Basal Flux Research Articles

Glucose-starved Cells Do Not Engage in Prosurvival Autophagy

In response to nutrient shortage or organelle damage, cells undergo macroautophagy. Starvation of glucose, an essential nutrient, is thought to promote autophagy in mammalian cells. We thus aimed to determine the role of autophagy in cell death induced by glucose deprivation. Glucose withdrawal induces cell death that can occur by apoptosis (in Bax, Bak-deficient mouse embryonic fibroblasts or HeLa cells) or by necrosis (in Rh4 rhabdomyosarcoma cells). Inhibition of autophagy by chemical or genetic means by using 3-methyladenine, chloroquine, a dominant negative form of ATG4B or silencing Beclin-1, Atg7, or p62 indicated that macroautophagy does not protect cells undergoing necrosis or apoptosis upon glucose deprivation. Moreover, glucose deprivation did not induce autophagic flux in any of the four cell lines analyzed, even though mTOR was inhibited. Indeed, glucose deprivation inhibited basal autophagic flux. In contrast, the glycolytic inhibitor 2-deoxyglucose induced prosurvival autophagy. Further analyses indicated that in the absence of glucose, autophagic flux induced by other stimuli is inhibited. These data suggest that the role of autophagy in response to nutrient starvation should be reconsidered.

Mitochondrial dynamics in mitochondrial optic neuropathies

AbstractOPA1 mutations are the commonest cause of dominantly inherited optic atrophy (DOA) and so‐called DOA plus. The Opa1 protein is essential for normal mitochondrial fusion. In mouse DOA, autophagy (recycling of spent cellular components) is dysregulated in retinal ganglion cells (RGCs), and mitophagy (mitochondrial recycling) has been implicated. We validated and used novel ImageStream technology to quantitate mitophagy in primary cells. Co‐localisation of mitochondria and autophagosomes was increased in fibroblasts in five patients from four families with severe OPA1 phenotypes indicating increased mitophagy. ImageStream also showed that basal mitophagy was increased when control cultures were depleted of Opa1 by siRNA. Western blotting confirmed increased basal autophagy and autophagic flux in the presence of activators. Increased mitochondrial fragmentation, mitophagy and failure of mitochondrial transport may together cause local depletion of mitochondria in critical regions of the retinal ganglion cells, such as axons or synapses. Fragmentation may also impair stress induced mitochondrial hyperfusion. Increased mitochondrial fragmentation, mislocalisation and mitophagy thus link low OPA1 to neurodegeneration.

Ets-1 global gene expression profile reveals associations with metabolism and oxidative stress in ovarian and breast cancers

BackgroundThe Ets-1 proto-oncogene is frequently upregulated in cancer cells, with known involvement in cancer angiogenesis, metastasis, and more recently energy metabolism. In this study we have performed various bioinformatic analyses on existing microarray data to further clarify the role of Ets-1 in ovarian cancer, and validated these results with functional assays.MethodsFunctional pathway analyses were conducted on existing microarray data comparing 2008 and 2008-Ets1 ovarian cancer cells. Methods included over-representation analysis, functional class scoring and pathway topology, and network representations were visualized in Cytoscape. Oxidative stress regulation was examined in ovarian cancer cells by measuring protein expression and enzyme activity of glutathione peroxidases, as well as intracellular reactive oxygen species using dichlorofluorescin fluorescence. A stable Ets-1 knockdown MDA-MB-231 cell line was created using short hairpin RNA, and glycolytic dependence of these cells was measured following treatment with 2-deoxy-D-glucose and Hoechst nuclear staining to determine cell number. High-resolution respirometry was performed to measure changes in basal oxygen flux between MDA-MB-231 cells and MDA-Ets1KD variants.ResultsEnrichments in oxidoreductase activity and various metabolic pathways were observed upon integration of the different analyses, suggesting that Ets-1 is important in their regulation. As oxidative stress is closely associated with these pathways, we functionally validated our observations by showing that Ets-1 overexpression resulted in decreased reactive oxygen species with increased glutathione peroxidase expression and activity, thereby regulating cellular oxidative stress. To extend our findings to another cancer type, we developed an Ets-1 knockdown breast cancer cell model, which displayed decreased glycolytic dependence and increased oxygen consumption following Ets-1 knockdown confirming our earlier findings.ConclusionsCollectively, this study confirms the important role of Ets-1 in the regulation of cancer energy metabolism in ovarian and breast cancers. Furthermore, Ets-1 is a key regulator of oxidative stress in ovarian cancer cells by mediating alterations in glutathione antioxidant capacity.

