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Chemical staining of biological specimens is commonly utilised to boost contrast in soft tissue structures, but unambiguous identification of staining location and distribution is difficult without confirmation of the elemental signature, especially for chemicals of similar density contrast. Hyperspectral X-ray computed tomography (XCT) enables the non-destructive identification, segmentation and mapping of elemental composition within a sample. With the availability of hundreds of narrow, high resolution (~ 1 keV) energy channels, the technique allows the simultaneous detection of multiple contrast agents across different tissue structures. Here we describe a hyperspectral imaging routine for distinguishing multiple chemical agents, regardless of contrast similarity. Using a set of elemental calibration phantoms, we perform a first instance of direct stain concentration measurement using spectral absorption edge markers. Applied to a set of double- and triple-stained biological specimens, the study analyses the extent of stain overlap and uptake regions for commonly used contrast markers. An improved understanding of stain concentration as a function of position, and the interaction between multiple stains, would help inform future studies on multi-staining procedures, as well as enable future exploration of heavy metal uptake across medical, agricultural and ecological fields. [ABSTRACT FROM AUTHOR]

The seal heartworm Acanthocheilonema spirocauda (Nematoda: Onchocercidae) parasitizes the heart and pulmonary arteries of various phocid seals of the Northern Hemisphere. Over many decades, potential vectors of this parasite have been discussed, and to this date, the life cycle is not fully known. The seal louse Echinophthirius horridus (Anoplura: Echinophthiriidae) is an obligatory, permanent and haematophagous ectoparasite of phocids that has been hypothesized to function as obligate intermediate host for A. spirocauda. We examined 11 adult E. horridus specimens collected from stranded harbour seals (Phoca vitulina) in rehabilitation at the Sealcentre Pieterburen by X-ray microCT imaging, aiming to illustrate larval A. spirocauda infection sites in situ. In three of these specimens, thread-like larvae were detected in insect organs. Detailed imaging of the most infected louse revealed a total of 54 A. spirocauda larvae located either in fat bodies or the haemocoel. Histological analysis of the same specimen illustrated nematode cross-sections, confirming X-ray microCT data. The current data strongly suggest that E. horridus is a natural intermediate host for A. spirocauda. Moreover, we demonstrate the potential of X-ray microCT-based imaging as a non-destructive method to analyze host-parasite interactions, especially in the neglected field of marine mammal parasitology. [ABSTRACT FROM AUTHOR]

The subfamily Drusinae (Limnephilidae, Trichoptera) comprises a range of species exhibiting differently shaped head capsules in their larval stages. These correspond to evolutionary lineages pursuing different larval feeding ecologies, each of which uses a different hydraulic niche: scraping grazers and omnivorous shredders sharing rounded head capsules and filtering carnivores with indented and corrugated head capsules. In this study, we assess whether changes in head capsule morphology are reflected by changes in internal anatomy of Drusinae heads. To this end, internal and external head morphology was visualized using µCT methods and histological sections in three Drusinae species—Drusus franzi, D. discolor and D. bosnicus—representing the three evolutionary lineages. Our results indicate that Drusinae head musculature is highly conserved across the evolutionary lineages with only minute changes between taxa. Conversely, the tentorium is reduced in D. discolor, the species with the most aberrant head capsule investigated here. Integrating previous research on Drusinae head anatomy, we propose a fundamental Drusinae blueprint comprising 29 cephalic muscles and discuss significance of larval head capsule corrugation in Trichoptera. [ABSTRACT FROM AUTHOR]

Here we apply hyperspectral bright field imaging to collect computed tomographic images with excellent energy resolution (~ 1 keV), applying it for the first time to map the distribution of stain in a fixed biological sample through its characteristic K-edge. Conventionally, because the photons detected at each pixel are distributed across as many as 200 energy channels, energy-selective images are characterised by low count-rates and poor signal-to-noise ratio. This means high X-ray exposures, long scan times and high doses are required to image unique spectral markers. Here, we achieve high quality energy-dispersive tomograms from low dose, noisy datasets using a dedicated iterative reconstruction algorithm. This exploits the spatial smoothness and inter-channel structural correlation in the spectral domain using two carefully chosen regularisation terms. For a multi-phase phantom, a reduction in scan time of 36 times is demonstrated. Spectral analysis methods including K-edge subtraction and absorption step-size fitting are evaluated for an ex vivo, single (iodine)-stained biological sample, where low chemical concentration and inhomogeneous distribution can affect soft tissue segmentation and visualisation. The reconstruction algorithms are available through the open-source Core Imaging Library. Taken together, these tools offer new capabilities for visualisation and elemental mapping, with promising applications for multiply-stained biological specimens. [ABSTRACT FROM AUTHOR]

