Na,K-ATPase α2 activity in mammalian skeletal muscle T-tubules is acutely stimulated by extracellular K+

The Na,K-ATPase α2 isoform is the predominant Na,K-ATPase in adult skeletal muscle and the sole Na,K-ATPase in the transverse tubules (T-tubules). In quiescent muscles, the α2 isozyme operates substantially below its maximal transport capacity. Unlike the α1 isoform, the α2 isoform is not required for maintaining resting ion gradients or the resting membrane potential, canonical roles of the Na,K-ATPase in most other cells. However, α2 activity is stimulated immediately upon the start of contraction and, in working muscles, its contribution is crucial to maintaining excitation and resisting fatigue. Here, we show that α2 activity is determined in part by the K⁺ concentration in the T-tubules, through its K⁺ substrate affinity. Apparent K⁺ affinity was determined from measurements of the K_1/2 for K⁺ activation of pump current in intact, voltage-clamped mouse flexor digitorum brevis muscle fibers. Pump current generated by the α2 Na,K-ATPase, Ip, was identified as the outward current activated by K⁺ and inhibited by micromolar ouabain. Ip was outward at all potentials studied (−90 to −30 mV) and increased with depolarization in the subthreshold range, −90 to −50 mV. The Q₁₀ was 2.1 over the range of 22–37°C. The K_1/2,K of Ip was 4.3 ± 0.3 mM at −90 mV and was relatively voltage independent. This K⁺ affinity is lower than that reported for other cell types but closely matches the dynamic range of extracellular K⁺ concentrations in the T-tubules. During muscle contraction, T-tubule luminal K⁺ increases in proportion to the frequency and duration of action potential firing. This K_1/2,K predicts a low fractional occupancy of K⁺ substrate sites at the resting extracellular K⁺ concentration, with occupancy increasing in proportion to the frequency of membrane excitation. The stimulation of preexisting pumps by greater K⁺ site occupancy thus provides a rapid mechanism for increasing α2 activity in working muscles.     

Tool for the environmental assessment of cranes based on parameterization

Purpose  

Information constitutes one of the main barriers for applying life cycle assessment (LCA) due to complexity and need for great amounts of it. However, most of the parameters that determine the data are defined early in the product development process. Knuckle boom cranes constitute a complex product which poses a particularly pressing need for simplification. This paper models the LCA inventory information out of design parameters. The paper also presents a tool implementing this.     

Method for Setting Environmental Targets in Product Development: Incorporating Use‐Phase Impact by Subsystem

In order to address environmental aspects during redesign, the product specification must include related targets that are reachable and challenging. To do so, this article presents a stepwise approach for combining benchmarking information and component impact, out of life cycle assessment (LCA) scaling. This approach requires allocating environmental impacts to each subsystem, which is not commonly done for some life cycle phases in LCAs, most particularly for use phases. This article includes a methodology for allocating such impacts. The underlying criterion is avoiding complex calculations, to make the method more agile. This methodology is presented in a full case study of a complex product: a knuckle boom crane. The case study results in the percentage of impact reduction needed to meet the market average or best competitors. In particular, the results show that the cylinders of the crane have a high contribution to environmental impact, not only because of their weight, but also because of the active power consumed to activate them.     

Adult vascular smooth muscle cells in culture express neural stem cell markers typical of resident multipotent vascular stem cells

Differentiation of resident multipotent vascular stem cells (MVSCs) or de-differentiation of vascular smooth muscle cells (vSMCs) might be responsible for the SMC phenotype that plays a major role in vascular diseases such as arteriosclerosis and restenosis. We examined vSMCs from three different species (rat, murine and bovine) to establish whether they exhibit neural stem cell characteristics typical of MVSCs. We determined their SMC differentiation, neural stem cell marker expression and multipotency following induction in vitro by using immunocytochemistry, confocal microscopy, fluorescence-activated cell sorting analysis and quantitative real-time polymerase chain reaction. MVSCs isolated from rat aortic explants, enzymatically dispersed rat SMCs and rat bone-marrow-derived mesenchymal stem cells served as controls. Murine carotid artery lysates and primary rat aortic vSMCs were both myosin-heavy-chain-positive but weakly expressed the neural crest stem cell marker, Sox10. Each vSMC line examined expressed SMC differentiation markers (smooth muscle α–actin, myosin heavy chain and calponin), neural crest stem cell markers (Sox10⁺, Sox17⁺) and a glia marker (S100β⁺). Serum deprivation significantly increased calponin and myosin heavy chain expression and decreased stem cell marker expression, when compared with serum-rich conditions. vSMCs did not differentiate to adipocytes or osteoblasts following adipogenic or osteogenic inductive stimulation, respectively, or respond to transforming growth factor-β1 or Notch following γ-secretase inhibition. Thus, vascular SMCs in culture express neural stem cell markers typical of MVSCs, concomitant with SMC differentiation markers, but do not retain their multipotency. The ultimate origin of these cells might have important implications for their use in investigations of vascular proliferative disease in vitro.     

Estimating Environmental Behavior Without Performing a Life Cycle Assessment

Many authors have agreed on the interest of considering environmental concerns in the early stages of product development. However, most eco‐design tools are based on life cycle assessment principles and require a model to give information about the product's environmental performance. This modeling can have negative effects on team performance and on the potential for innovation, not to mention on the project's duration. Additionally, the model requires information that is not available in the early design stages. This article analyzes the potential of inferring conclusions about the life cycle stages with the highest impact by using similar products. From a database of previous products, environmental profile estimations are carried out, that is, the assessments of the contribution of each life cycle stage to the total impact and the variability of this measure. It is then possible to discard—or ensure consideration of—life cycle stages. Furthermore, the level of the conclusions is assessed on a five‐point scale. The proposed approach is applied to four case studies with different levels of abstraction and the relevance of the conclusions is assessed. The article resolves the problems regarding potential for estimating the distribution of the environmental impacts along the life cycle.