Numerical simulations of CO2 migration during charnockite genesis

Charnockite is considered to be generated either through the dehydration of granitic magma by CO2 purging or by solid-state dehydration through CO2 metasomatism during granulite facies metamorphism. To understand the extent of dehydration, CO2 migration is quantitatively modeled in silicate melt and metasomatic fluid as a function of temperature, H2O wt%, pressure, basal CO2 flux and dynamic viscosity. Numerical simulations show that CO2 advection through porous and permeable high-grade metamorphic rocks can generate dehydrated patches close to the CO2 flow path, as illustrated by the occurrences of ``incipient charnockites.'' CO2 reaction-front velocity constrained by field observations is 0.69 km/m.y., a reasonable value, which matches well with other studies. On the other hand, temperature, rate of cooling, and basal CO2 flux are the critical parameters affecting CO2 diffusion through a silicate melt. CO2 diffusion through silicate melt can only occur at temperature greater than 840 degrees C and during slow cooling (<= 3.7 x 10(-5) degrees C/yr), features that are typical of magma emplacement in the lower crust. Stalling of CO2 fluxing at similar to 840 degrees C explains why some deep-level plutons contain both hydrous and anhydrous (charnockitic) mineral assemblages. CO2 diffusion through silicate melt is virtually insensitive to pressure. Addition of CO2 basal flux facilitates episodic dehydrated melt migration by generating fracture pathways.

Autophagy is required for exercise training‐induced skeletal muscle adaptation and improvement of physical performance

Pathological and physiological stimuli, including acute exercise, activate autophagy; however, it is unknown whether exercise training alters basal levels of autophagy and whether autophagy is required for skeletal muscle adaptation to training. We observed greater autophagy flux (i.e., a combination of increased LC3-II/LC3-I ratio and LC3-II levels and reduced p62 protein content indicating a higher rate of initiation and resolution of autophagic events), autophagy protein expression (i.e., Atg6/Beclin1, Atg7, and Atg8/LC3) and mitophagy protein Bnip3 expression in tonic, oxidative muscle compared to muscles of either mixed fiber types or of predominant glycolytic fibers in mice. Long-term voluntary running (4 wk) resulted in increased basal autophagy flux and expression of autophagy proteins and Bnip3 in parallel to mitochondrial biogenesis in plantaris muscle with mixed fiber types. Conversely, exercise training promoted autophagy protein expression with no significant increases of autophagy flux and mitochondrial biogenesis in the oxidative soleus muscle. We also observed increased basal autophagy flux and Bnip3 content without increases in autophagy protein expression in the plantaris muscle of sedentary muscle-specific Pgc-1α transgenic mice, a genetic model of augmented mitochondrial biogenesis. These findings reveal that endurance exercise training-induced increases in basal autophagy, including mitophagy, only take place if an enhanced oxidative phenotype is achieved. However, autophagy protein expression is mainly dictated by contractile activity independently of enhancements in oxidative phenotype. Exercise-trained mice heterozygous for the critical autophagy protein Atg6 showed attenuated increases of basal autophagy flux, mitochondrial content, and angiogenesis in skeletal muscle, along with impaired improvement of endurance capacity. These results demonstrate that increased basal autophagy is required for endurance exercise training-induced skeletal muscle adaptation and improvement of physical performance.

