A novel functional assessment of the differentiation of micropatterned muscle cells

Duchenne muscular dystrophy (DMD) is a lethal disease characterized by rapid, progressive atrophy of muscle tissues. Timely screening of therapeutic interventions is necessary for the development of effective treatment approaches for DMD. We have developed an in vitro model using a combination of micropatterning of C2C12 skeletal muscle cells and cell traction force microscopy (CTFM). In this model, C2C12 cells were micropatterned on a highly elongated adhesive island such that the cells assumed a shape typical of a myotube. During differentiation, these cells gradually fused together and began expressing dystrophin, a structural protein of myotubes, meanwhile, their contractile forces, represented by cell traction forces, continually increased until the myotubes reached maturation. In addition, the high-degree alignment of cells favored myotube differentiation and dystrophin expression. Since the fundamental structural unit of muscle tissue is myofiber, which is responsible for muscle contraction, such a technology that can directly quantify the contractile forces of the myotube, a precursor of myofiber, may constitute a fast and efficient screening approach for DMD therapies.     

犬、猫肾功能衰竭的中西医结合疗法

本文介绍了中医的肾脏生理功能理论、阐述了目前西兽医及中兽医对犬猫急性肾功能衰竭、慢性肾功能衰竭发病原因的认识,分析了肾功能衰竭的中医辨证分型和急性肾功能衰竭、慢性肾功能衰竭的治疗原则。     

Flavonoid C- and O-glycosides from the Mongolian medicinal plant Dianthus versicolor Fisch

Eighteen flavonoids were identified from an aqueous extract of the aerial parts of Dianthus versicolor, a plant used in traditional Mongolian medicine against liver diseases. The flavonoid C- and O-glycosides isoorientin-7-O-rutinoside, isoorientin-7-O-rhamnosyl-galactoside, isovitexin-7-O-rutinoside, isovitexin-7-O-rhamnosyl-galactoside, isoscoparin-7-O-rutinoside, isoscoparin-7-O-rhamnosyl-galactoside, isoscoparin-7-O-galactoside, and isoorientin-7-O-galactoside were isolated and structurally elucidated. Their structures were established on the basis of extensive spectroscopic techniques including LC–UV–DAD, LC–MSⁿ, LC–HRMS, 1D and 2D NMR spectroscopy, and by GC–MS analysis after hydrolysis. Flavonoids with such a high glycosylation pattern are rare within the genus Dianthus. Furthermore, isovitexin-7-O-glucoside (saponarin), isovitexin-2″-O-rhamnoside, apigenin-6-glucoside (isovitexin), luteolin-7-O-glucoside, apigenin-7-O-glucoside, as well as the aglycons luteolin, apigenin, chrysoeriol, diosmetin, and acacetin were identified by TLC and LC–DAD–MSⁿ in comparison to reference substances or literature data. The NMR data of seven structures have not been reported in the literature to date.     

Anti-acetylcholinesterase activities of traditional Chinese medicine for treating Alzheimer's disease

Alzheimer's disease (AD) is a neurodegenerative disease characterized by progressive memory loss and cognitive impairment. It is the most common type of dementia in the ageing population due to a severe loss of cholinergic neurons in selected brain area. At present, acetylcholinesterase inhibitors (AChEI) are the first group of drugs approved by the FDA to treat mild to moderate Alzheimer's disease. Most of these drugs such as huperzine and galanthamine are originally isolated from plants. In this study, the AChE inhibitory activities from extracts of Chinese medicinal herbs that have traditionally been prescribed to treat insomnia and brain function disorders were examined in a 96-well plate assay based on Ellman's method. Both ethanol and aqueous extracts of 26 traditional Chinese medicinal herbs were tested. Inhibitory effects were expressed as the percentage of inhibition. For the herbal extracts that were shown to exert a significant inhibition, dose-dependent inhibitory assays were also performed. Ethanol and aqueous extracts of six herbs were found to have high AChE inhibitory activities in a dose-dependent manner. The IC(50) of these herbal extracts on inhibition of AChE are at around 5-85mum/ml. The results of this study indicate that there is a great potential to search for novel usage of these medicinal herbs for the treatment of AD.     

Investigating the complexities of sustainable fishing at a prehistoric village on western Vancouver Island, British Columbia, Canada

Zooarchaeological data provide direct evidence of long-term human resource utilization relevant to the modern study of sustainability and conservation. On the northwest coast of North America, prehistoric aboriginal communities relied heavily on nearshore marine fish species, many of which have life history characteristics that make them vulnerable to overexploitation. This study examines temporal trends in the abundance and distribution of nearshore fish from skeletal remains recovered from three separate areas of a prehistoric coastal village on the west coast of Vancouver Island (ca. 1800–250 yr BP). Over the course of 1500 years, site inhabitants from these contemporaneously occupied areas regularly utilized Sebastes spp. (rockfish), with varying impacts on abundance and total length. Contrasting temporal trends in Sebastes spp. total length in separate village areas suggest different consequences to prehistoric harvesting practices, differences that can be linked to ethnographic and archaeological evidence of lineage-based territories. These long-term zooarchaeological data provide unique insights into the context and challenges of sustainable resource use in prehistory and can inform contemporary conservation efforts focused on similar species in this modern-day marine protected area.     

