ER localized bestrophin1 activates Ca2+ dependent ion channels TMEM16A and SK4

Bestrophins form Ca2+ activated Cl- channels and regulate intercellular Ca2+ signaling1. We demonstrate that bestrophin 1 is localized in the endoplasmic reticulum (ER), where it physically interacts with stromal interacting molecule 1 (Stim1), the ER-Ca2+ sensor2,3. Intracellular Ca2+ transients in HEK293 cells elicited by stimulation of purinergic P2Y2-receptors were augmented but more transient after expression of hBest1, in contrast to dominant negative hBest1-R218C, which attenuated Ca2+ increase. The p21-activated protein kinase Pak2 was found to phosphorylate hBest1, thereby enhancing Ca2+ signaling and activation of Ca2+ dependent Cl- (TMEM16A)4 and K+ (SK4)5 channels. Lack of bestrophin 1 expression in respiratory epithelial cells of mBest1 knockout mice caused expansion of ER cisterns and induced Ca2+ deposits. We propose that hBest1 is important for Ca2+ handling of the ER store, probably by controlling the function of Stim1 and by acting as a counter-ion channel to balance transient membrane potentials occurring through inositol trisphosphate (IP3) induced Ca2+ release and refill of the ER-Ca2+ store. Thus bestrophin 1 controls activation of Ca2+ dependent ion channels by regulation of compartmentalized Ca2+ signaling

On the Influence of Diurnal and Subdiurnal Signals in the Normal Vector on Large Ring Laser Gyroscope Observations

The ring laser gyroscope (RLG) technique has been investigated for over 20 years as a potential complement to space geodetic techniques in measuring Earth rotation. However, RLGs are also sensitive to changes in their terrestrial orientation. Therefore in this paper, we review how the high-frequency band (i.e. signals shorter than 0.5 cycle per day) of the known phenomena causing site deformation contribute to the RLG observable, the Sagnac frequency. We study the impact of solid Earth tides, ocean tidal loading and non-tidal loading phenomena (atmospheric pressure loading and continental hydrosphere loading). Also, we evaluate the differences between available models of the phenomena and the importance of the Love numbers used in modeling the impact of solid Earth tides. Finally, we compare modeled variations in the instrument orientation with the ones observed with a tiltmeter. Our results prove that at the present accuracy of the RLG technique, solid Earth tides and ocean tidal loading effects have significant effect on RLG measurements, and continental hydrosphere loading can be actually neglected. Regarding the atmospheric loading model, its application might introduce some undesired signals. We also show that discrepancies arising from the use of different models can be neglected, and there is almost no impact arising from the use of different Love numbers. Finally, we discuss differences between data reduced with tiltmeter observations and these reduced with modeled signal, and potential causes of this discrepancies

HIGH-OXYGEN MODIFIED ATMOSPHERE PACKAGING OF ORGANIC MEATS?

Abstract In the last few years, it became widespread to package fresh meat for retail under modified atmosphere containing 70-80% oxygen ("high O 2 -MAP"). Oxygen preserves the bright-red colour of oxymyoglobin while CO 2 (20-30%) inhibits obligatory aerobic bacteria such as psychrotrophic Pseudomonas spp., the common spoilage agents for unpackaged fresh meat. On the other hand, high oxygen partial pressure has been shown to cause some oxidative changes in the meat, resulting in sensory deviations. Use of high O 2 -MAP is also legally permitted for the packaging of organic meats. This may be regarded as being in conflict with the principle of "processing food with care" as stipulated by Article 6 of the Regulation (EU) 834/2007 on organic production and labelling of organic products. Therefore, the aim of our research was to provide information to decision-makers in the organic food chain on the necessity of high O 2 -MAP for organic meats. Literature data from market research show that even consumers of organic food seek "convenience" in purchasing and preparing it. Hence, of the major associations of organic agriculture in Germany, only Demeter e. V. prohibits the use of high O 2 -MAP. Literature data also indicate that undesirable changes of the meat could be delayed by reducing the oxygen content in the packages to about 40-50%, with only a slight reduction of shelf life. In our view, this would be a good compromise between "convenience" and eating quality

