Rapid tyrosine phosphorylation of neuronal proteins including tau and focal adhesion kinase in response to amyloid-beta peptide exposure: Involvement of src family protein kinases

The increased production of amyloid beta -peptide (A beta) in Alzheimer's disease is acknowledged to be a key pathogenic event. In this study, we examined the response of primary human and rat brain cortical cultures to A beta administration and found a marked increase in the tyrosine phosphorylation content of numerous neuronal proteins, including tau and putative microtubule-associated protein 2c (MAP2c). We also found that paired helical filaments of aggregated and hyperphosphorylated tau are tyrosine phosphorylated, indicating that changes in the phosphotyrosine content of cytoplasmic proteins in response to A beta are potentially an important process. Increased tyrosine phosphorylation of cytoskeletal and other neuronal proteins was specific to fibrillar A beta (25-35) and A beta (1-42). The tyrosine phosphorylation was blocked by addition of the Src family tyrosine kinase inhibitor 4-amino-5-( 4-chlorophenyl)- 7(t-butyl) pyrazol(3,4-D) pyramide (PP2) and the phosphatidylinositol 3-kinase inhibitor LY 294002. Tyrosine phosphorylation of tau and MAP2c was concomitant with an increase in the tyrosine phosphorylation and subsequent putative activation of the non-receptor kinase, focal adhesion kinase (FAK). Immunoprecipitation of Fyn, a member of the Src family, from A beta (25-35)-treated neurons showed an increased association of Fyn with FAK. A beta treatment of cells also stimulated the sustained activation of extracellular regulated kinase-2, which was blocked by addition of PP2 and LY 294002, suggesting that FAK/Fyn/PI3-kinase association is upstream of mitogen-activated protein (MAP) kinase signaling in A beta -treated neurons. This cascade of signaling events contains the earliest biochemical changes in neurons to be described in response to A beta exposure and may be critical for subsequent neurodegenerative changes

Plasma lipid profiles change with increasing numbers of mild traumatic brain injuries in rats

Mild traumatic brain injury (mTBI) causes structural, cellular and biochemical alterations which are difficult to detect in the brain and may persist chronically following single or repeated injury. Lipids are abundant in the brain and readily cross the blood-brain barrier, suggesting that lipidomic analysis of blood samples may provide valuable insight into the neuropathological state. This study used liquid chromatography-mass spectrometry (LC-MS) to examine plasma lipid concentrations at 11 days following sham (no injury), one (1×) or two (2×) mTBI in rats. Eighteen lipid species were identified that distinguished between sham, 1× and 2× mTBI. Three distinct patterns were found: (1) lipids that were altered significantly in concentration after either 1× or 2× F mTBI: cholesterol ester CE (14:0) (increased), phosphoserine PS (14:0/18:2) and hexosylceramide HCER (d18:0/26:0) (decreased), phosphoinositol PI(16:0/18:2) (increased with 1×, decreased with 2× mTBI); (2) lipids that were altered in response to 1× mTBI only: free fatty acid FFA (18:3 and 20:3) (increased); (3) lipids that were altered in response to 2× mTBI only: HCER (22:0), phosphoethanolamine PE (P-18:1/20:4 and P-18:0/20:1) (increased), lysophosphatidylethanolamine LPE (20:1), phosphocholine PC (20:0/22:4), PI (18:1/18:2 and 20:0/18:2) (decreased). These findings suggest that increasing numbers of mTBI induce a range of changes dependent upon the lipid species, which likely reflect a balance of damage and reparative responses

Use of a genome-wide haploid genetic screen to identify treatment predicting factors: a proof-of-principle study in pancreatic cancer.

The ability to develop a comprehensive panel of treatment predicting factors would significantly improve our ability to stratify patients for cytotoxic or targeted therapies, and prevent patients receiving ineffective treatments. We have investigated if a recently developed genome-wide haploid genetic screen can be used to reveal the critical mediators of response to anticancer therapy. Pancreatic cancer is known to be highly resistant to systemic therapy. Recently epigenetic changes have been shown to be a key determinant in the maintenance of subpopulations of cancer cells with high-level resistance to cytotoxic therapy. We show that in human pancreatic cancer cell lines, treatment with the potent class I histone deacetylase inhibitor, entinostat, synergistically enhances gemcitabine-induced inhibition of cell proliferation and apoptosis. Using a genome-wide haploid genetic screen, we identified deoxycytidine kinase (DCK) as one of the genes with the highest degree of insertional enrichment following treatment with gemcitabine and entinostat; DCK is already known to be the rate-limiting activating enzyme for gemcitabine. Immunoblotting confirmed loss of DCK protein expression in the resistant KBM7 cells. CRISPR/Cas-9 inactivation of DCK in pancreatic cancer cell lines resulted in resistance to gemcitabine alone and in combination with entinostat. We have identified gemcitabine and entinostat as a potential new combination therapy in pancreatic cancer, and in this proof-of-principle study we have demonstrated that a recently developed haploid genetic screen can be used as a novel approach to identify the critical genes that determine treatment response

