Site-specific genetic and functional signatures of aortic endothelial cells at aneurysm predilection sites in healthy and AngII ApoE-/- mice.

Aortic aneurysm is characterized by a pathological dilation at specific predilection sites of the vessel and potentially results in life-threatening vascular rupture. Herein, we established a modified "Häutchen method" for the local isolation of endothelial cells (ECs) from mouse aorta to analyze their spatial heterogeneity and potential role in site-specific disease development. When we compared ECs from aneurysm predilection sites of healthy mice with adjacent control segments we found regulation of genes related to extracellular matrix remodeling, angiogenesis and inflammation, all pathways playing a critical role in aneurysm development. We also detected enhanced cortical stiffness of the endothelium at these sites. Gene expression of ECs from aneurysms of the AngII ApoE-/- model when compared to sham animals mimicked expression patterns from predilection sites of healthy animals. Thus, this work highlights a striking genetic and functional regional heterogeneity in aortic ECs of healthy mice, which defines the location of aortic aneurysm formation in disease.

Controlling absence seizures from the cerebellar nuclei via activation of the Gq signaling pathway.

Absence seizures (ASs) are characterized by pathological electrographic oscillations in the cerebral cortex and thalamus, which are called spike-and-wave discharges (SWDs). Subcortical structures, such as the cerebellum, may well contribute to the emergence of ASs, but the cellular and molecular underpinnings remain poorly understood. Here we show that the genetic ablation of P/Q-type calcium channels in cerebellar granule cells (quirky) or Purkinje cells (purky) leads to recurrent SWDs with the purky model showing the more severe phenotype. The quirky mouse model showed irregular action potential firing of their cerebellar nuclei (CN) neurons as well as rhythmic firing during the wave of their SWDs. The purky model also showed irregular CN firing, in addition to a reduced firing rate and rhythmicity during the spike of the SWDs. In both models, the incidence of SWDs could be decreased by increasing CN activity via activation of the Gq-coupled designer receptor exclusively activated by designer drugs (DREADDs) or via that of the Gq-coupled metabotropic glutamate receptor 1. In contrast, the incidence of SWDs was increased by decreasing CN activity via activation of the inhibitory Gi/o-coupled DREADD. Finally, disrupting CN rhythmic firing with a closed-loop channelrhodopsin-2 stimulation protocol confirmed that ongoing SWDs can be ceased by activating CN neurons. Together, our data highlight that P/Q-type calcium channels in cerebellar granule cells and Purkinje cells can be relevant for epileptogenesis, that Gq-coupled activation of CN neurons can exert anti-epileptic effects and that precisely timed activation of the CN can be used to stop ongoing SWDs.

Estimating population density of insectivorous bats based on stationary acoustic detectors: A case study.

Automated recording units are commonly used by consultants to assess environmental impacts and to monitor animal populations. Although estimating population density of bats using stationary acoustic detectors is key for evaluating environmental impacts, estimating densities from call activity data is only possible through recently developed numerical methods, as the recognition of calling individuals is impossible.We tested the applicability of generalized random encounter models (gREMs) for determining population densities of three bat species (Common pipistrelle Pipistrellus pipistrellus, Northern bat Eptesicus nilssonii, and Natterer's bat Myotis nattereri) based on passively collected acoustical data. To validate the results, we compared them to (a) density estimates from the literature and to (b) Royle-Nichols (RN) models of detection/nondetection data.Our estimates for M. nattereri matched both the published data and RN-model results. For E. nilssonii, the gREM yielded similar estimates to the RN-models, but the published estimates were more than twice as high. This discrepancy might be because the high-altitude flight of E. nilssonii is not accounted for in gREMs. Results of gREMs for P. pipistrellus were supported by published data but were ~10 times higher than those of RN-models. RN-models use detection/nondetection data, and this loss of information probably affected population estimates of very active species like P. pipistrellus.gREM models provided realistic estimates of bat population densities based on automatically recorded call activity data. However, the average flight altitude of species should be accounted for in future analyses. We suggest including flight altitude in the calculation of the detection range to assess the detection sphere more accurately and to obtain more precise density estimates.

Foliar Roundup application has minor effects on the compositional and functional diversity of soil microorganisms in a short-term greenhouse experiment.

