Structural basis for membrane insertion by the human ER membrane protein complex

A defining step in the biogenesis of a membrane protein is the insertion of its hydrophobic transmembrane helices into the lipid bilayer. The nine-subunit endoplasmic reticulum (ER) membrane protein complex (EMC) is a conserved co- and posttranslational insertase at the ER. We determined the structure of the human EMC in a lipid nanodisc to an overall resolution of 3.4 angstroms by cryo–electron microscopy, permitting building of a nearly complete atomic model. We used structure-guided mutagenesis to demonstrate that substrate insertion requires a methionine-rich cytosolic loop and occurs via an enclosed hydrophilic vestibule within the membrane formed by the subunits EMC3 and EMC6. We propose that the EMC uses local membrane thinning and a positively charged patch to decrease the energetic barrier for insertion into the bilayer

Lifetime measurement of the ^3P_2 metastable state of strontium atoms

We have measured the lifetime of the 5s5p ^3P_2 metastable state of strontium atoms by magneto-optically trapping the decayed atoms to the ground state, which allowed sensitive detection of the rare decay events. We found that the blackbody radiation-induced decay was the dominant decay channel for the state at T = 300 K. The lifetime was determined to be 500^{+280}_{-130} s in the limit of zero temperature.Comment: 4 pages, 3 figures, submitted to Physical Review Letter

Regular dipyridamole therapy produces sustained protection against cardiac ischemia-reperfusion injury: is it time to revisit PARIS?

BACKGROUND: Increased activated Akt and eNOS expression coincide with this persistent cardioprotection. Emergent coronary reperfusion therapies are rarely carried out before considerable myocardial injury has occurred. Moreover, reperfusion after prolonged ischemia produces paradoxical ischemia-reperfusion injury, attenuating the efficacy of reperfusion therapies. This has provided impetus for identifying chronic therapies to protect against ischemia-reperfusion injury in those at risk. We previously found that regular dipyridamole therapy produces a chronic preconditioning-like effect mediated through adenosine A1 receptors. METHODS: To determine how long this chronic preconditioning effect of dipyridamole remains present after discontinuing therapy, guinea pigs received 4 mg/kg/day in their water for 6 weeks. Ischemia-reperfusion was performed at 0, 2, 3, and 4 days after dipyridamole discontinuation (0 day, 2 days, 3 days and 4 days; n=8 per group). Left ventricular developed pressure (LVDP), end-diastolic pressure (LVEDP), coronary flow (CF), infarct size, and western blot analyses for Akt and endothelial nitric oxide synthase (eNOS) were studied. RESULTS: After ischemia-reperfusion, 0 day, 2 days and 3 days, but not 4 days, had significantly higher LVDP and lower LVEDP compared to control. Myocardial infarct size was significantly reduced at 0 day, 2 days and 3 days, but not 4 days, compared to control. Western blot analyses demonstrated upregulation of phospho-Akt and phospho-eNOS expression at 0 day, 2 days, and 3 days, but not 4 days. CONCLUSIONS: A chronic preconditioning-like cardioprotection by regular dipyridamole treatment persists for 3 days after discontinuing therapy. Increased activated Akt and eNOS expression may play a role in this persistent cardioprotection

Cardioprotection by regular ethanol consumption: potential mechanisms and clinical application.