Ca II H+K fluxes from S-indices of large samples: a reliable and consistent conversion based on PHOENIX model atmospheres

Context. Historic stellar activity data based on chromospheric line emission using O.C. Wilson’s S-index reach back to the 1960ies and represent a very valuable data resource both in terms of quantity and time-coverage. However, these data are not flux-calibrated and are therefore difficult to compare with modern spectroscopy and to relate to quantitative physics. Aims. In order to make use of the rich archives of Mount Wilson and many other S-index measurements of thousands of main sequence stars, subgiants and giants in terms of physical Ca ii H+K line chromospheric surface fluxes and the related R-index, we seek a new, simple but reliable conversion method of the S-indices. A first application aims to obtain the (empirical) basal chromospheric surface flux to better characterise stars with minimal activity levels. Methods. We collect 6024 S-indices from six large catalogues from a total of 2530 stars with well-defined parallaxes (as given by the Hipparcos catalogue) in order to distinguish between main sequence stars (2133), subgiants (252) and giants (145), based on their positions in the Hertzsprung-Russell diagram. We use the spectra of a grid of PHOENIX model atmospheres to obtain the photospheric contributions to the S-index. To convert the latter into absolute Ca ii H+K chromospheric line flux, we first derive new, colour-dependent photospheric flux relations for, each, main sequence, subgiant and giant stars, and then obtain the chromospheric flux component. In this process, the PHOENIX models also provide a very reliable scale for the physical surface flux. Results. For very large samples of main sequence stars, giants and subgiants, we obtain the chromospheric Ca ii H+K line surface fluxes in the colour range of 0.44 < B −V < 1.6 and the related R-indices. We determine and parametrize the lower envelopes, which we find to well coincide with historic work on the basal chromospheric flux. There is good agreement in the apparently simpler cases of inactive giants and subgiants, and distinguishing different luminosity classes proves important. Main sequence stars, surprisingly, show a remarkable lack of inactive chromospheres in the B − V range of 1.1 to 1.5. Finally, we intoduce a new, “pure” and universal activity indicator: a derivative of the R-index based on the non-basal, purely activity-related Ca ii H+K line surface flux, which puts different luminosity classes on the same scale. Conclusions. The here presented conversion method can be used to directly compare historical S-indices with modern chromospheric Ca ii H+K line flux measurements, in order to derive activity records over long periods of time or to establish the long-term variability of marginally active stars, for example. The numerical simplicity of this conversion allows for its application to very large stellar samples.

Abstract 19297: Cardiac Specific Deletion of Mcl-1 Leads to Impaired Autophagy and Rapid Development of Heart Failure

Mcl-1 is an anti-apoptotic Bcl-2 protein which is expressed at high levels in the heart compared to other anti-apoptotic Bcl-2 proteins. However, very little is known with respect to how Mcl-1 regulates cell survival in cardiomyocytes. To investigate the functional role of Mcl-1 in the heart, we used a loxP/α-MHC-Cre-recombinase system to generate inducible, cardiomyocyte-specific Mcl-1 knockout mice. Mcl-1 deletion led to rapid contractile dysfunction, cardiac hypertrophy, and interstitial fibrosis in the absence of stress. Surprisingly, loss of Mcl-1 did not activate apoptosis. Instead, Mcl-1 deficiency led to necrotic cell death characterized by mitochondrial permeability transition pore (mPTP) opening, mitochondrial swelling, and cellular rupture. To clarify how mPTP opening and necrosis contribute to heart failure in Mcl-1-/- mice, we generated Mcl-1/Cyclophilin D (CypD) double knockouts. CypD is an essential component of the mPTP, and CypD deficiency has been shown to reduce cardiac necrosis. CypD deletion significantly delayed development of heart failure in Mcl-1-/- mice, but did not rescue the phenotype. Although autophagy is an important cellular quality control mechanism, electron microscopy revealed no autophagosome formation in Mcl-1-/- cardiomyocytes. Further analysis revealed that loss of Mcl-1 led to impaired basal autophagic flux. Western blot analysis for autophagy proteins revealed reduced LC3II/I ratios and accumulation of p62 in Mcl-1-/- hearts. Chloroquine injection, which inhibits the fusion between autophagosomes and lysosomes, led to accumulation of LC3II and p62 in WT hearts but not in Mcl-1-/- hearts. To investigate whether autophagy could be induced in Mcl-1-/- hearts, we subjected mice to acute exercise. Autophagy was rapidly enhanced in wild type hearts after 80 minutes of swimming, but no change in autophagy occurred in Mcl-1-/- hearts. Conversely, adenoviral overexpression of Mcl-1 in rat neonatal cardiomyocytes led to increased LC3 processing and autophagy. These data suggest that, in addition to its anti-apoptotic role, Mcl-1 is essential in initiating autophagy in cardiomyocytes. Increased understanding of Mcl-1 mechanisms in the myocardium may provide new therapeutic avenues for heart disease.

