Molecular manipulation of keratin 8/18 intermediate filaments: modulators of FAS-mediated death signaling in human ovarian granulosa tumor cells

Background: Granulosa cell tumors (GCT) are a rare ovarian neoplasm but prognosis is poor following recurrence. Keratin intermediate filaments expressed in these tumors are a diagnostic marker, yet paradoxically, may also constitute a target for therapeutic intervention. In the current study, we evaluated keratin 8/18 (K8/18) filament expression as a mechanism of resistance to apoptosis in GCT, specifically focusing on regulation of the cell surface death receptor, Fas (FAS). Methods: The GCT cell line, KGN, was transiently transfected with siRNA to KRT8 and KRT18 to reduce K8/18 filament expression. Expression of K8/18, FAS, and apoptotic proteins (PARP, cleaved PARP) were evaluated by fluorescence microscopy, flow cytometric analysis, and immunoblotting, respectively. The incidence of FAS-mediated apoptosis in KGN cells was measured by caspase 3/7 activity. All experiments were performed independently three to six times, using a fresh aliquot of KGN cells for each experiment. Quantitative data were analyzed by one- or two-way analysis of variance (ANOVA), followed by a Tukey’s post-test for multiple comparisons; differences among means were considered statistically significant at P \u3c 0.05. Results: Control cultures of KGN cells exhibited abundant K8/18 filament expression (~90 % of cells), and minimal expression of FAS (\u3c25 % of cells). These cells were resistant to FAS-activating antibody (FasAb)-induced apoptosis, as determined by detection of cleaved PARP and measurement of caspase 3/7 activity. Conversely, siRNA-mediated knock-down of K8/18 filament expression enhanced FAS expression (\u3e 70 % of cells) and facilitated FasAb-induced apoptosis, evident by increased caspase 3/7 activity (P \u3c 0.05). Additional experiments revealed that inhibition of protein synthesis, but not MEK1/2 or PI3K signaling, also prompted FasAb-induced apoptosis. Conclusions: The results demonstrated that K8/18 filaments provide resistance to apoptosis in GCT by impairing FAS expression. The abundance of keratin filaments in these cells and their role in apoptotic resistance provides a greater mechanistic understanding of ovarian tumorgenicity, specifically GCT, as well as a clinically-relevant target for potential therapeutic intervention

Statistical mechanics of transcription-factor binding site discovery using Hidden Markov Models

Hidden Markov Models (HMMs) are a commonly used tool for inference of transcription factor (TF) binding sites from DNA sequence data. We exploit the mathematical equivalence between HMMs for TF binding and the "inverse" statistical mechanics of hard rods in a one-dimensional disordered potential to investigate learning in HMMs. We derive analytic expressions for the Fisher information, a commonly employed measure of confidence in learned parameters, in the biologically relevant limit where the density of binding sites is low. We then use techniques from statistical mechanics to derive a scaling principle relating the specificity (binding energy) of a TF to the minimum amount of training data necessary to learn it.Comment: 25 pages, 2 figures, 1 table V2 - typos fixed and new references adde

Internal tree cycling and atmospheric archiving of mercury: examination with concentration and stable isotope analyses

Trees predominantly take up mercury (Hg) from the atmosphere via stomatal assimilation of gaseous elemental Hg (GEM). Hg is oxidised in leaves/needles and transported to other tree anatomy including bole wood where it can be stored long-term. Using Hg associated with growth rings facilitates archiving of historical GEM concentrations. Nonetheless, there are significant knowledge gaps on the cycling of Hg within trees. We investigate Hg archived in tree rings, internal tree Hg cycling, and differences in Hg uptake mechanisms in Norway spruce and European larch sampled within 1 km of a HgCl2 contaminated site using total Hg (THg) and Hg stable isotope analyses. Tree ring samples are indicative of significant increases in THg concentrations (up to 521&micro;g&middot;kg-1) from background period (BGP; facility closed; 1992&mdash;present) to secondary industrial period (2ndIP; no HgCl2 wood treatment; 1962&ndash;1992) to primary industrial period (1stIP; active HgCl2 wood treatment; &asymp;1900&ndash;1962). Mass dependent fractionation (MDF) Hg stable isotope data are shifted negative during industrial periods (&delta;202Hg: 1stIP: 4.32&plusmn;0.15 &permil;; 2ndIP: 4.04&plusmn;0.32 &permil;; BGP: 2.83&plusmn;0.74 &permil;; 1SD). Even accounting for a &asymp; 2.6 &permil; MDF shift associated with stomatal uptake, these data are indicative of emissions derived from industrial activity being enriched in lighter isotopes associated with HgCl2 reduction and Hg0 volatilisation. Similar MDF (&delta;202Hg: 3.90&plusmn;0.30 &permil;; 1SD) in bark Hg (137&plusmn;105&micro;g&middot;kg-1) suggests that stomatal assimilation and downward transport is also the dominant uptake mechanism for bark Hg (reflective of negative stomatal uptake MDF shift) rather than deposition to bark. THg was enriched in sapwood of all sampled trees across both tree species. This may indicate long-term storage of a fraction of Hg in sapwood or xylem solution. These data advance our understanding of the physiological processing of Hg within trees and provide critical direction to future research into the use of trees as archives for historical atmospheric Hg.</p

