Multi-target genome editing reduces polyphenol oxidase activity in wheat (Triticum aestivum L.) grains

Introduction: Polyphenol oxidases (PPO) are dual activity metalloenzymes that catalyse the production of quinones. In plants, PPO activity may contribute to biotic stress resistance and secondary metabolism but is undesirable for food producers because it causes the discolouration and changes in flavour profiles of products during post-harvest processing. In wheat (Triticum aestivum L.), PPO released from the aleurone layer of the grain during milling results in the discolouration of flour, dough, and end-use products, reducing their value. Loss-of-function mutations in the PPO1 and PPO2 paralogous genes on homoeologous group 2 chromosomes confer reduced PPO activity in the wheat grain. However, limited natural variation and the proximity of these genes complicates the selection of extremely low-PPO wheat varieties by recombination. The goal of the current study was to edit all copies of PPO1 and PPO2 to drive extreme reductions in PPO grain activity in elite wheat varieties. Results: A CRISPR/Cas9 construct with one single guide RNA (sgRNA) targeting a conserved copper binding domain was used to edit all seven PPO1 and PPO2 genes in the spring wheat cultivar ‘Fielder’. Five of the seven edited T1 lines exhibited significant reductions in PPO activity, and T2 lines had PPO activity up to 86.7% lower than wild-type. The same construct was transformed into the elite winter wheat cultivars ‘Guardian’ and ‘Steamboat’, which have five PPO1 and PPO2 genes. In these varieties PPO activity was reduced by >90% in both T1 and T2 lines. In all three varieties, dough samples from edited lines exhibited reduced browning. Discussion: This study demonstrates that multi-target editing at late stages of variety development could complement selection for beneficial alleles in cropbreeding programs by inducing novel variation in loci inaccessible to recombinatio

An Extension for Direct Gauge Mediation of Metastable Supersymmetry Breaking

We study the direct mediation of metastable supersymmetry breaking by a \Phi^2-deformation to the ISS model and extend it by splitting both Tr\Phi and Tr\Phi^2 terms in the superpotential and gauging the flavor symmetry. We find that with such an extension the enough long-lived metastable vacua can be obtained and the proper gaugino masses can be generated. Also, this allows for constructing a kind of models which can avoid the Landau pole problem. Especially, in our metastable vacua there exist a large region for the parameter m_3 which can satisfy the phenomenology requirements and allow for a low SUSY breaking scale (\sim 100 TeV).Comment: version in Europhys. Let

Two dimensional Sen connections in general relativity

The two dimensional version of the Sen connection for spinors and tensors on spacelike 2-surfaces is constructed. A complex metric $\gamma_{AB}$ on the spin spaces is found which characterizes both the algebraic and extrinsic geometrical properties of the 2-surface $\$$. The curvature of the two dimensional Sen operator $\Delta_e$ is the pull back to $\$$ of the anti-self-dual part of the spacetime curvature while its `torsion' is a boost gauge invariant expression of the extrinsic curvatures of $\$$. The difference of the 2 dimensional Sen and the induced spin connections is the anti-self-dual part of the `torsion'. The irreducible parts of $\Delta_e$ are shown to be the familiar 2-surface twistor and the Weyl--Sen--Witten operators. Two Sen--Witten type identities are derived, the first is an identity between the 2 dimensional twistor and the Weyl--Sen--Witten operators and the integrand of Penrose's charge integral, while the second contains the `torsion' as well. For spinor fields satisfying the 2-surface twistor equation the first reduces to Tod's formula for the kinematical twistor.Comment: 14 pages, Plain Tex, no report numbe

Configuring large high-performance clusters at lightspeed: A case study

Abstract Over a decade ago, the TOP500 list was started as a way to measure supercomputers by their sustained performance on a particular linear algebra benchmark. Once reserved for the exotic machines and extremely well-funded centers and laboratories, commodity clusters now make it possible for smaller groups to deploy and use highperformance machines in their own laboratories. This paper describes a weekend activity where two existing 128-node commodity clusters were fused into a single 256-node cluster for the specific purpose of running the benchmark used to rank the machines in the TOP500 supercomputer list. The resulting metacluster sits on the November 2002 list at position 233. A key differentiator for this cluster is that it was assembled, in terms of its software, from the NPACI Rocks open-source cluster toolkit as downloaded from the public website. The toolkit allows non-cluster experts to deploy and run supercomputerclass machines in a matter of hours instead of weeks or months. With the exception of recompiling the University of Tennessee's Automatically Tuned Linear Algebra Subroutines (ATLAS) library with a recommended version of the GNU C compiler, this metacluster ran a "stock" Rocks distribution. Successful first-time deployment of the fused cluster was completed in a scant 6 hours. Partitioning of the metacluster and restoration of the two 128-node clusters to their original configuration was completed in just over 40 minutes. This paper describes early (pre-weekend) benchmark activities to empirically determine reasonably good parameters for the High-Performance Linpack (HPL) code on both Ethernet and Myrinet interconnects. It fully describes the physical layout of the machine, the description-based installation methods used in Rocks to re-deploy two independent clusters as a single cluster, and gives the benchmark results that were gathered over the 40-hour period allotted for the complete experiment. In addition, we describe some of the online monitoring and measurement techniques that were employed during the experiment. Finally, we point out the issues uncovered with a commodity cluster of this size. The techniques presented in this paper truly bring supercomputers into the hands of the masses of computational scientists

Two-spinor Formulation of First Order Gravity coupled to Dirac Fields

Two-spinor formalism for Einstein Lagrangian is developed. The gravitational field is regarded as a composite object derived from soldering forms. Our formalism is geometrically and globally well-defined and may be used in virtually any 4m-dimensional manifold with arbitrary signature as well as without any stringent topological requirement on space-time, such as parallelizability. Interactions and feedbacks between gravity and spinor fields are considered. As is well known, the Hilbert-Einstein Lagrangian is second order also when expressed in terms of soldering forms. A covariant splitting is then analysed leading to a first order Lagrangian which is recognized to play a fundamental role in the theory of conserved quantities. The splitting and thence the first order Lagrangian depend on a reference spin connection which is physically interpreted as setting the zero level for conserved quantities. A complete and detailed treatment of conserved quantities is then presented.Comment: 16 pages, Plain TE

A One Health overview, facilitating advances in comparative medicine and translational research.

