A cycle of Vam7p release from and PtdIns 3-P–dependent rebinding to the yeast vacuole is required for homotypic vacuole fusion

Vacuole fusion requires a coordinated cascade of priming, docking, and fusion. SNARE proteins have been implicated in the fusion itself, although their precise role in the cascade remains unclear. We now report that the vacuolar SNAP-23 homologue Vam7p is a mobile element of the SNARE complex, which moves from an initial association with the cis-SNARE complex via a soluble intermediate to the docking site. Soluble Vam7p is specifically recruited to vacuoles and can rescue a fusion reaction poisoned with antibodies to Vam7p. Both the recombinant Vam7p PX domain and a FYVE domain construct of human Hrs block the recruitment of Vam7p and vacuole fusion, demonstrating that phosphatidylinositol 3-phosphate is a primary receptor of Vam7p on vacuoles. We propose that the Vam7p cycle is linked to the availability of a lipid domain on yeast vacuoles, which is essential for coordinating the fusion reaction prior to and beyond docking

Spatial and temporal variations of microorganisms in grassland soils : influences of land-use intensity, plants and soil properties

Grassland ecosystems provide a wide range of services to human societies (Allan et al., 2015) and plants and soil microorganisms have been identified as key drivers of ecosystem functioning (Soliveres et al., 2016). Therefore, understanding soil microbial distributions and processes in agricultural grassland soils is crucial for characterizing these ecosystems and for predicting how they may shift in a changing environment. Yet we are only beginning to understand these complex ecosystems, which account for about 26% of the worlds terrestrial surface (FAOSTATS, 2018), making it especially urgent to gain better insights into the effects of land-use intensity on soil microbial properties and plant-microbe interactions. This thesis was conducted to evaluate the impact land-use intensity has on soil microbial biogeography of grasslands with respect to both spatial patterns and temporal changes in soil microbial abundance, function (in terms of enzyme activities), and community composition. It also investigated the relationships between plants and the spatial and temporal distributions of soil microorganisms. Thereby both, land-use intensity effects and plant-microbe interactions, were assessed in light of ecological niche and neutral theory. This thesis is based on three observational studies conducted on from one to 150 continuously farmed, un-manipulated grassland sites in three regions of Germany within the Biodiversity Exploratories project (DFG priority program 1374). The first study assessed the effects of land-use intensity and physico-chemical soil properties on the spatial biogeography of soil microbial abundance and function in 18 grasslands sites from two of the three regions, sampled at one time point. The second study analyzed spatial and temporal distributions of alpha- and beta-diversity of arbuscular mycorrhizal fungi in a low land-use intensity grassland with six sampling time points across one season. The third study investigated both legacy and short-term change effects of land-use intensity, soil physico-chemical properties, plant functional traits, and plant biomass properties on temporal changes in soil microbial abundance, function, and community composition in 150 grassland sites across three regions, with particular regard to direct and indirect land-use intensity effects. Although the three studies used different approaches and assessed different soil microbial properties, general patterns were detectable. Abiotic soil properties, namely pH, nitrogen content, texture, and bulk density played fundamental roles for spatial and temporal microbial biogeography. Since these factors were specific and unique for each investigated site, they formed the background based on which other processes occurred. In addition to abiotic soil properties, impacts of land-use intensity and plants were detected, though to various degrees in the three studies. Land-use intensity played a much smaller role than anticipated in the first and third study. No influence on the spatial distribution of soil microbial abundance and function could be detected in the first study. In the third study, short-term changes in and legacy effects of land-use intensity played a minor role with respect to short-term changes in soil microbial abundance, function, and community composition. Where detected, changes in land-use intensity had a direct and negative effect on soil microbial properties in structural equation modelling; i.e., increases in land-use intensity reduced, e.g., soil microbial enzyme activities, while legacy effects of land-use intensity were shown to act both directly and indirectly on soil microbial properties. Thereby indirect legacy effects were mediated via plant functional traits. Only one of the three studies detected minor plant diversity effects on soil microbial properties. Instead, functional properties of the plant communities, i.e., plant functional traits, biomass, and nutritional quality, were significantly related to spatial and temporal distributions of soil microorganisms. Finally, the findings of the three studies suggest that processes related to niche and neutral theory both drive spatial and temporal patterns of soil microbial properties at the investigated plot scale (up to 50 m × 50 m). This thesis concluded that in order to gain deeper