Basal magnetic flux and the local solar dynamo

The average unsigned magnetic flux density in magnetograms of the quiet Sun is generally dominated by instrumental noise. Due to the entirely different scaling behavior of the noise and the solar magnetic pattern it has been possible to determine the standard deviation of the Gaussian noise distribution and remove the noise contribution from the average unsigned flux density for the whole 15-yr SOHO/MDI data set and for a selection of SDO/HMI magnetograms. There is a very close correlation between the MDI disk-averaged unsigned vertical flux density and the sunspot number, and regression analysis gives a residual level of 2.7 G when the sunspot number is zero. The selected set of HMI magnetograms, which spans the most quiet phase of solar activity, has a lower limit of 3.0 G to the noise-corrected average flux density. These apparently cycle-independent levels may be identified as a basal flux density, which represents an upper limit to the possible flux contribution from a local dynamo, but not evidence for its existence. The 3.0 G HMI level, when scaled to the Hinode spatial resolution, translates to 3.5 G, which means that the much higher average flux densities always found by Hinode in quiet regions do not originate from a local dynamo. The contributions to the average unsigned flux density come almost exclusively from the extended wings of the probability density function (PDF), also in the case of HMI magnetograms with only basal-level magnetic flux. These wings represent intermittent magnetic flux. While the global dynamo appears to dominate the magnetic energy spectrum at all the resolved spatial scales, there are indications from the observed Hanle depolarization in atomic lines that the local dynamo may dominate the spectrum at scales of order 1-10 km and below.

Inhibitory Effect of mTOR Activator MHY1485 on Autophagy: Suppression of Lysosomal Fusion

Autophagy is a major degradative process responsible for the disposal of cytoplasmic proteins and dysfunctional organelles via the lysosomal pathway. During the autophagic process, cells form double-membraned vesicles called autophagosomes that sequester disposable materials in the cytoplasm and finally fuse with lysosomes. In the present study, we investigated the inhibition of autophagy by a synthesized compound, MHY1485, in a culture system by using Ac2F rat hepatocytes. Autophagic flux was measured to evaluate the autophagic activity. Autophagosomes were visualized in Ac2F cells transfected with AdGFP-LC3 by live-cell confocal microscopy. In addition, activity of mTOR, a major regulatory protein of autophagy, was assessed by western blot and docking simulation using AutoDock 4.2. In the result, treatment with MHY1485 suppressed the basal autophagic flux, and this inhibitory effect was clearly confirmed in cells under starvation, a strong physiological inducer of autophagy. The levels of p62 and beclin-1 did not show significant change after treatment with MHY1485. Decreased co-localization of autophagosomes and lysosomes in confocal microscopic images revealed the inhibitory effect of MHY1485 on lysosomal fusion during starvation-induced autophagy. These effects of MHY1485 led to the accumulation of LC3II and enlargement of the autophagosomes in a dose- and time- dependent manner. Furthermore, MHY1485 induced mTOR activation and correspondingly showed a higher docking score than PP242, a well-known ATP-competitive mTOR inhibitor, in docking simulation. In conclusion, MHY1485 has an inhibitory effect on the autophagic process by inhibition of fusion between autophagosomes and lysosomes leading to the accumulation of LC3II protein and enlarged autophagosomes. MHY1485 also induces mTOR activity, providing a possibility for another regulatory mechanism of autophagy by the MHY compound. The significance of this study is the finding of a novel inhibitor of autophagy with an mTOR activating effect.

