Unexpected cell type-dependent effects of autophagy on polyglutamine aggregation revealed by natural genetic variation in C. elegans.

BACKGROUND: Monogenic protein aggregation diseases, in addition to cell selectivity, exhibit clinical variation in the age of onset and progression, driven in part by inter-individual genetic variation. While natural genetic variants may pinpoint plastic networks amenable to intervention, the mechanisms by which they impact individual susceptibility to proteotoxicity are still largely unknown. RESULTS: We have previously shown that natural variation modifies polyglutamine (polyQ) aggregation phenotypes in C. elegans muscle cells. Here, we find that a genomic locus from C. elegans wild isolate DR1350 causes two genetically separable aggregation phenotypes, without changing the basal activity of muscle proteostasis pathways known to affect polyQ aggregation. We find that the increased aggregation phenotype was due to regulatory variants in the gene encoding a conserved autophagy protein ATG-5. The atg-5 gene itself conferred dosage-dependent enhancement of aggregation, with the DR1350-derived allele behaving as hypermorph. Surprisingly, increased aggregation in animals carrying the modifier locus was accompanied by enhanced autophagy activation in response to activating treatment. Because autophagy is expected to clear, not increase, protein aggregates, we activated autophagy in three different polyQ models and found a striking tissue-dependent effect: activation of autophagy decreased polyQ aggregation in neurons and intestine, but increased it in the muscle cells. CONCLUSIONS: Our data show that cryptic natural variants in genes encoding proteostasis components, although not causing detectable phenotypes in wild-type individuals, can have profound effects on aggregation-prone proteins. Clinical applications of autophagy activators for aggregation diseases may need to consider the unexpected divergent effects of autophagy in different cell types

Well-defined hyperstar copolymers based on a thiol–yne hyperbranched core and a poly(2-oxazoline) shell for biomedical applications

Well defined ‘hyperstar’ copolymers were synthesized by combining hyperbranched polymers produced by thiol–yne chemistry with poly(oxazoline)s. The hyperbranched core was prepared using an AB2 monomer and a trifunctional alkene, applying a monomer feeding approach. The degree of branching was high (0.9) while maintaining low dispersities (1.3). Poly(2-ethyl-2-oxazoline) (PEtOx) functionalized with a thiol end group was coupled to the surface of the hyperbranched structure accessing terminal alkyne units. PEtOx-SH was produced by the termination of the living polymerization with ethyl xanthate and subsequent conversion to thiol under alkaline conditions. The degree of polymerization was varied producing PEtOx with 23 or 42 repeating units, respectively with a dispersity of around 1.1. After conjugation of the polymer arms, hyperstar copolymers were characterized by SEC, NMR spectroscopy, light scattering, and AFM. The polymers were able to encapsulate the hydrophobic dye Nile red within the core of the structure with loading efficiencies between 0.3 and 0.9 wt%. Cytotoxicity of the hyperstars was assessed using A2780 human ovarian carcinoma cells resulting in IC50 values of around 0.7 mg ml−1. Successful internalization and colocalization with lysosomal compartments was observed by confocal microscopy studies

Evidence for Intergalactic Absorption in the TeV Gamma-Ray Spectrum of Mkn 501

The recent HEGRA observations of the blazar Mkn 501 show strong curvature in the very high energy gamma-ray spectrum. Applying the gamma-ray opacity derived from an empirically based model of the intergalactic infrared background radiation field (IIRF), to these observations, we find that the intrinsic spectrum of this source is consistent with a power-law: dN/dE~ E^-alpha with alpha=2.00 +/- 0.03 over the range 500 GeV - 20 TeV. Within current synchrotron self-Compton scenarios, the fact that the TeV spectral energy distribution of Mkn 501 does not vary with luminosity, combined with the correlated, spectrally variable emission in X-rays, as observed by the BeppoSAX and RXTE instruments, also independently implies that the intrinsic spectrum must be close to alpha=2. Thus, the observed curvature in the spectrum is most easily understood as resulting from intergalactic absorption.Comment: 7 pages, 1 figure, accepted in ApJ Letters 1999 April