insights into the complex functions and processes occurring in grassland ecosystems, a multidisciplinary approach investigating fundamental physico-chemical site characteristics, microbial soil properties, and plants is necessary. The results of the thesis suggest that focus be turned to functional properties of plant and microbial communities, as they are closely intermingled, provide more detailed insights into plant-microbe interactions, and are able to reflect effects of human impacts on grassland soils better than diversity measures.Die vorliegende Dissertation diente dem Zweck, die Einflüsse der Landnutzungsintensität von Grünländern auf die Biogeographie von Bodenmikroorganismen in Bezug auf die räumliche Verteilung und zeitliche Veränderung von mikrobieller Biomasse, Funktion und Gemeinschaftsstrukturen aufzudecken und die Beziehungen zwischen Pflanzen und den räumlichen und zeitlichen Verteilungsmustern von Bodenmikroorganismen zu untersuchen. Dabei wurde auch betrachtet, inwieweit die Verteilung der Mikroorganismen den Konzepten der ökologischen Nischen- und Neutraltheorie entspricht. Die Dissertation basiert auf drei beobachtenden Studien, die auf einer bis 150 dauerhaft bewirtschafteten und nicht manipulierten Grünlandflächen durchgeführt wurden. Die Flächen sind verteilt auf drei Regionen in Deutschland und Teil des Schwerpunktforschungsprogramms Biodiversitäts-Exploratorien der Deutschen Forschungsgemeinschaft (DFG 1374). Die erste Studie untersuchte die Effekte von Landnutzungsintensität und physikalisch-chemischen Bodeneigenschaften auf die räumliche Biogeographie von mikrobieller Biomasse und Funktion zu einem einzelnen Zeitpunkt in 18 Grünlandflächen, die auf zwei Regionen verteilt sind. Die zweite Studie analysierte die räumliche und zeitliche Verteilung der Alpha- und Betadiversität arbuskulärer Mykorrhizapilze (AMF) in einem extensiv genutzten Grünland mittels sechs Probennahmezeitpunkten verteilt über die Vegetationsperiode eines Jahres. Die dritte Studie untersuchte sogenannte Legacy und kurzfristige Effekte von veränderter Landnutzungsintensität, physikalisch-chemischen Bodeneigenschaften, funktionellen Pflanzeneigenschaften und Charakteristika der pflanzlichen Biomasse auf zeitliche Veränderungen von mikrobieller Biomasse, Funktion und Gemeinschaftsstruktur in 150 Grünlandböden verteilt auf drei Regionen, sowie direkte und indirekte Effekte der Landnutzungsintensität. Obwohl alle drei Studien unterschiedliche Ansätze verfolgten und verschiedene bodenmikrobielle Eigenschaften untersuchten, sind generelle Muster erkennbar: abiotische Bodeneigenschaften, namentlich pH-Wert, Stickstoffgehalt, Textur und Lagerungsdichte, waren für die räumliche und zeitliche Biogeographie der Bodenmikroorganismen von fundamentaler Bedeutung. Sie waren spezifisch für jede der untersuchten Flächen und bildeten den Hintergrund, vor dem sich andere Prozesse abspielten. Zusätzlich zu den abiotischen Bodeneigenschaften wurden, wenn auch in unterschiedlichem Maße, Einflüsse von Landnutzungsintensität und Pflanzen in den drei Studien detektiert. Die Landnutzungsintensität spielte dabei eine wesentlich geringere Rolle als ursprünglich angenommen. Sie hatte keinen Einfluss auf die räumliche Verteilung der bodenmikrobiellen Biomasse oder Funktion in der ersten Studie. In der dritten Studie waren kurzfristige Veränderungen und Legacy-Effekte nur in geringem Maße mit kurzfristigen Veränderungen von bodenmikrobieller Biomasse, Funktion und Gemeinschaftsstruktur assoziiert. Die Strukturgleichungsmodelle zeigen, dass sich die Effekte von kurzfristigen Veränderung der Landnutzungsintensität, dort wo sie auftraten, direkt und negative auf die kurzfristigen Veränderungen bodenmikrobieller Eigenschaften auswirkten. Dahingegen wirkten sich Legacy-Effekte der Landnutzungsintensität sowohl direkt als auch indirekt auf bodenmikrobielle Eigenschaften aus. Die indirekten Legacy-Effekte wurden dabei über funktionelle Pflanzeneigenschaften auf die Mikroorganismen übertragen. Nur eine der drei Studien fand einen marginalen Einfluss der Pflanzendiversität auf die bodenmikrobiellen Eigenschaften. Stattdessen waren funktionelle Eigenschaften der Pflanzengemeinschaften sowie deren Biomasse und Futterqualität signifikant mit der räumlichen und zeitlichen Verteilung von Bodenmikroorganismen verbunden. In Bezug auf die Nischen- und Neutraltheorie sprechen die Ergebnisse der drei Studien dafür, dass sowohl Prozesse, die mit der Nischentheorie zusammenhängen, als auch solche, die mit der Neutraltheorie in Verbindung stehen, die räumliche und zeitliche Verteilung von bodenmikrobiellen Eigenschaften auf der untersuchten Plotskala (bis 50 m × 50 m) steuern. Diese Dissertation zieht die Schlussfolgerung, dass multidisziplinäre Forschung notwendig ist, um die komplexen Funktionen und Prozesse von Grünlandökosystemen zu erforschen. Diese müssen sowohl die fundamentalen physikalischen und chemischen Eigenschaften der Böden, als auch die Eigenschaften der Bodenmikroorganismen und Pflanzengemeinschaften umfassen. Dabei sprechen die Ergebnisse dieser Arbeit dafür, dass ein besonderes Augenmerk auf die funktionellen Eigenschaften von Pflanzen- und Mikrobengemeinschaften gelegt werden sollte, da diese eng miteinander verflochten sind und bessere Einblicke in die Interaktionen zwischen Pflanzen und Mikroorganismen gewähren sowie besser in der Lage sind die Effekte von menschlichen Einflüssen auf Grünlandböden wieder zu spiegeln, als die bisher oft üblichen Diversitätsmessungen