Hydraulic niche descriptors of final instar larvae of nine Drusus species (Trichoptera) were studied in small, spring-fed, first-order headwaters located in the Mühlviertel (Upper Austria), Koralpe (Carinthia, Austria), and in the Austrian and Italian Alps. The species investigated covered all three clades of Drusinae: the shredder clade (Drusus franzi, D. alpinus), the grazer clade (D. biguttatus, D. chauvinianus, D. dudor, D. monticola), and the filtering carnivore clade (D. chrysotus, D. katagelastos, D. muelleri). Flow velocity was measured at front center of 68 larvae, head upstream, on the top of mineral substrate particles at water depths of 10–30 mm, using a tripod-stabilized Micro propeller meter (propeller diameter = 10 mm). Each data series consisted of a sampled measurement lasting 30 s (measuring interval = 1 s). In total, 2040 single velocity measurements were taken. Instantaneous flow velocities and drag at the sites of the 68 larvae varied from 0 to 0.93 m s−1 and 0 to 8346 *10−6 N, respectively. Flow velocities and drag between the three clades were highly significantly different (p < 0.001); mean velocity (+ 95% confidence limits) for the three clades were 0.09 + 0.00 m s−1 for the shredder, 0.25 + 0.00 m s−1 for the grazer, and 0.31 + 0.01 m s−1 for the filtering carnivore clade; the corresponding data for drag were (85 + 18)*10−6 N, (422 + 61)*10−6 N and (1125 + 83)*10−6 N, respectively. Adhesive friction ranged from (41.07 + 53.03)*10−6 N in D. franzi to (255.24 + 216.87)*10−6 N in D. chrysotus. Except in D. franzi and D. dudor adhesive friction was always well below drag force, indicating that submerged weight alone was not sufficient to stabilize the larvae in their hydraulic environment. Reynolds numbers varied between 0 in D. franzi and D. alpinus, and 12,634 in D. katagelastos, with 7% of the total in the laminar (R < 500), 30% in the transitional (R = 500–2000), and 61% in the fully turbulent stage (R > 2000). Froude numbers (Fr) varied from 0 to 2.97. The two Drusus species of the shredder clade and three out of four species of the grazer clade were exposed to subcritical Fr < 1, one species of the grazer clade and two out of three species of the filtering clade to supercritical Froude numbers >1. [ABSTRACT FROM AUTHOR]

By studying hydraulic stress parameters of larvae of the cased caddisfly Drusus biguttatus (Pictet, 1834) in a tributary of the Schwarze Sulm (Carinthia, Austria), we aimed on (1) detecting the flow properties of the spatio-temporally filtered velocity measurements taken, and (2) on defining the hydraulic niche of this caddisfly larva. For this, we took 31 measurement series lasting 30 to 300 s, yielding 2176 single velocity measurements. The probability density functions of the 31 data series were Gaussian or sub-Gaussian, and the mean recurrent interval between velocity maxima within a data series was only 15.00 s. As a consequence, the Trichoptera larvae studied have to face strong flow accelerations in short intervals which is a much higher stress than conventional mean velocity measurements would suggest. The hydraulic niche of Drusus biguttatus is defined by instantaneous flow velocities ranging from 0.04 to 0.69 m s−1, by drag forces from 13 × 10−6 to 3737 × 10−6 N, by Froude numbers from 0.13 to 1.20, and mostly by Reynolds numbers > 2000. Under such conditions, only 5.1% of the drag force is compensated by submerged weight, whereas the remainder has to be counterbalanced by the active efforts of the larvae to remain attached to the substrate. [ABSTRACT FROM AUTHOR]

Garra barreimiae (Fowler and Steinitz, Bull Res Counc Isr 5B:262-289, 1956) is a freshwater fish that lives throughout the Hajar Mountains of Oman and the United Arab Emirates. Previously, four different genetic clades (West clade, Central Clade, North clade and East clade) have been identified within this species. This study observes morphological differences between these clades using morphometrics and meristics as well as micro CT imaging, further strengthening the assumption that the taxon of G. barreimiae should be restricted to one of the genetic clades detected. Although many morphometric and meristic characteristics are highly variable within the clades, the West clade fits the original description of G. barreimiae and is distinguishable by its higher number of fifteen to seventeen gill rakers on the lower limb of the first gill arch (compared to the other clades with ten to fourteen gill rakers), its seven branched pelvic fin rays (in contrary to the other clades with eight branched pelvic fin rays) and its number of vertebrae, with typically nineteen precaudal and one intermediate vertebrae, compared to eighteen precaudal and two intermediate vertebrae in the Central and North clade and seventeen precaudal and two intermediate vertebrae in the East clade. [ABSTRACT FROM AUTHOR]

