Post-natal induction of PGC-1α protects against severe muscle dystrophy independently of utrophin

Background: Duchenne muscle dystrophy (DMD) afflicts 1 million boys in the US and has few effective treatments. Constitutive transgenic expression of the transcriptional coactivator peroxisome proliferator-activated receptor gamma coactivator (PGC)-1α improves skeletal muscle function in the murine “mdx” model of DMD, but how this occurs, or whether it can occur post-natally, is not known. The leading mechanistic hypotheses for the benefits conferred by PGC-1α include the induction of utrophin, a dystrophin homolog, and/or induction and stabilization of the neuromuscular junction. Methods: The effects of transgenic overexpression of PGC-1β, a homolog of PGC-1α in mdx mice was examined using different assays of skeletal muscle structure and function. To formally test the hypothesis that PGC-1α confers benefit in mdx mice by induction of utrophin and stabilization of neuromuscular junction, PGC-1α transgenic animals were crossed with the dystrophin utrophin double knock out (mdx/utrn-/-) mice, a more severe dystrophic model. Finally, we also examined the effect of post-natal induction of skeletal muscle-specific PGC-1α overexpression on muscle structure and function in mdx mice. Results: We show here that PGC-1β does not induce utrophin or other neuromuscular genes when transgenically expressed in mouse skeletal muscle. Surprisingly, however, PGC-1β transgenesis protects as efficaciously as PGC-1α against muscle degeneration in dystrophin-deficient (mdx) mice, suggesting that alternate mechanisms of protection exist. When PGC-1α is overexpressed in mdx/utrn-/- mice, we find that PGC-1α dramatically ameliorates muscle damage even in the absence of utrophin. Finally, we also used inducible skeletal muscle-specific PGC-1α overexpression to show that PGC-1α can protect against dystrophy even if activated post-natally, a more plausible therapeutic option. Conclusions: These data demonstrate that PGC-1α can improve muscle dystrophy post-natally, highlighting its therapeutic potential. The data also show that PGC-1α is equally protective in the more severely affected mdx/utrn-/- mice, which more closely recapitulates the aggressive progression of muscle damage seen in DMD patients. The data also identify PGC-1β as a novel potential target, equally efficacious in protecting against muscle dystrophy. Finally, the data also show that PGC-1α and PGC-1β protect against dystrophy independently of utrophin or of induction of the neuromuscular junction, indicating the existence of other mechanisms

WFPC2 Observations of the Hubble Deep Field-South

The Hubble Deep Field-South observations targeted a high-galactic-latitude field near QSO J2233-606. We present WFPC2 observations of the field in four wide bandpasses centered at roughly 300, 450, 606, and 814 nm. Observations, data reduction procedures, and noise properties of the final images are discussed in detail. A catalog of sources is presented, and the number counts and color distributions of the galaxies are compared to a new catalog of the HDF-N that has been constructed in an identical manner. The two fields are qualitatively similar, with the galaxy number counts for the two fields agreeing to within 20%. The HDF-S has more candidate Lyman-break galaxies at z > 2 than the HDF-N. The star-formation rate per unit volume computed from the HDF-S, based on the UV luminosity of high-redshift candidates, is a factor of 1.9 higher than from the HDF-N at z ~ 2.7, and a factor of 1.3 higher at z ~ 4.Comment: 93 pages, 25 figures; contains very long table

The Hubble Deep Field: Observations, Data Reduction, and Galaxy Photometry

The Hubble Deep Field (HDF) is a Director's Discretionary program on HST in Cycle 5 to image an undistinguished field at high Galactic latitude in four passbands as deeply as reasonably possible. These images provide the most detailed view to date of distant field galaxies and are likely to be important for a wide range of studies in galaxy evolution and cosmology. In order to optimize observing in the time available, a field in the northern continuous viewing zone was selected and images were taken for ten consecutive days, or approximately 150 orbits. Shorter 1-2 orbit images were obtained of the fields immediately adjacent to the primary HDF in order to facilitate spectroscopic follow-up by ground-based telescopes. The observations were made from 18 to 30 December 1995, and both raw and reduced data have been put in the public domain as a community service. We present a summary of the criteria for selecting the field, the rationale behind the filter selection and observing times in each band, and the strategies for planning the observations to maximize the exposure time while avoiding earth-scattered light. Data reduction procedures are outlined, and images of the combined frames in each band are presented. Objects detected in these images are listed in a catalog with their basic photometric parameters.Comment: 37 pages, XX PostScript figures, uses aaspp4.sty astrobib.sty. (Astrobib is available from http://www.stsci.edu/software/TeX.html .) To appear the Astronomical Journal. More info on the Hubble deep field can be found at http://www.stsci.edu/../ftp/observer/hdf/hdf.html . More figures (images) can be found at http://www.stsci.edu/../ftp/observer/hdf/references/williams/ and the full source catalog is available at http://www.stsci.edu/../ftp/observer/hdf/archive/v2catalog