The herbicide Roundup (and glyphosate, its active ingredient) is extensively used for weed control on a worldwide scale. It is absorbed after foliar application and quickly translocated inside the plant. In this study, we investigated the effects of Roundup speed, a commercial glyphosate formulation, on the structural composition (dominance of microbial groups, phospholipid fatty acid analysis - PLFA) and functional diversity (use of carbon sources, Multiple Substrate Induced Respiration - MSIR) of soil microorganisms. We specifically aimed at understanding the potential impact of biotic interactions on herbicide effects and included plants, earthworms, and endomycorrhizal fungi in the experimental setup. For this, we grew clover (Trifolium repens) in the greenhouse and added mycorrhizal inoculum (Glomus mosseae) and earthworms (Lumbricus terrestris) to the pots. Two weeks after foliar Roundup application and subsequent plant death, the pots were destructively sampled. The application resulted in a significant increase of microbial respiration (SIR) by approximately 30%. A multivariate analysis of the MSIR data exhibited small but significant differences between the microbial communities of treated and untreated pots, while no significant difference was apparent for the PLFA data. Bacterial PLFAs generally decreased following herbicide application, while mycorrhizal and fungal PLFAs were not affected. We did not find a consistent difference between the fatty acid markers of gram negative and gram positive bacteria. For all investigated parameters, there were highly significant differences between the upper (0-5 cm depth) and lower (5-10 cm) soil layers. The fact that rooting density differed by a factor of 3.5 between the two layers indicated that herbicide effects were especially pronounced in the clover rhizosphere and were likely due to changes in root exudate composition. We found significant, though very small, interactions between Roundup and other experimental factors (especially mycorrhizal inoculum).

Insectivorous bats respond to vegetation complexity in urban green spaces.

Structural complexity is known to determine habitat quality for insectivorous bats, but how bats respond to habitat complexity in highly modified areas such as urban green spaces has been little explored. Furthermore, it is uncertain whether a recently developed measure of structural complexity is as effective as field-based surveys when applied to urban environments. We assessed whether image-derived structural complexity (MIG) was as/more effective than field-based descriptors in this environment and evaluated the response of insectivorous bats to structural complexity in urban green spaces. Bat activity and species richness were assessed with ultrasonic devices at 180 locations within green spaces in Vienna, Austria. Vegetation complexity was assessed using 17 field-based descriptors and by calculating the mean information gain (MIG) using digital images. Total bat activity and species richness decreased with increasing structural complexity of canopy cover, suggesting maneuverability and echolocation (sensorial) challenges for bat species using the canopy for flight and foraging. The negative response of functional groups to increased complexity was stronger for open-space foragers than for edge-space foragers. Nyctalus noctula, a species foraging in open space, showed a negative response to structural complexity, whereas Pipistrellus pygmaeus, an edge-space forager, was positively influenced by the number of trees. Our results show that MIG is a useful, time- and cost-effective tool to measure habitat complexity that complemented field-based descriptors. Response of insectivorous bats to structural complexity was group- and species-specific, which highlights the need for manifold management strategies (e.g., increasing or reinstating the extent of ground vegetation cover) to fulfill different species' requirements and to conserve insectivorous bats in urban green spaces.

Operator bias in software-aided bat call identification.

Software-aided identification facilitates the handling of large sets of bat call recordings, which is particularly useful in extensive acoustic surveys with several collaborators. Species lists are generated by "objective" automated classification. Subsequent validation consists of removing any species not believed to be present. So far, very little is known about the identification bias introduced by individual validation of operators with varying degrees of experience. Effects on the quality of the resulting data may be considerable, especially for bat species that are difficult to identify acoustically. Using the batcorder system as an example, we compared validation results from 21 volunteer operators with 1-26 years of experience of working on bats. All of them validated identical recordings of bats from eastern Austria. The final outcomes were individual validated lists of plausible species. A questionnaire was used to enquire about individual experience and validation procedures. In the course of species validation, the operators reduced the software's estimate of species richness. The most experienced operators accepted the smallest percentage of species from the software's output and validated conservatively with low interoperator variability. Operators with intermediate experience accepted the largest percentage, with larger variability. Sixty-six percent of the operators, mainly with intermediate and low levels of experience, reintroduced species to their validated lists which had been identified by the automated classification, but were finally excluded from the unvalidated lists. These were, in many cases, rare and infrequently recorded species. The average dissimilarity of the validated species lists dropped with increasing numbers of recordings, tending toward a level of ˜20%. Our results suggest that the operators succeeded in removing false positives and that they detected species that had been wrongly excluded during automated classification. Thus, manual validation of the software's unvalidated output is indispensable for reasonable results. However, although application seems easy, software-aided bat call identification requires an advanced level of operator experience. Identification bias during validation is a major issue, particularly in studies with more than one participant. Measures should be taken to standardize the validation process and harmonize the results of different operators.

Differentiation among cave populations of the Eukoenenia spelaea species-complex (Arachnida: Palpigradi) in the southwestern Alps.