Nature of the solar dynamo at small scales

AbstractIt is often claimed that there is not only one, but two different types of solar dynamos: the one that is responsible for the appearance of sunspots and the 11-yr cycle, frequently referred to as the “global dynamo”, and a statistically time-invariant dynamo, generally referred to as the “local dynamo”, which is supposed to be responsible for the ubiquitous magnetic structuring observed at small scales. Here we examine the relative contributions of these two qualitatively different dynamos to the small-scale magnetic flux, with the following conclusion: The local dynamo does not play a significant role at any of the spatially resolved scales, nearly all the small-scale flux, including the flux revealed by Hinode, is supplied by the global dynamo. This conclusion is reached by careful determination of the Sun's noise-corrected basal magnetic flux density while making use of a flux cancellation function determined from Hinode data. The only allowed range where there may be substantial or even dominating contributions from a local dynamo seems to be the scales below about 10 km, as suggested by observations of the Hanle depolarization effect in atomic spectral lines. To determine the fraction of the Hanle depolarization that may be due to the action of a local dynamo, a synoptic program is being initiated at IRSOL (Istituto Ricerche Solari Locarno).

Noble gases and coupled heat/fluid flow modeling for evaluating hydrogeologic conditions of volcanic island aquifers

Understanding groundwater conditions in the upland parts of volcanic island aquifers is critical for sustainable groundwater development in these resource-limited environments. Yet groundwater conditions in such settings are generally difficult to characterize because of sparse well drilling (high cost and/or limited access). Information needed for resource evaluation includes upland depth to water, recharge rate, and aquifer permeability. In this study, noble-gas recharge temperatures and coupled heat/fluid flow modeling are used to indirectly infer these groundwater conditions. Mosteiros Basin on Fogo Island of the Cape Verde archipelago was selected as a representative volcanic island aquifer. Simulation results are calibrated to water-table altitude and temperature data, along with indirect information provided by noble and dissolved gases. Results of numerical modeling are most sensitive to recharge rates and hydraulic conductivity, less sensitive to basal heat flux, and not sensitive to porosity and thermal conductivity. Simulation results show that only a relatively narrow range of combined recharge values (12–25% of precipitation) and hydraulic conductivity (10−8–10−7 m s−1) is consistent with observed data. The simulated recharge of 3–6 million cubic meters per year (Mm3 yr−1) is much higher than measured discharge from the basin (0.25 Mm3 yr−1), indicating the occurrence of significant amounts of submarine groundwater discharge. The modeling results suggest a very deep upland water table located 600–1000 m beneath the floor of the central caldera, although the water table becomes more shallow and accessible towards the coast. Perhaps most significant is the dominant role that modest amounts of recharge (mean rates of 70–140 mm yr−1) plays in decreasing the geothermal gradient, resulting in relatively cool temperatures in the deep vadose and at the water table. Noble-gas recharge temperatures, constrained by numerical simulation results, range from about 15 to 22 °C and indicate a large water-table temperature lapse of about −15 °C/km, much steeper than typical adiabatic lapse rates.

Beclin 1 and autophagy are required for the tumorigenicity of breast cancer stem-like/progenitor cells

Malignant breast tissue contains a rare population of multi-potent cells with the capacity to self-renew; these cells are known as cancer stem-like cells (CSCs) or tumor-initiating cells. Primitive mammary CSCs/progenitor cells can be propagated in culture as floating spherical colonies termed ‘mammospheres'. We show here that the expression of the autophagy protein Beclin 1 is higher in mammospheres established from human breast cancers or breast cancer cell lines (MCF-7 and BT474) than in the parental adherent cells. As a result, autophagic flux is more robust in mammospheres. We observed that basal and starvation-induced autophagy flux is also higher in aldehyde dehydrogenase 1-positive (ALDH1+) population derived from mammospheres than in the bulk population. Beclin 1 is critical for CSC maintenance and tumor development in nude mice, whereas its expression limits the development of tumors not enriched with breast CSCs/progenitor cells. We found that decreased survival in autophagy-deficient cells (MCF-7 Atg7 knockdown cells) during detachment does not contribute to an ultimate deficiency in mammosphere formation. This study demonstrates that a prosurvival autophagic pathway is critical for CSC maintenance, and that Beclin 1 plays a dual role in tumor development.