The mitochondrial Ca2+ channel MCU is critical for tumor growth by supporting cell cycle progression and proliferation

Introduction: The mitochondrial uniporter (MCU) Ca2+ ion channel represents the primary means for Ca2+ uptake by mitochondria. Mitochondrial matrix Ca2+ plays critical roles in mitochondrial bioenergetics by impinging upon respiration, energy production and flux of biochemical intermediates through the TCA cycle. Inhibition of MCU in oncogenic cell lines results in an energetic crisis and reduced cell proliferation unless media is supplemented with nucleosides, pyruvate or α-KG. Nevertheless, the roles of MCU-mediated Ca2+ influx in cancer cells remain unclear, in part because of a lack of genetic models.Methods: MCU was genetically deleted in transformed murine fibroblasts for study in vitro and in vivo. Tumor formation and growth were studied in murine xenograft models. Proliferation, cell invasion, spheroid formation and cell cycle progression were measured in vitro. The effects of MCU deletion on survival and cell-death were determined by probing for live/death markers. Mitochondrial bioenergetics were studied by measuring mitochondrial matrix Ca2+ concentration, membrane potential, global dehydrogenase activity, respiration, ROS production and inactivating-phosphorylation of pyruvate dehydrogenase. The effects of MCU rescue on metabolism were examined by tracing of glucose and glutamine utilization for fueling of mitochondrial respiration.Results: Transformation of primary fibroblasts in vitro was associated with increased MCU expression, enhanced MCU-mediated Ca2+ uptake, altered mitochondrial matrix Ca2+ concentration responses to agonist stimulation, suppression of inactivating-phosphorylation of pyruvate dehydrogenase and a modest increase of mitochondrial respiration. Genetic MCU deletion inhibited growth of HEK293T cells and transformed fibroblasts in mouse xenograft models, associated with reduced proliferation and delayed cell-cycle progression. MCU deletion inhibited cancer stem cell-like spheroid formation and cell invasion in vitro, both predictors of metastatic potential. Surprisingly, mitochondrial matrix [Ca2+], membrane potential, global dehydrogenase activity, respiration and ROS production were unaffected. In contrast, MCU deletion elevated glycolysis and glutaminolysis, strongly sensitized cell proliferation to glucose and glutamine limitation, and altered agonist-induced cytoplasmic Ca2+ signals.Conclusion: Our results reveal a dependence of tumorigenesis on MCU, mediated by a reliance on MCU for cell metabolism and Ca2+ dynamics necessary for cell-cycle progression and cell proliferation

Modeling Insertional Mutagenesis Using Gene Length and Expression in Murine Embryonic Stem Cells

Background. High-throughput mutagenesis of the mammalian genome is a powerful means to facilitate analysis of gene function. Gene trapping in embryonic stem cells (ESCs) is the most widely used form of insertional mutagenesis in mammals. However, the rules governing its efficiency are not fully understood, and the effects of vector design on the likelihood of genetrapping events have not been tested on a genome-wide scale. Methodology/Principal Findings. In this study, we used public gene-trap data to model gene-trap likelihood. Using the association of gene length and gene expression with gene-trap likelihood, we constructed spline-based regression models that characterize which genes are susceptible and which genes are resistant to gene-trapping techniques. We report results for three classes of gene-trap vectors, showing that both length and expression are significant determinants of trap likelihood for all vectors. Using our models, we also quantitatively identifie

V838 Monocerotis: A Geometric Distance from Hubble Space Telescope Polarimetric Imaging of its Light Echo

Following the outburst of the unusual variable star V838 Monocerotis in 2002, a spectacular light echo appeared. A light echo provides the possibility of direct geometric distance determination, because it should contain a ring of highly linearly polarized light at a linear radius of ct, where t is the time since the outburst. We present imaging polarimetry of the V838 Mon light echo, obtained in 2002 and 2005 with the Advanced Camera for Surveys onboard the Hubble Space Telescope, which confirms the presence of the highly polarized ring. Based on detailed modeling that takes into account the outburst light curve, the paraboloidal echo geometry, and the physics of dust scattering and polarization, we find a distance of 6.1+-0.6 kpc. The error is dominated by the systematic uncertainty in the scattering angle of maximum linear polarization, taken to be theta_{max}=90^o +- 5^o. The polarimetric distance agrees remarkably well with a distance of 6.2+-1.5 kpc obtained from the entirely independent method of main-sequence fitting to a sparse star cluster associated with V838 Mon. At this distance, V838 Mon at maximum light had M_V\simeq-9.8, making it temporarily one of the most luminous stars in the Local Group. Our validation of the polarimetric method offers promise for measurement of extragalactic distances using supernova light echoes.Comment: 43 pages, 17 figures, 3 tables; accepted for publication in the Astronomical Journal. Version with high-quality figures available at http://www.stsci.edu/~bond/v838monpolariz.pd