Current knowledge of the taxonomy and distribution of European cave palpigrades largely rests upon incidental records. Samples seldom comprise more than one or two specimens, and many regions have only been unevenly explored, if at all. The present study is the first to investigate morphological variability within, and differentiation between, subterranean palpigrade populations in a small, geographically coherent area. It is based on relatively abundant material from six localities in the SW Alps (Piedmont, Italy) and two non-Italian comparative localities. Discrete and continuous characters reveal, on the one hand, a close grouping within the Eukoenenia spelaea species-complex, and, on the other hand, a remarkable short-range endemism. The occurrence of five distinct morphologies in a SW Alpine area of just 2000 square kilometres or so indicates that subterranean biodiversity parallels the rich above-ground biodiversity there. The Cottian Alps in the North harbour E. roscia Christian sp. n., E. lanai Christian sp. n. and the widespread E. spelaea. Two caves in the Maritime and Ligurian Alps are populated by E. bonadonai, which was originally described from the Provence Alps, while the Bossea cave in the Ligurian Alps remains the only known locality for E. strinatii. We evaluate the traditionally employed morphological characters and those recently proposed for use in Eukoenenia systematics. Further, we provide data on the formerly undescribed females of E. strinatii and E. bonadonai, and provide comprehensive pictorial information on the SW Alpine cave palpigrades to set new benchmarks for the taxonomy of the E. spelaea species-complex.

The role of microarthropods in terrestrial decomposition: a meta-analysis of 40 years of litterbag studies.

Litterbags have been utilized in soil ecology for about 50 years. They are useful because they confine organic material and thus enable the study of decomposition dynamics (mass loss and/or nutrient loss through time, colonization by soil biota) in situ, i.e. under field conditions. Researchers can easily restrict or permit access to certain size classes of soil fauna to determine their contribution to litter mass loss by choosing adequate mesh size or applying specific biocides. In particular, the mesofauna has received much attention since it comprises two very abundant and diverse microarthropod groups, the Collembola (springtails) and Acari (mites). We comprehensively searched the literature from the mid-1960s to the end of 2005 for reports on litterbag experiments investigating the role of microarthropods in terrestrial decomposition. Thirty papers reporting 101 experiments satisfied our selection criteria and were included in the database. Our meta-analysis revealed that microarthropods have a moderate but significant effect on mass loss. We discuss in detail the interactions of the microarthropod effect with study characteristics such as experimental design (e.g. number of bags, duration of experiment), type of exposed organic matter, climatic zone and land use of the study site. No publication bias was detected; however, we noticed a significant decrease in the microarthropod effect with publication year, indicating that, in the first decades of litterbag use, soil zoologists may have studied "promising" sites with a higher a priori probability of positive microarthropod effects on litter mass loss. A general weakness is that the treatments differ not only with respect to the presence or absence of microarthropods, but also with regard to mesh size (small to exclude microarthropods, wide to permit their access) or presence (to exclude microarthropods) and absence (to permit their access) of an insecticide. Consequently, the difference between the decomposition rates in the treatments is not a pure microarthropod effect but will be influenced by the additive effects of mesh size and insecticide. The relative contribution of the "true" microarthropod effect remains unknown without additional treatments controlling for the differential mesh size/insecticide effect. A meta-analysis including only those studies using different mesh size and for which the data were corrected by subtracting an estimated mesh size effect based on data from the literature yielded a significantly negative microarthropod effect on litter decomposition. These results cast doubt on the widely accepted hypothesis that microarthropods generally exert a positive effect on litter mass loss. We conclude that after 40 years of litterbag studies our knowledge on the role of microarthropods in litter mass loss remains limited and that the inclusion of a third treatment in future studies is a promising way to retain litterbags as a meaningful tool of soil biological studies.

Efficacy and side effects of five sampling methods for soil earthworms (Annelida, Lumbricidae).

In this study, carried out on an experimental meadow in Austria, in non calceric cambisol, five common methods for sampling earthworms were jointly compared for their efficacy (handsorting, formalin, and allyl isothiocyanate (AITC) application, heat extraction in Kempson apparatus, and electrical octet method). Additionally, short- and long-term effects of the non-destructive of these methods (formalin and AITC application, octet method) on soil organisms (microarthropod abundance, phospholipid fatty acids) and shoot and root biomass were analysed. The Kempson extraction yielded the greatest number of individuals, followed by the octet method and handsorting. Formalin and AITC showed lower efficacy, but expelled high numbers of adult earthworms. Whereas AITC scarcely had nontarget effects on soil organisms, formalin negatively affected soil microorganisms and vegetation on the treated plots. The octet method seems to be well applicable especially in protected areas, since it is efficient, non-destructive and does not adversely affect soil organisms. The recommendations for method application are given, depending strongly on the scope of studies, as well as on ecological conditions and legal study site limitations.

Earthworm expulsion by formalin has severe and lasting side effects on soil biota and plants.

The application of formalin to the ground is a commonly used method to estimate the abundance and biomass of earthworms in soil. From April to September 2003, we investigated potential negative side effects of this method on soil organisms and plants in the sampling soil area to which formalin has been applied. The application of 0.4% formalin (40 L per 0.25 m(2)) severely decreased abundances of Collembola and Nematoda, soil respiration, dehydrogenase activity, fatty acid contents, vegetation cover, and shoot and root biomass. These effects lasted for the whole study period. The impact of formalin may have been aggravated due to unusually hot and dry weather conditions. However, owing to its harmful consequences, the formalin method cannot be recommended as a standard for earthworm extraction.