THE ELEVENTH AND TWELFTH DATA RELEASES OF THE SLOAN DIGITAL SKY SURVEY: FINAL DATA FROM SDSS-III

Citation: Alam, S., Albareti, F. D., Prieto, C. A., Anders, F., Anderson, S. F., Anderton, T., . . . Zhu, G. T. (2015). THE ELEVENTH AND TWELFTH DATA RELEASES OF THE SLOAN DIGITAL SKY SURVEY: FINAL DATA FROM SDSS-III. Astrophysical Journal Supplement Series, 219(1), 27. doi:10.1088/0067-0049/219/1/12The third generation of the Sloan Digital Sky Survey (SDSS-III) took data from 2008 to 2014 using the original SDSS wide-field imager, the original and an upgraded multi-object fiber-fed optical spectrograph, a new near-infrared high-resolution spectrograph, and a novel optical interferometer. All of the data from SDSS-III are now made public. In particular, this paper describes Data Release 11 (DR11) including all data acquired through 2013 July, and Data Release 12 (DR12) adding data acquired through 2014 July (including all data included in previous data releases), marking the end of SDSS-III observing. Relative to our previous public release (DR10), DR12 adds one million new spectra of galaxies and quasars from the Baryon Oscillation Spectroscopic Survey (BOSS) over an additional 3000 deg(2) of sky, more than triples the number of H-band spectra of stars as part of the Apache Point Observatory (APO) Galactic Evolution Experiment (APOGEE), and includes repeated accurate radial velocity measurements of 5500 stars from the Multi-object APO Radial Velocity Exoplanet Large-area Survey (MARVELS). The APOGEE outputs now include the measured abundances of 15 different elements for each star. In total, SDSS-III added 5200 deg(2) of ugriz imaging; 155,520 spectra of 138,099 stars as part of the Sloan Exploration of Galactic Understanding and Evolution 2 (SEGUE-2) survey; 2,497,484 BOSS spectra of 1,372,737 galaxies, 294,512 quasars, and 247,216 stars over 9376 deg(2); 618,080 APOGEE spectra of 156,593 stars; and 197,040 MARVELS spectra of 5513 stars. Since its first light in 1998, SDSS has imaged over 1/3 of the Celestial sphere in five bands and obtained over five million astronomical spectra.Additional Authors: Berlind, A. A.;Beutler, F.;Bhardwaj, V.;Bird, J. C.;Bizyaev, D.;Blake, C. H.;Blanton, M. R.;Blomqvist, M.;Bochanski, J. J.;Bolton, A. S.;Bovy, J.;Bradley, A. S.;Brandt, W. N.;Brauer, D. E.;Brinkmann, J.;Brown, P. J.;Brownstein, J. R.;Burden, A.;Burtin, E.;Busca, N. G.;Cai, Z.;Capozzi, D.;Rosell, A. C.;Carr, M. A.;Carrera, R.;Chambers, K. C.;Chaplin, W. J.;Chen, Y. C.;Chiappini, C.;Chojnowski, S. D.;Chuang, C. H.;Clerc, N.;Comparat, J.;Covey, K.;Croft, R. A. C.;Cuesta, A. J.;Cunha, K.;da Costa, L. N.;Da Rio, N.;Davenport, J. R. A.;Dawson, K. S.;De