Molecular evolution and functional divergence of the bestrophin protein family

<p>Abstract</p> <p>Background</p> <p>Mutations in human bestrophin 1 are associated with at least three autosomal-dominant macular dystrophies including Best disease, adult onset vitelliform macular dystrophy and autosomal dominant vitreo-retinochoroidopathy. The protein is integral to the membrane and is likely involved in Ca²⁺-dependent transport of chloride ions across cellular membranes. Bestrophin 1 together with its three homologues forms a phylogenetically highly conserved family of proteins.</p> <p>Results</p> <p>A bioinformatics study was performed to investigate the phylogenetic relationship among the bestrophin family members and to statistically evaluate sequence conservation and functional divergence. Phylogenetic tree assembly with all available eukaryotic bestrophin sequences suggests gene duplication events in the lineage leading to the vertebrates. A common N-terminal topology which includes four highly conserved transmembrane domains is shared by the members of the four paralogous groups of vertebrate bestrophins and has been constrained by purifying selection. Pairwise comparison shows that altered functional constraints have occurred at specific amino acid positions after phylogenetic diversification of the paralogues. Most notably, significant functional divergence was found between bestrophin 4 and the other family members, as well as between bestrophin 2 and bestrophin 3. Site-specific profiles were established by posterior probability analysis revealing significantly divergent clusters mainly in two hydrophilic loops and a region immediately adjacent to the last predicted transmembrane domain. Strikingly, codons 279 and 347 of human bestrophin 4 reveal high divergence when compared to the paralogous positions strongly indicating the functional importance of these residues for the bestrophin 4 protein. None of the functionally divergent amino acids were found to reside within obvious sequences patterns or motifs.</p> <p>Conclusion</p> <p>Our study highlights the molecular evolution of the bestrophin family of transmembrane proteins and indicates amino acid residues likely relevant for distinct functional properties of the paralogues. These findings may provide a starting point for further experimental verifications.</p

Paneth Cell Secretion in vivo Requires Expression of Tmem16a and Tmem16f

Background and Aims Paneth cells play a central role in intestinal innate immune response. These cells are localized at the base of small intestinal crypts of Lieberkuhn. The calcium-activated chloride channel TMEM16A and the phospholipid scramblase TMEM16F control intracellular Ca2+ signaling and exocytosis. We analyzed the role of TMEM16A and TMEM16F for Paneth cells secretion. Methods Mice with intestinal epithelial knockout of Tmem16a (Tmem16a−/−) and Tmem16f (Tmem16f−/−) were generated. Tissue structures and Paneth cells were analyzed, and Paneth cell exocytosis was examined in small intestinal organoids in vitro. Intracellular Ca2+ signals were measured and were compared between wild-type and Tmem16 knockout mice. Bacterial colonization and intestinal apoptosis were analyzed. Results Paneth cells in the crypts of Lieberkuhn from Tmem16a−/− and Tmem16f−/− mice demonstrated accumulation of lysozyme. Tmem16a and Tmem16f were localized in wild-type Paneth cells but were absent in cells from knockout animals. Paneth cell number and size were enhanced in the crypt base and mucus accumulated in intestinal goblet cells of knockout animals. Granule fusion and exocytosis on cholinergic and purinergic stimulation were examined online. Both were strongly compromised in the absence of Tmem16a or Tmem16f and were also blocked by inhibition of Tmem16a/f. Purinergic Ca2+ signaling was largely inhibited in Tmem16a knockout mice. Jejunal bacterial content was enhanced in knockout mice, whereas cellular apoptosis was inhibited. Conclusion The present data demonstrate the role of Tmem16 for exocytosis in Paneth cells. Inhibition or activation of Tmem16a/f is likely to affect microbial content and immune functions present in the small intestine

Tracing the sites of obscured star formation in the Antennae galaxies with Herschel-PACS

FIR imaging of interacting galaxies allows locating even hidden sites of star formation and measuring of the relative strength of nuclear and extra-nuclear star formation. We want to resolve the star-forming sites in the nearby system of the Antennae. Thanks to the unprecedented sharpness and depth of the PACS camera onboard ESA's Herschel Space Observatory, it is possible for the first time to achieve a complete assessment of individual star-forming knots in the FIR with scan maps at 70, 100, and 160 um. We used clump extraction photometry and SED diagnostics to derive the properties related to star-forming activity. The PACS 70, 100, and 160 um maps trace the knotty structure of the most recent star formation along an arc between the two nuclei in the overlap area. The resolution of the starburst knots and additional multi-wavelength data allow their individual star formation history and state to be analysed. In particular, the brightest knot in the mid-infrared (K1), east of the southern nucleus, exhibits the highest activity by far in terms of dust heating and star formation rate, efficiency, and density. With only 2 kpc in diameter, this area has a 10-1000 um luminosity, which is as high as that of our Milky Way. It shows the highest deficiency in radio emission in the radio-to-FIR luminosity ratio and a lack of X-ray emission, classifying it as a very young complex. The brightest 100 and 160 um emission region (K2), which is close to the collision front and consists of 3 knots, also shows a high star formation density and efficiency and lack of X-ray emission in its most obscured part, but an excess in the radio-to-FIR luminosity ratio. This suggests a young stage, too, but different conditions in its interstellar medium. Our results provide important checkpoints for numerical simulations of interacting galaxies when modelling the star formation and stellar feedback.Comment: 4 pages, 4 figures, 2 tables (A&A Herschel special issue