Mechanical spectroscopy of retina explants at the protein level employing nanostructured scaffolds

Development of neuronal tissue, such as folding of the brain, and formation of the fovea centralis in the human retina are intimately connected with the mechanical properties of the underlying cells and the extracellular matrix. In particular for neuronal tissue as complex as the vertebrate retina, mechanical properties are still a matter of debate due to their relation to numerous diseases as well as surgery, where the tension of the retina can result in tissue detachment during cutting. However, measuring the elasticity of adult retina wholemounts is difficult and until now only the mechanical properties at the surface have been characterized with micrometer resolution. Many processes, however, such as pathological changes prone to cause tissue rupture and detachment, respectively, are reflected in variations of retina elasticity at smaller length scales at the protein level. In the present work we demonstrate that freely oscillating cantilevers composed of nanostructured TiO2 scaffolds can be employed to study the frequency-dependent mechanical response of adult mammalian retina explants at the nanoscale. Constituting highly versatile scaffolds with strong tissue attachment for long-term organotypic culture atop, these scaffolds perform damped vibrations as fingerprints of the mechanical tissue properties that are derived using finite element calculations. Since the tissue adheres to the nanostructures via constitutive proteins on the photoreceptor side of the retina, the latter are stretched and compressed during vibration of the underlying scaffold. Probing mechanical response of individual proteins within the tissue, the proposed mechanical spectroscopy approach opens the way for studying tissue mechanics, diseases and the effect of drugs at the protein level

Progress in Landslide Susceptibility Mapping over Europe Using Tier-Based Approaches

The European Thematic Strategy for Soil Protection aims, among other objectives, to ensure a sustainable use of soil. The legal instrument of the strategy, the proposed Framework Directive, suggests identifying priority areas of several soil threats including landslides using a coherent and compatible approach based on the use of common thematic data. In a first stage, this can be achieved through landslide susceptibility mapping using geographically nested, multi-step tiered approaches, where areas identified as of high susceptibility by a first, synoptic-scale Tier (Tier 1) can then be further assessed and mapped at larger scale by successive Tiers. In order to identify areas prone to landslides at European scale (Tier 1), a number of thematic terrain and environmental data sets already available for the whole of Europe can be used as input for a continental scale susceptibility model. However, since no coherent landslide inventory data is available at the moment over the whole continent, qualitative heuristic zonation approaches are proposed. For Tier 1 a preliminary, simplified model has been developed. It consists of an equally weighting combination of a reduced, continent-wide common dataset of landslide conditioning factors including soil parent material, slope angle and land cover, to derive a landslide susceptibility index using raster mapping units consisting of 1 x 1 km pixels. A preliminary European-wide susceptibility map has thus been produced at 1:1 Million scale, since this is compatible with that of the datasets used. The map has been validated by means of a ratio of effectiveness using samples from landslide inventories in Italy, Austria, Hungary and United Kingdom. Although not differentiated for specific geomorphological environments or specific landslide types, the experimental model reveals a relatively good performance in many European regions at a 1:1 Million scale. An additional Tier 1 susceptibility map at the same scale and using the same or equivalent thematic data as for the one above has been generated for six French departments using a heuristic, weighting-based multi-criteria evaluation model applied also to raster-cell mapping units. In this experiment, thematic data class weights have been differentiated for two stratification areas, namely mountains and plains, and four main landslide types. Separate susceptibility maps for each landslide type and a combined map for all types have been produced. Results have been validated using BRGM's BDMvT landslide inventory. Unlike Tier 1, Tier 2 assessment requires landslide inventory data and additional thematic data on conditioning factors which may not be available for all European countries. For the Tier 2, a nation-wide quantitative landslide susceptibility assessment has been performed for Italy by applying a statistical model. In this assessment, multivariate analysis was applied using bedrock, soil and climate data together with a number of derivatives from SRTM90 DEM. In addition, separate datasets from a historical landslide inventory were used for model training and validation respectively. The mapping units selected were based on administrative boundaries (municipalities). The performance of this nation-wide, quantitative susceptibility assessment has been evaluated using multi-temporal landslide inventory data. Finally, model limitations for Tier 1 are discussed, and recommendations for enhanced Tier 1 and Tier 2 models including additional thematic data for conditioning factors are drawn. This project is part of the collaborative research carried out within the European Landslide Expert Group coordinated by JRC in support to the EU Soil Thematic Strategy. It is also supported by the International Programme on Landslides of the International Consortium on Landslides.JRC.DDG.H.7-Land management and natural hazard