Chandra Observations of 3C Radio Sources with z<0.3: Nuclei, Diffuse Emission, Jets and Hotspots

We report on our Chandra Cycle 9 program to observe half of the 60 (unobserved by Chandra) 3C radio sources at z<0.3 for 8 ksec each. Here we give the basic data: the X-ray intensity of the nuclei and any features associated with radio structures such as hot spots and knots in jets. We have measured fluxes in soft, medium and hard bands and are thus able to isolate sources with significant intrinsic column density. For the stronger nuclei, we have applied the standard spectral analysis which provides the best fit values of X-ray spectral index and column density. We find evidence for intrinsic absorption exceeding a column density of 10^{22} cm^{-2} for one third of our sources.Comment: 12 pages, 37 figures (the complete version of the paper with all figures is available on line, see appendix for details), ApJ accepte

Interactions of ultrahigh-energy cosmic rays with photons in the galactic center

Ultrahigh-energy cosmic rays passing through the central region of the Galaxy interact with starlight and the infrared photons. Both nuclei and protons generate secondary fluxes of photons and neutrinos on their passage through the central region. We compute the fluxes of these secondary particles, the observations of which can be used to improve one's understanding of origin and composition of ultrahigh-energy comic rays, especially if the violation of the Greisen--Zatespin--Kuzmin cutoff is confirmed by the future data.Comment: 8 pages, 2 figure

Episodic Star Formation Coupled to Reignition of Radio Activity in 3C 236

We present Hubble Space Telescope UV and optical imaging of the radio galaxy 3C 236, whose relic 4 Mpc radio jet lobes and inner 2 kpc CSS radio source are evidence of multiple epochs of AGN activity. Our data confirm the presence of four bright knots of FUV emission in an arc along the edge of the inner circumnuclear dust disk in the galaxy's nucleus, as well as FUV emission cospatial with the nucleus itself. We interpret these to be sites of recent or ongoing star formation. We present photometry of these knots, as well as an estimate for the internal extinction in the source using the Balmer decrement from SDSS spectroscopy. We estimate the ages of the knots by comparing our extinction-corrected photometry with stellar population synthesis models. We find the four knots cospatial with the dusty disk to be young, of order 10^7 yr old. The FUV emission in the nucleus is likely due to an episode of star formation triggered ~10^9 yr ago. We argue that the young 10^7 yr old knots stem from an episode of star formation that was roughly coeval with the event resulting in reignition of radio activity, creating the CSS source. The 10^9 yr old stars in the nucleus may be associated with the previous epoch of activity that generated the 4 Mpc relic source, before it was cut off by exhaustion or interruption. The ages of the knots, considered in context with the disturbed morphology of the nuclear dust and the double-double morphology of the "old" and "young" radio sources, present evidence for an episodic AGN/starburst connection. We suggest that the AGN fuel supply was interrupted for ~10^7 yr due to a minor merger event and has now been restored, and the resultant non-steady flow of gas toward the nucleus is likely responsible for both the new episode of infall-induced star formation and also the multiple epochs of radio activity.Comment: 15 pages, 7 figures. Accepted for publication in ApJ

First Steps towards Underdominant Genetic Transformation of Insect Populations

The idea of introducing genetic modifications into wild populations of insects to stop them from spreading diseases is more than 40 years old. Synthetic disease refractory genes have been successfully generated for mosquito vectors of dengue fever and human malaria. Equally important is the development of population transformation systems to drive and maintain disease refractory genes at high frequency in populations. We demonstrate an underdominant population transformation system in Drosophila melanogaster that has the property of being both spatially self-limiting and reversible to the original genetic state. Both population transformation and its reversal can be largely achieved within as few as 5 generations. The described genetic construct {Ud} is composed of two genes; (1) a UAS-RpL14.dsRNA targeting RNAi to a haploinsufficient gene RpL14 and (2) an RNAi insensitive RpL14 rescue. In this proof-of-principle system the UAS-RpL14.dsRNA knock-down gene is placed under the control of an Actin5c-GAL4 driver located on a different chromosome to the {Ud} insert. This configuration would not be effective in wild populations without incorporating the Actin5c-GAL4 driver as part of the {Ud} construct (or replacing the UAS promoter with an appropriate direct promoter). It is however anticipated that the approach that underlies this underdominant system could potentially be applied to a number of species. Figure