Borehole temperatures reveal details of 20th century warming at Bruce Plateau, Antarctic Peninsula

Abstract. Two ice core boreholes of 143.18 m and 447.73 m (bedrock) were drilled during the 2009–2010 austral summer on the Bruce Plateau at a location named LARISSA Site Beta (66°02' S, 64°04' W, 1975.5 m a.s.l.). Both boreholes were logged with thermistors shortly after drilling. The shallow borehole was instrumented for 4 months with a series of resistance thermometers with satellite uplink. Surface temperature proxy data derived from an inversion of the borehole temperature profiles are compared to available multi-decadal records from weather stations and ice cores located along a latitudinal transect of the Antarctic Peninsula to West Antarctica. The LARISSA Site Beta profiles show temperatures decreasing from the surface downward through the upper third of the ice, and warming thereafter to the bed. The average temperature for the most recent year is −14.78°C (measured at 15 m depth, abbreviated T15). A minimum temperature of −15.8°C is measured at 173 m depth, and basal temperature is estimated to be −10.2°C. Current mean annual temperature and the gradient in the lower part of the measured temperature profile have a best fit with an accumulation rate of 1.9×103 kg m−2 a−1 and basal heat flux (q) of 88 mW m−2, if steady-state conditions are assumed. However, the mid-level temperature variations show that recent temperature has varied significantly. Reconstructed surface temperatures (Ts=T15) over the last 200 yr are derived by an inversion technique (Tikhonov and Samarskii, 1990). From this, we find that cold temperatures (minimum Ts=−16.2°C) prevailed from ~1920 to ~1940, followed by a gradual rise of temperature to −14.2°C around 1995, then cooling over the following decade and warming in the last few years. The coldest period was preceded by a relatively warm 19th century at T15≥−15°C. To facilitate regional comparisons of the surface temperature history, we use our T15 data and nearby weather station records to refine estimates of lapse rates (altitudinal, adjusted for latitude: Γa(l)). Good temporal and spatial consistency of Γa(l) over the last 35 yr are observed, implying that the climate trends observed here are regional and consistent over a broad altitude range.

Acute experimental stress evokes a differential gender‐determined increase in human intestinal macromolecular permeability

Intestinal epithelial dysfunction is a common pathophysiologic feature in irritable bowel syndrome (IBS) patients and might be the link to its clinical manifestations. We previously showed that chronic psychosocial stress induces jejunal epithelial barrier dysfunction; however, whether this epithelial response is gender-specific and might thus explain the enhanced female susceptibility to IBS remains unknown. Intestinal responses to acute stress were compared in age-matched groups of healthy women and men (n = 10 each) experiencing low background stress. A 20-cm jejunal segment, was perfused with an isosmotic solution, and intestinal effluents were collected under basal conditions, for 15 min during cold pain stress and for a 45-min recovery period. Epithelial function (net water flux and albumin output), changes in stress hormones, and cardiovascular and psychologic responses to cold stress were measured. Heart rate and blood pressure significantly increased during cold pain stress with no differences between men and women. Adrenocorticotropic hormone and cortisol levels during cold pain stress were significantly higher in men. Basal net water flux and epithelial permeability were similar in men and women. Cold pain stress increased water flux in both groups (72 ± 23 and 107 ± 18 μL min(-1) cm(-1) , respectively; F(5, 90) = 5.5; P = 0.003 for Time) and, interestingly, this was associated with a marked increase of albumin permeability in women but not in men (0.8 ± 0.2 vs.-0.7 ± 0.2 mg/15 min; P < 0.0001). Intestinal macromolecular permeability in response to acute experimental stress is increased in healthy women, a mechanism that may contribute to female oversusceptibility to IBS.

