Microsatellite Diversity of the Agriculturally Important Alpine Grass Poa alpina in Relation to Land Use and Natural Environment

Background and Aims The Alpine Meadow Grass Poa alpina is common in subalpine and alpine natural sites and agriculturally used land, where it is an important fodder grass. Natural factors and human land use are supposed to have been shaping its genetic diversity for hundreds of years. The species comprises sexually and vegetatively reproducing plants. The aim of this study was to investigate the effects of agricultural land use, environmental factors and the mode of reproduction on the distribution of its microsatellite diversity within and among populations and to analyse whether its genetic diversity is correlated with plant species diversity in grassland parcels. Methods Genetic diversity of P. alpina was assessed with five microsatellite markers for 569 plants originating from 20 natural sites and from 54 grassland parcels of different cultural tradition, land use and altitude in the Swiss Alps. Due to polyploidy and frequent aneuploidy of the species, data analyses were based on the presence of microsatellite bands. Key Results A low but significant differentiation was found in microsatellite bands among natural sites and agriculturally used parcels, while their microsatellite band diversity within populations did not differ. An increased differentiation was found in microsatellite bands with increasing geographic distance among parcels, and a differentiation among grazed and mown parcels, and among sexually and vegetatively reproducing populations. Band richness of sampled plants per village was higher for villages where parcels represented more different land-use types. Within populations, microsatellite band diversity was higher in grazed than in mown parcels. Conclusions The diversity of human land use in the Alps was associated with genetic diversity of P. alpina. Therefore, the ongoing socio-economically motivated land-use changes, which reduce the number of different land-use types, will affect the genetic diversity of P. alpina negativel

Adaptation of Poa alpina to altitude and land use in the Swiss Alps

Current land use and climate change are prompting questions about the ability of plants to adapt to such environmental change. Therefore, we experimentally addressed plant performance and quantitative-genetic diversity of the common Alpine Meadow Grass Poa alpina. We asked how land use and altitude affect the occurrence of P. alpina in the field and whether its common-garden performance suggests adaptation to conditions at plant origin and differences in quantitative genetic diversity among plant origins. Among 216 candidate grassland sites of different land use and altitude from 12 villages in the Swiss Alps, P. alpina occurred preferentially in fertilized and grazed sites and at higher elevations. In a common garden at 1,500m asl, we grew two plants of >600 genotypes representing 78 grassland sites. After 2years, nearly 90% of all plants had reproduced. In agreement with adaptive advantages of vegetative reproduction at higher altitudes, only 23% of reproductive plants from lower altitudes reproduced via vegetative bulbils, but 55% of plants from higher altitudes. In agreement with adaptive advantages of reproduction in grazed sites, allocation to reproductive biomass was higher in plants from grazed grasslands than from mown ones. For 53 grasslands, we also investigated broad-sense heritability H2, which was significant for all studied traits and twice as high for grazed as for mown grasslands. Moreover, possibly associated with their higher landscape diversity, H2 was higher for sites of villages of Romanic cultural tradition than for those of Germanic and Walser traditions. We suggest promoting diverse land use regimes to conserve not only landscape and plant species diversity, but also adaptive genetic differentiation and heritable genetic variatio

Omnipresence of leaf herbivory by invertebrates and leaf infection by fungal pathogens in agriculturally used grasslands of the Swiss Alps, but low plant damage

Agriculturally used grasslands in the Alps are characterised by a trade-off between high fodder production in some and high plant species richness in others. In contrast to plant species richness and production, however, little is known on the relevance of biological interactions between plants, invertebrate herbivores, and fungal pathogens for grasslands in the Alps. At the time when the vegetation was fully developed, but prior to agricultural use, we examined whether leaf damage by herbivory and fungal pathogen infection, and their diversity, are affected by plant functional group, land use, and altitude. Moreover, we studied whether extent and diversity of leaf damage are related to each other, to plant species richness, and to standing crop. We recorded the leaf area damaged by ten types of herbivory and five types of fungal pathogen infection on 12,054 plant leaves of legumes, other forbs, and graminoids collected in 215 grassland parcels in 12 valleys in the Swiss Alps. With 83% of all leaves infested, herbivory and fungal pathogen infection were omnipresent. However, only 2.7% leaf area was damaged by herbivory and 1.2% by fungal pathogens. Damage by herbivory was highest on legumes, and damage by fungal pathogens was highest on graminoids. More leaf damage by herbivory occurred in traditionally mown sites and at lower altitudes, while damage by fungal pathogen infection was independent of land use and altitude. Most types of herbivory were found on legumes and on leaves from fertilised sites, whereas the number of fungal pathogen types was highest on graminoids and in unfertilised sites. Larger standing crop was associated with higher leaf damage and diversity of herbivory types per leaf. Neither damage by herbivory nor by fungal pathogens was correlated with plant species diversity. In more plant species rich parcels, the number of herbivory types was lower at the leaf level, but tended to be higher at the parcel level. Our results highlight the omnipresence of plant-herbivore and plant-pathogen interactions. They suggest that current land use changes from mowing to grazing or to abandonment decrease the diversity of herbivory, and that fertilisation decreases pathogen diversity. As our results did not reveal conservation conflicts between diversities of plants, herbivores, and fungal pathogens, and as the damage from herbivory and pathogens is generally low, we conclude that for protecting the high diversity of plant-herbivore and plant-pathogen interactions a diverse low-intensity land use should be maintaine

