Components required for in vitro cleavage and polyadenylation of eukaryotic mRNA

We have studied in vitro cleavage/polyadenylation of precursor RNA containing herpes simplex virus type 2 poly A site sequences and have analysed four RNA/protein complexes which form during in vitro reactions. Two complexes, A and B, form extremely rapidly and are then progressively replaced by a third complex, C which is produced following cleavage and polyadenylation of precursor RNA. Substitution of ATP with cordycepin triphosphate prevents polyadenylation and the formation of complex C however a fourth complex, D, results which contains cleaved RNA. A precursor RNA lacking GU-rich downstream sequences required for efficient cleavage/ polyadenylation fails to form complex B and produces a markedly reduced amount of complex A. As these GU-rich sequences are required for efficient cleavage, this establishes a relationship between complex B formation and cleavage/polyadenylation of precursor RNA in vitro. The components required for in vitro RNA processing have been separated by fractionation of the nuclear extract on Q-Sepharose and Biorex 70 columns. A Q-Sepharose fraction forms complex B but does not process RNA. Addition of a Biorex 70 fraction restores cleavage activity at the poly A site but this fraction does not appear to contribute to complex formation. Moreover, in the absence of polyethylene glycol, precursor RNA is not cleaved and polyadenylated, however, complexes A and B readily form. Thus, while complex B is necessary for in vitro cleavage and polyadenylation, it may not contain all the components required for this processing

Economic evaluation of alternative farm water sources in the claypan area of Illinois

The claypan area of Illinois is characterized by unreliable water supplies. In addition, water quality problems have recently been recognized in the form of widespread nitrate coliform contamination of water in private wells in Washington County. A ten-square mile area in this county was selected for study. The nature of present water supplies was described and the costs of six alternative supply systems were estimated: (1) present on-farm sources, (2) treatment of pond water, (3) combination of present on-farm sources plus hauling, (4) hauling all water, (5) purchasing water from nearby municipality via pipeline, (6) combination pipeline plus on-farm sources. The six alternatives were examined in terms of the net present value of costs associated with each alternative, considering a 40-year planning period. Although the present sources proved to be the least costly they are unsatisfactory for reasons of health and also reliability. Treatment of pond water was the least-cost source of meeting the quality requirements for water for the total area. However, the reliability of this source and questions of personal preference against drinking pond water may indicate a preference among some users for connection with a municipal system which was somewhat more expensive.U.S. Geological SurveyU.S. Department of the InteriorOpe

Critical points in edge tunneling between generic FQH states

A general description of weak and strong tunneling fixed points is developed in the chiral-Luttinger-liquid model of quantum Hall edge states. Tunneling fixed points are a subset of `termination' fixed points, which describe boundary conditions on a multicomponent edge. The requirement of unitary time evolution at the boundary gives a nontrivial consistency condition for possible low-energy boundary conditions. The effect of interactions and random hopping on fixed points is studied through a perturbative RG approach which generalizes the Giamarchi-Schulz RG for disordered Luttinger liquids to broken left-right symmetry and multiple modes. The allowed termination points of a multicomponent edge are classified by a B-matrix with rational matrix elements. We apply our approach to a number of examples, such as tunneling between a quantum Hall edge and a superconductor and tunneling between two quantum Hall edges in the presence of interactions. Interactions are shown to induce a continuous renormalization of effective tunneling charge for the integrable case of tunneling between two Laughlin states. The correlation functions of electronlike operators across a junction are found from the B matrix using a simple image-charge description, along with the induced lattice of boundary operators. Many of the results obtained are also relevant to ordinary Luttinger liquids.Comment: 23 pages, 6 figures. Xiao-Gang Wen: http://dao.mit.edu/~we

A note on the convergence of parametrised non-resonant invariant manifolds

Truncated Taylor series representations of invariant manifolds are abundant in numerical computations. We present an aposteriori method to compute the convergence radii and error estimates of analytic parametrisations of non-resonant local invariant manifolds of a saddle of an analytic vector field, from such a truncated series. This enables us to obtain local enclosures, as well as existence results, for the invariant manifolds

A Berger type normal holonomy theorem for complex submanifolds

We prove a kind of Berger-Simons' Theorem for the normal holonomy group of a complex submanifold of the projective spac

Edge Dynamics in Quantum Hall Bilayers II: Exact Results with Disorder and Parallel Fields

We study edge dynamics in the presence of interlayer tunneling, parallel magnetic field, and various types of disorder for two infinite sequences of quantum Hall states in symmetric bilayers. These sequences begin with the 110 and 331 Halperin states and include their fractional descendants at lower filling factors; the former is easily realized experimentally while the latter is a candidate for the experimentally observed quantum Hall state at a total filling factor of 1/2 in bilayers. We discuss the experimentally interesting observables that involve just one chiral edge of the sample and the correlation functions needed for computing them. We present several methods for obtaining exact results in the presence of interactions and disorder which rely on the chiral character of the system. Of particular interest are our results on the 331 state which suggest that a time-resolved measurement at the edge can be used to discriminate between the 331 and Pfaffian scenarios for the observed quantum Hall state at filling factor 1/2 in realistic double-layer systems.Comment: revtex+epsf; two-up postscript at http://www.sns.ias.edu/~leonid/ntwoup.p