Sustainability of a subsurface ocean within Triton’s interior

We present a study of coupled thermal and structural evolution of Neptune’s moon, Triton, driven by tidal dissipation and radiogenic heating. Triton’s orbital history likely involves capture from a binary system by Neptune, followed by a period of circularization. This work investigates Triton’s evolution past its circularization. We examine the rate of ice shell growth as a function of different orbital eccentricities, in the presence of radiogenic heating. Tidal dissipation in the ice shell, proportional to orbital eccentricity squared, concentrates heating near the base, reducing the basal heat flux. As the growth of the ice shell is proportional to the basal heat flux, increased tidal heating creates a blanketing effect, reducing the rate of ice shell growth. Radiogenic heating from Triton’s core is the other, more dominant, source of heat to the shell. Despite being several orders of magnitude higher than the tidal dissipation, radiogenic heating alone fails to sustain an ocean within Triton over 4.5Ga. For orbital eccentricities of 5×10−7 and 3×10−5 it takes approximately 2Ga and 3Ga, respectively, to completely freeze the ocean. For higher values of orbital eccentricities, an ocean can be sustained in Triton’s interior over 4.5Ga. If Triton’s history past circularization involves a slow decrease in orbital eccentricity to the current value, a thin, possibly NH3-rich ocean exists beneath Triton’s icy shell.

Basal chromospheric flux and Maunder Minimum-type stars: the quiet-Sun chromosphere as a universal phenomenon

Aims: We demonstrate the universal character of the quiet-Sun chromosphere among inactive stars (solar-type and giants). By assessing the main physical processes, we shed new light on some common observational phenomena. Methods: We discuss measurements of the solar Mt. Wilson S-index, obtained by the Hamburg Robotic Telescope around the extreme minimum year 2009, and compare the established chromospheric basal Ca II K line flux to the Mt. Wilson S-index data of inactive ("flat activity") stars, including giants. Results: During the unusually deep and extended activity minimum of 2009, the Sun reached S-index values considerably lower than in any of its previously observed minima. In several brief periods, the Sun coincided exactly with the S-indices of inactive ("flat", presumed Maunder Minimum-type) solar analogues of the Mt. Wilson sample; at the same time, the solar visible surface was also free of any plages or remaining weak activity regions. The corresponding minimum Ca II K flux of the quiet Sun and of the presumed Maunder Minimum-type stars in the Mt. Wilson sample are found to be identical to the corresponding Ca II K chromospheric basal flux limit. Conclusions: We conclude that the quiet-Sun chromosphere is a universal phenomenon among inactive stars. Its mixed-polarity magnetic field, generated by a local, "fast" turbulent dynamo finally provides a natural explanation for the minimal soft X-ray emission observed for inactive stars. Given such a local dynamo also works for giant chromospheres, albeit on larger length scales, i.e., l ~ R/g, with R and g as stellar radius and surface gravity, respectively, the existence of giant spicular phenomena and the guidance of mechanical energy toward the acceleration zone of cool stellar winds along flux-tubes have now become traceable.

Geometry and dynamics of an East Antarctic Ice Sheet outlet glacier, under past and present climates

[1] The behavior of East Antarctic Ice Sheet (EAIS) outlet glaciers both at present and during the recent geological past is of considerable interest both from the point of view of understanding the mechanics of contemporary glacier dynamics, and also with regard to epoch-scale ice sheet stability during Plio-Pleistocene climate transitions. Here we use a glacier flowline model that incorporates the effects of longitudinal stresses to numerically simulate Ferrar Glacier, first under present-day environmental conditions, and subsequently under both colder and warmer climate regimes representing the Last Glacial Maximum (LGM) and mid-Pliocene peak warmth respectively. Using airborne radar profiles, InSAR-derived surface velocities, ice core and geological data for empirical constraint, we present a diagnostic simulation that uses an iterative method to closely reproduce observed dynamics. Our model suggests that the glacier is largely cold-based under present conditions, flows predominantly by way of internal deformation, and ‘cascades’ over bedrock ridges due to the combined action of changes in cross-sectional valley geometry, local steepening of the glacier surface, and the non-local effects of longitudinal coupling. Time-dependent (evolutionary) simulation of a lower-profile glacier under a colder, drier, LGM climate, predicts flow velocities lower than present with minimal bedrock erosion or basal till flux. Conversely, the warmer-than-present mid-Pliocene climate produces a more dynamic glacier that is warm-based and sliding along much of its bed. We propose that EAIS outlet glaciers, such as the Ferrar, respond dynamically along their length in response to changing environmental forcings, with most significant changes taking place in their lower reaches. Adjustment to perturbations in upper catchments is more muted.

