Store-operated Ca(2+) channels and Stromal Interaction Molecule 1 (STIM1) are targets for the actions of bile acids on liver cells

Cholestasis is a significant contributor to liver pathology and can lead to primary sclerosis and liver failure. Cholestatic bile acids induce apoptosis and necrosis in hepatocytes but these effects can be partially alleviated by the pharmacological application of choleretic bile acids. These actions of bile acids on hepatocytes require changes in the release of Ca(2+) from intracellular stores and in Ca(2+) entry. However, the nature of the Ca(2+) entry pathway affected is not known. We show here using whole cell patch clamp experiments with H4-IIE liver cells that taurodeoxycholic acid (TDCA) and other choleretic bile acids reversibly activate an inwardly-rectifying current with characteristics similar to those of store-operated Ca(2+) channels (SOCs), while lithocholic acid (LCA) and other cholestatic bile acids inhibit SOCs. The activation of Ca(2+) entry was observed upon direct addition of the bile acid to the incubation medium, whereas the inhibition of SOCs required a 12 h pre-incubation. In cells loaded with fura-2, choleretic bile acids activated a Gd(3+)-inhibitable Ca(2+) entry, while cholestatic bile acids inhibited the release of Ca(2+) from intracellular stores and Ca(2+) entry induced by 2,5-di-(tert-butyl)-1,4-benzohydro-quinone (DBHQ). TDCA and LCA each caused a reversible redistribution of stromal interaction molecule 1 (STIM1, the endoplasmic reticulum Ca(2+) sensor required for the activation of Ca(2+) release-activated Ca(2+) channels and some other SOCs) to puncta, similar to that induced by thapsigargin. Knockdown of Stim1 using siRNA caused substantial inhibition of Ca(2+)-entry activated by choleretic bile acids. It is concluded that choleretic and cholestatic bile acids activate and inhibit, respectively, the previously well-characterised Ca(2+)-selective hepatocyte SOCs through mechanisms which involve the bile acid-induced redistribution of STIM1.     

IL-6 Promotes compensatory liver regeneration in cirrhotic rat after partial hepatectomy

Major hepatic resection in cirrhotic patients is associated with impaired liver regeneration and failure, leading to high peri-operative mortality. In this work, the causes of defective regeneration in cirrhotic liver and the utility of IL-6 treatment were investigated in an experimental model combining cirrhosis and partial hepatectomy in the rat. Relative to normal controls, decompensated cirrhotic animals showed decreased survival, while compensated cirrhotic animals showed similar survival but reduced hepatic DNA synthesis and newly regenerated liver mass amount. Defective liver regeneration was associated with a decrease in STAT3 and NF-kB activation, consistent with an increased accumulation of their respective inhibitors PIAS3 and IkBα, and with a decreased induction of Bcl-xL. Treatment with recombinant IL-6 enhanced survival of decompensated cirrhotic animals, while it did not affect survival of compensated cirrhotic animals but sustained liver regeneration, by restoring STAT3 and NF-kB activation and Bcl-xL induction to the levels found in normal controls. The pro-growth effects exerted by IL-6 treatment in cirrhotic liver were attained also at low, pharmacologically acceptable doses. In conclusion, our results suggest that IL-6 treatment may be therapeutic in major resection of cirrhotic liver.     

The war of tools: how can NMR spectroscopists detect errors in their structures?

Protein structure determination by NMR methods has started in the mid-eighties and has been growing steadily since then. Ca. 14% of the protein structures deposited in the PDB have been solved by NMR. The evaluation of the quality of NMR structures however is still lacking a well-established practice. In this work, we examined various tools for the assessment of structural quality to ascertain the extent to which these tools could be applied to detect flaws in NMR structures. In particular, we investigated the variation in the scores assigned by these programs as a function of the deviation of the structures induced by errors in assignments or in the upper distance limits used. These perturbations did not distort radically the protein fold, but resulted in backbone RMS deviations up to 3 Å, which is in line with errors highlighted in the available literature. We found that it is quite difficult to discriminate the structures perturbed because of misassignments from the original ones, also because the spread in score over the conformers of the original bundle is relatively large. ?–ψ distributions and normality scores related to the backbone conformation and to the distribution of side-chain dihedral angles are the most sensitive indicators of flaws.     

Another Lesson from Plants: The Forward Osmosis-Based Actuator

Osmotic actuation is a ubiquitous plant-inspired actuation strategy that has a very low power consumption but is capable of generating effective movements in a wide variety of environmental conditions. In light of these features, we aimed to develop a novel, low-power-consumption actuator that is capable of generating suitable forces during a characteristic actuation time on the order of a few minutes. Based on the analysis of plant movements and on osmotic actuation modeling, we designed and fabricated a forward osmosis-based actuator with a typical size of 10 mm and a characteristic time of 2–5 minutes. To the best of our knowledge, this is the fastest osmotic actuator developed so far. Moreover, the achieved timescale can be compared to that of a typical plant cell, thanks to the integrated strategy that we pursued by concurrently addressing and solving design and material issues, as paradigmatically explained by the bioinspired approach. Our osmotic actuator produces forces above 20 N, while containing the power consumption (on the order of 1 mW). Furthermore, based on the agreement between model predictions and experimental observations, we also discuss the actuator performance (including power consumption, maximum force, energy density and thermodynamic efficiency) in relation to existing actuation technologies. In light of the achievements of the present study, the proposed osmotic actuator holds potential for effective exploitation in bioinspired robotics systems.     

Piante nuove o notevoli per la flora delle Marche rinvenute nel Bacino montano dell'Esino

Abstract

New or interesting plants in the mountainous basin of the Esino (Marche region).—The Authors point out a discovery of 46 species of remarkable importance concerning the flora of the Marche region in the mountainous basin of the Esino. Among these, Poa chaixi and Monotropa hypopithis have been discovered in this region for the first time.