Unraveling spatiotemporal variability of arbuscular mycorrhizal fungi in a temperate grassland plot

© The Author(s), 2019. This article is distributed under the terms of the Creative Commons Attribution License. The definitive version was published in Goldmann, K., Boeddinghaus, R. S., Klemmer, S., Regan, K. M., Heintz-Buschart, A., Fischer, M., Prati, D., Piepho, H., Berner, D., Marhan, S., Kandeler, E., Buscot, F., & Wubet, T. Unraveling spatiotemporal variability of arbuscular mycorrhizal fungi in a temperate grassland plot. Environmental Microbiology, 22(3),(2020): 873-888, doi:10.1111/1462-2920.14653.Soils provide a heterogeneous environment varying in space and time; consequently, the biodiversity of soil microorganisms also differs spatially and temporally. For soil microbes tightly associated with plant roots, such as arbuscular mycorrhizal fungi (AMF), the diversity of plant partners and seasonal variability in trophic exchanges between the symbionts introduce additional heterogeneity. To clarify the impact of such heterogeneity, we investigated spatiotemporal variation in AMF diversity on a plot scale (10 × 10 m) in a grassland managed at low intensity in southwest Germany. AMF diversity was determined using 18S rDNA pyrosequencing analysis of 360 soil samples taken at six time points within a year. We observed high AMF alpha‐ and beta‐diversity across the plot and at all investigated time points. Relationships were detected between spatiotemporal variation in AMF OTU richness and plant species richness, root biomass, minimal changes in soil texture and pH. The plot was characterized by high AMF turnover rates with a positive spatiotemporal relationship for AMF beta‐diversity. However, environmental variables explained only ≈20% of the variation in AMF communities. This indicates that the observed spatiotemporal richness and community variability of AMF was largely independent of the abiotic environment, but related to plant properties and the cooccurring microbiome.We thank the managers of the three Exploratories, Kirsten Reichel‐Jung, Swen Renner, Katrin Hartwich, Sonja Gockel, Kerstin Wiesner, and Martin Gorke for their work in maintaining the plot and project infrastructure; Christiane Fischer and Simone Pfeiffer for giving support through the central office, Michael Owonibi and Andreas Ostrowski for managing the central data base, and Eduard Linsenmair, Dominik Hessenmöller, Jens Nieschulze, Ernst‐Detlef Schulze, Wolfgang W. Weisser and the late Elisabeth Kalko for their role in setting up the Biodiversity Exploratories project. The work has been funded by the DFG Priority Program 1374 ‘Infrastructure‐Biodiversity‐Exploratories’ (BU 941/22‐1, BU 941/22‐3, KA 1590/8‐2, KA 1590/8‐3). Field work permits were issued by the responsible state environmental office of Baden‐Württemberg (according to § 72 BbgNatSchG). Likewise, we kindly thank Beatrix Schnabel, Melanie Günther and Sigrid Härtling for 454 sequencing in Halle. AHB gratefully acknowledges the support of the German Centre for Integrative Biodiversity Research (iDiv) Halle‐Jena‐Leipzig funded by the German Research Foundation (FZT 118). Authors declare no conflict of interests