Update of the European Landslide Susceptibility Map (ELSUS Version 2)

We present an update of the initial version of the European Landslide Susceptibility Map (ELSUS Version 1) that was released in 2012 through the EU Joint Research Centre (JRC) European Soil Data Centre (ESDAC). The susceptibility evaluation methodology employed for the updated map ELSUS Version 2 presented in this paper is identical to the previous approach, and comprises the differentiation of the analyzed European area into seven climate-physiographical model zones, the use of a reduced set of spatial susceptibility predictors (shallow subsurface lithology, slope angle, and land cover), and model zone-specific heuristic spatial multicriteria evaluations (SMCE) for susceptibility mapping. The most important improvement for ELSUS version 2 is the replacement of the original “lithology” data set consisting of soil parent material information derived from the European Soil Database (ESDB) by new information derived from the digital version of the International Hydrogeological Map of Europe at scale 1:1.5 Million (IHME 1500). IHME lithology describes both consolidated and unconsolidated shallow geological materials over Europe and can be shown to have a higher significance for landslide susceptibility evaluation than the soil parent material derived from ESDB. Other improvements consist in the change of the mapping unit from 1 km to 200 m grid size and the incorporation of terrains not covered by ELSUS version 1 (e.g., Iceland, the Faroers, the Shetlands, and Cyprus). Additionally, the new ELSUS version 2 was calibrated and validated with an updated pan-European landslide inventory now containing more than 155,000 landslides (30% more than used for ELSUS version 1). The enhanced and updated landslide inventory and the higher quality of the “lithology” data enabled us to establish more consistent SMCE-schemes for the individual model zones. The enhancements of ELSUS Version 2 result in an overall increase of the predictive power of the map for about 10%, as indicated by ROC curve metrics obtained with the updated landslide inventory. However, the assessment still suffers from missing landslide information in many European terrains. It can be suspected that more distributed landslides information in specific model zones will further enhance the accuracy of ELSUS in the future.JRC.H.5-Land Resources Managemen

Involvement of A1 adenosine receptors in osmotic volume regulation of retinal glial cells in mice

PURPOSE: Osmotic swelling of Müller glial cells has been suggested to contribute to retinal edema. We determined the role of adenosine signaling in the inhibition of Müller cell swelling in the murine retina. METHODS: The size of Müller cell somata was recorded before and during perfusion of retinal sections and isolated Müller cells with a hypoosmolar solution. Retinal tissues were freshly isolated from wild-type mice and mice deficient in A(1) adenosine receptors (A(1)AR(-/-)), or cultured as whole-mounts for three days. The potassium conductance of Müller cells was recorded in isolated cells, and retinal slices were immunostained against Kir4.1. RESULTS: Hypotonic exposure for 4 min induced a swelling of Müller cell bodies in retinal slices from A(1)AR(-/-) mice but not wild-type mice. Pharmacological inhibition of A(1) receptors or of the ecto-5'-nucleotidase induced hypoosmotic swelling of Müller cells from wild-type mice. Exogenous adenosine prevented the swelling of Müller cells from wild-type but not A(1)AR(-/-) mice. The antiinflammatory corticosteroid, triamcinolone acetonide, inhibited the swelling of Müller cells from wild-type mice; this effect was blocked by an antagonist of A(1) receptors. The potassium conductance of Müller cells and the Kir4.1 immunolabeling of retinal slices were not different between A(1)AR(-/-) and wild-type mice, both in freshly isolated tissues and retinal organ cultures. CONCLUSIONS: The data suggest that autocrine activation of A(1) receptors by extracellularly generated adenosine mediates the volume homeostasis of Müller cells in the murine retina. The swelling-inhibitory effect of triamcinolone is mediated by enhancement of endogenous adenosine signaling

Müller cell activation, proliferation and migration following laser injury.