Geometric effects on T-breaking in p+ip and d+id superconductors

Superconducting order parameters that change phase around the Fermi surface modify Josephson tunneling behavior, as in the phase-sensitive measurements that confirmed $d$ order in the cuprates. This paper studies Josephson coupling when the individual grains break time-reversal symmetry; the specific cases considered are $p \pm ip$ and $d \pm id$, which may appear in Sr$_2$RuO$_4$ and Na$_x$CoO$_2 \cdot$(H$_2$O)$_y$ respectively. $T$-breaking order parameters lead to frustrating phases when not all grains have the same sign of time-reversal symmetry breaking, and the effects of these frustrating phases depend sensitively on geometry for 2D arrays of coupled grains. These systems can show perfect superconducting order with or without macroscopic $T$-breaking. The honeycomb lattice of superconducting grains has a superconducting phase with no spontaneous breaking of $T$ but instead power-law correlations. The superconducting transition in this case is driven by binding of fractional vortices, and the zero-temperature criticality realizes a generalization of Baxter's three-color model.Comment: 8 page

Curved Flats, Pluriharmonic Maps and Constant Curvature Immersions into Pseudo-Riemannian Space Forms

We study two aspects of the loop group formulation for isometric immersions with flat normal bundle of space forms. The first aspect is to examine the loop group maps along different ranges of the loop parameter. This leads to various equivalences between global isometric immersion problems among different space forms and pseudo-Riemannian space forms. As a corollary, we obtain a non-immersibility theorem for spheres into certain pseudo-Riemannian spheres and hyperbolic spaces. The second aspect pursued is to clarify the relationship between the loop group formulation of isometric immersions of space forms and that of pluriharmonic maps into symmetric spaces. We show that the objects in the first class are, in the real analytic case, extended pluriharmonic maps into certain symmetric spaces which satisfy an extra reality condition along a totally real submanifold. We show how to construct such pluriharmonic maps for general symmetric spaces from curved flats, using a generalised DPW method.Comment: 21 Pages, reference adde

Assembly of a Three-Dimensional Multitype Bronchiole Coculture Model Using Magnetic Levitation

A longstanding goal in biomedical research has been to create organotypic cocultures that faithfully represent native tissue environments. There is presently great interest in representative culture models of the lung, which is a particularly challenging tissue to recreate in vitro. This study used magnetic levitation in conjunction with magnetic nanoparticles as a means of creating an organized three-dimensional (3D) coculture of the bronchiole that sequentially layers cells in a manner similar to native tissue architecture. The 3D coculture model was assembled from four human cell types in the bronchiole: endothelial cells, smooth muscle cells (SMCs), fibroblasts, and epithelial cells (EpiCs). This study represents the first effort to combine these particular cell types into an organized bronchiole coculture. These cell layers were first cultured in 3D by magnetic levitation, and then manipulated into contact with a custom-made magnetic pen, and again cultured for 48 h. Hematoxylin and eosin staining of the resulting coculture showed four distinct layers within the 3D coculture. Immunohistochemistry confirmed the phenotype of each of the four cell types and showed organized extracellular matrix formation, particularly, with collagen type I. Positive stains for CD31, von Willebrand factor, smooth muscle a-actin, vimentin, and fibronectin demonstrate the maintenance of the phenotype for endothelial cells, SMCs, and fibroblasts. Positive stains for mucin-5AC, cytokeratin, and E-cadherin after 7 days with and without 1% fetal bovine serum showed that EpiCs maintained the phenotype and function. This study validates magnetic levitation as a method for the rapid creation of organized 3D cocultures that maintain the phenotype and induce extracellular matrix formation

Gamma rays from dark matter annihilation in the Draco and observability at ARGO

The CACTUS experiment recently observed a gamma ray excess above 50 GeV from the direction of the Draco dwarf spheroidal galaxy. Considering that Draco is dark matter dominated the gamma rays may be generated through dark matter annihilation in the Draco halo. In the framework of the minimal supersymmetric extension of the standard model we explore the parameter space to account for the gamma ray signals at CACTUS. We find that the neutralino mass is constrained to be approximately in the range between 100 GeV ~ 400 GeV and a sharp central cuspy of the dark halo profile in Draco is necessary to explain the CACTUS results. We then discuss further constraints on the supersymmetric parameter space by observations at the ground based ARGO detector. It is found that the parameter space can be strongly constrained by ARGO if no excess from Draco is observed above 100 GeV.Comment: 15 pages, 4 figure