Epidemiological studies demonstrate that excessive drinking is associated with hypertension, cerebral bleeding and loss of cardiac contractility. Conversely, studies have shown that mortality rates for people who regularly drink ethanol in moderation are lower than in abstainers, primarily due to decreased fatal ischemic heart disease. Further, moderate ethanol consumers have lower rates of myocardial infarction compared with abstainers. These beneficial cardiac effects may be due to pleiotropic effects of ethanol on lipids, platelets, and fibrinolytic activity. During the past decade, studies conducted in several animal models have revealed that light to moderate regular ethanol consumption renders hearts more tolerant to myocardial ischemia-reperfusion injury; to a degree similar to cardiac ischemic preconditioning (brief episodes of ischemia dramatically limit infarct size following prolonged ischemia). Recent clinical evidence suggests that light to moderate ethanol consumption in the year prior to myocardial infarction is associated with reduced mortality following myocardial infarction. These findings suggest that light to moderate ethanol consumption not only prevents myocardial infarction but also improves survival after myocardial infarction. Proposed mechanisms of cardioprotection by regular ethanol consumption include activation of adenosine A1 receptors, alpha(1)-adrenoceptors, protein kinase C-delta and epsilon, adenosine triphosphate-dependent potassium (K(ATP)) channels, nitric oxide synthase and reduced leukocyte-endothelial cell adhesive interactions. In this review, we focus on the recent progress in elucidating the endogenous myocyte signaling mediating cardioprotection by light to moderate ethanol consumption

Direct evidence for inhibition of mitochondrial permeability transition pore opening by sevoflurane preconditioning in cardiomyocytes: comparison with cyclosporine A.

To assess whether sevoflurane preconditioning is associated with inhibition of mitochondrial permeability transition pore (MPTP), the effects of sevoflurane were compared with those of cyclosporine A, a known inhibitor of MPTP opening. Isolated perfused guinea pig hearts underwent 30 min global ischemia and 120 min reperfusion (control). Sevoflurane preconditioning was elicited by administration of 2% sevoflurane for 10 min with 10 min washout before ischemia (sevoflurane). A preconditioning-like cardioprotection was also induced by administering cyclosporine A (0.2 μM) for 15 min, starting 5 min before ischemia and for 10 min after the onset of reperfusion (cyclosporine A). Left ventricular developed and end-diastolic pressures, coronary flow and infarct size were measured. Expressions of Akt and glycogen synthase kinase 3β (GSK3β), known mediators of inhibition of MPTP opening, were determined by Western blot analysis. GSK3β inhibition was achieved with LY294002. The effects of sevoflurane and cyclosporine A on calcium-induced MPTP opening in isolated calcein-loaded mitochondria were assessed. After ischemia-reperfusion, sevoflurane and cyclosporine A had higher left ventricular developed pressure. Infarct size was significantly reduced in sevoflurane and cyclosporine A vs. control. This was abolished by LY294002 in sevoflurane, but not in cyclosporine A. Akt and GSK3β phosphorylation after reperfusion were significantly increased in sevoflurane and cyclosporine A. Ca²⁺-induced reduction in calcein fluorescence was significantly attenuated in sevoflurane and cyclosporine A. Preconditioning agents, sevoflurane and cyclosporine A increase the threshold of calcium-induced MPTP opening to a similar extent. This effect by sevoflurane, but not cyclosporine A is at least partially mediated by GSK3β inactivation

Primary palliative care in Japan: needs estimation and projections – national database study with international comparisons

Objectives We aimed to estimate the potential population that requires palliative care, clarify the relationship between this population and the rate of ageing in Japan, and compare these trends with those of other countries.Design We used the national death registration data and population projections for Japan to estimate the population in need of palliative care using the minimal estimate method developed by Murtagh et al. Linear regression was used to create a model of mortality using sex, age at intervals of 5 years, and each major disease classification. We calculated the future population in need of palliative care until 2040 and compared the ageing data to those of other countries.Setting/participants All adults in Japan who died from 1980 to 2040 at intervals of 5 years.Results The number of people who might need palliative care from 2020 to 2040 will also increase linearly from 1 059 000 to 1 405 000. The proportion of Alzheimer’s, dementia and senility of the total need for palliative care will increase to 43.4% in 2040. The correlation coefficient between the proportion of the population in need of palliative care and the rate of ageing was 0.24 in developed countries.Conclusion In Japan, the population requiring palliative care in 2040 will be 1.5 times that in 2015. Palliative care needs to be provided urgently for people with Alzheimer’s disease, dementia and senility. The proportion of patients in need of palliative care may not change, although the number of patients requiring such gradually increases in developed countries