Abstract: The RANK (receptor activator of nuclear factor κB)/RANKL (RANK ligand)/OPG (osteoprotegerin) axis is activated after myocardial infarction (MI), but its pathophysiological role is not well understood. Here, we investigated how global and cell compartment-selective inhibition of RANKL affects cardiac function and remodeling after MI in mice. Global RANKL inhibition was achieved by treatment of human RANKL knock-in (huRANKL-KI) mice with the monoclonal antibody AMG161. huRANKL-KI mice express a chimeric RANKL protein wherein part of the RANKL molecule is humanized. AMG161 inhibits human and chimeric but not murine RANKL. To dissect the pathophysiological role of RANKL derived from hematopoietic and mesenchymal cells, we selectively exchanged the hematopoietic cell compartment by lethal irradiation and across-genotype bone marrow transplantation between wild-type and huRANKL-KI mice, exploiting the specificity of AMG161. After permanent coronary artery ligation, mice were injected with AMG161 or an isotype control antibody over 4 weeks post-MI. MI increased RANKL expression mainly in cardiomyocytes and scar-infiltrating cells 4 weeks after MI. Only inhibition of RANKL derived from hematopoietic cellular sources, but not global or mesenchymal RANKL inhibition, improved post-infarct survival and cardiac function. Mechanistically, hematopoietic RANKL inhibition reduced expression of the pro-inflammatory cytokine IL-1ß in the cardiac cellular infiltrate. In conclusion, inhibition of RANKL derived from hematopoietic cellular sources is beneficial to maintain post-ischemic cardiac function by reduction of pro-inflammatory cytokine production.Key messages: Experimental myocardial infarction (MI) augments cardiac RANKL expression in mice.RANKL expression is increased in cardiomyocytes and scar-infiltrating cells after MI.Global or mesenchymal cell RANKL inhibition has no influence on cardiac function after MI.Inhibition of RANKL derived from hematopoietic cells improves heart function post-MI.Hematopoietic RANKL inhibition reduces pro-inflammatory cytokines in scar-infiltrating cells. [ABSTRACT FROM AUTHOR]

Re- Engineering Philosophy for Limited Beings is about new approaches to many of the big topics in philosophy of science today, but with a very different take. To begin with, we are urged to reject the received Cartesian-Laplacean myths: Descartes' certainty and Laplace's computational omniscience. Instead, Wimsatt re-engineers a philosophy for human beings with all their cognitive limitations. His approaches find their starting point in the actual practices of scientists themselves, which he strongly identifies with engineering practices as the source of researchers' solutions for dealing with a complex world. He aims to construct an understanding of scientific methodology around the central role of reduction. But he dismisses eliminative reductionism in favor of a heuristic-based realist view. Wimsatt's world is a complex one, and this means that science needs to do away with all the absolute and simple answers, because they do not reflect the world we are living in. A complex world requires the mindset and tinkering of an engineer to uncover its reality. The appropriate response must be heuristics all the way down as we constantly seek out reliable inferences on often shifting ground. To this end, we aim for models and theories that are robust, just as engineers aim to build robust machines. And although errors occur and approaches are fallible, they allow us to continually adapt the heuristics applied and sharpen our perceptions so as to develop more refined tools for investigating and understanding the world. [ABSTRACT FROM AUTHOR]

A well-developed suction pump in the head represents an important adaptation for nectar-feeding insects, such as Hymenoptera, Lepidoptera and Diptera. This pumping organ creates a pressure gradient along the proboscis, which is responsible for nectar uptake. The extremely elongated proboscis of the genus Prosoeca (Nemestrinidae) evolved as an adaptation to feeding from long, tubular flowers. According to the functional constraint hypothesis, nectar uptake through a disproportionately elongated, straw-like proboscis increases flower handling time and consequently lowers the energy intake rate. Due to the conspicuous length variation of the proboscis of Prosoeca, individuals with longer proboscides are hypothesised to have longer handling times. To test this hypothesis, we used field video analyses of flower-visiting behaviour, detailed examinations of the suction pump morphology and correlations of proboscis length with body length and suction pump dimensions. Using a biomechanical framework described for nectar-feeding Lepidoptera in relation to proboscis length and suction pump musculature, we describe and contrast the system in long-proboscid flies. Flies with longer proboscides spent significantly more time drinking from flowers. In addition, proboscis length and body length showed a positive allometric relationship. Furthermore, adaptations of the suction pump included an allometric relationship between proboscis length and suction pump muscle volume and a combination of two pumping organs. Overall, the study gives detailed insight into the adaptations required for long-proboscid nectar feeding, and comparisons with other nectar-sucking insects allow further considerations of the evolution of the suction pump in insects with sucking mouthparts. [ABSTRACT FROM AUTHOR]