Inhibition of G1 cyclin-dependent kinase activity during growth arrest of human breast carcinoma cells by prostaglandin A2

Prostaglandin A2 (PGA2) potently inhibits cell proliferation and suppresses tumor growth in vivo, but little is known regarding the molecular mechanisms mediating these effects. Here we demonstrate that treatment of breast carcinoma MCF-7 cells with PGA2 leads to G1 arrest associated with a dramatic decrease in the levels of cyclin D1 and cyclin-dependent kinase 4 (cdk4) and accompanied by an increase in the expression of p21. We further show that these effects occur independent of cellular p53 status. The decline in cyclin D and cdk4 protein levels is correlated with loss in cdk4 kinase activity, cdk2 activity is also significantly inhibited in PGA2-treated cells, an effect closely associated with the upregulation of p21. Immunoprecipitation experiments verified that p21 was indeed complexed with cdk2 in PGA2-treated cells. Additional experiments with synchronized MCF-7 cultures stimulated with serum revealed that treatment with PGA2 prevents the progression of cells from G1 to S. Accordingly, the kinase activity associated with cdk4, cyclin E, and cdk2 immunocomplexes, which normally increases following serum addition, was unchanged in PGA2-treated cells. Furthermore, the retinoblastoma protein (Rb), a substrate of cdk4 and cdk2 whose phosphorylation is necessary for cell cycle progression, remains underphosphorylated in PGA2-treated serum-stimulated cells. These findings indicate that PGA2 exerts its growth-inhibitory effects through modulation of the expression and/or activity of several key G1 regulatory proteins. Our results highlight the chemotherapeutic potential of PGA2, particularly for suppressing growth of tumors lacking p53 function.     

A Framework for Interactive t-SNE Clustering

In this paper, we describe our progress in creating the framework for an interactive application that allows humans to actively participate in a t-SNE clustering process. t-SNE (t-Distributed Stochastic Neighbor Embedding) is a dimensionality reduction technique that maps high dimensional data sets to lower dimensions that can then be visualized for human interpretation. By prompting users to monitor outlying points during the t-SNE clustering process, we hypothesize that users may be able to make clustering faster and more accurate than purely algorithmic methods. Further research would test these hypotheses directly. We would also attempt to decrease the lag time between the various components of our application and develop an intuitive approach for humans to aid in clustering unlabeled data. Research into human assisted clustering can combine the strengths of both humans and computer programs to improve the results of data analysis.     

The structural consequences of exchanging tryptophan and tyrosine residues in B.stearothermophilus lactate dehydrogenase

A mutant Bacillus stearothermophilus lactate dehydrogenase hasbeen prepared in which all three tryptophan residues in thewild-type enzyme have been replaced by tyrosines. In addition,a tyrosine residue has been mutated to a tryptophan, which actsas a fluorescence probe to monitor protein folding. The mutantenzyme crystallizes in the same crystal form as the wild-type.The crystal structure of the mutant has been determined at 2.8Å resolution. Solution studies have suggested that thereis little effect upon the mutant enzyme as judged by its kineticproperties. Comparison of the crystal structures of the mutantand wild-type enzymes confirms this conclusion, and revealsthat alterations in structure in the region of these mutationsare of a similar magnitude to those observed throughout thestructure, and are not significant when compared with the errorsin atomic positions expected for a structure at this resolution.     

Foraging mechanisms of age-0 lake trout (Salvelinus namaycush)

Reaction distances under various light intensities (0-19 uE/m2/s), angles of attack, swimming speeds, and percentage of overall foraging success were measured. Extensive efforts have been invested in restoring lean lake trout (Salvelinus namaycush) populations in the Laurentian Great Lakes, but successful natural recruitment of lake trout continues to be rare outside of Lake Superior and parts of Lake Huron. There is evidence of high mortality during the first several months after eggs hatch in the spring, but little is known about the foraging mechanisms of this age-0 life stage. We developed a foraging model for age-0 lake trout (S. namaycush) in response to amphipods (Hyalella azteca) and mysids (Mysis diluviana) by simulating underwater environmental conditions in the Great Lakes using a temperature-controlled chamber and spectrally matched lighting. Reaction distances under various light intensities (0–19 uE/m²/s), angles of attack, swimming speeds, and percentage of overall foraging success were measured. Intake rates under different light intensities and prey densities were also measured. Age-0 lake trout were non-responsive in the dark, but were equally responsive under all light levels tested. Age-0 lake trout also demonstrated a longer reaction distance in response to moving prey, particularly mysids, which had an escape response that reduced overall foraging success. We determined that prey intake rate (numeric or biomass) could be modeled most accurately as a function of prey density using a Michaelis–Menton equation and that even under low mysid densities (3 individuals/m²), age-0 lake trout could quickly satisfy their energetic demands in a benthic setting.     

Castles and Pavilions: Creating New Hybrid Places of Exchange

Tom Holbrook , Director at architecture and urban design practice 5th Studio, describes two projects in the south of England that respond spatially to the needs of ‘the knowledge economy’. The first is the River Lea Park, to the east of London, which promotes open connectivity between people in a post- industrial landscape; and the second is the Creative Exchange in St Neots, Cambridgeshire, a building designed to accommodate local enterprise. Both are designed to redirect social and economic activities, supporting creative swarming, and establishing what Stafford Beer has coined as ‘platforms for change’.     