Time- and temperature-dependent postmortem concentration changes of the (synthetic) cannabinoids JWH-210, RCS-4, as well as ∆9-tetrahydrocannabinol following pulmonary administration to pigs

In forensic toxicology, interpretation of postmortem (PM) drug concentrations might be complicated due to the lack of data concerning drug stability or PM redistribution (PMR). Regarding synthetic cannabinoids (SC), only sparse data are available, which derived from single case reports without any knowledge of dose and time of consumption. Thus, a controlled pig toxicokinetic study allowing for examination of PMR of SC was performed. Twelve pigs received a pulmonary dose of 200 µg/kg BW each of 4-ethylnaphthalene-1-yl-(1-pentylindole-3-yl)methanone (JWH-210), 2-(4-methoxyphenyl)-1-(1-pentyl-indole-3-yl)methanone (RCS-4), and Δ9-tetrahydrocannabinol via an ultrasonic nebulizer. Eight hours after, the pigs were put to death with T61 and specimens of relevant tissues and body fluids were collected. Subsequently, the animals were stored at room temperature (n = 6) or 4 °C (n = 6) and further samples were collected after 24, 48, and 72 h each. Concentrations were determined following enzymatic cleavage and solid-phase extraction by liquid-chromatography tandem mass spectrometry applying the standard addition approach. High concentrations of the parent compounds were observed in lung, liver, kidney and bile fluid/duodenum content as well as brain. HO-RCS-4 was the most prevalent metabolite detected in PM specimens. In general, changes of PM concentrations were found in every tissue and body fluid depending on the PM interval as well as storage temperature

Is adipose tissue suitable for detection of (synthetic) cannabinoids? A comparative study analyzing antemortem and postmortem specimens following pulmonary administration of JWH-210, RCS-4, as well as ∆9-tetrahydrocannabinol to pigs

Examining fatal poisonings, chronic exposure may be refected by the concentration in tissues known for long-term storage of drugs. Δ9-tetrahydrocannabinol (THC) persists in adipose tissue (AT), but sparse data on synthetic cannabinoids (SC) are available. Thus, a controlled pig study evaluating antemortem (AM) disposition and postmortem (PM) concentration changes of the SC 4-ethylnaphthalene-1-yl-(1-pentylindole-3-yl)methanone (JWH-210) and 2-(4-methoxyphenyl)-1-(1-pentyl-indole3-yl)methanone (RCS-4) as well as THC in AT was performed. The drugs were administered pulmonarily (200 µg/kg body weight) to twelve pigs. Subcutaneous (s.c.) AT specimens were collected after 15 and 30 min and then hourly up to 8 h. At the end, pigs were sacrifced and s.c., perirenal, and dorsal AT specimens were collected. The carcasses were stored at room temperature (RT; n=6) or 4 °C (n=6) and specimens were collected after 24, 48, and 72 h. After homogenization in acetonitrile and standard addition, LC–MS/MS was performed. Maximum concentrations were reached 0.5–2 h after administration amounting to 21±13 ng/g (JWH-210), 24±13 ng/g (RCS-4), and 22±20 ng/g (THC) and stayed at a plateau level. Regarding the metabolites, very low concentrations of N-hydroxypentyl-RCS-4 (HO-RCS-4) were detected from 0.5 to 8 h. PM concentrations of parent compounds did not change signifcantly (p>0.05) over time under both storage conditions. Concentrations of HO-RCS-4 signifcantly (p<0.05) increased in perirenal AT during storage at RT. These results suggest a rapid distribution and persistence in s.c. AT. Furthermore, AT might be resistant to PM redistribution of parent compounds. However, signifcant PM increases of metabolite concentrations might be considered in perirenal AT

A global inventory of mountains for bio-geographical applications

Mountains are hotspots of biodiversity. Yet, evaluating their importance in global biodiversity inventories requires the adoption of a pertinent definition of mountains. Here, we first compare the well-established WCMC and GMBA definitions, which both use geographical information systems. We show that the WCMC approach arrives at twice the global mountain area and much higher human population numbers than the GMBA one, which is explained by the inclusion of (mostly) low latitude hill country below 600 m elevation. We then present an inventory of the world’s mountains based on the GMBA definition. In this inventory, each of the 1003 entries corresponds to a polygon drawn around a mountain or a mountain range and includes the name of the delineated object, the area of mountainous terrain it covers stratified into different bioclimatic belts (all at 2.5′ resolution), and demographic information. Taken together, the 1003 polygons cover 13.8 Mio km2 of mountain terrain, of which 3.3 Mio km2 are in the alpine and nival belts. This corresponds to 83.7% of the global mountain area sensu GMBA, and 94% of the alpine/nival area. The 386 Mio people inhabiting mountainous terrain within polygons represent 75% of the people globally inhabiting mountains sensu GMBA. This inventory offers a robust framework for the integration of mountain biota in regional and larger scale biodiversity assessments, for biogeography, bioclimatology, macroecology, and conservation research, and for the exploration of a multitude of socio-ecological and climate change-related research questions in mountain biota, including the potential pressure on alpine ecosystems

Sensing the (digital) pulse. Future steps for improving the secondary use of data for research in Switzerland.