Table of contentsA1 One health advances and successes in comparative medicine and translational researchCheryl StroudA2 Dendritic cell-targeted gorilla adenoviral vector for cancer vaccination for canine melanomaIgor Dmitriev, Elena Kashentseva, Jeffrey N. Bryan, David T. CurielA3 Viroimmunotherapy for malignant melanoma in the companion dog modelJeffrey N. Bryan, David Curiel, Igor Dmitriev, Elena Kashentseva, Hans Rindt, Carol Reinero, Carolyn J. HenryA4 Of mice and men (and dogs!): development of a commercially licensed xenogeneic DNA vaccine for companion animals with malignant melanomaPhilip J. BergmanA5 Successful immunotherapy with a recombinant HER2-expressing Listeria monocytogenes in dogs with spontaneous osteosarcoma paves the way for advances in pediatric osteosarcomaNicola J. Mason, Josephine S. Gnanandarajah, Julie B. Engiles, Falon Gray, Danielle Laughlin, Anita Gaurnier-Hausser, Anu Wallecha, Margie Huebner, Yvonne PatersonA6 Human clinical development of ADXS-HER2Daniel O'ConnorA7 Leveraging use of data for both human and veterinary benefitLaura S. TremlA8 Biologic replacement of the knee: innovations and early clinical resultsJames P. StannardA9 Mizzou BioJoint Center: a translational success storyJames L. CookA10 University and industry translational partnership: from the lab to commercializationMarc JacobsA11 Beyond docking: an evolutionarily guided OneHealth approach to drug discoveryGerald J. Wyckoff, Lee Likins, Ubadah Sabbagh, Andrew SkaffA12 Challenges and opportunities for data applications in animal health: from precision medicine to precision husbandryAmado S. GuloyA13 A cloud-based programmable platform for healthHarlen D. HaysA14 Comparative oncology: One Health in actionAmy K. LeBlancA15 Companion animal diseases bridge the translational gap for human neurodegenerative diseaseJoan R. Coates, Martin L. Katz, Leslie A. Lyons, Gayle C. Johnson, Gary S. Johnson, Dennis P. O'BrienA16 Duchenne muscular dystrophy gene therapyDongsheng DuanA17 Polycystic kidney disease: cellular mechanisms to emerging therapiesJames P. CalvetA18 The domestic cat as a large animal model for polycystic kidney diseaseLeslie A. Lyons, Barbara GandolfiA19 The support of basic and clinical research by the Polycystic Kidney Disease FoundationDavid A. BaronA20 Using naturally occurring large animal models of human disease to enable clinical translation: treatment of arthritis using autologous stromal vascular fraction in dogsMark L. WeissA21 Regulatory requirements regarding clinical use of human cells, tissues, and tissue-based productsDebra A. WebsterA22 Regenerative medicine approaches to Type 1 diabetes treatmentFrancis N. KaranuA23 The zoobiquity of canine diabetes mellitus, man's best friend is a friend indeed-islet transplantationEdward J. RobbA24 One Medicine: a development model for cellular therapy of diabetesRobert J. Harman

The Status of GMSB After 1/fb at the LHC

We thoroughly investigate the current status of supersymmetry in light of the latest searches at the LHC, using General Gauge Mediation (GGM) as a well-motivated signature generator that leads to many different simplified models. We consider all possible promptly-decaying NLSPs in GGM, and by carefully reinterpreting the existing LHC searches, we derive limits on both colored and electroweak SUSY production. Overall, the coverage of GGM parameter space is quite good, but much discovery potential still remains even at 7 TeV. We identify several regions of parameter space where the current searches are the weakest, typically in models with electroweak production, third generation sfermions or squeezed spectra, and we suggest how ATLAS and CMS might modify their search strategies given the understanding of GMSB at 1/fb. In particular, we propose the use of leptonic $M_{T2}$ to suppress $t{\bar t}$ backgrounds. Because we express our results in terms of simplified models, they have broader applicability beyond the GGM framework, and give a global view of the current LHC reach. Our results on 3rd generation squark NLSPs in particular can be viewed as setting direct limits on naturalness.Comment: 44 pages, refs added, typos fixed, improved MC statistics in fig 1

Twistor Strings with Flavour

We explore the tree-level description of a class of N=2 UV-finite SYM theories with fundamental flavour within a topological B-model twistor string framework. In particular, we identify the twistor dual of the Sp(N) gauge theory with one antisymmetric and four fundamental hypermultiplets, as well as that of the SU(N) theory with 2N hypermultiplets. This is achieved by suitably orientifolding/orbifolding the original N=4 setup of Witten and adding a certain number of new topological 'flavour'-branes at the orientifold/orbifold fixed planes to provide the fundamental matter. We further comment on the appearance of these objects in the B-model on CP(3|4). An interesting aspect of our construction is that, unlike the IIB description of these theories in terms of D3 and D7-branes, on the twistor side part of the global flavour symmetry is realised geometrically. We provide evidence for this correspondence by calculating and matching amplitudes on both sides.Comment: 38+12 pages; uses axodraw.sty. v2: References added, minor clarification