The role of whole brain radiation therapy in the management of melanoma brain metastases

Background: Brain metastases are common in patients with melanoma, and optimal management is not well defined. As melanoma has traditionally been thought of as “radioresistant,” the role of whole brain radiation therapy (WBRT) in particular is unclear. We conducted this retrospective study to identify prognostic factors for patients treated with stereotactic radiosurgery (SRS) for melanoma brain metastases and to investigate the role of additional up-front treatment with whole brain radiation therapy (WBRT). Methods: We reviewed records of 147 patients who received SRS as part of initial management of their melanoma brain metastases from January 2000 through June 2010. Overall survival (OS) and time to distant intracranial progression were calculated using the Kaplan-Meier method. Prognostic factors were evaluated using the Cox proportional hazards model. Results: WBRT was employed with SRS in 27% of patients and as salvage in an additional 22%. Age at SRS > 60 years (hazard ratio [HR] 0.64, p = 0.05), multiple brain metastases (HR 1.90, p = 0.008), and omission of up-front WBRT (HR 2.24, p = 0.005) were associated with distant intracranial progression on multivariate analysis. Extensive extracranial metastases (HR 1.86, p = 0.0006), Karnofsky Performance Status (KPS) ≤ 80% (HR 1.58, p = 0.01), and multiple brain metastases (HR 1.40, p = 0.06) were associated with worse OS on univariate analysis. Extensive extracranial metastases (HR 1.78, p = 0.001) and KPS (HR 1.52, p = 0.02) remained significantly associated with OS on multivariate analysis. In patients with absent or stable extracranial disease, multiple brain metastases were associated with worse OS (multivariate HR 5.89, p = 0.004), and there was a trend toward an association with worse OS when up-front WBRT was omitted (multivariate HR 2.56, p = 0.08). Conclusions: Multiple brain metastases and omission of up-front WBRT (particularly in combination) are associated with distant intracranial progression. Improvement in intracranial disease control may be especially important in the subset of patients with absent or stable extracranial disease, where the competing risk of death from extracranial disease is low. These results are hypothesis generating and require confirmation from ongoing randomized trials

Mergers, AGN, and 'Normal' Galaxies: Contributions to the Distribution of Star Formation Rates and Infrared Luminosity Functions

We use a novel method to predict the contribution of normal star-forming galaxies, merger-induced bursts, and obscured AGN, to IR luminosity functions (LFs) and global SFR densities. We use empirical halo occupation constraints to populate halos with galaxies and determine the distribution of normal and merging galaxies. Each system can then be associated with high-resolution hydrodynamic simulations. We predict the distribution of observed luminosities and SFRs, from different galaxy classes, as a function of redshift from z=0-6. We provide fitting functions for the predicted LFs, quantify the uncertainties, and compare with observations. At all redshifts, 'normal' galaxies dominate the LF at moderate luminosities ~L* (the 'knee'). Merger-induced bursts increasingly dominate at L>>L*; at the most extreme luminosities, AGN are important. However, all populations increase in luminosity at higher redshifts, owing to increasing gas fractions. Thus the 'transition' between normal and merger-dominated sources increases from the LIRG-ULIRG threshold at z~0 to bright Hyper-LIRG thresholds at z~2. The transition to dominance by obscured AGN evolves similarly, at factor of several higher L_IR. At all redshifts, non-merging systems dominate the total luminosity/SFR density, with merger-induced bursts constituting ~5-10% and AGN ~1-5%. Bursts contribute little to scatter in the SFR-stellar mass relation. In fact, many systems identified as 'ongoing' mergers will be forming stars in their 'normal' (non-burst) mode. Counting this as 'merger-induced' star formation leads to a stronger apparent redshift evolution in the contribution of mergers to the SFR density.Comment: 16 pages, 9 figures (+appendices), accepted to MNRAS. A routine to return the galaxy merger rates discussed here is available at http://www.cfa.harvard.edu/~phopkins/Site/mergercalc.htm