INTRODUCTION Ensuring that the health data infrastructure and governance permits an efficient secondary use of data for research is a policy priority for many countries. Switzerland is no exception and many initiatives have been launched to improve its health data landscape. The country now stands at an important crossroad, debating the right way forward. We aimed to explore which specific elements of data governance can facilitate - from ethico-legal and socio-cultural perspectives - the sharing and reuse of data for research purposes in Switzerland. METHODS A modified Delphi methodology was used to collect and structure input from a panel of experts via successive rounds of mediated interaction on the topic of health data governance in Switzerland. RESULTS First, we suggested techniques to facilitate data sharing practices, especially when data are shared between researchers or from healthcare institutions to researchers. Second, we identified ways to improve the interaction between data protection law and the reuse of data for research, and the ways of implementing informed consent in this context. Third, we put forth ideas on policy changes, such as the steps necessary to improve coordination between different actors of the data landscape and to win the defensive and risk-adverse attitudes widespread when it comes to health data. CONCLUSIONS After having engaged with these topics, we highlighted the importance of focusing on non-technical aspects to improve the data-readiness of a country (e.g., attitudes of stakeholders involved) and of having a pro-active debate between the different institutional actors, ethico-legal experts and society at large

Protocol for German trial of Acyclovir and corticosteroids in Herpes-simplex-virus-encephalitis (GACHE): a multicenter, multinational, randomized, double-blind, placebo-controlled German, Austrian and Dutch trial [ISRCTN45122933]

Background The treatment of Herpes-simplex-virus-encephalitis (HSVE) remains a major unsolved problem in Neurology. Current gold standard for therapy is acyclovir, a drug that inhibits viral replication. Despite antiviral treatment, mortality remains up to 15%, less than 20% of patients are able to go back to work, and the majority of patients suffer from severe disability. This is a discouraging, unsatisfactory situation for treating physicians, the disabled patients and their families, and constitutes an enormous burden to the public health services. The information obtained from experimental animal research and from recent retrospective clinical observations, indicates that a substantial benefit in outcome can be expected in patients with HSVE who are treated with adjuvant dexamethasone. But currently there is no available evidence to support the routine use of adjuvant corticosteroid treatment in HSVE. A randomized multicenter trial is the only useful instrument to address this question. Design GACHE is a multicenter, randomized, double-blind, placebo-controlled, parallel group clinical trial of treatment with acyclovir and adjuvant dexamethasone, as compared with acyclovir and placebo in adults with HSVE. The statistical design will be that of a 3-stage-group sequential trial with potential sample size adaptation in the last stage. Conclusion 372 patients with proven HSVE (positive HSV-DNA-PCR), aged 18 up to 85 years; with focal neurological signs no longer than 5 days prior to admission, and who give informed consent will be recruited from Departments of Neurology of academic medical centers in Germany, Austria and The Netherlands. Sample size will potentially be extended after the second interim analysis up to a maximum of 450 patients. Trial Registration Current Controlled Trials ISRCTN4512293

Detrimental proarrhythmogenic interaction of Ca2+/calmodulin-dependent protein kinase II and NaV1.8 in heart failure

An interplay between Ca2+/calmodulin-dependent protein kinase IIδc (CaMKIIδc) and late Na+ current (INaL) is known to induce arrhythmias in the failing heart. Here, we elucidate the role of the sodium channel isoform NaV1.8 for CaMKIIδc-dependent proarrhythmia. In a CRISPR-Cas9-generated human iPSC-cardiomyocyte homozygous knock-out of NaV1.8, we demonstrate that NaV1.8 contributes to INaL formation. In addition, we reveal a direct interaction between NaV1.8 and CaMKIIδc in cardiomyocytes isolated from patients with heart failure (HF). Using specific blockers of NaV1.8 and CaMKIIδc, we show that NaV1.8-driven INaL is CaMKIIδc-dependent and that NaV1.8-inhibtion reduces diastolic SR-Ca2+ leak in human failing cardiomyocytes. Moreover, increased mortality of CaMKIIδc-overexpressing HF mice is reduced when a NaV1.8 knock-out is introduced. Cellular and in vivo experiments reveal reduced ventricular arrhythmias without changes in HF progression. Our work therefore identifies a proarrhythmic CaMKIIδc downstream target which may constitute a prognostic and antiarrhythmic strategy