Phosphoproteins associated with cyclic nucleotide stimulation of ciliary motility in Paramecium

Permeabilized, MgATP-reactivated cells of Paramecium (models) respond to cyclic AMP and cyclic GMP by increasing forward swimming speed. In association with the motile response, cyclic AMP and 8-bromo-cyclic GMP (8-Br-cyclic GMP) stimulated protein phosphorylation. Cyclic AMP addition to permeabilized cells reproducibly stimulated the phosphorylation of 10 proteins, ranging in molecular weight from 15 to 110K (K = 10^3 M_r). 8-Br-cyclic GMP, which selectively activates the cyclic GMP-dependent protein kinase of Paramecium, stimulated the phosphorylation of a subset of the proteins phosphorylated by cyclic AMP. Ca^(2+) addition caused backward swimming and stimulated the phosphorylation of four substrates, including a 25K target that may also be phosphorylated in response to cyclic nucleotide addition. Ba^(2+) and Sr^(2+) also induced backward swimming, but did not cause detectable phosphorylation. To identify ciliary targets of cyclic nucleotide-dependent protein kinase activity, permeabilized cells were deciliated following reactivation of motility with Mg-[y-^(32)P]ATP in the presence or absence of cyclic nucleotide. Soluble proteins of the deciliation supernatant were enriched in 15 cyclic AMP-stimulated phosphoproteins, ranging in molecular weight from 15 to 95K. Most of the ciliary substrates were axonemal and could be released by high salt solution. A 29K protein that copurified in sucrose gradients with the 22S dynein, and a high molecular weight protein (greater than 300K) in the 19 S region were phosphorylated when cyclic AMP was added to permeabilized, motile cells. These data suggest that regulation of ciliary motility by cyclic AMP may include phosphorylation of dynein-associated proteins

Critical parameters for efficient sonication and improved chromatin immunoprecipitation of high molecular weight proteins

Solubilization of cross-linked cells followed by chromatin shearing is essential for successful chromatin immunoprecipitation (ChIP). However, this task, typically accomplished by ultrasound treatment, may often become a pitfall of the process, due to inconsistent results obtained between different experiments under seemingly identical conditions. To address this issue we systematically studied ultrasound-mediated cell lysis and chromatin shearing, identified critical parameters of the process and formulated a generic strategy for rational optimization of ultrasound treatment. We also demonstrated that whereas ultrasound treatment required to shear chromatin to within a range of 100–400 bp typically degrades large proteins, a combination of brief sonication and benzonase digestion allows for the generation of similarly sized chromatin fragments while preserving the integrity of associated proteins. This approach should drastically improve ChIP efficiency for this class of proteins

Development and Monitoring of Revegetation Methods: Connecting Students with Restoration Activities at Awcomin Marsh

Five classes in a local elementary school participated in an effort to grow and plant high marsh and upper border vegetation at a salt marsh restoration site in spring 2005. Seeds of six marsh upper edge species were successfully germinated and grown into seedlings by third graders. The seedlings were planted by the students in late spring 2005, but only switchgrass and quackgrass plants appeared to have established and survived after one year. Mature shoots of three high marsh species planted by the third graders (salt hay, salt grass and black grass) established successfully and continue to proliferate. In addition, we assessed an experiment of cordgrass plantings performed by community volunteers in 2002. The experiment was designed to test the effectiveness of three planting techniques at a salt marsh restored by the excavation of old dredge spoil that had been colonized by common reed. After four growing seasons, Plug, Bare Root Shoot, and Seed Head planting techniques exhibited greater cover of cordgrass and total cover of vascular plants when compared with unplanted areas. Cover of perennial plants (e.g., cordgrass), which contributes directly to belowground soil development in salt marshes, dominated the planted plots. Cover of annual species dominated the unplanted plots. Planting cordgrass in areas where dredge spoils and common reed had been excavated from a historic marsh accelerated the development of native vegetation compared with unplanted areas. Performance and evaluation of the two sets of plantings has provided information about appropriate planting techniques for our region and has involved and educated the local community about the values of salt marsh to promote stewardship. Recommendations included the use of bare root shoot and seed head planting techniques where cordgrass is desired. Outside plots or a greenhouse may be needed for successful propagation of upper edge marsh species from seed, and a planting program that includes mature plants as well as seedlings is recommended to ensure success

Localization transitions in non-Hermitian quantum mechanics

We study the localization transitions which arise in both one and two dimensions when quantum mechanical particles described by a random Schr\"odinger equation are subjected to a constant imaginary vector potential. A path-integral formulation relates the transition to flux lines depinned from columnar defects by a transverse magnetic field in superconductors. The theory predicts that the transverse Meissner effect is accompanied by stretched exponential relaxation of the field into the bulk and a diverging penetration depth at the transition.Comment: 4 pages (latex) with 3 figures (epsf) embedded in the text using the style file epsf.st

The Police Powers: A Pretext For Protectionism?

The first part of this note will define administrative and technical regulations and discuss recent trends toward these regulations in the European Community. The second part will concentrate on protectionism and analyze the negative effects of protectionism on consumers and producers. This part will set forth four arguments against protectionism. These four rationales can be divided into: (1) economic rationale; (2) retaliation rationale; (3) legitimacy of international law; and (4) purpose of the World Trade Organization (WTO). Part three will examine the current legal atmosphere surrounding health and safety regulations. This part will examine the Agreement on Sanitary and Phytosanitary Measures (hereinafter SPS Agreement), and the Agreement on Technical Barriers to Trade, (hereinafter TBT Agreement). Part four will address health and safety regulations adopted by the European Union. This part will discuss four of the major directives that have been put in place. These four directives are: Council Directive 90/220/EEC, Commission Directive 97/35/EC, the Novel Foods Regulation (EC) No. 258/97 and Directive 88/146/EEC. Part five of this note will analyze the current standards used under the SPS Agreement and then look at the standards under the TBT Agreement. Part six will review the U.S. Supreme Court\u27s dormant commerce clause jurisprudence and articulate a new standard to be used when analyzing protectionist policies in the area of international trade. Part seven will apply the newly articulated test to the EU directives

Structure and dynamics of topological defects in a glassy liquid on a negatively curved manifold

We study the low-temperature regime of an atomic liquid on the hyperbolic plane by means of molecular dynamics simulation and we compare the results to a continuum theory of defects in a negatively curved hexagonal background. In agreement with the theory and previous results on positively curved (spherical) surfaces, we find that the atomic configurations consist of isolated defect structures, dubbed "grain boundary scars", that form around an irreducible density of curvature-induced disclinations in an otherwise hexagonal background. We investigate the structure and the dynamics of these grain boundary scars

A case of large bowel obstruction secondary to twin pregnancy

No abstract available