Lee, N.;Delubac, T.;Deshpande, R.;Dhital, S.;Dutra-Ferreira, L.;Dwelly, T.;Ealet, A.;Ebelke, G. L.;Edmondson, E. M.;Eisenstein, D. J.;Ellsworth, T.;Elsworth, Y.;Epstein, C. R.;Eracleous, M.;Escoffier, S.;Esposito, M.;Evans, M. L.;Fan, X. H.;Fernandez-Alvar, E.;Feuillet, D.;Ak, N. F.;Finley, H.;Finoguenov, A.;Flaherty, K.;Fleming, S. W.;Font-Ribera, A.;Foster, J.;Frinchaboy, P. M.;Galbraith-Frew, J. G.;Garcia, R. A.;Garcia-Hernandez, D. A.;Perez, A. E. G.;Gaulme, P.;Ge, J.;Genova-Santos, R.;Georgakakis, A.;Ghezzi, L.;Gillespie, B. A.;Girardi, L.;Goddard, D.;Gontcho, S. G. A.;Hernandez, J. I. G.;Grebel, E. K.;Green, P. J.;Grieb, J. N.;Grieves, N.;Gunn, J. E.;Guo, H.;Harding, P.;Hasselquist, S.;Hawley, S. L.;Hayden, M.;Hearty, F. R.;Hekker, S.;Ho, S.;Hogg, D. W.;Holley-Bockelmann, K.;Holtzman, J. A.;Honscheid, K.;Huber, D.;Huehnerhoff, J.;Ivans, II;Jiang, L. H.;Johnson, J. A.;Kinemuchi, K.;Kirkby, D.;Kitaura, F.;Klaene, M. A.;Knapp, G. R.;Kneib, J. P.;Koenig, X. P.;Lam, C. R.;Lan, T. W.;Lang, D. T.;Laurent, P.;Le Goff, J. M.;Leauthaud, A.;Lee, K. G.;Lee, Y. S.;Licquia, T. C.;Liu, J.;Long, D. C.;Lopez-Corredoira, M.;Lorenzo-Oliveira, D.;Lucatello, S.;Lundgren, B.;Lupton, R. H.;Mack, C. E.;Mahadevan, S.;Maia, M. A. G.;Majewski, S. R.;Malanushenko, E.;Malanushenko, V.;Manchado, A.;Manera, M.;Mao, Q. Q.;Maraston, C.;Marchwinski, R. C.;Margala, D.;Martell, S. L.;Martig, M.;Masters, K. L.;Mathur, S.;McBride, C. K.;McGehee, P. M.;McGreer, I. D.;McMahon, R. G.;Menard, B.;Menzel, M. L.;Merloni, A.;Meszaros, S.;Miller, A. A.;Miralda-Escude, J.;Miyatake, H.;Montero-Dorta, A. D.;More, S.;Morganson, E.;Morice-Atkinson, X.;Morrison, H. L.;Mosser, B.;Muna, D.;Myers, A. D.;Nandra, K.;Newman, J. A.;Neyrinck, M.;Nguyen, D. C.;Nichol, R. C.;Nidever, D. L.;Noterdaeme, P.;Nuza, S. E.;O'Connell, J. E.;O'Connell, R. W.;O'Connell, R.;Ogando, R. L. C.;Olmstead, M. D.;Oravetz, A. E.;Oravetz, D. J.;Osumi, K.;Owen, R.;Padgett, D. L.;Padmanabhan, N.;Paegert, M.;Palanque-Delabrouille, N.;Pan, K. K.;Parejko, J. K.;Paris, I.;Park, C.;Pattarakijwanich, P.;Pellejero-Ibanez, M.;Pepper, J.;Percival, W. J.;Perez-Fournon, I.;Perez-Rafols, I.;Petitjean, P.;Pieri, M. M.;Pinsonneault, M. H.;de Mello, G. F. P.;Prada, F.;Prakash, A.;Price-Whelan, A. M.;Protopapas, P.;Raddick, M. J.;Rahman, M.;Reid, B. A.;Rich, J.;Rix, H. W.;Robin, A. C.;Rockosi, C. M.;Rodrigues, T. S.;Rodriguez-Torres, S.;Roe, N. A.;Ross, A. J.;Ross, N. P.;Rossi, G.;Ruan, J. J.;Rubino-Martin, J. A.;Rykoff, E. S.;Salazar-Albornoz, S.;Salvato, M.;Samushia, L.;Sanchez, A. G.;Santiago, B.;Sayres, C.;Schiavon, R. P.;Schlegel, D. J.;Schmidt, S. J.;Schneider, D. P.;Schultheis, M.;Schwope, A. D.;Scoccola, C. G.;Scott, C.;Sellgren, K.;Seo, H. J.;Serenelli, A.;Shane, N.;Shen, Y.;Shetrone, M.;Shu, Y. P.;Aguirre, V. S.;Sivarani, T.;Skrutskie, M. F.;Slosar, A.;Smith, V. V.;Sobreira, F.;Souto, D.;Stassun, K. G.;Steinmetz, M.;Stello, D.;Strauss, M. A.;Streblyanska, A.;Suzuki, N.;Swanson, M. E. C.;Tan, J. C.;Tayar, J.;Terrien, R. C.;Thakar, A. R.;Thomas, D.;Thomas, N.;Thompson, B. A.;Tinker, J. L.;Tojeiro, R.;Troup, N. W.;Vargas-Magana, M.;Vazquez, J. A.;Verde, L.;Viel, M.;Vogt, N. P.;Wake, D. A.;Wang, J.;Weaver, B. A.;Weinberg, D. H.;Weiner, B. J.;White, M.;Wilson, J. C.;Wisniewski, J. P.;Wood-Vasey, W. M.;Yeche, C.;York, D. G.;Zakamska, N. L.;Zamora, O.;Zasowski, G.;Zehavi, I.;Zhao, G. B.;Zheng, Z.;Zhou, X.;Zhou, Z. M.;Zou, H.;Zhu, G. T