Mechanical competition triggered by innate immune signaling drives the collective extrusion of bacterially infected epithelial cells

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Habitat Use of Booted Eagles (Hieraaetus pennatus) in a Special Protection Area: Implications for Conservation

We examined the patterns of habitat use of six radio-tagged booted eagles (Hieraaetus pennatus) in a Special Protection Area (SPA) in southeastern Spain. Variable percentages of radio locations (20.8–72.0 %) and home range areas (45.2–81.3 %) lay outside the SPA limits. A model selection procedure showed that habitat use was mainly influenced by habitat type and the distance to the nest. Edges and open lands were the most selected habitat types at long distances, while the probability of forest use strongly decreased with the distance. We conclude that effective conservation strategies for this species should encourage not only the protection of the forest nesting habitats, but also the traditional agricultural practices of the surrounding agroecosystems.     

Impact of baculoviral transduction of fluorescent actin on cellular forces

Live staining of actin brings valuable information in the field of mechanobiology. Gene transfer of GFP-actin has been reported to disturb cell rheological properties while gene transfer of fluorescent actin binding proteins was not. However the influence of gene transfer on cellular forces in adhered cells has never been investigated. This would provide a more complete picture of mechanical disorders induced by actin live staining for mechanobiology studies. Indeed, most of these techniques were shown to alter cell morphology. Change in cell morphology may in itself be sufficient to perturb cellular forces. Here we focus on quantifying the alterations of cellular stresses that result from baculoviral transduction of GFP-actin in MDCK cell line. We report that GFP-actin transduction increases the proportion of cells with large intracellular or surface stresses, especially in epithelia with low cell density. We show that the enhancement of the mechanical stresses is accompanied by small perturbations of cell shape, but not by a significant change in cell size. We thus conclude that this live staining method enhances the cellular forces but only brings subtle shape alterations.     

The Large Blue butterfly, Phengaris [Maculinea] arion, as a conservation umbrella on a landscape scale: The case of the Czech Carpathians

Conservation umbrellas are charismatic species, the conservation of which also conserves the high diversity of associated plants and animals. The Large Blue butterfly, Phengaris [Maculinea] arion (Lepidoptera, Lycaenidae), is a textbook example of a charismatic endangered invertebrate, intensively studied throughout Europe and protected by the EU Habitat Directive. While surveying P. arion at the westernmost outskirts of the Carpathians (Javorniky and Vsetinske Mts.), within a stronghold of the species in the Czech Republic, we asked whether occupied sites differed from unoccupied ones in the composition of vascular plants and butterfly assemblages. The occupied sites (n = 65) were small pastures, including abandoned ones, with S to W exposure, located on rugged terrain and displaying a high microtopographic heterogeneity; the unoccupied sites (n = 101) were typically mown or intensively grazed. The vegetation of occupied sites was characteristic for non-intensive submountain pasture, butterfly assemblages were species richer, contained more specialised species, and significantly higher proportion of red-listed species. P. arion thus may act as an umbrella for a high number of species associated with traditional land use in the study area and elsewhere. Its survival will depend on the continuation of small-scale land use varying in space and time, and can be threatened by uniformisation of management, even if practised under the guise of agri-environmental payments.     

Tissue engineering science: Consequences of cell traction force

Blood and tissue cells mechanically interact with soft tissues and tissue-equivalent reconstituted collagen gels in a variety of situations relevant to biomedicine and biotechnology. A key phenomenon in these interactions is the exertion of traction force by cells on local collagen fibers which typically constitute the solid network of these tissues and gels and impart gross mechanical integrity. Two important consequences of cells exerting traction on such collagen networks are first, when the cells co-ordinate their traction, resulting in cell migration, and second, when their traction is sufficient to deform the network. Such cell-collagen network interactions are coupled in a number of ways. Network deformation, for example, can result in net alignment of collagen fibers, eliciting contact guidance, wherein cells move with bidirectional bias along an axis of fiber alignment, potentially leading to a nonuniform cell distribution. This may govern cell accumulation in wounds and be exploited to control cell infiltration of bioartificial tissues and organs. Another consequence of cell traction is the resultant stress and strain in the network which modulate cell protein and DNA synthesis and differentiation. We summarize, here, relevant mathematical theories which we have used to describe the inherent coupling of cell dynamics and tissue mechanics in cell-populated collagen gels via traction. The development of appropriate models based on these theories, in an effort to understand how events in wound healing govern the rate and extent of wound contraction, and to measure cell traction forces in vitro, are described. Relevant observations and speculation from cell biology and medicine that motivate or serve to critique the assumptions made in the theories and models are also summarized.Abbreviations ECM Extracellular Matrix - FPCL Fibroblast-Populated Collagen Lattice - FPCM Fibroblast-Populated Collagen Microsphere