Polyampholyte acrylic latexes for tablet coating applications

Polyampholyte latexes can exist within a certain pH range as low‐viscosity aqueous dispersions, while upon a pH shift to the vicinity of the isoelectric point they undergo ionic coacervation. Three classes of coacervation latexes were synthesized and evaluated for their suitability for use in tablet coating applications. Pharmaceutical tablet coatings are commonly based on hydroxypropyl methyl cellulose, poly(vinyl alcohol), and acrylic polymers. Because of the high viscosity of their aqueous solutions, and to the consequent required low concentrations of the tablet coating polymers in the coating solutions to enable sufficiently low viscosity for effective spray application, the current commercial pharmaceutical tablet coating technology requires the removal of large amounts of water during the manufacturing process. In this work, films prepared from high‐solids, low‐viscosity coacervated acrylic latexes showed good hardness, very low tackiness, an excellent combination of optical properties, and very low water vapor permeability. © 2013 Wiley Periodicals, Inc. J. Appl. Polym. Sci. 2014 , 131, 40049.     

Tumor promoter arsenite activates extracellular signal-regulated kinase through a signaling pathway mediated by epidermal growth factor receptor and Shc

Although arsenite is an established carcinogen, the mechanisms underlying its tumor-promoting properties are poorly understood. Previously, we reported that arsenite treatment leads to the activation of the extracellular signal-regulated kinase (ERK) in rat PC12 cells through a Ras-dependent pathway. To identify potential mediators of the upstream signaling cascade, we examined the tyrosine phosphorylation profile in cells exposed to arsenite. Arsenite treatment rapidly stimulated tyrosine phosphorylation of several proteins in a Ras-independent manner, with a pattern similar to that seen in response to epidermal growth factor (EGF) treatment. Among these phosphorylated proteins were three isoforms of the proto-oncoprotein Shc as well as the EGF receptor (EGFR). Tyrosine phosphorylation of Shc allowed for enhanced interactions between Shc and Grb2 as identified by coimmunoprecipitation experiments. The arsenite-induced tyrosine phosphorylation of Shc, enhancement of Shc and Grb2 interactions, and activation of ERK were all drastically reduced by treatment of cells with either the general growth factor receptor poison suramin or the EGFR-selective inhibitor tyrphostin AG1478. Down-regulation of EGFR expression through pretreatment of cells with EGF also attenuated ERK activation and Shc tyrosine phosphorylation in response to arsenite treatment. These results demonstrate that the EGFR and Shc are critical mediators in the activation of the Ras/ERK signaling cascade by arsenite and suggest that arsenite acts as a tumor promoter largely by usurping this growth factor signaling pathway.     

Edge dislocation climb over non-deformable circular inclusions

Edge dislocation climb over non-deformable circular inclusions is studied as a possible rate limiting mechanism for creep of dispersion strengthened metals. The inclusion and matrix are assumed to exhibit no misfit and to be elastically isotropic, with the inclusion, more rigid than the matrix. The climb motion of the dislocation is assumed to be diffusion controlled and very much slower than glide. We calculate the path taken by the dislocation as it climbs past the inclusion. Special attention is given to the equilibrium separation distance between the dislocation and inclusion, the total climb distance required to pass the inclusion and the total lag time imposed on the dislocation motion by the presence of the inclusion. We discuss the results of these calculations and their implications in connection with the problem of creep of dispersion strengthened metals.     

Optimality of the Münch mechanism for translocation of sugars in plants

Plants require effective vascular systems for the transport of water and dissolved molecules between distal regions. Their survival depends on the ability to transport sugars from the leaves where they are produced to sites of active growth; a flow driven, according to the Münch hypothesis, by osmotic gradients generated by differences in sugar concentration. The length scales over which sugars are produced (L_leaf) and over which they are transported (L_stem), as well as the radius r of the cylindrical phloem cells through which the transport takes place, vary among species over several orders of magnitude; a major unsettled question is whether the Münch transport mechanism is effective over this wide range of sizes. Optimization of translocation speed predicts a scaling relation between radius r and the characteristic lengths as r ∼ (L_leaf L_stem)^1/3. Direct measurements using novel in vivo techniques and biomimicking microfluidic devices support this scaling relation and provide the first quantitative support for a unified mechanism of sugar translocation in plants spanning several orders of magnitude in size. The existence of a general scaling law for phloem dimensions provides a new framework for investigating the physical principles governing the morphological diversity of plants.     

Security and Software Quality: An Interview with Frank Perry

The ramifications of failing to completely and correctly address security can devastate an organization, not only in compromised data and financial cost but also in the time and energy spent to recover. One of us, Jane Hayes, sat down with an expert in the field, Frank Perry, to discuss the state of the art and future directions of system security and particularly how it interacts with software quality. Perry is chief engineer of the System and Network Solutions Group at SAIC. The group of almost 10,000 people focuses on network technologies and applications, primarily in the US federal market.