PurposeMüller cells are well known for their critical role in normal retinal structure and function, but their reaction to retinal injury and subsequent role in retinal remodeling is less well characterized. In this study we used a mouse model of retinal laser photocoagulation to examine injury-induced Müller glial reaction, and determine how this reaction was related to injury-induced retinal regeneration and cellular repopulation.MethodsExperiments were performed on 3-4-week-old C57BL/6 mice. Retinal laser photocoagulation was used to induce small, circumscribed injuries; these were principally confined to the outer nuclear layer, and surrounded by apparently healthy retinal tissue. Western blotting and immunohistochemical analyses were used to determine the level and location of protein expression. Live cell imaging of green fluorescent protein (GFP)-infected Müller cells (AAV-GFAP-GFP) were used to identify the rate and location of retinal Müller cell nuclear migration.ResultsUpon injury, Müller cells directly at the burn site become reactive, as evidenced by increased expression of the intermediate filament proteins glial fibrillary acidic protein (GFAP) and nestin. These reactive cells re-enter the cell cycle as shown by expression of the markers Cyclin D1 and D3, and their nuclei begin to migrate toward the injury site at a rate of approximately 12 microm/hr. However, unlike other reports, evidence for Müller cell transdifferentiation was not identified in this model.ConclusionsRetinal laser photocoagulation is capable of stimulating a significant glial reaction, marked by activation of cell cycle progression and retinal reorganization, but is not capable of stimulating cellular transdifferentiation or neurogenesis

A new framework for assessing subject-specific whole brain circulation and perfusion using MRI-based measurements and a multi-scale continuous flow model

A large variety of severe medical conditions involve alterations in microvascular circulation. Hence, measurements or simulation of circulation and perfusion has considerable clinical value and can be used for diagnostics, evaluation of treatment efficacy, and for surgical planning. However, the accuracy of traditional tracer kinetic one-compartment models is limited due to scale dependency. As a remedy, we propose a scale invariant mathematical framework for simulating whole brain perfusion. The suggested framework is based on a segmentation of anatomical geometry down to imaging voxel resolution. Large vessels in the arterial and venous network are identified from time-of-flight (ToF) and quantitative susceptibility mapping (QSM). Macro-scale flow in the large-vessel-network is accurately modelled using the Hagen-Poiseuille equation, whereas capillary flow is treated as two-compartment porous media flow. Macro-scale flow is coupled with micro-scale flow by a spatially distributing support function in the terminal endings. Perfusion is defined as the transition of fluid from the arterial to the venous compartment. We demonstrate a whole brain simulation of tracer propagation on a realistic geometric model of the human brain, where the model comprises distinct areas of grey and white matter, as well as large vessels in the arterial and venous vascular network. Our proposed framework is an accurate and viable alternative to traditional compartment models, with high relevance for simulation of brain perfusion and also for restoration of field parameters in clinical brain perfusion applications.publishedVersio

Is Internal Rotation Measurement of the Hip Useful for Ruling in Cam or Pincer Morphology in Asymptomatic Males? A Diagnostic Accuracy Study.