Modulation of enhancer looping and differential gene targeting by Epstein-Barr virus transcription factors directs cellular reprogramming

Epstein-Barr virus (EBV) epigenetically reprogrammes B-lymphocytes to drive immortalization and facilitate viral persistence. Host-cell transcription is perturbed principally through the actions of EBV EBNA 2, 3A, 3B and 3C, with cellular genes deregulated by specific combinations of these EBNAs through unknown mechanisms. Comparing human genome binding by these viral transcription factors, we discovered that 25% of binding sites were shared by EBNA 2 and the EBNA 3s and were located predominantly in enhancers. Moreover, 80% of potential EBNA 3A, 3B or 3C target genes were also targeted by EBNA 2, implicating extensive interplay between EBNA 2 and 3 proteins in cellular reprogramming. Investigating shared enhancer sites neighbouring two new targets (WEE1 and CTBP2) we discovered that EBNA 3 proteins repress transcription by modulating enhancer-promoter loop formation to establish repressive chromatin hubs or prevent assembly of active hubs. Re-ChIP analysis revealed that EBNA 2 and 3 proteins do not bind simultaneously at shared sites but compete for binding thereby modulating enhancer-promoter interactions. At an EBNA 3-only intergenic enhancer site between ADAM28 and ADAMDEC1 EBNA 3C was also able to independently direct epigenetic repression of both genes through enhancer-promoter looping. Significantly, studying shared or unique EBNA 3 binding sites at WEE1, CTBP2, ITGAL (LFA-1 alpha chain), BCL2L11 (Bim) and the ADAMs, we also discovered that different sets of EBNA 3 proteins bind regulatory elements in a gene and cell-type specific manner. Binding profiles correlated with the effects of individual EBNA 3 proteins on the expression of these genes, providing a molecular basis for the targeting of different sets of cellular genes by the EBNA 3s. Our results therefore highlight the influence of the genomic and cellular context in determining the specificity of gene deregulation by EBV and provide a paradigm for host-cell reprogramming through modulation of enhancer-promoter interactions by viral transcription factors

Subnational climate entrepreneurship: innovative climate action in California and São Paulo

The distinct role of subnational governments such as states and provinces in addressing climate change has been increasingly acknowledged. But while most studies investigate the causes and consequences of particular governments’ actions and networking activities, this article argues that subnational governments can develop climate action as a collective entrepreneurial activity. Addressing many elements explored in this special issue, it focuses on the second question and identifies climate entrepreneurship in two subnational governments—the states of California (USA) and São Paulo (Brazil). Examining internal action, as well as interaction with local authorities, national governments and the international regime, entrepreneurial activities are identified in the invention, diffusion and evaluation of subnational climate policy in each case. The article draws from the recent scholarship on policy innovation, entrepreneurship and climate governance. It contributes to the literature by exploring entrepreneurial subnational government activity in addressing climate change and expanding the understanding of the effects of policy innovation at the subnational level

Correction of tau mis-splicing caused by FTDP-17 MAPT mutations by spliceosome-mediated RNA trans-splicing