Baseline high sensitivity cardiac troponin I level below limit of quantitation rules out acute myocardial infarction in the emergency department

The objective of our study was to determine the utility of a baseline high sensitivity cardiac troponin (hs-cTnI) value below the limit of quantitation to rule out acute myocardial infarction (AMI) in patients presenting to the emergency department with any suspicious symptoms of a cardiac etiology. We enrolled subjects presenting to the Emergency Department with symptoms suspicious for AMI. Blood specimens were collected within one hour after a triage electrocardiogram. Cardiac troponin I was measured using the Beckman Coulter Access hs-cTnI assay. The diagnosis of AMI was adjudicated by two cardiologists using the Third Universal Definition of AMI and Roche Diagnostics Troponin T Generation 5 assay with all available clinical data at 30 days after presentation. A total of 567 subjects had all data required for data analyses. AMI was diagnosed in 46 (8.1%) patients. 232 (40.9%) individuals had presentation hs-cTnI results \u3c 4.0 ng/L. None of the patients with baseline hs-cTnI \u3c 4.0 ng/L had an AMI, yielding a negative predictive value of 100.0% and a sensitivity of 100%, and a good prognosis (no AMIs or cardiac-related deaths at 30 days). In this single center emergency department study, a baseline presenting novel hs-cTnI value of \u3c 4.0 ng/L effectively ruled out AMI in 40.9% of all patients presenting to the emergency department and having any symptoms suspicious for AMI. Importantly all patients, not only those with chest pain, and those having symptoms for any duration or those with end-stage renal disease requiring dialysis were included

Update on high-sensitivity cardiac troponin in patients with suspected myocardial infarction

High-sensitivity cardiac troponin (hs-cTn) assays have been used clinically in many countries throughout the world for years and complement detailed clinical assessment and the electrocardiogram in the early diagnosis of myocardial infarction (MI). High-sensitivity cardiac troponin assays for the first time allow to precisely quantify cardiomyocyte injury around the 99th percentile and thereby substantially increase the accuracy for MI already for blood draws obtained at presentation to the emergency department (ED). Higher diagnostic accuracy at ED presentation enabled the development and extensive validation of early hs-cTn-based diagnostic algorithms, which substantially reduced the time required for safe rule-out or rule-in of MI

Divergent drivers of the microbial methane sink in temperate forest and grassland soils

Aerated topsoils are important sinks for atmospheric methane (CH4) via oxidation by CH4‐oxidizing bacteria (MOB). However, intensified management of grasslands and forests may reduce the CH4 sink capacity of soils. We investigated the influence of grassland land‐use intensity (150 sites) and forest management type (149 sites) on potential atmospheric CH4 oxidation rates (PMORs) and the abundance and diversity of MOB (with qPCR) in topsoils of three temperate regions in Germany. PMORs measurements in microcosms under defined conditions yielded approximately twice as much CH4 oxidation in forest than in grassland soils. High land‐use intensity of grasslands had a negative effect on PMORs (−40%) in almost all regions and fertilization was the predominant factor of grassland land‐use intensity leading to PMOR reduction by 20%. In contrast, forest management did not affect PMORs in forest soils. Upland soil cluster (USC)‐α was the dominant group of MOBs in the forests. In contrast, USC‐γ was absent in more than half of the forest soils but present in almost all grassland soils. USC‐α abundance had a direct positive effect on PMOR in forest, while in grasslands USC‐α and USC‐γ abundance affected PMOR positively with a more pronounced contribution of USC‐γ than USC‐α. Soil bulk density negatively influenced PMOR in both forests and grasslands. We further found that the response of the PMORs to pH, soil texture, soil water holding capacity and organic carbon and nitrogen content differ between temperate forest and grassland soils. pH had no direct effects on PMOR, but indirect ones via the MOB abundances, showing a negative effect on USC‐α, and a positive on USC‐γ abundance. We conclude that reduction in grassland land‐use intensity and afforestation has the potential to increase the CH4 sink function of soils and that different parameters determine the microbial methane sink in forest and grassland soils.Deutsche Forschungsgemeinschaft http://dx.doi.org/10.13039/501100001659ESFMinistry of Education, Science and Culture of Mecklenburg‐Western PomeraniaPeer Reviewe

Trends in Coronary and Structural Heart Interventions in Switzerland over the Last 16 Years and Impact of COVID-19: Insights from the National Swiss PCI Survey.