Assimilation of satellite images into a sediment transport model of Lake Michigan

Peer Reviewedhttp://deepblue.lib.umich.edu/bitstream/2027.42/95634/1/wrcr11678.pd

Ultra-Sensitive Hot-Electron Nanobolometers for Terahertz Astrophysics

The background-limited spectral imaging of the early Universe requires spaceborne terahertz (THz) detectors with the sensitivity 2-3 orders of magnitude better than that of the state-of-the-art bolometers. To realize this sensitivity without sacrificing operating speed, novel detector designs should combine an ultrasmall heat capacity of a sensor with its unique thermal isolation. Quantum effects in thermal transport at nanoscale put strong limitations on the further improvement of traditional membrane-supported bolometers. Here we demonstrate an innovative approach by developing superconducting hot-electron nanobolometers in which the electrons are cooled only due to a weak electron-phonon interaction. At T<0.1K, the electron-phonon thermal conductance in these nanodevices becomes less than one percent of the quantum of thermal conductance. The hot-electron nanobolometers, sufficiently sensitive for registering single THz photons, are very promising for submillimeter astronomy and other applications based on quantum calorimetry and photon counting.Comment: 19 pages, 3 color figure

Seismic volcanostratigraphy of the western Indian rifted margin: The pre-Deccan igneous province

The Indian Plate has been the focus of intensive research concerning the flood basalts of the Deccan Traps. Here we document a volcanostratigraphic analysis of the offshore segment of the western Indian volcanic large igneous province, between the shoreline and the first magnetic anomaly (An 28 ∼63 Ma). We have mapped the different crustal domains of the NW Indian Ocean from stretched continental crust through to oceanic crust, using seismic reflection and potential field data. Two volcanic structures, the Somnath Ridge and the Saurashtra High, are identified, extending ∼305 km NE-SW in length and 155 km NW-SE in width. These show the internal structures of buried shield volcanoes and hyaloclastic mounds, surrounded by mass-wasting deposits and volcanic sediments. The structures observed resemble seismic images from the North Atlantic and northwest Australia, as well as volcanic geometries described for Runion and Hawaii. The geometry and internal seismic facies within the volcanic basement suggest a tholeiitic composition and subaerial to shallow marine emplacement. At the scale of the western Indian Plate, the emplacement of this volcanic platform is constrained by structural lineations associated with rifting. By reviewing the volcanism in the Indian Ocean and plate reconstruction of the area, the timing of the volcanism can be associated with eruption of a pre-Deccan continental flood basalt (∼75-65.5 Ma). The volcanic platform in this study represents an addition of 19-26.5% to the known volume of the West Indian Volcanic Province. Copyright 2011 by the American Geophysical Union

Cooperation between STAT3 and c-Jun Suppresses Fas Transcription

Decreased Fas expression during tumor progression often results in a loss of Fas-ligand (FasL)-mediated apoptosis. Human and mouse melanoma exhibit an inverse correlation between the degree of Fas cell surface expression, tumorigenicity, and metastatic capacity. The expression of dominant negative Stat3 or c-Jun in melanoma cells efficiently increased Fas expression and sensitized cells to FasL-induced apoptosis. Stat3+/− as well as c-Jun−/− cells exhibited increased Fas cell surface expression and higher sensitivity to FasL-mediated apoptosis. Suppression of Fas expression by Stat3 and c-Jun is uncoupled from Stat3-mediated transcriptional activation. Our findings indicate that Stat3 oncogenic activities could also be mediated through its cooperation with c-Jun, resulting in downregulation of Fas surface expression, which is implicated in the tumor's ability to resist therapy and metastasize

Missouri 2011 Soft Red Winter Wheat Performance Tests

This report is published by the MU Variety Testing Program, Division of Plant Sciences, University of Missouri. The work was supported by fees from companies and organizations submitting varieties for evaluation. The large number of varieties available makes selection of a superior variety difficult. To select intelligently, producers need a reliable, unbiased, up-to-date source of information that will permit valid comparisons among available varieties. The objective of the MU Variety Testing Program is to provide this information. Tests are conducted under as close to uniform conditions as possible. Small plots are used to reduce the chance of soil and other variations occurring among variety plots. Results obtained should aid individual growers in judging the relative merits of many of the commercial wheat varieties available in Missouri