Mediterranean cyclones and windstorms in a changing climate

Changes in the frequency and intensity of cyclones and associated windstorms affecting the Medi-terranean region simulated under enhanced Greenhouse Gas forcing conditions are investigated. The analysis is based on 7 climate model integrations performed with two coupled global models (ECHAM5 MPIOM and INGV CMCC), comparing the end of the twentieth century and at least the first half of the twenty-first century. As one of the models has a considerably enhanced resolution of the atmosphere and the ocean, it is also investigated whether the climate change signals are influenced by the model resolution. While the higher resolved simulation is closer to reanalysis climatology, both in terms of cyclones and windstorm distributions, there is no evidence for an influence of the resolution on the sign of the climate change signal. All model simulations show a reduction in the total number of cyclones crossing the Mediterranean region under climate change conditions. Exceptions are Morocco and the Levant region, where the models predict an increase in the number of cyclones. The reduction is especially strong for intense cyclones in terms of their Laplacian of pressure. The influence of the simulated positive shift in the NAO Index on the cyclone decrease is restricted to the Western Mediterranean region, where it explains 10–50 % of the simulated trend, depending on the individual simulation. With respect to windstorms, decreases are simulated over most of the Mediterranean basin. This overall reduction is due to a decrease in the number of events associated with local cyclones, while the number of events associated with cyclones outside of the Mediterranean region slightly increases. These systems are, however, less intense in terms of their integrated severity over the Mediterranean area, as they mostly affect the fringes of the region. In spite of the general reduction in total numbers, several cyclones and windstorms of intensity unknown under current climate conditions are identified for the scenario simulations. For these events, no common trend exists in the individual simulations. Thus, they may rather be attributed to long-term (e.g. decadal) variability than to the Greenhouse Gas forcing. Nevertheless, the result indicates that high-impact weather systems will remain an important risk in the Mediterranean Basin

Intracranial Aneurysm Classifier Using Phenotypic Factors: An International Pooled Analysis

Intracranial aneurysms (IAs) are usually asymptomatic with a low risk of rupture, but consequences of aneurysmal subarachnoid hemorrhage (aSAH) are severe. Identifying IAs at risk of rupture has important clinical and socio-economic consequences. The goal of this study was to assess the effect of patient and IA characteristics on the likelihood of IA being diagnosed incidentally versus ruptured. Patients were recruited at 21 international centers. Seven phenotypic patient characteristics and three IA characteristics were recorded. The analyzed cohort included 7992 patients. Multivariate analysis demonstrated that: (1) IA location is the strongest factor associated with IA rupture status at diagnosis; (2) Risk factor awareness (hypertension, smoking) increases the likelihood of being diagnosed with unruptured IA; (3) Patients with ruptured IAs in high-risk locations tend to be older, and their IAs are smaller; (4) Smokers with ruptured IAs tend to be younger, and their IAs are larger; (5) Female patients with ruptured IAs tend to be older, and their IAs are smaller; (6) IA size and age at rupture correlate. The assessment of associations regarding patient and IA characteristics with IA rupture allows us to refine IA disease models and provide data to develop risk instruments for clinicians to support personalized decision-making

PGC-1β regulates angiogenesis in skeletal muscle

Aerobic metabolism requires oxygen and carbon sources brought to tissues via the vasculature. Metabolically active tissues such as skeletal muscle can regulate blood vessel density to match metabolic needs; however, the molecular cues that coordinate these processes remain poorly understood. Here we report that the transcriptional coactivator peroxisome proliferator-activated receptor-γ coactivator-1β (PGC-1β), a potent regulator of mitochondrial biology, induces angiogenesis in skeletal muscle. PGC-1β induces the expression of vascular endothelial growth factor (VEGF) in cell culture and in vivo. The induction of VEGF by PGC-1β requires coactivation of the orphan nuclear receptor estrogen-related receptor-α (ERRα) and is independent of the hypoxia-inducible factor (HIF) pathway. In coculture experiments, overexpression of PGC-1β in skeletal myotubes increases the migration of adjacent endothelial cells, and this depends on VEGF signaling. Transgenic expression of PGC-1β in skeletal myocytes dramatically increases muscular vessel density. Taken together, these data indicate that PGC-1β is a potent regulator of angiogenesis, thus providing a novel link between the regulations of oxidative metabolism and vascular density