BACKGROUND Considering the global burden of cardiovascular disease, we analysed trends in interventional coronary and structural procedures over the past 16 years (2005-2021), using continuous data from the Swiss national registry. METHODS Based on a standardised questionnaire, data on coronary and structural interventions in Switzerland were assessed by the Working Group Interventional Cardiology of the Swiss Society of Cardiology (SSC). Here, we analysed the trend of annually performed interventions from 2005 to 2021 in Switzerland and the impact of the COVID-19 pandemic. RESULTS We observed a constant increase in the total number of cases (including coronary angiographies (CA) and percutaneous coronary interventions (PCI)) from 36,436 cases in 2005 to 56,555 cases in 2021 (+55%). With 18 cases in 2007, TAVI procedures have increased to 2004 cases in 2021. During the early phase of the COVID-19 pandemic in 2020, a slight decrease in CAs and PCIs of 9.15% was observed. In contrast, we did not observe an impact of the COVID-19 pandemic on the number of no TAVI procedures. Most importantly, all cause in-hospital mortality for coronary interventions before and during the peak of the COVID-19 pandemic was comparable (1.4% vs. 1.3%). CONCLUSION Over a 16-year period, we observed an upward trend in diagnostic and therapeutic procedures for coronary as well as structural heart disease, with only a small short-term impact of the COVID-19 pandemic on interventions and a similar procedure-related in-hospital-mortality in Switzerland

Effects of warming and drought on potential N2O emissions and denitrifying bacteria abundance in grasslands with different land-use

Increased warming in spring and prolonged summer drought may alter soil microbial denitrification. We measured potential denitrification activity and denitrifier marker gene abundances (nirK, nirS, nosZ) in grasslands soils in three geographic regions characterized by site-specific land-use indices (LUI) after warming in spring, at an intermediate sampling and after summer drought. Potential denitrification was significantly increased by warming, but did not persist over the intermediate sampling. At the intermediate sampling, the relevance of grassland land-use intensity was reflected by increased potential N2O production at sites with higher LUI. Abundances of total bacteria did not respond to experimental warming or drought treatments, displaying resilience to minor and short-term effects of climate change. In contrast, nirS- and nirK-type denitrifiers were more influenced by drought in combination with LUI and pH, while the nosZ abundance responded to the summer drought manipulation. Land-use was a strong driver for potential denitrification as grasslands with higher LUI also had greater potentials for N2O emissions. We conclude that both warming and drought affected the denitrifying communities and the potential denitrification in grassland soils. However, these effects are overruled by regional and site-specific differences in soil chemical and physical properties which are also related to grassland land-use intensit

Confounding Factors Affecting the Marginal Quality of an Intra-Oral Scan

Objectives: To assess the effect of clinical factors on the quality of intra-oral scans of crown margins. These factors are; presence of adjacent teeth, proximity to gingivae, encumbrance of wand positioning within oral cavity. Methods: A typodont lower molar (Frasaco, Germany) was prepared for an all-ceramic crown with 1.5 mm supraginigival (lingual) and equigingival (buccal) margins. The tooth was scanned in a model scanner, creating a master scan. An intra-oral scanner (IOS) (Omnicam, Sirona Dental) was used to acquire sets of 5 scans each, under varying conditions; (1) the presence/absence of adjacent teeth, (2) model mounted in manikin head/hand-held, (3) with/without a 1 mm shim to elevate the margin. Every combination was investigated, yielding 40 scans (8 groups of 5). The master scan margin was identified by selecting the highest curvature region (>1.8). The master was aligned to each IOS scan, and 4 regions of each IOS scan margin were extracted, lying within 100 μm of predefined mesial, distal, buccal and lingual sections of the master margin. The mean curvature of each margin section was calculated using Meshlab. The effect of each confounding factor on margin curvature was analysed using ANOVA. Results: Lingual margin curvature remained consistent regardless of scanning conditions. Buccal margin curvature was significantly affected when located equigingivally. Mesial margin curvature was significantly affected in the presence of adjacent teeth and proximity to the gingivae. Distal margin curvature was significantly affected by all three confounding factors. Conclusions: The curvature (sharpness) of the margin recorded by a commercial IOS is significantly affected by clinical factors obscuring visibility