Characterization of macroautophagic flux in vivo using a leupeptin-based assay

Macroautophagy is a highly conserved catabolic process that is crucial for organ homeostasis in mammals. However, methods to directly measure macroautophagic activity (or flux) in vivo are limited. In this study we developed a quantitative macroautophagic flux assay based on measuring LC3b protein turnover in vivo after administering the protease inhibitor leupeptin. Using this assay we then characterized basal macroautophagic flux in different mouse organs. We found that the rate of LC3b accumulation after leupeptin treatment was greatest in the liver and lowest in spleen. Interestingly we found that LC3a, an ATG8/LC3b homologue and the LC3b-interacting protein p62 were degraded with similar kinetics to LC3b. However, the LC3b-related proteins GABARAP and GATE-16 were not rapidly turned over in mouse liver, implying that different LC3b homologues may contribute to macroautophagy via distinct mechanisms. Nutrient starvation augmented macroautophagic flux as measured by our assay, while refeeding the animals after a period of starvation significantly suppressed flux. We also confirmed that beclin 1 heterozygous mice had reduced basal macroautophagic flux compared to wild-type littermates. These results illustrate the usefulness of our leupeptin-based assay for studying the dynamics of macroautophagy in mice.

Bone Cell Autophagy Is Regulated by Environmental Factors

The goal of this investigation was to ascertain whether bone cells undergo autophagy and to determine if this process is regulated by environmental factors. We showed that osteocytes in both murine and human cortical bone display a punctuate distribution of microtubule-associated protein light chain 3, indicative of autophagy. In addition, we noted a basal level of autophagy in preosteocyte-like murine long bone-derived osteocytic (MLO)-A5 cells. Autophagy was upregulated following nutrient deprivation and hypoxic culture, stress conditions that osteocytes encounter in vivo. Furthermore, in response to calcium stress, the transcription factor hypoxia inducible factor 1 regulated MLO-A5 autophagy. Finally, we showed that the more differentiated MLO-Y4 osteocyte-like cells exhibited a significant basal autophagic flux. Based on these findings, we suggest that raising the level of autophagic flux is a mechanism by which differentiated bone cells survive in a stressful environment.

Tuning flux: autophagy as a target of heart disease therapy

Despite maximum medical and mechanical support therapy, heart failure remains a relentlessly progressive disorder with substantial morbidity and mortality. Autophagy, an evolutionarily conserved process of cellular cannibalization, has been implicated in virtually all forms of cardiovascular disease. Indeed, its role is context dependent, antagonizing or promoting disease depending on the circumstance. Here, we review current understanding of the role of autophagy in the pathogenesis of heart failure and explore this pathway as a target of therapeutic intervention. In preclinical models of heart disease, cardiomyocyte autophagic flux is activated; indeed, its role in disease pathogenesis is the subject of intense investigation to define mechanism. Similarly, in failing human heart of a variety of etiologies, cardiomyocyte autophagic activity is upregulated, and therapy, such as with mechanical support systems, elicits declines in autophagy activity. However, when suppression of autophagy is complete, rapid and catastrophic cell death occurs, consistent with a model in which basal autophagic flux is required for proteostasis. Thus, a narrow zone of 'optimal' autophagy seems to exist. The challenge moving forward is to tune the stress-triggered autophagic response within that 'sweet spot' range for therapeutic benefit. Whereas we have known for some years of the participation of lysosomal mechanisms in heart disease, it is only recently that upstream mechanisms (autophagy) are being explored. The challenge for the future is to dissect the underlying circuitry and titrate the response into an optimal, proteostasis-promoting range in hopes of mitigating the ever-expanding epidemic of heart failure.