Frontotemporal dementia with parkinsonism linked to chromosome 17 (FTDP-17) is caused by mutations in the MAPT gene, encoding the tau protein that accumulates in intraneuronal lesions in a number of neurodegenerative diseases. Several FTDP-17 mutations affect alternative splicing and result in excess exon 10 (E10) inclusion in tau mRNA. RNA reprogramming using spliceosome-mediated RNA trans-splicing (SMaRT) could be a method of choice to correct aberrant E10 splicing resulting from FTDP-17 mutations. SMaRT creates a hybrid mRNA through a trans-splicing reaction between an endogenous target pre-mRNA and a pre-trans-splicing RNA molecule (PTM). However, FTDP-17 mutations affect the strength of cis-splicing elements and could favor cis-splicing over trans-splicing. Excess E10 inclusion in FTDP-17 can be caused by intronic mutations destabilizing a stem-loop protecting the 5′ splice site at the E10/intron 10 junction. COS cells transfected with a minigene containing the intronic +14 mutation produce exclusively E10+ RNA. Generation of E10− RNA was restored after co-transfection with a PTM designed to exclude E10. Similar results were obtained with a target containing the exonic N279K mutation which strengthens a splicing enhancer within E10. Conversely, increase or decrease in E10 content was achieved by trans-splicing from a target carrying the Δ280K mutation, which weakens the same splicing enhancer. Thus E10 inclusion can be modulated by trans-splicing irrespective of the strength of the cis-splicing elements affected by FTDP-17 mutations. In conclusion, RNA trans-splicing could provide the basis of therapeutic strategies for impaired alternative splicing caused by pathogenic mutations in cis-acting splicing elements

Sheep Updates 2005 - Part 4

This session covers twelve papers from different authors: REPRODUCTION 1. Is it worth increasing investment to increase lambing percentages? Lucy Anderton Department of Agriculture Western Australia. 2. What value is a lamb? John Young, Farming Systems Analysis Service, Kojonup, WA 3. Providing twin-bearing ewes with extra energy at lambing produces heavier lambs at marking. Rob Davidson WAMMCO International,, formerly University of Western Australia; Keith Croker, Ken Hart, Department of Agriculture Western Australia, Tim Wiese, Chuckem , Highbury, Western Australia. GENETICS 4. Underlying biological cause of trade-off between meat and wool. Part 1. Wool and muscle glycogen, BM Thomson, I Williams, University of WA, Crawley, JRBriegel, CSIRO Livestock Industries, Floreat Park WA &CRC for the Australian Sheep Industry, JC Greeff, Department of Agriculture Western Australia &CRC for the Australian Sheep Industry. 5. Underlying biological cause of trade-off between meat and wool. Part 2. Wool and fatness, NR Adams1,3, EN Bermingham1,3, JR Briegel1,3, JC Greeff2,3 1CSIRO Livestock Industries, Floreat Park WA 2Department of Agriculture Western Australia, 3CRC for the Australian Sheep Industry 6. Genetic trade-offs between lamb and wool production in Merino breeding programs, Johan Greeff, Department of Agriculture, Western Australia. 7. Clean fleece weight is no phenotypically independent of other traits. Sue Hatcherac and Gordon Refshaugebc aNSWDPI Orange Agricultural Institute, Orange NSW 2800 bUNE c/- NSWDPI Cowra AR&AS Cowra NSW 2794 cAustralian Sheep Industry CRC. 8. When you\u27re on a good thing, do it better: An economic analysis of sheep breed profitability. Emma Kopke, Ross Kingwell, Department of Agriculture, Western Australia, John Young, Farming Systems Analysis Service, Kojonup, WA. 9. Selection Demonstration Flocks: Demonstrating improvementsin productivity of merinos, K.E. Kemper, M.L. Hebart, F.D. Brien, K.S. Jaensch, R.J. Grimson, D.H. Smith South Australian Research and Development Institute 10. You are compromising yield by using Dust Penetration and GFW in breeding programs, Melanie Dowling, Department of Agriculture, Western Australia, A. (Tony) Schlink, CSIRO Livestock Industries, Wembley, Johan Greeff, Department of Agriculture Western Australia. 11. Merino Sheep can be bred for resistance to breech strike. Johan Greeff , John Karlsson, Department of Agriculture Western Australia 12. Parasite resistant sheep and hypersensitivity diarrhoea, L.J.E. Karlsson & J.C. Greeff, Department of Agriculture Western Australi