BACKGROUND Cam and pincer morphologies are associated with limited internal rotation. However, the routine clinical examination for hip rotation has limited reliability. A more standardized method of measuring hip rotation might increase test-retest and interobserver reliability and might be useful as a screening test to detect different hip morphologies without the need for imaging. We developed an examination chair to standardize the measurement of internal hip rotation, which improved interobserver reliability. However, the diagnostic test accuracy for this test is unknown. QUESTION/PURPOSE Is a standardized method of determining internal hip rotation using an examination chair useful in detecting cam and pincer morphology with MRI as a reference standard? METHODS A diagnostic test accuracy study was conducted in a sample of asymptomatic males. Using an examination chair with a standardized seated position, internal rotation was measured in 1080 men aged 18 to 21 years who had been conscripted for the Swiss army. The chair prevents compensatory movement by stabilizing the pelvis and the thighs with belts. The force to produce the internal rotation was standardized with a pulley system. Previous results showed that the measurements with the examination chair are similar to clinical assessment but with higher interobserver agreement. A random sample of 430 asymptomatic males was invited to undergo hip MRI. Of those, 244 White European males responded to the invitation and had a mean age of 20 ± 0.7 years and a mean internal rotation of the hip of 33° ± 8.5°. Using MRI as the reference standard, 69% (169 of 244) had a normal hip, 24% (59 of 244) a definite cam morphology (Grades 2 and 3), 3% (8 of 244) an increased acetabular depth, and 3% (8 of 244) a combination of both. One experienced radiologist graded cam morphology as follows: 0 = normal, 1 = mild, 2 = moderate, and 3 = severe. Pincer morphology was defined by increased acetabular depth (≤ 3 mm distance between the center of the femoral neck and the line connecting the anterior and posterior acetabular rims). The intraobserver agreement was substantial (weighted κ of 0.65). A receiver operating characteristic (ROC) curve was fitted, and sensitivity, specificity, and likelihood ratios were estimated for different internal rotation cutoffs. RESULTS For cam morphology, the area under the ROC curve was 0.75 (95% CI 0.67 to 0.82). Internal hip rotation of less than 20° yielded a positive likelihood ratio of 9.57 (sensitivity 0.13, specificity 0.99), and a value of 40° or more resulted in a negative likelihood ratio of 0.36 (sensitivity 0.93, specificity 0.20). The area under the curve for detecting the combination of cam and pincer morphologies was 0.87 (95% CI 0.74 to 1.0). A cutoff of 20° yielded a positive likelihood ratio of 9.03 (sensitivity 0.33, specificity 0.96). CONCLUSION This examination chair showed moderate-to-good diagnostic value to rule in hip cam morphology in White European males. However, at the extremes of the 95% confidence intervals, diagnostic performance would be poor. Nonetheless, we believe this test can contribute to identifying cam morphologies, and we hope that future, larger studies-ideally in more diverse patient populations-will seek to validate this to arrive at more precise estimates of the diagnostic performance of this test. LEVEL OF EVIDENCE Level III, diagnostic study

Failure of potassium siphoning by Muller cells: a new hypothesis of perfluorocarbon liquid-induced retinopathy,”

PURPOSE. To determine the effect of perfluorocarbon liquid (PFCL)-induced abolition of potassium siphoning by the vitreal end feet of Müller cells. METHODS. Porcine eyecups were filled with stained balanced salt solution and PFCLs (perfluorodecalin, perfluorooctane, perfluoroperhydrophenanthrene or the semifluorocarbon perfluorohexylhexane). With optical coherence tomography, the distance between PFCL and retina was determined, and the size of the aqueous space covering the retinal surface was estimated. The data were used to calculate the retinal potassium siphoning into small aqueous volumes. RESULTS. The distance between PFCL and retinal surface was found to be less than 5 to 10 m with any PFCL tested. The resultant volume of the aqueous space was too small to act as a sufficient sink for K ϩ ion siphoning. CONCLUSIONS. A certain threshold volume of vitreal fluid seems to be necessary for efficient buffering of intraretinal increases of K ϩ and perhaps other (e.g., H ϩ ) ions through the Müller cells. When the aqueous fluid is replaced by a PFCL (or by silicone oil) for longer periods, the outer retina becomes subject to long-lasting K ϩ accumulation, and consequent neurodegeneration and reactive gliosis occurs. The authors propose to search for new vitreous-substituting fluids with the capability to dissolve ions. (Invest Ophthalmol Vis Sci. 2000;41:256 -261