Image segmentation techniques for granular materials

To improve understanding of the mechanical behavior of granular materials it is important to be able to quantify the relative arrangement of the grains, i.e. the fabric. This can be done, for example, by measuring the orientations of the particles (e.g. the long axis orientation) or by considering the orientations of the vectors normal to each grain‐grain contact. In two dimensional (2D) analyses this information can be obtained by digital image analysis of images of thin sections obtained from an optical microscope. While such data is useful, granular materials of engineering interest are three dimensional (3D) materials and quantification of the 3D fabric is necessary. Micro Computed‐Tomography (μCT) together with 3D image analysis has emerged as a promising technique for obtaining the 3D data required. This paper aims to highlight the challenges associated with using image analysis to provide quantitative information on fabric. While automated image segmentation has proved to produce reasonable results in some cases, it is sometimes less successful when dealing with highly irregular and angular soil grains. This paper evaluates the effectiveness of 2D and 3D segmentation techniques that rely on the watershed segmentation algorithm. The primary material considered is Reigate Silver Sand, a natural quartzitic sand with grain diameters in the range of 150–300 μm. While the sand considered is primarily of interest to geotechnical engineers, the results of this study will be of interest to anyone seeking to quantify granular material fabric using either 2D microscopy data or μCT 3D data sets

Epoxy-functionalised 4-vinylguaiacol for the synthesis of bio-based, degradable star polymers via a RAFT/ROCOP strategy

An epoxy derivative of a naturally occuring vinylphenolic compound, 4-vinylguaiacol (4VGEP), was used for the synthesis of a well-defined (Mw/Mn = 1.08–1.14), bio-based styrene-type polymer. Block copolymers of 4VGEP with styrene and diethylacrylamide were also prepared and used as macro-monomers in metal, and metal-free, ring-opening copolymerisation (ROCOP) with cis-4-cyclohexene-1,2-dicarboxylic anhydride to form ester cross-linked star polymers in high yields (82–90%), with narrow dispersity (Mw/Mn = 1.27–1.40). Finally, the selective degradation of the ester core of the styrene-based star was achieved under acidic conditions

The Large-scale and Small-scale Clustering of Lyman-Break Galaxies at 3.5 < z< 5.5 from the GOODS survey

We report on the angular correlation function of Lyman-break galaxies (LBGs) at z~4 and 5 from deep samples obtained from the Great Observatories Deep Origins Survey (GOODS). Similar to LBGs at z~3, the shape of w(theta) of the GOODS LBGs is well approximated by a power-law with slope beta~0.6 at angular separation theta > 10 arcsec. The clustering strength of z~4, 5 LBGs also depends on the rest-frame UV luminosity, with brighter galaxies more strongly clustered than fainter ones, implying a general correlation between halos' mass and LBGs' star-formation rate. At smaller separations, w(theta) of deep samples significantly exceeds the extrapolation of the large-scale power-law fit, implying enhanced spatial clustering at scales r < 1 Mpc. We also find that bright LBGs statistically have more faint companions on scales theta < 20 arcsec than fainter ones, showing that the enhanced small-scale clustering is very likely due to sub-structure, namely the fact that massive halos can host multiple galaxies. A simple model for the halo occupation distribution and the CDM halo mass function reproduce well the observed w(theta). The scaling relationship of the clustering strength with volume density and with redshift is quantitatively consistent with that of CDM halos. A comparison of the clustering strength of three samples of equal luminosity limit at z ~ 3, 4 and 5 shows that the LBGs at z~5 are hosted in halos about one order of magnitude less massive than those in the lower redshift bins, suggesting that star-formation was more efficient at higher-redshift.Comment: replaced with the version accepted for publication in ApJ. 46 pages, 10 figures; minor changes to text, one subsection adde