Guidelines for the use and interpretation of assays for monitoring autophagy (3rd edition)

In 2008 we published the first set of guidelines for standardizing research in autophagy. Since then, research on this topic has continued to accelerate, and many new scientists have entered the field. Our knowledge base and relevant new technologies have also been expanding. Accordingly, it is important to update these guidelines for monitoring autophagy in different organisms. Various reviews have described the range of assays that have been used for this purpose. Nevertheless, there continues to be confusion regarding acceptable methods to measure autophagy, especially in multicellular eukaryotes. For example, a key point that needs to be emphasized is that there is a difference between measurements that monitor the numbers or volume of autophagic elements (e.g., autophagosomes or autolysosomes) at any stage of the autophagic process versus those that measure fl ux through the autophagy pathway (i.e., the complete process including the amount and rate of cargo sequestered and degraded). In particular, a block in macroautophagy that results in autophagosome accumulation must be differentiated from stimuli that increase autophagic activity, defi ned as increased autophagy induction coupled with increased delivery to, and degradation within, lysosomes (inmost higher eukaryotes and some protists such as Dictyostelium ) or the vacuole (in plants and fungi). In other words, it is especially important that investigators new to the fi eld understand that the appearance of more autophagosomes does not necessarily equate with more autophagy. In fact, in many cases, autophagosomes accumulate because of a block in trafficking to lysosomes without a concomitant change in autophagosome biogenesis, whereas an increase in autolysosomes may reflect a reduction in degradative activity. It is worth emphasizing here that lysosomal digestion is a stage of autophagy and evaluating its competence is a crucial part of the evaluation of autophagic flux, or complete autophagy. Here, we present a set of guidelines for the selection and interpretation of methods for use by investigators who aim to examine macroautophagy and related processes, as well as for reviewers who need to provide realistic and reasonable critiques of papers that are focused on these processes. These guidelines are not meant to be a formulaic set of rules, because the appropriate assays depend in part on the question being asked and the system being used. In addition, we emphasize that no individual assay is guaranteed to be the most appropriate one in every situation, and we strongly recommend the use of multiple assays to monitor autophagy. Along these lines, because of the potential for pleiotropic effects due to blocking autophagy through genetic manipulation it is imperative to delete or knock down more than one autophagy-related gene. In addition, some individual Atg proteins, or groups of proteins, are involved in other cellular pathways so not all Atg proteins can be used as a specific marker for an autophagic process. In these guidelines, we consider these various methods of assessing autophagy and what information can, or cannot, be obtained from them. Finally, by discussing the merits and limits of particular autophagy assays, we hope to encourage technical innovation in the field

Messung der Energieverlustfunktion von 18.6 keV Elektronen an Deuterium Gas und Bestimmung des Q-Werts von Tritium am KATRIN Experiment

Das Karlsruher Tritium Neutrino Experiment KATRIN hat das Ziel, die Masse des Elektron-Antineutrinos mit einer Sensitivität von 0.2 eV/c^2 zu bestimmen. Ein Hauptziel dieser Arbeit ist die Untersuchung der absoluten Energieskala von ATRIN um den Q-Wert des β-Zerfalls von Tritium zu bestimmen. Um das Potenzial in der Analysierebene untersuchen zu können, wurde eine winkelselektive, monoenergetische Photoelektronenquelle (‘e-gun’) in dieser Arbeit zusammengebaut, kommissioniert und charakterisiert. Die Fowler Methode wird erklärt und verwendet um die Austrittsarbeit der e-gun und der ‘rear wall’ zu bestimmen. Der Tritium-Q-Wert wird mit den Daten der ersten Tritium Kommissionierungsphase (FT) von KATRIN und für die erste Neutrino-Messkampagne (KNM1) bestimmt. Die e-gun wird zudem zur Bestimmung der Energieverlustfunktion von Deuterium verwendet. Basierend auf integralen- und Flugzeitmessungen wird eine neue Parametrisierung der Energieverlustfunktion bestimmt.The Karlsruhe tritium neutrino experiment KATRIN aims to measure the mass of the electron antineutrino with a sensitivity of 0.2 eV/c 2 with a model-independent approach. A main goal of this thesis is to investigate the absolute energy scale of the KATRIN experiment and to determine the Q-value of tritium β-decay. To investigate the potential in the analyzing plane an angular selective, monoenergetic photoelectron source, referred to as ’e-gun’, is assembled, commissioned, and characterized during this work. The Fowler method is introduced and used to determine the work function of the e-gun and the ‘rear wall’. The e-gun is also used to measure the energy loss function, which describes the energy loss of signal electrons passing the gaseous source. The measurement is performed on deuterium gas with an integrating and a time of flight method. A new model parametrization of the energy loss function is created and fitted to the data

Alien plant species with a wider global distribution are better able to capitalize on increased resource availability

A high ability of alien plant species to capitalize on increases in resource availability has been suggested as an explanation for being globally successful. Here, we tested this hypothesis meta-analytically using existing data from experiments manipulating plant resources (light, water and nutrients).From these studies we extracted the response to resource increase of biomass, as an indicator of plant performance, and the responses of two traits related to resource capture: root : shoot ratio and specific leaf area (SLA). For 211 species recorded in the Global Compendium of Weeds, we assessed the relationship between effect sizes from such studies and the number of global regions where a species was established.We found that globally widespread species exhibited greater biomass responses to increases in resources overall, compared to less widespread species. Root : shoot ratio and SLA responses to increased resource availability were not related to species global distribution.In general, globally widespread alien plant species were better able to capitalize on increased availability of resources, through achieving increased growth and biomass accumulation, while greater plasticity of key resource-capture traits per se did not appear to be related to greater success